Genesi dellinformazione.

556

Unit IX

88 and Sensory Physiology

The Nervous System: A. General Principles
Somesthetic areas

Dendrites
DENDRITI

5. ELECTROTONIC PROPERTIES OF AXONS AND DE

Motor cortex

2
Thalamus

ENCEFALO
Brain

Pons

Cell body

Bulboreticular
formation

Medulla

Cerebellum

1
Spinal cord

Skin
3. Integration

Pain, cold,
3. INTEGRAZIONE
warmth (Free
nerve ending)
Pressure

1. Intrinsic
(Pacinian corpuscle)
1. INTRINSECO

(Expanded tip
receptor)
Touch
(Meissner's corpuscle)
4. Encoding
4. CODIFICA

ASSONE
Axon
Synapses
INPUT SINAPTICI

MIDOLLO
Spinal cord
SPINALE

Muscle spindle

Golgi tendon
apparatus

Muscle

Kinesthetic receptor
5. Output
5. USCITA

Joint

NEURONI DI

Second-order
SECONDO
neurons

ORDINE

Figure 451

GENERAZIONE
DELLINFORMAZIONE
Generation

2.2.RICEZIONE
Reception

INPUT
SINAPTICI

Figure 452
Somatosensory axis of the nervous system.

FIGURE 5.1 Nerve cells have four main regions and five main
functions. Electrotonic potential spread is fundamental for coorditrolling
muscles,
natingsmooth
the regions
and their glands,
functions.and other internal

of current sp
development
mission of ele
floor. The ele
often are ref
theory was fir
system in th
electric curren
1940s, it was
squid) axons
of the Hodgk
action potent
Mathemati
theory to co
1960s, Wilfrid
solved by the
mental mode
herd, 1968). T
a theory of de
bined with m
synaptic pote
the basis for a
ronal activity
A variety o
sible for even
tional propert
These tools a
NEURON; G
therefore pre
the context
models. Exp
student great
are present in
of current in

Sinapsi.
The Nobel Prize in Physiology
and Medicine 1932

DEFINIZIONE Dal greco syn aptein, vale a dire connettere.

C O N TAT T O F U N Z I O N A L E
INSTAURATO DAI NEURONI
Attraverso la trasmissione
sinaptica, l'impulso nervoso pu
viaggiare da un neurone all'altro
o da un neurone ad una fibra
neuromuscolare, in questo caso
si parla di giunzione
(neuromuscolare: motoneuroni
alfa o gamma).

Charles Scott Sherrington (1857-1952)

L i m p u l s o v i a g g i a d a u n
NEURONE PRESINAPTICO ad
uno POSTSINAPTICO.
Textbook of Physiology, part three:
The Central Nervous System

Sinapsi
S. Ramon y Cajal

C. Golgi

The war of
the Soup and
In Memory of J. David Robertson
Sparks

by John E. Heuser, M.D., Department of Cell Biology, Washington University School of Medicine, St. Louis, MO 63110

>`,Li]iiiiiVVVvViiL>i]
`i`>i
}]xv>V>Liii>i>
i>`1iiLivi`i>]i>>Vii}>}i`
Li`V>ii>V>`>>*vi iv iL}>
i1i-Vvi`Vi>]
ii`L
viv{i>]`]>`LiiV`i]>i]> }
vi>6>> >Li]> }vi>1iv

>`>`>i]]>>
>L`}i]>>Vi

Otto

,Li>iviii>iLiv/i
iV>-Vi
v
i }]}i>>vi>v`i`]>`
>>>i>VLi>v
i }]L
Loewi
}ii>iwiii>]L>Vi>>i
>v V>>` ViV>
}iV>iiiV
VV>>i`V>L>V}`

J. D. Robertson
>`,Li]ii}iiVVV]}>i`
>}Li`x

D. Potter
B. Katz
>`>i`/V>>]
>L>>]iv>VViVi>>`>}>`iVi>Vi],Li>
i`V>i`>i>ii}i/ii]ii>Vi`wi>vi`V>V>7`7>Li}>
iii`i1- >>`>iwi`V>V>>>`
viiVi}>`>}Liy

Trasmissione dellinformazione: segnali

chimici ed elettrici.
WIRING (SINAPSI)

VOLUME

Endocrina & Neuroendocrina

Sinapsi
elettriche
paracrina

autocrina

ELECTRICAL
Effetto di
campo
elettrico

Sinapsi
chimiche

Classificazione funzionale delle sinapsi.

WIRING (SINAPSI)

La trasmissione del segnale

avviene direttamente
mediante accoppiamento
elettrico delle cellule.
Sinapsi elettriche

La trasmissione del segnale

avviene per liberazione
localizzata di sostanze
chimiche.
Sinapsi chimiche

Due domande nelle Neuroscenze

Come comunicano i neuroni?

CANALI IONICI-POTENZIALE
DAZIONE-SINAPSI

Come possiamo ascoltarli?

TECNICHE DINDAGINE
ELETTROFISIOLOGICA

Due domande nelle Neuroscenze

Come comunicano i neuroni?

CANALI IONICI-POTENZIALE
DAZIONE-SINAPSI

Come possiamo ascoltarli?

TECNICHE DINDAGINE
ELETTROFISIOLOGICA

Classificazione strutturale delle sinapsi.

WIRING (SINAPSI)
In relazione agli elementi neuronali che
entrano in contatto nella sinapsi, si
possono distinguere sinapsi:
(1) asso-detritiche, in cui l'assone
di un neurone contatta l'albero
dendritico di un altro neurone.
(1)

(2) asso-assoniche, in cui sono a

contatto due assoni.
(3)

(2)

( 3 ) a s s o - s o m a t i c h e , c h e s i
stabiliscono tra l'assone di un
neurone ed il corpo cellulare
(soma) di un secondo neurone.

Sinapsi.

Sinapsi elettriche.
Poco diffuse, ma presenti nel SN di tutti gli
animali.
Sono presenti tra neuroni e tra neuroni e
cellule gliali.
Le membrane di cellule adiacenti sono
collegate mediante giunzioni comunicanti
(gap junction): struttura intercellulare
specializzata.
Permettono una rapida comunicazione tra
neuroni adiacenti, consentendo un flusso
passivo diretto di corrente elettrica tra una
cellula e laltra.

Sinapsi Elettriche vs. Sinapasi chimiche.

PROPIETA

SIN. ELETTRICHE

SIN. CHIMICHE

Meccanismo
Velocit
Consumo E
Direzione
Messaggio
Tipologia
Sensibilit farmaci

Puramente biofisico
Alta
Basso
Bidirezionale
Eccitatorio
Stereotipato
Bassa

Biochimico
Bassa
Alto
Unidirezionale
Ecc. / inib. / metab.
Plastico
Alto

Flusso della corrente in una sinapsi elettrica.

Le cariche
positive iniettate
tendono a
depolarizzare la
m e m b r a n a
postsinaptica.

Flusso della corrente in una sinapsi chimica.

La depolarizzazione
postsinaptica dipende
dal rilascio di
neurotrasmettitore.

Sinapsi elettriche.
Cellula presinaptica
CONNESSONE

CARATTERISTICHE DELLE SINAPSI ELETTRICHE

1. Connessioni di neuroni in rete.
2. Trasmissione segnali elettrotonici e metabolici.
3. Networks neuronali, networks gliali e networks neuro-gliali.
4. Importanti per lo sviluppo (transizione da multipla innervazione a singola)
5. Non permettono lintegrazione di pi segnali.
USATE :
1. Quando serve rapidit nella trasmissione (es. sistemi di fuga).
2. Quando serve sincronizzazione di pi cellule (es. nel muscolo cardiaco)

Cellula postsinaptica
CONNESSONE

connessina

Sinapsi elettriche: sincronizzazione.

Una funzione pi generale delle sinapsi elettriche quella di sincronizzare
lattivit elettrica tra varie popolazioni di neuroni.

Membrane Potential (mV)

Sincronizzazione dellattivit
elettrica in interneuroni
ippocampali.
La generazione di un
potenziale dazione in un
neurone spesso da origine alla
scarica sincronizzata di un
potenziale dazione nellaltro
neurone.

Time (ms)

demonstration of the ubiquitous presence of electrical synapses in the mammalian

brain led to the indisputable conclusion that chemical transmission and electrical
transmission coexist in all nervous systems.

in vertebrate neurons, including connexin 36 (CX36; also

known as GJD2), CX45 (also known as GJC1), CX50
(also known as GJA8), CX30.2 (also known as GJC3)
and CX31.1 (also known as GJB5) (see REFS1618 for
Are chemical synapses more sophisticated?
reviews). To date, the retina is the neuronal tissue in
Chemical versus electrical synapses. From an evolution- which the diversity of neuronal connexins has been most
ary point of view, chemical communication between cells investigated (see REF.19 for a review). Because of its widepreceded electrical communication9. Communication spread distribution, CX36 is considered to be the main
with chemical signals occurs in unicellular organisms synaptic connexin in the mammalian brain20. However,
such as bacteria10, in which it mediates important phe- although electrical transmission is dramatically reduced
nomena such as quorum sensing 11 (the mechanism in CX36-knockout mice, some level of electrical and
underlying the detection of bacterial population den- tracer coupling can be detected in these animals21,22, sugsity). More-specific communication between individual gesting that synaptic gap junctions may be composed
cellsE.Pereda
requires complex cellular specializations between of additional connexins. Several of the 25 members of
Alberto
the interacting partners. The presence of suchbcellular
the
innexin family have been identified in invertebrate
a Chemical synapse
Electrical
synapse
Abstract
|
Brain
function
relies
on
the
ability
of
neurons
to
communicate
with
each other.
specializations is not exclusive to neurons and is a char- neurons,
mainly
in fly 23, leech24 and worm neurons25.
Interneuronal
communication
primarily
takes
place
at
synapses,
where
information
from one
acteristic that
neuronal chemical and electrical synapses
Although electrical
synapses can act, to some extent,
Action
CS
ES
neuron
is
rapidly
conveyed
to
a
second
neuron.
There
are
two
main
modalities
of
synaptic
potential
share with immunological
synapses between a lympho- in a metabotropic manner by allowing the passage of
transmission:
chemical
and electrical.cell
Far12,from
functioning
independently
and serving(a critical property of electrical syncyte and an
antigen-presenting
which
dynamically
small metabolites
unrelated
functions,
mounting
evidence
indicates
that
these
two
modalities
of synaptic
form to facilitate the transfer of information between apses during
development; see below), they, unlike
Ca2+
transmission
closely
interact,
both
during
development
and
in
the
adult
brain.
Rather
than
these two cells. Chemical transmission requires sophis- chemical synapses,
lack the ability to amplify and
Action
Presynaptic
conceiving
synaptic
transmission
as
either
chemical
or
electrical,
this
article
emphasizes
the
ticated presynaptic molecular machinery that regulates transform presynaptic
signals. However, this does not
potential
terminal
notion
that synapticSynaptic
transmission
is both
chemical andmanner
electrical, and
that
interactions
neurotransmitter
release in
a probabilistic
mean
that
electrical synapses are less sophisticated;
1
between
twovesicle
forms
of interneuronal
might
be required
for normal
whenthese
an action
potential
invades the communication
synaptic terminal
their
sophistication
just relies on a different functional
Gap
junction
brain(although
development
and
function.
at some contacts, release can also be propor- property their bidirectionality which enables them
channel
Neurotransmitter
tional to changes in
resting membrane potential13). Such to coordinate the activity of large groups of intercontransmission also requires a similarly complex
post- nected neurons2. Based on this bidirectional and anaCommunication between neurons is required for
Electrical and chemical synapses are now known to
Ionotropic
synaptic molecular machinery,Metabotropic
including
ionotropic
logical
(that
is, theybutdo
not require an action potential)
coexist
in most organisms
and brain
structures,
details
Lateral excitation
receptor brain function, and the quality of such communicareceptor
(ligand-gated
ion
channels)
and
metabotropic
receptors
nature,
electrical
synapses
tion
enables
hardwired
neural
networks
to
act
in
a
of
the
properties
and
distribution
of
these
two
modalities
of are particularly efficient at
The ability of an excited neuron
dynamic
fashion.
Functional
interactions
between
transmission
are
still
emerging.
Most
research
efforts
(G protein-coupled receptors that act indirectly through detecting the coincidence of simultaneous subthresh(or sensory afferent) to excite
occur at anatomically identifiable cellular have focused on exploring the mechanisms of chemiits neighbours. Although it neurons
a secondary
messenger) (FIG.1a), which are capable of old depolarizations within a group of coupled neurons,
regions
called
synapses. Although the nature of synaptic cal transmission, and considerably less is known
reduces discrimination, lateral
detecting
and
translating
messages
into various
postsyna phenomenon
that increases
neuronal excitability and
has been an
area of enormous
controversy
about those
underlying
electrical transmission.
It was
Membrane
Gene
excitation greatly enhances transmission
22,2629
events,
ranging
fromtransmission
changes in
potential
promotes
synchronous
firing
. Electrical synapses
(BOX1)
,aptic
two main
modalities
of synaptic
resting
thought
that electrical
synapses were
moreCoupling
abundant
potential
expression
input sensitivity. It is a
1 in invertebrates and cold-blooded vertebrates than
namely,
chemical
and
electrical

are
now
recognized.
At
(synaptic potential) to gene expression . These features are also highly effective
potentialat mediating lateral excitation
less-appreciated property of
Biochemical
chemical
synapses,
information
is transferred
through
in mammals.
However,
a wealth
of data the
now sensitivity
indicate
Postsynaptic
sensory and
cortical networks.
enable
chemical
synapses
to
adapt
to
diverse
functional
and
increasing
of sensory systems, as it
cascades

Sinapsi elettriche vs. Sinapsi chimiche.

Electrical synapses and their
functional interactions with chemical
synapses

VIEWS

the release of a neurotransmitter from one neuron that electrical synapses are widely distributed in the
and detection of the neurotransmitter by an adjacent mammalian brain5. In addition to the retina, inferior
1
Figure 1 | Thecell
two
main modalities
of synaptic
transmission.
a | Chemical
transmission
sophisticated
, whereas
in electrical synapses,
the cytoplasm
olive and
olfactory bulb,
structures in requires
which electriNATUREpresynaptic
REVIEWS | NEUROSCIENCE
VOLUME 15 | APRIL 2014 | 251
Nature
Reviews
| Neuroscience
of
adjacent
cells
is
directly
connected
by
clusters
of
cal
transmission
has
been
known
to
occur
for
some
molecular machinery that regulates neurotransmitter release in a probabilistic manner upon
2
5
intercellular
channels
called
gap
junctions
.
Although
time
,
electrical
synapses
have
been
found
in
dispadepolarization of the presynaptic terminal
in this case,
by the
arrival of
an reserved
action potential which leads to the
2014
Publishers
cellular specializations for both
formsMacmillan
of transmisrate Limited.
regions All
of rights
the mammalian
CNS and have been
activation of voltage-gated
calcium channels (VGCCs). Similarly complex postsynaptic
molecular machinery is also
sion can be found at various neuronal sites (dendrites, shown to constitute a distinct phenotypic feature

terminal

demonstration of the ubiquitous presence of electrical synapses in the mammalian

brain led to the indisputable conclusion that chemical transmission and electrical
transmission coexist in all nervous systems.

Hetero andAremixed
synapses.
chemical synapses
more sophisticated?

Lateral excitation
The ability of an excited neuron
(or sensory afferent) to excite
its neighbours. Although it
reduces discrimination, lateral
excitation greatly enhances
input sensitivity. It is a
less-appreciated property of
sensory and cortical networks.

Chemical versus electrical synapses. From an evolutionary point of view, chemical communication between cells
preceded electrical communication9. Communication
with chemical signals occurs in unicellular organisms
such as bacteria10, in which it mediates important phenomena such as quorum sensing 11 (the mechanism
underlying the detection of bacterial population density). More-specific communication between individual
cells requires complex cellular specializations between
the interacting partners. The presence of such cellular
specializations is not exclusive to neurons and is a characteristic that neuronal chemical and electrical synapses
share with immunological synapses between a lymphocyte and an antigen-presenting cell12, which dynamically
form to facilitate the transfer of information between
these two cells. Chemical transmission requires sophisticated presynaptic molecular machinery that regulates
neurotransmitter release in a probabilistic manner
when an action potential invades the synaptic terminal1
(although at some contacts, release can also be proportional to changes in resting membrane potential13). Such
transmission also requires a similarly complex postsynaptic molecular machinery, including ionotropic
(ligand-gated ion channels) and metabotropic receptors
(G protein-coupled receptors that act indirectly through
a secondary messenger) (FIG.1a), which are capable of
detecting and translating messages into various postsynaptic events, ranging from changes in resting potential
(synaptic potential) to gene expression1. These features
enable chemical synapses to adapt to diverse functional

in vertebrate neurons, including connexin 36 (CX36; also

known as GJD2), CX45 (also known as GJC1), CX50
(also known as GJA8), CX30.2 (also known as GJC3)
and CX31.1 (also known as GJB5) (see REFS1618 for
reviews). To date, the retina is the neuronal tissue in
which the diversity of neuronal connexins has been most
investigated (see REF.19 for a review). Because of its widespread distribution, CX36 is considered to be the main
synaptic connexin in the mammalian brain20. However,
although electrical transmission is dramatically reduced
in CX36-knockout mice, some level of electrical and
tracer coupling can be detected in these animals21,22, suggesting that synaptic gap junctions may be composed
of additional connexins. Several of the 25 members of
the innexin family have been identified in invertebrate
neurons, mainly in fly 23, leech24 and worm neurons25.
Although electrical synapses can act, to some extent,
in a metabotropic manner by allowing the passage of
small metabolites (a critical property of electrical synapses during development; see below), they, unlike
chemical synapses, lack the ability to amplify and
transform presynaptic signals. However, this does not
mean that electrical synapses are less sophisticated;
their sophistication just relies on a different functional
property their bidirectionality which enables them
to coordinate the activity of large groups of interconnected neurons2. Based on this bidirectional and analogical (that is, they do not require an action potential)
nature, electrical synapses are particularly efficient at
detecting the coincidence of simultaneous subthreshold depolarizations within a group of coupled neurons,
a phenomenon that increases neuronal excitability and
promotes synchronous firing 22,2629. Electrical synapses
are also highly effective at mediating lateral excitation
and increasing the sensitivity of sensory systems, as it

NATURE REVIEWS | NEUROSCIENCE

VOLUME 15 | APRIL 2014 | 251

2014 Macmillan Publishers Limited. All rights reserved

Sinapsi.

Sinapsi chimiche.
Sono le pi diffuse nei mammiferi. I tipi di sinapsi
pi diffuse nel SNC e SNP.
Il ritardo sinaptico di qualche ms, anche se in
alcuni casi pu essere di 0.3 ms (0.3 5 ms).
Lo spazio pre- e post- sinaptico maggiore
rispetto a quello di una sinapsi elettrica: fessura
sinaptica (20-40 nm), quindi non c continuit
anatomica tra il neurone pre- e post-sinaptico.
Caratteristica fondamentale: presenza di piccoli
organelli delimitati dalla membrana noti come
vescicole sinaptiche nella terminazione sinaptica.

Sinapsi chimiche.

Le vescicole sinaptiche sono

organelli sferici contenenti uno o
pi neurotrasmettitori. Questi
sono sostanze chimiche
funzionanti da messaggeri fra
neuroni comunicanti, legandosi ai
propri recettori post sinaptici.

Sinapasi chimiche
POSTSINAPSI
specializzata in RICEZIONE
trasduzione CHIMICA-ELETTRICA E/O METABOLICA
1. Densit postsinaptica
2. Recettori per NT
3. Canali ionici ed altri effettori intracellulari

FESSURA SINAPTICA
PRESINAPSI

1. Matrice extracellulare
2. Sistemi di catabolismo del NT

specializzata in TRASMISSIONE
trasduzione ELETTRICA-CHIMICA
1. Vescicole sinaptiche
2. Citoscheletrodi actina
3. Memb. Pres. con trasduttori (canali Ca2+ VD)
4. Proteine di fusione e recettore per il Ca2+
5. Mitocondri
6. Sist. di sintesi, trasporto e catabolismo del NT

R E PO R TS

Ricostruzione 3D di un bottone sinaptico.

Wilhelm et al., SCIENCE, VOL 344 ISSUE 6187.

Ricostruzione 3D di un bottone sinaptico, particolari.

Fig. 3. A 3D model of synaptic architecture. (A) A section through the synaptic bouton, indicating 60 proteins. The proteins are shown in the copy numbers
indicated in tables S1 and S2 and in positions determined according to the imaging data (Fig. 2 and fig. S6) and to the literature (see fig. S6 for details). (B) Highzoom view of the active zone area. (C) High-zoom view of one vesicle within the vesicle cluster. (D) High-zoom view of a section of the plasma membrane in the
vicinity of the active zone. Clusters of syntaxin (yellow) and SNAP 25 (red) are visible, as well as a recently fused synaptic vesicle (top). The graphical legend indicates
the different proteins (right). Displayed synaptic vesicles have a diameter of 42 nm.

1026

30 MAY 2014 VOL 344 ISSUE 6187

sciencemag.org SCIENCE

Wilhelm et al., SCIENCE, VOL 344 ISSUE 6187.

Le proteine delle vescicole sinaptiche (SV).

Le proteine delle vescicole sinaptiche (SV).

Trasporto Attivo vescicolare: esocitosi.

Assume caratteristiche specifiche dipendendo dal tipo di cellule in cui avviene.
Richiedendo energia considerato trasporto attivo. Richiede la formazione di
vescicole di membrana che spostano il loro contenuto attraverso la membrana.
Esocitosi, se le molecole passano dallambiente interno all'esterno
Esocitosi costitutiva: comune a
tutte le cellule eucariotiche. Non
richiede un segnale. Funzione:
mezzo per rilasciare sostanze
allesterno della cellula e per
modificare le componenti della
membrana plasmatica.
Esocitosi regolata: propria delle
cellule specializzate a
secernere i prodotti che
elaborano (es. neuroni, cellule
esocrine ed endocrine). E
attivata da uno stimolo
extracellulare specifico. Media
la comunicazione cell.-cell.

Published online: July 9, 2015

Ricostruzione 3D della zona attiva presinaptica.

Frauke Ackermann et al

EMBO reports

Presynaptic active zones

AP2
RIM

Syntaxin1

Endophilin

Piccolo

Munc13

Bassoon

Dynamin

VAMP2

Amphiphysin

Clathrin

SNAP 25

Ackermann et al., EMBO reports Vol. 16 | No 8 | 2015.

Figure 4. Model of presynaptic bouton and active zone organization.

A section through the active and endocytic zones of a vertebrate synapse indicating the spatial distribution and copy number of presynaptic proteins that help define the
presynaptic AZ as the site of SV exocytosis and clathrin-mediated endocytosis. Panel at the bottom, a graphical legend of the predicted structures of presynaptic proteins

Proteine specifiche organizzano la zona

Presynaptic attiva.
active zones
Frauke Ackermann et al

Published online: July 9, 2015

EMBO reports

B
Actin

SV

SYNAPTIC VESICLE
POOL
Rab3a
SV

SV

Synapsin
SV

Piccolo

Bassoon
ERC2

RIM
ACTIVE
ZONE
Munc13

SV

RBP

Ca2+ channel

Synaptotagmin

SNARE
complex

Figure 2. Cytoskeletal matrix proteins organize synaptic vesicle release sites at presynaptic active zones.
(A) 3D reconstruction of filaments (pink) and SVs (yellow) tethered near the active zone from a vertebrate (rat) hippocampal synapse. Docked vesicles are in blue.
Reproduced with permission from [11]. (B) Schematic diagram of CAZ molecules mediating the capture
(synapsin, et
actin),
complex),
Ackermann
al.,docking
EMBO(SNARE
reports
Vol priming
16 | No 8 | 2015.
(RIM, Munc13, Rab3) and fusion (synaptotagmin) of SVs and VGCCs (Bassoon, RBP, RIM, ELKS) at presynaptic active zones. At present, the spatial relationship of these
molecules within the cryo-fixed EM image in (A) is not well resolved.

Synaptic Strenght and Quantal Parameters of NT

release.
Reserve Pool
(RsP)

Readily Releasable Pool

(RRP)

Recycling Pool
(RcP)

PRE SYN

POST SYN

Ciclo eso- endocitotico.

reserve pool

RR pool

Recycling pool

Published online: July 9, 2015

EMBO reports

Presynaptic active zones

Synaptic vesicle cycle at presynaptic zone.

A

Clathrin-mediated endocytosis

Bulk endocytosis

Frauke Ackermann et al

Ultrafast endocytosis

MBV

Endosome
B

SV

RRP

PSD

Ackermann et al., EMBO reports Vol 16 | No 8 | 2015.

Figure 5. Synaptic vesicle cycle at the presynaptic terminal.
The presynaptic AZ functionally defines the space within boutons where upon calcium influx synaptic vesicle fusion and neurotransmission takes place (lower panel). It is the
center of the SV life cycle. Vesicles are recruited from the vesicle cluster toward the AZ where they undergo maturation steps such as docking and priming and finally

Receptors
Receptors

Postsynaptic density

Postsynaptic
spine

Recycling vesicles

Endosomes

Considerazioni temporali sulle sinapsi chimiche.

Postsynaptic
spine

Recycling vesicles

Endosomes

B
B

POST-STIM

Presynaptic action potential (APpre)

Presynaptic action potential (APpre)

Gating

Ca2+-channels

Gating Ca2+-channels

PRE-STIM

ded from http://cshperspectives.cshlp.org/ on March 18, 2015 - Published by Cold Spring Harbor Laboratory Press

0.3 msec

APpre

Ca2+-Current (ICa2+)

Ca2+-Current (ICa2+)
2+

Ca -triggered
Ca2+-triggered
0.5 msec
Fusion
pore opening
Fusion pore
opening

T.C. Sudhof

0.3 msec

ICa2+

APpre

ICa2+

25 m
25 mV

0.5 nA

0.5

0.5 msec

Exocytosis (plotted
Exocytosis (plotted
as release rate)
as release rate)
Neurotransmitter
Neurotransmitter
Mitochondria
binding to
0.4 msec
0.4 msec
postsynaptic receptors binding to

A
Synapse
Presynaptic
terminal
Synaptic
vesicles

Endosomes

Release
rate

Release
rate

0.5 ms1
2 nA

postsynaptic receptors

Evoked postsynaptic current (EPSC)

0.5 m

2 n

Evoked postsynaptic current (EPSC)

Summation of EPSCs
0.1 msec
triggers action
potential of EPSCs
Summation

triggers action potential

Ca2+

EPSC

0.1 msec

EPSC

Postsynaptic action
Potential (APpost)

Postsynaptic action
Potential (APpost)

Postsynaptic density
Receptors
Postsynaptic
spine

Recycling vesicles

Endosomes

25 mV

25 m
APpost

APpost

0.5 msec

Figure 1. Principle and time course of Ca2-triggered

synaptic
transmission.
Schematic diagram
of a synapse
Cold Spring
Harb
Perspect Biol(A)
2012;4:a011353,
modified.
0.5 m

B
Presynaptic action potential (APpre)
Gating Ca2+-channels

0.3 msec

Ca2+-Current (ICa2+)
Ca2+-triggered
Fusion pore opening

0.5 msec

APpre

ICa2+

illustrating the localized influx of Ca2 at the active zone (red secreted neurotransmitters). (B) Schematic
illustration
of the
sequence and
transmission
as measured
by simultaneous
pre- anddia
Figure
1. Principle
andtime
timecourse
courseofofsynaptic
Ca2-triggered
synaptic
transmission.
(A) Schematic
2
postsynaptic patch-clamp recordings at the calyx of2
Held synapse. Note that Ca currents and EPSC are shown
25 mV

0.5 nA

illustrating the localized influx of Ca

at the active zone (red secreted neurotransmitter

(4, 5)
and then
culturing
them
a de- and
nervous
system
(CNS)
neurons
by in
sevenfold
lia, and there was little colo-functional synapses on centralpurity

are functionally
immature.
Astrocytes
increase is
the little
number overlap
of mature,
of
glia there
sponseform
of RGCs
tothe
pulsesabsence
of L-glutamate

ap

threefold
participate
in synaptic
plasticity.
blot
analysis
of extracts
of RGCs
culturedsynapses on central nervous system
neous
synaptic
currents
are larger
(CNS)
neurons
by sevenfold
and in RGCspn
fined serum-free
Using
ining
(Fig.
3E). Glia
increased
thesynaptic
number
appliedfunctional
to change
the cell somas.
Regardless
of theand frequently
are required for
maintenance
in vitro.medium
We also(5).
show
thatthese
most
synbetween
prepostsynaptic
markers,
the few
presynaptic
puncta
observed
(B).
All
without
glia
or
with
glia
indicates
no
ap
areof
required
for synaptic
maintenance
We also show
thatglia
most
syn-1D), consist
methods,
have
previously shown
thatinRGCs
with
(Fig.
presence
glia, RGCs
responded
with in- in vitro.
apses arepuncta
generatedby
concurrently
withwethe
development
of in
glia
Thesesynaptotagmin
synaptic
currents
were CNQXtotalvivo.
synaptic
and synapGluR2/3-positive
synaptic
six-cultured
ese
data show that presynapwhereas
in
the
presence
of
glia
there
is
a
high
pre
apses
are
generated
concurrently
with
the
development
of
glia
in
vivo.
These
in
the
presence
of
astrocyte-condiApplicationamplitude of spon
ward
(Fig.
that were
blocked
by or sensitive
data demonstrate a previously unknown function
for gliatophysin
in
inducing
andlevels.
creased
Astrocytes,
which
ensheathe
synapses
throughrified
RGCs
in the
presence
absence(10).
of (C)
protein
p115
is acurrents
control
for 1C)
markers
colocalize
in number of immu-tioned medium have 10 times higher
wa
data
demonstrate
a previously
unknownBar,
function
for
gliaof (E)
in100
inducing
and
levels
of
degree
of
overlap.
50
#m.
Double
of
50-ms
pulses
!M
daptic
(Fig.
3F). The
average
receptor
antagonists
(10).
The cultured
size
stabilizing CNS synapses, show that CNS synapseout
number
can
be
profoundly
total
protein.
(B) Synaptic
protein
immunominiature
events (minia
the
CNS,
are
presently
thought ofglutamate
as synastrocytes.
RGCs
were
1 week
glu
stabilizing
CNS synapses,
show
that CNS synapse
number
can
profoundly
Lfor
-glutamate
to RGC
cellbe
bodies
synaptic
activity
(6).
This
glial
effect
is
not
tructural counterparts of syn-regulated by nonneuronal signals,
postsynaptic
response,
was
the possibility that
glia may actively
labelinghowever,
of autaptic
neurons
indicates
that
reactivity
in RGCs culturedofinthe
the
presence
eactive
on RGCs in thesimplyanddueraise
at apossibility
holding
potential
ofmay
"70actively
of
regulated
by nonneuronal
signals, and
raise
the
thatpostsynaptic
glia
to enhancement
of RGC
maturation,
currents,
or
mE
aptic
support
cells,
clearing
ions
and
neurofollowed
by
the
addition
of
astrocytes
either
esence
of puncta
glia, but that
rarelyformed
inparticipate
threefold larger
RGC neurons
culturedpuncta
in synaptic plasticity.
absence
werein
mV results
sustained inward
synaptic
arein present
in the
thr
participate
in fewer
synaptic plasticity.
in theand
presence
or of glia. Cultured RGCs
sence
and absence
of glia was 35 ! 4 andbecause RGCs cultured either
he
glial-dependent
clustering
in densities
the presence of glia (Fig
transmitters
from
theparaformaldehyde
synaptic cleft, with
but and
accucontact
with RGCs
a feeding
layer(D)
forCurrent
gliaimmuno(Fig.in1D),
consistent
with or
theasinfixed
with
currents.
wi
absence of glia are electrically excitable, develabsence
of
glia
than
in
the
presence
of
glia.
of RGCs cultured in the absynaptic markers
was comparison,
creased of
amplitude
of spontaneous synaptic
Astrocytes,
which ensheathethe
synapses
throughrified RGCs in thestained
presencewith
or absence
of against
antibodies
preandensheathe
!on 1,of respectively.
By
av-op
cre
Astrocytes,
which
synapses
throughrified
RGCs
in
the
presence
or
absence
of
comparable numbersmulating
of dendritesevidence
and axons,suggests the possibility
sence six
or presence
ofasglia.
Bar,
50 #m.
(F) About
times
many
miniature
events
(miniature
excitatory
out the CNS, are presently thought of as synastrocytes. RGCs were
cultured for
1 week
postsynaptic
markers.
Staining
of RGCs
withpresently
when RGCs were cocultured
mi
out
the
CNS,
are
thought
of
as
synastrocytes.
RGCs
were
cultured
for
1
week
and
have
identical
survival
rates.
In
addition,
Whole-cell
currents:
RGCs
more
active
roles
as
well
(13).
The
role
of
glia
number
quanta
released
response
an antibody
against
the presynaptic
marker
postsynaptic
or mEPSCs)
aptic
cells, in
clearing
ions and neurofollowed by the addition
of astrocytes
either
synaptic
present
in15.7RGCs
cultured
yge
purified
target of
neurons
fromsupport
po
aptic support currents,
cells, clearing
ions puncta
andrecorded
neuro-arefollowed
addition
of astrocytes either
withoutby
glia,the
# 4.7 pA/pF;
RGCs cultured under both conditions synthesynaptotagmin
shows
that intothe
absence of glia (Fig. 2G). To deterFig. 1. Glia
atwith
CNS
synapses
has been
study
the
presence
transmitters
from the synaptic
cleft, but
RGCs
or as
feeding
layerdifficult
for
a strong
neurons
RGCsas
with
glia,
45 #
lliculus
(24),stimulation
indicating
that in autaptic
in
transmitters
from the in
synaptic
cleft,
but accu- of inglia
contact
with
RGCs
or8.6
as apA/
feeding
layer for
the
presence
in
the
absence
of
size,accusecrete, in
andcontact
respond
to glutamate
(6)aand
glia cells
presynaptic
markers
arecultures
diffuse (left),
mulating
evidence suggests the possibility
ofnearly identical because
enhance syn
pF (P $ 0.015, Students t
glial
are
present
in
CNS
mulating
evidence
suggests
the
possibility
of
express
levels
of
mRNAs
for
a
RGCs
to
form
synapses
with4.3(EGluR2/3
puncta
tured in the presence
androles
absence
of The
glia
whereas in the presence of glia presynaptic glia: with glia, 34.5 !test).
and F) Autaptic
EPSCs per both presyn
more active
as well (13).
role
of glia
more active
as well (13). The role of glia
large
battery of neurotransmitter
receptors,
volt- for neuronal survival.
R
E PRGCs
O Rcultured
T S in the abbe accounted for by the aband are often
necessary
In roles(C)
from
markers
are discretely clustered
(right).
cell;
without
glia,
6.1
!
0.9
GluR2/3
puncta aptic mecha
Fig.
1.
Glial
astrocytes
(glia)
at
CNS
synapses
has
been
difficult
to
study
snormal
47 !target
10 and
7 !
Theage-dependent ion channels, and synaptic
CNS synapses has been difficult to study
prosence (E) or presence (F) of
neurons
[see4,glialrespectively.
Staining
of
RGCs
with
anofatantibody
to the
enhance
synaptic
efficacy
by
this
study,
we
have
taken
advantage
recently
because
cells are present in CNSteins,
cultures
(34), co
Fig.
4.
Glial
astrocytes
(glia)
incell.
because
glialthat
cellsinareper
present
in CNS cultures
as assessed by gene chip analysis
(7). In marker PSD-95
glia. (G and H) Higher synaptic
the absence
postsynaptic
reveals
both presynaptic and postsynmilarity
the
ta (25)]. of these values
albumin
and are oftensuggests
necessary for that
neuronal
survival.
In we examinedeveloped
crease
the number
oftype
synapses
methods
to
isolate
a
defined
of
and
are
often
necessary
for
neuronal
survival.
In
failure
rates
are
seen
in
RGCs
this study,
whether the
increased
aptic mechanisms. (A and B) In
frequency
o
the absence
of glia there
few
transfer
e if these puncta correspond
this study,
have taken
advantage
ofsynaptic
recentlyactivity in glia is due to an
per
neuron.
(A andare
B)relatively
Electron
without
glia
(G)
than
in
RGCs
munoreactive
puncta
thatweform
in the
presthis
study,
we
have
taken
advantage
of
recently
increase
in
the absence of glia, only a low
CNS
neuron,
retinal
ganglion
cells
(RGCs),
by
PSD-95
puncta,
whereas
in
the
presence
of
tamine
ous synapti
with glia (H). (I) Without glia,
napses, we counted synaptic
developed methods to isolate a definedthetype
of of synapses or to the efficacymicrographs
of synapses
be- frequency
number
of
developed
methods
type of
small spontaneglia numerous
puncta
arethan
apparent.
(D)
In to isolateofa defined
ce
glia using
correspond
to retinal
functional
syn44 # 18% autaptic neurons
served(10
(A),ng
immunopanning
the
cells
to
greater
99.5%
tween
RGC
neurons
cultured
in
CNS neuron,
ganglion cells
(RGCs),
by
ptic of
neurons
immunoous
synaptic
currents
is
obsynaptic transmission.
CNS
neuron,
retinal ganglion cells (RGCs), by
(BDNF)
the absence
of
glia
there
is
little
overlap
have
detectable
EPSCs,
wherethe
absence ofthem
gliaimmunopanning
(A,
and the
served
(A),
whereas
in the
immunopanning
cellssynapses
to greater than 99.5%
presence
(10 of
"M
To address this question,
cultured
cells to
greater
than 99.5%
purity we
(4,between
5) and puthen
in aleft
de-markers,
es.Glia
They
alsotheindicate
that the
few
E).
increased
number
as 100% have EPSCS in the
pre-culturing
andpresence
postsynaptic
presence
of gliathem
largeinspontaR
EPO
R pr
T
and
of glia
(B,and then
R E P O R T S whereasright)
purity
(4, 5) and then culturing them in a dewere
neous
syna
purity
(4,
5)
culturing
a
depresence
of
glia
(Mann-Whititive
synaptic
puncta
by
sixinmedium
the
of
glia
there
fined serum-free left
(5). No
Using
theseis aishighneous synaptic currents are
m between RGCsfinedin serum-free
the absence
andpresence
right).
difference
trodoto
medium of
(5). glia
Using these
ney U test, P % 0.05). (J)
fined
serum-free
medium un(5). Using these
frequently
degree of
overlap.
Bar,
#m.
(E)Fig.
Double
he average number of immuRGCs
(8). 50
Similar
results
were
obtained
observed(glia)
(B). inAll
Antibod
4. frequently
Glial
astrocytes
inshown
synaptic
ultrastrucweDepartment
haveapparent
previously
that
RGCs
Stanford
of Medicine,
Quantal content of RGCs culmethods,
we total
have previously
shownof
that
RGCsUniversity Schoolmethods,
methods,
we 2have
previously
shownwere
that CNQXRGCs
da that
thatformed
gliaonincrease
the
number
labeling
of
autaptic
neurons
indicates
that
synaptic
cur
synaptic
currents
der
both
conditions.
After
weeks
in
culture,
tophysi
RGCscultured
in the
of
Neurobiology,
Fairchild
Science
Building,
Stanford,
ture.
Bar,
200
nm.
(C)
The
numcrease
the
number
of
synapses
tured
in
the
absence
of
glia
is
Control
of
Synapse
Number
in the presence
of astrocyte-condicultured
in in
thethe
presence
oftoastrocyte-condicultured
in
the
presence
of
astrocyte-condisensitive
(10).
(C)
Application
fewer
synaptic
puncta
are
present
and
ant
we
used
whole-cell
patch-clamp
recording
CA
943055125,
USA.
sensitive
(10
7
#
4
compared
with
47
#
10
sence of per
glia was
35 !by
4 and
napses
RGC
sixto sevenfold.
Thelevels of
ber of synapses detected
byper
EM neuron.
(A pulses
and B)of levels
Electron
tioned
medium
have 10 times higher
tioned
medium
have
10oftimes
of
of 50-ms
100 !M
aptotag
absence
ofat glia
than
inthe
thesignificant
presence
of glia.
measure
enhancement
spon-higher
for
RGCs
cultured
with
glia
(intioned
medium
have
10
times
higher
levels
of
*To
whom
correspondence
should
be
addressed
increased
sevenfold
in
the
presof
50-ms
p
micrographs
ofglial
beBy comparison,
the
av- activity
L-glutamate
tosynapses
RGC
cellisbodies
synaptic
(6). absence
This glial
effect
is not
activity
(6).
This(Fig.
effect
not
rabbit w
by
wvely.
synapses
that do
form
in the
ofGlia
Bar,
50 ence
#m. of
(F)
About
sixsynaptic
times
as
many
cluding failures, P $ 0.004).
taneous
synaptic
activity
by
astrocytes
Stanford University School of Medicine,
Department
glia:
without
glia,
0.4
%
tween
RGC
neurons
cultured
in
L-glutamate
synaptic synaptic
activity
(6).
This
glial
effect
is
not
atofa astroglia,
holding
potential
of "70
f quanta released in response
termina
simply due to enhancement of RGC maturation,
simply
duecultured
to enhancement
of RGC
maturation,
Quantal content was deterpuncta
present
in0.7
RGCs
of Neurobiology, Fairchild Science Building,
Stanford,
1,are
A
and2.8
B) %
(9).
In per
the
absence
0.4;
with
glia,
cell absence
a are functionally
immature,
as either
indicated
mV results
in
inward
the
of
glia
left
and
All othe
M. Ullian,*
Stephanie
K.the
Sapperstein,
Christopherson,
at a holding
mined with the direct method.
CA 943055125,
USA.S.E-mail:
emu@stanford.edu
because
RGCs
cultured
either
insustained
the(A,
presence
or
because
RGCs cultured
in
presence
or Karen
mulation
in autaptic Erik
neurons
presence
of
glia
as
in
the
absence
of
simply
duein tothe
enhancement
of
RGC
maturation,
little
synaptic
activity
occurred
even
when
body and proximal neurite (P
& and
currents.
(D)
Current
densities
6.
F. W. Pf
right)
presence
of
glia
(B,
absencepuncta
of gliatheir
are
electrically
excitable, develBen
A. Barres
absence
glia are
electrically
excitable,
develglia: with
glia, either
34.5
!
4.3
GluR2/3
per
mV7.results
in
RGCs
cultured
with
presence
and absence
glia ofand
their high
failureof
rates
low
uptake
of
oftectal
RGCstarget
cultured
in the abbecause RGCs
cultured
inwere
the0.9
presence
0.013,
Students
t test).or
(D) and
K. Chris
left
right).
No
difference
is
op
comparable
numbers
of
dendrites
and
axons,
op
comparable
numbers
of
dendrites
and
axons,
cell;
without
glia,
6.1
!
GluR2/3
puncta
cells
(6).
This
difference
in
synaptic
activity
www.sciencemag.org
SCIENCE
VOL
291
26
JANUARY
2001
657
currents.
(D)
nd
7
!
4,
respectively.
The
sence
or
presence
of
glia.
R
E
P
O
R
T
S
are noexcitable,
differences
in apparent
the
B. A. Ba
M1-43. Although Although
functional
synaptic
activin synaptic
ultrastrucofperglia
areThere
electrically
develand
have
identical
survival
rates.
In
addition,
cell.
constitute
nearly
half of the cells in absence
our brain,
their
persisted
after
a month
culture,
indiand the
haveastrocytes
identical
survival
rates.
In addition,
Whole-cell
currents:
RGCs
of 8.RGCs
cu
ese values suggests that
total number
ofeven
vesicles
(34
%
6 in
Prepara
ture.
Bar,
200
nm.
(C)
The
numRGCs
cultured
under
conditions
synthefunction
is a long-standing
mystery.
Hereastrocytes
we op
show
by
quantal
that
itand
is
accounted
for
byboth
a matuwithout
glia,
15.7 # 4.7
pA/pF;
cultured
under bothneurobiological
conditions
(34), containing Neurobasal (Gibco), bovine
sera
comparable
ofcating
dendrites
axons,
Fig.
4. Glial
(glia)
in-numbers
has
reported
within
minutes
of
two synthepared
without
glia,
42 %
13not
with
glia)
punctabeen
that form
in theRGCs
pressence
or
p
ber of synapses detected by EM

Sinapsi e cellule gliali.

analyses,
FM1-43
and electron
microscopy
that few
size, secrete,
andimaging,
respond immunostaining,
to glutamate (6)crease
and
the number
of synapses

RGCsglia
with
glia, 45 #(6)
8.6and
pA/
secrete, and
respond
toregglutamate
rational delay.size,
To examine
whether

albumin, selenium, putrescine, triiodo-thyroni

gested w

orsurvival
docked vesicles
% 2 with- pF (P $ 0.015, Students t
and havethat
identical
rates. In(6.8
addition,
orrespond to functional
syn- nearly
Whole-cell
transferrin, progesterone, pyruvate
(1 (Falcon)
mM), g
increased
sevenfold
in the for
pressynapses
form inidentical
the absence
cellsper
andaneuron.
that the (A
fewand
synapses
do out
activity
by
enhancing
postsynexpress
nearly
identical
levels of mRNAs
a
express
levelsofofglial
mRNAs
for
B) Electron
glia,ulate
8.7 synaptic
% 0.8 with
glia)
per
tamine
(2
mM),
ciliary
neurotrophic
factor
(CN
test).
(E
and
F)
Autaptic
EPSCs
o indicate that few synapses
ence
of
glia:
without
glia,
0.4
%
without
glia,
RGCs
both
synthesurvive
form
functionally
immature.receptors,
Astrocytes
increase the of
number
ofcultured
mature,
apticconditions
responsiveness,
we of
measured
the re- receptors, voltlarge
battery
neurotransmitter
micrographs
synapses
be- under
largeare
battery
of neurotransmitter
voltsynapse.
(E) Double
labeling
of from
(10 ng/ml), brain-derived neurotrophic
fac
RGCs
cultured
inper
theproabwww.sciencemag.org
SCIENCE
VOL
291
26
JANUARY
2001
659
bovine
L
-glutamate
synapses
on
central
nervous
system
(CNS)
neurons
by
sevenfold
and
sponse
of
RGCs
to
pulses
of
0.4;
with
glia,
2.8
%
0.7
cell
RGCs in the absence functional
of
glia
age-dependent
ion
channels,
and
synaptic
RGCs
with
g
in respond
age-dependent ion channels, and synaptictween
pro- RGC neurons
size,cultured
secrete, and
glutamate(SC)
(6) and
superior tocolliculus
reveals sence (E) or presence (F) of
(BDNF) (50 ng/ml), insulin (5 "g/ml),
andmycin
forsko
(
applied to theteins,
cell somas.
Regardless
ofchip
the analysis
are
required
for synaptic
vitro.
We also of
show
body
and
proximal
neurite
(PIn&
as assessed
by
gene
(7).
theInabsence
gliathat
(A,most
left synand that synaptic
ncrease the total number
ofas assessed
teins,
by genemaintenance
chip analysisin(7).
pF
(P
$
0
(10
"M).
Recombinant
human
BDNF
and
CN
puncta
(green)
apglia.
(G
and
H)
Higher
synaptic
vate (1
express
nearly
levels
of mRNAs
for a0.013,
presence
of glia,
RGCs
responded
with
inapses are generated concurrently with theright)
development
of glia
inof
vivo.
These
Students
test).
(D)
this study,
whether
increased
and presence
glia
(B,identical
were provided by Regeneron Pharmaceuticals.
failure
rates aretthe
seen
in RGCs
thisThe
study, we examine whether the increased
pear around
postnatal
day we
6 examine
GC by six- to sevenfold.
test). shaken
(E and
ward
currents
(Fig.
1C)
that
were
blocked
by
data demonstrate a previously unknown left
function
for
glia
in
inducing
and
large
battery
of
neurotransmitter
receptors,
voltThere
are
no
differences
in
the
trodotoxin (TTX) and picrotoxin were enzyma
from R
synaptic
glia is due
increase
in
glia to
(G)anthan
in RGCs
theactivity
ap- inwithout
synaptic
activity in glia is due to an increase inand right). No difference is (P6), concurrently with
at do form in the absence
of
from
RGCs
c
glutamate receptor
antagonists
(10).
size
stabilizing CNS synapses, show that CNS synapse
number
can be ultrastrucprofoundly
Antibodies were obtained as follows: Anti-syn
apparent
in synaptic
total
number
of(H).
vesicles
(34 %
the number
of synapses
to
the
efficacy
of6
with The
gliaor
(I) Without
glia,
unless o
pearance
of
S100!-positive
asthe
number
of
synapses
or
to
the
efficacy
of
age-dependent
ion
channels,
and
synaptic
pronally immature, as indicated
sence
(E)
o
tophysin
(Sigma),
anti-GluR2/3
(Upstate
Biotec
of
the
postsynaptic
response,
however,
was
regulated by nonneuronal signals, and raise ture.
the possibility
that
glia
may
actively
# 18%
neurons
Bar, 200 nm. (C) The num- trocytes (red, arrows).
synaptic
without44glia,
42 %autaptic
13 with
glia)
9. Electrop
Bar,transmission.
synaptic
and antiPSD-95 (Chemicon). An antibody
to sH
teins, as assessed
by genethreefold
chip analysis
Inor50docked
larger (7).
in address
RGC
neurons
culturedwe
participate
in synaptic plasticity.
ilure rates and low uptake
of transmission.
have
detectable
EPSCs,
whereglia.
(G
and
ber
of
synapses
detected
by
EM
corded
To
this
question,
cultured
puvesicles
(6.8
%
2
with"m.
To address this question, we cultured puaptotagmin was generated by immunization
o
aswith
100%
have
EPSCS in the
with glia the
(Fig.increased
1D), consistent
the
in- with
peratur
gh functional synaptic activincreased sevenfold
instudy,
the presfailure
rates
this
we
examine
whether
out glia,
8.7
%
0.8
glia)
per
rabbit with a peptide corresponding to #70
the NH
presence
of
glia
(Mann-Whitcreased
amplitude
of
spontaneous
synaptic
Astrocytes,
which
ensheathe
synapses
throughrified
RGCs
in
the
presence
or
absence
of
m
ence of glia: without glia, 0.4 %
ported within minutes of two
(E)U Double
labeling
of
terminal lumenal portion of synaptotagmin
without
glia(21
synaptic
activity
in glia isminiature
due to an increase
insynapse.
ney
test, P %
0.05). (J)
(3 to
events
excitatory
out the CNS, are presently thought of as synastrocytes.
were
for
Stanford (miniature
University
School
of Medicine,
Department
0.4; withRGCs
glia, 2.8
%cultured
0.7 per
cell1 week
All other reagents were obtained from
Sigma.
superior
colliculus
(SC)
reveals
Stanford University School of Medicine, Department
Quantal
content
of
RGCs
culwith glia
(H
capillar
postsynaptic
currents,
mEPSCs)
aptic support cells, clearing ions and neurofollowed
by the
addition
of astrocytes
either
of Neurobiology,
Fairchildrecorded
Science Building, Stanford,
theneurite
number
or to the efficacy
oforthat
body and
proximal
(P of
& synapses
6. F. W. Pfrieger, B. A. Barres, Science 277, 1684
(199
of Neurobiology, Fairchild Science Building, Stanford,
synaptic
puncta
(green)
tured
indeterthe absence
of gliaapis
current
CA of
943055125,
USA.
in
the
presence
glia
(Fig.
2G).
To
transmitters
from
the
synaptic
cleft,
but
accuin
contact
with
RGCs
or
as
a
feeding
layer
for
44
#
18%
www.sciencemag.org
SCIENCE
VOL
291
26
JANUARY
2001
659
0.013,
Students
t
test).
(D)
CA 943055125, USA.
7. K. Christopherson, C. Harrington, S. Venkatapat
7 # 4 compared
with 47
# 106
synaptic transmission.
NaCl, 3
pear around
postnatal
day
mulating evidence suggests the possibility of
*To whom correspondence
be addressed
at
have detecta
There
B. A. Barres, data not shown.
for RGCsshould
cultured
withthe
glia (in7.3). Th
*To whom correspondence should be addressed
at are no differences in the
(P6),
concurrently
with
apStanfordpuUniversity School of Medicine, Department
more active roles as well (13). The role of glia
To address
total number of vesicles
(34 % 6 this question, we cultured
8. Preparation of astrocytes. Collicular
wereK-g
cluding
failures, P $ 0.004).
asglia
100%
hp
100
Stanford University School of Medicine, Department
pearance
of
S100!-positive
asof
Neurobiology,
Fairchild
Science
Building,
Stanford,
Fig. 1. Glial astrocytes
(glia)
at CNS ofsynapses
has been
difficult
to Building,
study Stanford,
#6
pared as described (23). Briefly, p1-2 SCstowere
without glia, 42 % 13 with glia)
Quantal
content was deterNeurobiology,
Fairchild
Science
10of
presence
CA 943055125,
USA.
E-mail:
emu@stanford.edu
(red,
50
enhancetrocytes
synaptic
efficacy
by
because CA
glial943055125,
cells are present
in CNSemu@stanford.edu
cultures
gested with trypsin and plated in tissue culture
fla
mined
witharrows).
the direct Bar,
method.
USA. E-mail:
autapti
or docked vesicles (6.8 % 2 withboth presynaptic
and postsynney
U
test
"m.
(Falcon)
in
a
medium
that
does
not
allow
neurons
and are often necessary for neuronal survival. In
tained
out glia, 8.7 % 0.8 with glia) per
aptic mechanisms. (A and B) In
survive SCIENCE
[Dulbuccos minimum
medium,
fe
Stanford
University
School of Medicine, Department
www.sciencemag.org
VOL 291essential
26 Quantal
JANUARY
2
this study, we have taken advantage of recently
con
Hepes,
synapse. (E) Double
labeling
of
the absence of glia, only a low
www.sciencemag.org
SCIENCE
VOL
291
26
JANUARY
2001
657
bovine serum (10%), penicillin (100 U/ml),
strep
creatine
developed methods to isolate a defined type of
of
Neurobiology,
Fairchild
Science
Building,
Stanford,
tured in the
superior colliculus (SC) reveals
frequency
spontaneThese findings raise
theof small
question
of
synapses and are necessary
maintain
synmycin (100 to
mg/ml),
glutamine
(2 mM) and Na-py

(50 U/m

Recettori dei Neurotrasmettitori (NT).

Proteine immerse nella membrana

plasmatica delle cellule
postsinaptiche e presinaptiche.
Il legame tra NT e recettore pu
causare direttamente o
indirettamente lapertura o la
chiusura dei canali ionici presenti
nella membrana postsinaptica o
presinaptica.

RECETTORI IONOTROPICI o canali ionici ligando-dipendenti.

RECETTORI METABOTROPICI o recettori accoppiati alle proteine G.

Recettori ionotropici.
poro

NEUROTRASMETTITORE

Legati direttamente al poro di selettivit.

Contengono due domini funzionali:
-Sito extracellulare che lega i trasmettitori.
-Regione che attraversa la membrana e
forma il canale ionico (poro).
Multipolimeri: quattro o cinque subunit
proteiche che contribuiscono a formare il
poro del canale ionico.
Producono una risposta veloce.

Recettori metabotropici.
NEURO
TRASMETTITORE

NEUROTRASMETTITORE

Il movimento dello ione attraverso il

canale dipende da pi tappe
metaboliche intermedie.
La loro struttura non comprende
direttamente canali ionici.
Agiscono sui canali ionici attivando
molecole intermedie: le proteine G.
I recettori sono proteine monomeriche
provviste di una regione:
- extracellulare dove si trova il sito di
legame del NT.
- intracellulare che lega le proteine G.
Producono una risposta lenta.

Trasmissione dellinformazione.
A. Recettori ionotropici

Acetilcolina (Rec. Nicotinici)

Glutammato (AMPA, Kainato, NMDA)
GABA (GABAA , GABAC)
Glicina
Serotonina (5-HT3)
ATP

B. Recettori metabotropici

Acetilcolina (Rec. Muscarinici)

Adrenalina e Noradrenalina
Dopomina
Serotonina (5-HT1, 5-HT2, 5-HT4)
Glutamato (mGluR1-4, mGluR6-8)
GABA (GABAB)

Rec. ionotropici vs. Rec. Metabotropici.

A. Recettori
ionotropici

B. Recettori
metabotropici

Tipologie di Sinapsi.
ECCITATORIE

INIBITORIE

Sinapsi che porta il potenziale della

membrana postsinaptica a valori pi
vicini al valore soglia in modo da
generare un PA.

Tipo di sinapsi che funziona in modo

tale da iperpolarizzare la membrana del
neurone postsinaptico o da
stabilizzarla al valore di riposo.

PPSE (pot. postsinaptico eccitatorio):

potenziale che fa aumentare la
probabilit di insorgenza di un PA
postsinaptico.

PPSI (pot. postsinaptico inibitorio): La

probabilit che un neurone
postsinaptico a livello del monticolo
assonale possa generare un PA
diminuisce.

Canali Na+ e K+.

Canali per K+ o Cl-.

Entrambi i tipi di potenziali sono graduati:

- Lampiezza dipende dal numero di molecole di NT che si legano al recettore.
- Decrescono allallontanarsi dal punto dove si sono formati.

Tipologie di Sinapsi chimiche.

ECCITATORIE

INIBITORIE

Allarrivo del PA il NT liberato nello spazio sinaptico per esocitosi ed il suo legame
con i recettori post sinaptici genera correnti in entrata (eccitatorie) od in uscita
(inibitorie) che modificano il Vm postsinaptico.

Inhibitory-Excitatory balance.
Homeostatic regulation of INHIB-EXC
balance.

INHIB

INHIB
INHIB

EXC
EXC

EXC

Physiological regulation of INHIB-EXC

imbalance.
Pathophysiological INHIB-EXC imbalance
VGLUT1
VGAT

GLU

GABA

AMPA
KAINATE
NMDA
mGLRUR

GABAA
GABAB
NERVOUS SYSTEM

VGLUT1

EPSP e IPSP.
I potenziali di inversione (Erev) e la soglia determinano leccitazione e linibizione
post sinaptica.

Se il Erev di un PSP pi positivo del valore di soglia leffetto di un NT di tipo

eccitatorio (EPSP), se pi negativo il NT inibitorio (IPSP). Gli IPSP possono in
ogni caso depolarizzare la cellula postsinaptica se il loro Erev compreso tra il
potenziale di riposo e il valore soglia.

Sinapsi eccitatorie rapide e lente.

A seconda che il NT eccitatorio attivi un recettore postsinaptico ionotroico o
metabotropico la risposta pu essere veloce o lenta.

Sinapsi eccitatorie rapide

Sinapsi eccitatorie
lente

Livelli di organizzazione sinaptica.

I microcircuiti sono modelli organizzativi costituiti dalle sinapsi di pochi elementi nervosi che possono
produrre operazioni funzionalmente rilevanti come:

CONVERGENZA SINAPTICA

F o n d a m e n t a l e
soprattutto nel SNC dove
i l p o t e n z i a l e
postsinaptico non
raggiunge il valore di
soglia e quindi non
riesce a generare da solo
un PA.
S.Glutammatergiche (GluR1),
GABAergiche (GABA-R) e
dendriti (MAP-2).

Integrazione: propagazione,
sommazione.
Sommazione spaziale

Potenziali post- sinaptici che

si generano simultaneamente in
diverse regioni della membrana
recettiva, si sommano.

Sommazione temporale

Potenziali post- sinaptici

prodotti in rapida successione a
livello della stessa sinapsi
possono sommarsi.
Sin. Ecc.

Vm (mV)

s.temp

Soma

s.spaz.

soglia
pot. rip.

Segm.
Iniz.
Sin. Inib.

s.spaz.

t (ms)

Lintegrazione sinaptica operata dal neurone fa s che quando la sommazione dei PPE prevale su
quelli inibitori ed in grado di portare il Vm del monticolo assonale al valore soglia, nasce un PA.

Modulazione Presinaptica.

Le sinapsi assoassoniche funzionano

come sinapsi modulatorie.
Nelle sinapsi assoassoniche il NT
rilasciato dal neurone presinaptico non
genera segnali elettrici nel neurone
postsinaptico.
Induce modificazioni del quantitativo di
Ca2+ che entra nel terminale assonico in
risposta ad un potenziale dazione
modificando il quantitativo di NT
rilasciato.

- Facilitazione presinaptica
- Inibizione presinaptica

Modulazione Presinaptica nelle sinapsi

assoassoniche.
1. Facilitazione presinaptica

2. Inibizione presinaptica

Neurotrasmettitori (NT).

Identificati circa 100 tipi diversi.

Un neurone pu sintetizzare e
rilasciare due o pi NT che possono
produrre effetti diversi sulla cellula
postsinaptica.
Due categorie, suddivise per loro
natura e processo di sintesi e
immagazzinamento:
- NT A BASSO PESO MOLECOLARE.
- NT PEPTIDERGICI.

Criteri per definire una molecola NT.

1.

Sintetizzati nel terminale presinaptico (o nel soma come

precursore e poi trasportati al terminale).

2.

Impacchettati in vescicole e rilasciati per esocitosi.

3.

La molecola deve essere rilasciata in risposta ad una

depolarizzazione presinaptica ed il rilascio deve essere Ca2+
dipendente.

4.

Diffondono nello spazio sinaptico legando selettivamente

recettori specifici postsinaptici, alterandone transitoriamente le
propriet biochimiche, o presinaptici dove esercitano una
modulazione autocrina.

5.

La loro azione bloccata da antagonisti recettoriali specifici.

6.

La loro azione termina per degradazione enzimatica e/o

ricaptazione nel terminale presinaptico o nelle cellule gliali
tramite trasportatori.

Natura chimica dei NT.

NT a peso basso molecolare:

NT Peptidergici:

Possono agire come cotrasmettitori allinterno della stessa sinapsi. Poich sono
immagazzinati da vescicole di tipo diverso il loro rilascio non avviene
contemporaneamente ed determinato dal tipo di attivit sinaptica:

Bassa frequenza di stimolazione rilascio di NT a basso peso molecolare.

Frequenza di stimolazione elevata rilascio anche di NT peptidici.

Sintesi e trasporto dei NT a basso peso molecolare.

terminale
presinaptico
VESCICOLE PICCOLE
a centro chiaro
(40-60 nm).

VESCICOLE
dendrite

La sintesi dei NT a basso peso molecolare si ha nelle

terminazioni dove poi vengono immagazzinati in vescicole
attraverso proteine trasportatori.
Gli enzimi necessari per la loro sintesi sono nel corpo
cellulare e sono trasportati nel citoplasma delle terminazioni
nervose a velocit di circa 0.5-5 mm/die (trasporto assonico
lento).
Per formare nuove molecole di NT i precursori vengono
assunti dalla terminazione nervosa presinaptica attraverso
proteine trasportatrici presenti nella membrana plasmatica
della terminazione.

Sintesi e trasporto dei NT peptidergici.

terminale
presinaptico
VESCICOLE

VESCICOLE GRANDI
a centro denso
(90-250 nm).

dendrite

I precursori dei NT peptidergici come gli enzimi che li

trasformano sono sintetizzati nel soma.
Il peptide prodotto a lunga distanza dal sito di rilascio.
Per questo motivo le vescicole che contengono il peptide
vengono trasportate nella terminazione presinaptica con
trasporto assonico rapido (velocit > 400 mm/die) durante
il quale le vescicole scivolano lungo i microtubuli spinti da
motori proteici ATP-dipendenti.

Rilascio dei NT.

nucleo chiaro.
(B): Neuropeptidi immagazzinati
in vescicole pi grandi (90 250
nm) a nucleo denso.

La stimolazione a bassa
frequenza provoca un aumento
preferenziale di ioni Ca2+ in
prossimit della membrana,
favorendo il rilascio del NT delle
vescicole piccole a centro
chiaro ancorate a strutture
presinaptiche specializzate.
La stimolazione ad alta
frequenza determina un
aumento pi generalizzato della
c o n c e n t r a z i o n e d i C a2+,
causando sia il rilascio di NT
peptidici da vescicole grandi a
centro denso, sia di NT a basso
peso molecolare da vescicole
piccole a centro chiaro.

Zone
attive

Le te
conte
vesci
trasm
ogget
trasm
I pep
pi le
effet
funzi

Principali sistemi neurotrasmettitoriali.

1.

DERIVATI DELLA COLINA: acetilcolina.

2.

AMMINE BIOGENE: catecolamine (dopamina,

adrenalina, noradrenalina), serotonina, istamina.

3.

AMINOACIDI: glutammato, aspartato, GABA,

glicina.

4.

NERUOPEPTIDI: vasopressina, ossitocina,

sostanza P.

Acetilcolina.

Prima sostanza identificata come NT

NT a basso peso molecolare
Dove agisce?
-Giunzioni neuromuscolari (unico NT)
-Sinapsi neuromuscolari tra nervo vago e le
fibrocellule muscolari cardiache.
-Sinapsi dei gangli della base del sistema
nervoso autonomo.
-Altre sinapsi del SNC.

Sistemi colinergici centrali implicati

in memoria e apprendimento.

Recettori dellAcetilcolina.
Recettori colinergici nicotinici.

rapidi

Recettori colinergici muscarinici.

lenti

Ammine biogene.
Regolano molte funzioni encefaliche sono anche attive a livello del SNP.
Sono coinvolte nella regolazione di diversi tipi di processi, dalle funzioni
omeostatiche centrali ai fenomeni cognitivi come lattenzione.
Alterazione della loro fisiologia causa disturbi psichiatrici.
Cinque ammine biogene hanno funzione di NT:
- Tre catecolammine: dopamina, noradrenalina, adrenalina.
- Istamina.
- Serotonina.

Dopamina.

La dopamina presente in diverse

regioni encefaliche. La zona pi ricca
il corpo striato che svolge un ruolo
fondamentale nella coordinazione del
movimento. Si ritiene coinvolta
anche negli stati di motivazione, di
ricompensa e di rinforzo.
I suoi recettori sono accoppiati a
proteine G (metabotropi).
E il NT carente nel morbo di
Parkinson (dove si ha una
degenerazione della sostanza nera),
e forse sovrabbondante in alcune
malattie mentali come la
schizofrenia.

Distribuzione della dopamina nellencefalo umano.

Noradrenalina.

La noradrenalina (o norepinefrina)
presente nel locus coeruleus, un
nucleo del tronco dellencefalo.
Influenza il sonno, lo stato di
veglia, lattenzione, il
comportamento alimentare e i
meccanismi di reazione.
Attiva due tipi di recettori
accoppiati alle proteine G (- e adrenergici).

Distribuzione della noradrenalina nellencefalo umano.

Adrenalina.

Ladrenalina (o epinefrina)
presente in minor quantit rispetto
alle altre catecolammine. Si trova
soprattutto nel sistema segmentale
laterale e nel bulbo, che proiettano
allipotalamo ed al talamo.
Due tipi di recettori accoppiati alle
proteine G (- e - adrenergici).

Distribuzione della adrenalina nellencefalo umano.

Istamina.

Listamina presente nellipotalamo.

Controlla lo stato di veglia e di
attenzione in maniera simile alle
proiezioni colinergiche e
noradrenergiche. Controlla anche le
attivit del sistema vestibolare.
Reazioni allergiche rilasciano
istamina dai mastociti: questo stretto
contatto, insieme alle potenti azioni
sui vasi, suggerisce che possa
influenzare il flusso cerebrale.
Tre tipi di recettori accoppiati alle
proteine G.
Distribuzione dellistamina nellencefalo umano.

Serotonina.

La serotonina generata a livello

dei nuclei del rafe e dai neuroni
mediani del ponte e del tronco
encefalico alto che inviano diffuse
proiezioni al proencefalo regolando
il sonno e la veglia. E il NT carente
negli stati depressivi e ansiogeni
(fluoxetina). Un eccesso provoca la
sindrome serotoninergica.
La maggior parte dei suoi recettori
sono metabotropi.

Distribuzione della serotonina nellencefalo umano.

Aminoacini eccitatori.
Il NT eccitatorio pi diffuso il glutamato, con molti ruoli a livello centrale (10 mM) e in
moltissime patologie nervose. Met circa delle sinapsi centrali lo libera. Il ciclo releaseuptake costituisce il meccanismo cellulare fondamentale di elaborazione rapida
dellinformazione, anche se pu avere effetti pi ritardati (centri glutamatergici bulbopontini: controllo liberazione di glicina durante il sonno; a livello ippocampale:
apprendimento e memoria).
I suoi recettori prendono il nome da agonisti esogeni che li attivano e sono ionotropi e
metabotropi.
IONOTROPI

METABOTROPI

Aminoacini inibitori.
I NT inibitori pi diffusi sono il GABA e la GLICINA. Un terzo delle sinapsi encefaliche
utilizza GABA come NT. Si trova prevalentemente in circuiti locali di interneuroni ma
anche nelle cellule di Purkinje nel cervelletto (coordinamento motorio). Controllo fine di
un network neuronale. La Glicina il NT inibitorio del midollo allungato e del midollo
spinale.
I recettori del GABA, tre tipi, sono inotropi, GABAA e GABAC, e metabotropi, GABAB.
Quello della glicina funzionalmente analogo al GABAA.

Agonista dei GABAA sono largamente

usati in neurofarmacologia. I
barbiturici sono agenti ipnotici ed
antiepilettici, le benzodiazepine
ansiolitici, sedativi, miorilassanti ed
anticonvulsivanti. Altri agonisti sono,
neurosteroidi, letanolo e zinco.
Antagonista sono la bicucullina e la
picrotoissina, entrambi agenti
convulsivanti.

Neuropeptidi.
Si trovano in tutte le regioni del SN e sono spesso corilasciati con altri
neurotrasmettitori. Possono agire anche come neuromodulatori (sostanza non in
grado, di per s, di generare un segnale nervoso o di arrestare un segnale, come un NT,
ma che modifica leffetto di un NT).

Sequenza aminoacidica dei peptidi oppioidi e altri neuropeptidi.

Trasmissione del dolore nel midollo spinale. a. In risposta a stimoli
dolorifici le DRG liberano sugli interneuroni del corno d. sostanza P
e GLU. Gli interneuroni della sost. gelatinosa liberano encefaline
(ENK) che bloccano la trasmissione eccitatoria delle cellule
gangliari. b. Nelle cellule gangliari delle radici dorsali, le ENK
riducono la durata dei PA al Ca2+.

Giunzione Neuromuscolare.

Sinapsi
NEURONE

NEURONE

Giunzione Neuromuscolare
MOTONEURONE

FIBRA
NEUROMUSCOLARE

Unit motoria e Giunzione Neuromuscolare.

La giunzione neuromuscolare (o placca motrice) la sinapsi periferica che
si instaura tra il motoneurone e la fibra muscolare dando origine allunit
motoria.
GIUNZIONE NEUROMUSCOLARE: ANATOMIA
E una delle sinapsi pi grandi del corpo.
Terminazione presinaptica con alto numero di zone attive.
Microdomini di canali del Ca2+ aumentano lefficienza del
meccanismo di rilascio.
Superficie postsinaptica a pieghe: ampia superficie
recettoriale.
Allineamento zone attive presinaptiche-zone recettoriali post
sinaptiche.
Ha una sola grande unit densa post sinaptica.

Unit motoria e Giunzione Neuromuscolare.

UNITA MOTORIA

GIUNZIONE NEUROMUSCOLARE

Motoneuroni.

accoppiamento elettromeccanico di eccitazione-contrazione

Prof. Davide Cervia - Fisiologia Fisiologia del muscolo scheletrico

11

Giunzione Neuromuscolare.

BOTTONE
SINAPTICO

membrana presinaptica nervosa

mitocondrio
zona attiva
vescicole sinaptiche
(acetilcolina)

canale del Ca2+

voltaggio dipendente

fessura sinaptica
(con acetilcolinesterasi)

membrana postsinaptica
muscolare

plica o cresta
(doccia sinaptica)

invaginazione delle creste

recettori ACh nicotinico

FIBRA
MIOFIBRILLARE

Na+

canali del
voltaggio dipendenti

Recettore nicotinico.

Ha una conduttanza di 25pS per 1.5 ms (entrano 35.000 cationi) e genera un dVm= 0.3 uV. Il contenuto
di una vescicola genera un dVm=0.5mV. Si aprono per ogni quanto rilasciato 1000-2000 canali.

(SMALL)

NON
AUTORIGENERATIVO

(LARGE)

AUTORIGENERATIVO

CORRENTE DI PLACCA

- Il canale attivato dallACh il solo responsabile della genesi

del potenziale di placca.
- Flusso dipendente dalla quantit di Ach.

Accoppiamento eccitazione-contrazione: generalit.

Il SNC controlla la contrazione delle fibre muscolari striate
attraverso i motoneuroni.
I segnali che partono dai motoneuroni hanno un effetto
eccitatorio sulle cellule muscolari (trasmissione colinergica).
Le cellule muscolari sono eccitabili.
Quando una cellula muscolare riceve un segnale da un
motoneurone la membrana si depolarizza e genera un PA che a
sua volta induce la contrazione.
Mentre un singolo motoneurone innerva pi fibre muscolari, una
singola cellula muscolare innervata da un solo motoneurone
che raggiunge il muscolo scheletrico.