Cells in Tissues

Cells in Tissues

Christopher J. Paradise, PhD

A. Malcolm Campbell, PhD
Cells in Tissues
Copyright Christopher J. Paradise and A. Malcolm Campbell. 2016.

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 Abstract
Two systems illustrate how individual cells of an organ system function,
communicate, and coordinate activities. The digestive system breaks
down and absorbs nutrients, and some specialized cells break down and
absorb nutrients. The case of parietal cells in the stomach and epithelial
cells in the small intestine are used to describe how cells function as a
unit within organ systems, coordinating activities and communicating
with one another. The endocrine system of insects affects molting and
metamorphosis, and specialized cells are also important in each of these
processes within that organ system. The experiments that were devised to
determine the role of hormones in insect molting and metamorphosis are
described. Finally, stem cells are healthy components of several different
systems in animal bodies and are described in relation to a disruption in
function. In this breakdown of function, cancer cells, in contrast to stem
cells, can abnormally affect cell cycle regulation.

Keywords
multicellular organisms, undifferentiated cells, stem cells, tissues,
muscle cells, neurons, alimentary canal, information, ingestion, diges-
tion, assimilation, homestasis, stomach, small intestines, epithelial
cells, epithelium, smooth muscle, gastric pits, parietal cells, histamine,
proton pump inhibitor, tubulovesicles, pepsinogen, pepsin, peristalsis,
endocrine system, hormones, exoskeleton, molting, epidermis, hemo-
lymph, corpus allatum, metamorphosis, protocerebrum, prothoracic
gland, neurosecretory cells, stem cells, cancer cells, tumors, multipo-
tent, tumor suppressors, mitogen, central nervous system, peripheral
nervous system, carcinoma
 Contents
Preface...................................................................................................ix
Acknowledgments....................................................................................xi
Introduction.........................................................................................xiii
Chapter 1 The Digestive System Breaks Down and Absorbs
Nutrients from Ingested Food............................................1
Chapter 2 Populations of Endocrine Cells in Animals Affect the
Whole Organism.............................................................17
Chapter 3 Similarities and Differences Between Stem Cells and
Cancer Cells.....................................................................27
Ethical, Legal, Social Implications: Decisions
Aboutto Whom and When Medical Interventions
are Given......................................................................38
Conclusion............................................................................................43
Glossary................................................................................................45
Index....................................................................................................49
 Preface
This book about cells and how they are part of tissues and organisms
is part of a thirty book series that collectively surveys all of the major
themes in biology. Rather than just present information as a collection
of facts, the reader is treated more like a scientist, which means the
data behind the major themes are presented. Reading any of the thirty
books by Paradise and Campbell provides readers with biological con-
text and comprehensive perspective so that readers can learn important
information from a single book with the potential to see how the major
themes span all size scales: molecular, cellular, organismal, population
and ecologic systems. The major themes of biology encapsulate the
entire discipline: information, evolution, cells, homeostasis and emer-
gent properties.
In the twentieth century, biology was taught with a heavy emphasis
on long lists of terms and many specific details. All of these details were
presented in a way that obscured a more comprehensive understanding.
In this book, readers will learn about cells in tissues and some of the sup-
porting evidence behind our understanding. The historic and more recent
experiments and data will be explored. Instead of believing or simply
accepting information, readers of this book will learn about the science
behind cells in the context of tissues the way professional scientists do
with experimentation and data analysis. In short, data are put back into the
teaching of biological sciences.
Readers of this book who wish to see the textbook version of this
content can go to www.bio.davidson.edu/icb where they will find
pedagogically-designed and interactive Integrating Concepts in Biology
for introductory biology college courses or a high school AP Biology
course.
 Acknowledgments
Publishing this book would not have been possible without the gener-
ous gift of Dr. David Botstein who shared some of his Breakthrough
Prize with co-author AMC. Davids gift allowed us to hire talented artists
(Tom Webster and his staff at Lineworks, Inc.) and copyeditor Laura
Loveall. Thanks go to Kristen Mandava of Mandava Editorial Services for
project management and guidance. In particular, we are indebted to Katie
Noble and Melissa Hayban for their many hours and attention to detail.
Kristen Eshleman, Paul Brantley, Bill Hatfield and Olivia Booker
helped us with technology at Davidson College. We are grateful to
administrators Tom Ross, Clark Ross, Carol Quillen, Wendy Raymond,
Verna Case, and Barbara Lom who had confidence in us and encouraged
us to persist despite setbacks along the way.
Thanks to my wife Amy Brooks for her constant support during the
development of this textbook, and my daughter Evelyn for her endless
energy. Thanks to Malcolm Campbell for his steadfast resolve and opti-
mism. Without him, this book would not exist. Thanks to collaborator
Laurie Heyer for taking my sometimes half-baked math ideas and turn-
ing them into powerful and elegant Bio-Math Explorations. I learned a
lot from both of them. While the math is largely absent from this book,
our collaboration with her made this a better book. Nancy Stamp at
Binghamton University, and Bill Dunson and Richard Cyr at The Penn-
sylvania State University influenced me greatly in how I think as a scientist
and approach my teaching. Finally, I thank my students in Integrated
Concepts in Biology II, who enthusiastically participated in our experi-
ment to redesign introductory biology, starting with the text and ending
with a new approach to teaching biology.
 Introduction

Individual cells are affected by a variety of factors and can disrupt the
function of other cells. Despite the focus on the cell as an individual
entity, no cell exists in isolation and many biologists study cells in the
context of the whole organism. In this book, the focus is on the popula-
tion, but cells are viewed as populations within multicellular organisms.
Over evolutionary time, some cell populations of certain species evolved
into colonies of interdependent individuals. From those colonies, mul-
ticellular organisms evolved. Within a multicellular organism, popula-
tions of similar cells exist and function as a unit. They make up tissues
and organs. Each cell type exists as a population of cells, and each has a
particular structure related to function. Some cells remain undifferenti-
ated, waiting for the signal to become active and develop into a particular
type of cell. These stem cells have some properties that are similar to
cancer cells. The description of these phenomena reflect the themes of the
concept of cells: All cells come from preexisting cells, cells maintain inter-
nal environments that differ from external environments, cell structure
defines cell function, and cells communicate with other cells.
 CHAPTER 1

The Digestive System

Breaks Down and
Absorbs Nutrients from
Ingested Food

In most multicellular organisms, cells make up tissues, which make up

organs, which make up organ systems. There may be various populations
of cells, or tissues, within one organ, and all the cells in the population
have similar structure and function. For instance, populations of muscle
cells respond similarly when neurons stimulate them. But the muscle
itself is made up of multiple types of cells. There are populations of con-
nective tissue cells, neurons, and other cells. Some of the cells within a
tissue are actually part of other systems. Neurons are part of the nervous
system and yet are found in all other systems in most animals. The cells,
tissues, organs, and systems are all highly interconnected, and the impact
of one population of cells upon an entire organism will be examined in
this book.
Even though cells at the tissue and organismal levels are investigated
in this book, whole organ systems will be considered to examine how
populations of cells may affect those systems. The digestive system is
an organ system that illustrates this. Specialized populations of cells
exist along the internal lining of the alimentary canal that are critical
in secreting digestive enzymes and absorbing nutrients. The alimentary
canal is a long tube in the body through which food passes after it is
ingested. Molecular information is detected by those cells, which trig-
gers release of enzymes and performs other functions as needed after a
meal is ingested. The processes of ingestion, digestion, and assimilation
2 CELLS IN TISSUES

are critical for maintaining homeostasis. Ingestion is the process of

taking food into the body. Digestion is the process of breaking down
food. Assimilation is the process of absorbing nutrients and incorporat-
ing them into the body. The components of the digestive system have
evolved in different animals for the particular diet of each animal. The
entire system is an emergent property that arises from the cell, tissue,
and organ components of the system.
The particular components of digestive systems of different animals
vary quite widely. Here, the human digestive system will be examined,
although we will examine data from closely related species in order to
understand the function of our own system. Early anatomists in the elev-
enth to fifteenth centuries (although Greek physicians began the practice
of dissection as early as 300 BCE), generated fairly accurate descriptions
and diagrams of the main structural components of the digestive system.
Often these anatomists worked on bodies of executed criminals, or they
worked in secret on cadavers due to the belief that souls of dissected bod-
ies could not go to heaven. Although many anatomists came before him,
Leonardo da Vinci provided detailed drawings of human digestive system
anatomy.
In large part, da Vinci did pretty well diagramming the positions of
the major organs of the human digestive system. The alimentary canal
and associated organs of the digestive system are all connected in many
ways to produce a functioning system that is an emergent property of its
component parts. The positions of the accessory glands and organs were
well-described by da Vinci.
Today the level of detail and knowledge of the anatomy, physiology,
and functions of the human digestive system is much greater than it was
500 years ago. The basic anatomy is well known (Figure 1). In general,
the mammalian digestive system consists of the alimentary canalalso
known as the gut, is the complete digestive tract through which ingested
food flows, is broken down, nutrients are assimilated, and waste is
expelled.
From mouth to anus, the alimentary canal of most mammals con-
sists of the esophagus, stomach, small intestine, and large intestine. In
humans, food (changing along the way) enters the mouth then passes to
the esophagus, stomach, small intestine, large intestine, and finally exits
 THE DIGESTIVE SYSTEM BREAKS DOWN FOOD 3

salivary gland

pharanx tongue

salivary gland

esophagus

liver
stomach
gall bladder
pancreas
duodenum

large intestine jejunum

ileum

rectum
anus

Figure 1 Anatomy of the human digestive system.

the anus as solid waste, or feces. Accessory glands (also part of the diges-
tive system) secrete digestive juices into the canal through ducts. These
accessory glands consist of salivary glands, pancreas, liver, and its stor-
age organthe gallbladder. Data will be analyzed that were collected in
order to learn about the function of the specialized cells mentioned previ-
ously that aid in digestion and assimilation of nutrients.
Various organs within the digestive system function to assist with
digestion and assimilation. For instance, the esophagus is a tube that
primarily functions to transport food to the stomach, the stomach does
some digestion but is also a food storage organ, the small intestine is
where most assimilation occurs, and the large intestine is where the solid
waste is formed and stored until defecation. Some of these organs will
be examined at the cellular level to discover how populations of cells in
these organs function in the digestion and assimilation processes. The
digestive system of animals is quite complex with organs of the system all
interconnected and with the system connected to other systems, such as
4 CELLS IN TISSUES

the circulatory and nervous systems. In this chapter only a small portion
of the system will be examined in order to understand how cells function
within larger systems.
The process of digestion follows ingestion of food and begins with
the simple act of chewing. Food is then swallowed; it travels down the
esophagus and enters the stomach. For centuries, scientists struggled with
determining whether the stomach secreted an acid and if so what kind
of acid. In 1823, William Prout definitively answered this question for
several species of mammal. Prout quantified the acid by feeding rabbits,
horses, cows, and dogs and then immediately removing the contents of
their stomachs. (He had to kill the animals to do this.) He then repeatedly
exposed the contents to distilled water and combined the mixtures. Prout
divided the mixture into four equal portions. The first was evaporated
to dryness, burned at high temperature, and dissolved again in distilled
water. He used silver nitrate to react with the chloride salts in the solution
to determine the amount of chloride ion (Table 1). To the second portion
Prout used a known amount of potassium hydroxide to exactly neutralize
the solution, allowing him to determine the amount of free acid present.
In the third fraction, Prout added a large quantity of potassium hydroxide,
which will react and neutralize all hydrochloric acid (HCl) to potassium
chloride and water. He then determined total chloride in the same manner
as before, with silver nitrate. The final fraction was used to determine if
any other acids were found.
Prout did not find any other acid in stomach secretions. He was only
able to find HCl. Of course, as soon as it is known that the stomach pro-
duces HCl, scientists wondered where the HCl was made and how it was
produced given that HCl is so destructive and could potentially eat away
the stomach itself.

Table 1 Results from chloride analysis of stomach contents of three

rabbits.
fraction of solution rabbit 1 rabbit 2 rabbit 3
chloride salts (first fraction) (g) 0.12 0.95 1.71
exact amount of base to neutralize acid (second fraction) (g) 1.56 0.76 0.40
chloride salts after neutralization (third fraction) (g) 1.59 2.22 2.72
total amount of chloride (g) 3.27 3.93 4.83
other acids (fourth fraction) 0 0 0

Source: From Prout, 1823.

 THE DIGESTIVE SYSTEM BREAKS DOWN FOOD 5

To address this question, something about the inner lining of the

stomach must be known. Careful microscopic examination led to knowl-
edge of the cellular makeup of the stomach wall. Like other organs, the
stomach consists of several types of cells. Most importantly, consider the
wall of the stomach to have several layers (Figure 2). The layer closest to
the lumen of the stomach, the inner open space, is called the mucosa.
Epithelial cells reside on the surface of the mucosa. Epithelial cells make
up the epithelium that encloses and protects organs and internal sur-
faces that have direct contact with the external environment. Inner to
the epithelium is connective tissue, which helps hold tissues and organs
together, and a thin layer of smooth muscle. The other layers, although
important to the functioning of the stomach, are not involved directly in
producing HCl. They consist of a submucosa with more connective tissue;
the muscularis externa beneath the submucosa, which consists of layers of
muscles arranged in different directions to help churn and move food;
and the serosa with more connective tissue.
Scientists discovered several cell types along the epithelium and that
the entire epithelium is arranged in convoluted pits called gastric pits.

stomach surface epithelium

gastric pit

mucosa
gastric gland

submucosa parietal
cell
muscularis
externa chief cell

serosa

enteroendocrine cell

Figure 2 Layers and cell types of the stomach. In the epithelium,

gastric pits lead to gastric glands that secrete gastric juice. The
gastric glands (one gland is shown enlarged on the right) contain
different types of cells that secrete a variety of enzymes, including
HCl, which activates the protein-digesting enzyme, pepsin.
6 CELLS IN TISSUES

These gastric pits are indentations that lead to the gastric glands from
which the secretions come. The human stomach has several million of
these pits, and further into each pit scientists discovered other types of
cells, which have different functions; one of which will be examined.
Figure 2 shows parietal cells, located in pits in the upper part of the
stomach, which are the cells that produce HCl. Any macromolecule that
is affected by acidic conditions can begin to break down in the stomach
after HCl is secreted. The parietal cells thus have an important role in the
digestion of food.
When a person eats, their stomach becomes distended and proteins
begin to be broken down into component amino acids; substances are
released that activate parietal cells (Figure 3). These substances include
gastrin, histamine, and acetylcholine. Activation leads to a series of events
at the molecular level within parietal cells.
Muallem and colleagues performed a study to determine how stim-
ulation of the parietal cells affected parietal cell internal pH. If pH of
individual cells goes up, then it can be concluded that hydrogen ions are
being excreted from the cell and pH of the surrounding environment goes
down. The scientists used a centrifugation technique to isolate parietal
cells from the epithelium of the upper part of a rabbits stomach. Keep
in mind the structure of the parietal cell, especially the deep infolding on
the lumen end of the cell, which increases the surface area for secretion.
The measurement of internal cell pH in parietal cell suspensions was
achieved through use of a dye that can permeate cells. This dye can be used

neuron

A parietal cell

A
G cell
ECL
neuron
cell HCl
H

G G
circ
ula
tion

Figure 3 Activation of parietal cells.

 THE DIGESTIVE SYSTEM BREAKS DOWN FOOD 7

to estimate pH inside the cells. The scientists measured the emission inten-
sity of the dye at two different wavelengths; the ratio of the intensities is
pH-dependent. Knowledge of the intensity ratio in solutions of known pH
allows researchers to estimate intracellular pH. The scientists incubated cells
with dye for 20 minutes and then washed the cells so that any dye not taken
up by cells was washed away. Keep in mind that relatively small changes in
pH, which is measured on a logarithmic scale, in single cell suspensions
could actually lead to large decreases in pH in the stomach lumen.
Muallem and colleagues exposed cells to a medium that contained no
sodium ions (Na+) and found that intracellular pH went down (Table 2).
When they added Na+, the pH rapidly went up. Addition of a chemical
that inhibited the Na+/H+ exchange protein at the same time that Na+
was added completely inhibited the rise in pH, illustrating that Na+/H+
exchange is important for maintaining intracellular pH in the resting cell.
Next, Muallem and colleagues exposed cells to histamine, a known
activator of parietal cells (Table 2). The Na+/H+ exchange inhibitor was
added in one treatment before exposure to histamine. Histamine binds

Table 2 Intracellular pH changes from experiments on

parietal cells. Negative values indicate declines in pH.
treatment pH
no Na+ 0.58
Na+ added after exposed to no Na+ 0.56
+ + +
Na and Na /H exchange inhibitor added after
0
exposed to no Na+
histamine 0.130 0.038
no Na+, then histamine 0.04
+ +
Na /H exchange inhibitor added, then histamine 0.04
histamine receptor antagonist, then histamine 0.01
histamine receptor antagonist, then histamine,
0.125 0.027
then chemical that increases cAMP
histamine receptor antagonist, then histamine,
then Na+/H+ exchange inhibitor, then chemical 0.01
that increases cAMP
H+/K+ATPase inhibitor, then histamine 0.09
no Na+, H+/K+ATPase inhibitor, then histamine 0
Na+/H+ exchange inhibitor added, then
0.02
H+/K+ATPase inhibitor, then histamine
Source: From Muallem et al., 1988, text and Figures 2 to 8.
8 CELLS IN TISSUES

to a receptor on parietal cells. A chemical known to block histamine-

receptor was added (an antagonist), after which histamine was added.
Binding of a ligand (such as, histamine) to a receptor often activates the
second messenger cyclic adenosine monophosphate (cAMP). A chemical
that raises cAMP levels in cells was added after action of histamine was
blocked by its antagonist to determine if histamine increased cAMP in
parietal cells (Table 2). If the Na+/H+ exchange inhibitor was also added,
there was essentially no change in intracellular pH.
Finally, an H+/K+-ATPase inhibitor was found to not substantially
affect histamine activation, but if there was no Na+ in the cell medium,
there was no effect of histamine. If the cells were exposed to the Na+/
H+ exchange inhibitor, there was no activation (Table 2). The H+/K+-
ATPase inhibitor is also called a proton pump inhibitor (PPI), and these
drugs are used to treat acid-related diseases, such as peptic ulcer disease.
In the absence of extracellular Na+ or in the presence of the Na+/H+
exchange inhibitor, intracellular pH increased only a very small amount.
The major contribution to the rise in intracellular pH is activity of the
Na+/H+ exchange protein. Prior to stimulation by histamine, gastrin,
or acetylcholine, parietal cells use Na+/H+ exchange to maintain pH
homeostasis. Most eukaryotic cells do this. Na+/H+ exchange proteins
are located on the side of the cell away from the lumen, and this should
make sense for a protein that is simply maintaining pH and not excret-
ing it into the stomach lumen. That end of the parietal cell also contains
Cl/HCO3 exchange proteins. These will become important when the
parietal cell is stimulated.
Upon stimulation, parietal cells secrete more acid than any other type
of cell in the body. Each activating chemical, histamine, gastrin, and ace-
tylcholine has a receptor on the parietal cell. Gastrin, which is released
from G cells in response to stretching of the stomach, nervous system
impulses, and presence of amino acids, also has a receptor on the special-
ized cells that secrete histamine and is what causes release of histamine.
The data suggest that histamine by itself produces a large increase in intra-
cellular pH and that it is in fact histamine having the effect, because
presence of the histamine receptor antagonist negates the effect of hista-
mine. Histamine has the effect of increasing cAMP levels in the parietal
cells, and this is because the effect of histamine receptor antagonist, then
 THE DIGESTIVE SYSTEM BREAKS DOWN FOOD 9

histamine, and then the chemical that increases cAMP leads to a large
increase in intracellular pH. This suggests that the histamine-histamine
receptor interaction leads to an increase in cAMP levels in the cell.
The increase in cAMP levels within the parietal cell leads to other
changes. The entire scheme is complex and not of it all is shown here, but
ultimately acid secretion at the lumen end of the parietal cell occurs. The
Cl/HCO3 exchange of proteins at the end of the cell away from the
lumen is responsible for expelling HCO3 and bringing in Cl.
Recall that in the absence of Na+, intracellular pH dropped dramati-
cally because the parietal cell could not rid itself of excess H+. In the
presence of Na+, the resting, non-stimulated cell has an intracellular
pH of about 7.1. In the absence of Na+ but the presence of the H+/
K+-ATPase inhibitor and histamine, there is a slight rise in intracellular
pH. This rise is larger when there is just Na+ and histamine but only
after a lag period. The researchers concluded that the H+/K+-ATPase
was responsible for the further rise in pH to about 7.3. The action of
this pump expelling H+ into the lumen is what leads to production
of HCO3. Expelling HCO3 brings Cl into the parietal cell, which
then diffuses to the lumen end and is then secreted into the lumen. The
K+ also leaks back out through a separate channel protein after being
exchanged with H+.
The acid secretion at the lumen end of the parietal cell results
in higher activity of anion exchange, which is caused ultimately by
the higher intracellular pH due to higher HCO3. This leads to main-
tenance of cell homeostasis; researchers have found that after the initial
increase in intracellular pH, if the parietal cell continues to secrete H+
and Cl, intracellular pH does not continue to rise. The combined
function of both exchangers is necessary for optimal acid secretion.
The parietal cell clearly has specialized proteins to facilitate the pro-
duction of acid. It also has a specialized morphology (Figure 4). The rest-
ing state looks very different from the stimulated state.
In the resting state, deep infoldings on the lumen end of the parietal
cell are seen. The H+/K+-ATPase pumps are sequestered within vesicles
called tubulovesicles. Activation of acid secretion leads to two func-
tional changes: tubulovesicles fuse with the membrane of the infolded
region that opens to the stomach lumen, and this membrane acquires
10 CELLS IN TISSUES

tubulovesicles

at rest

mitochondria

infolding

stimulated
nucleus
infolding

Figure 4 Schematic representation of parietal

cells at rest and under stimulation.

permeability to KCl, which allows both K+ and Cl to enter the lumen,

as shown earlier.
The stomach produces other substances, including the protein pepsino-
gen, which breaks down to pepsin in the presence of acid. Pepsin cleaves
proteins in ingested food. We have known about pepsin since 1836 when The-
odor Schwann described a water-soluble factor in gastric juice that digested
egg white. Mucus is also secreted by other cells. HCl, pepsinogen, and mucus
together make up gastric juice. The stomach lining is protected from gas-
tric juice by the epithelial cells, which produce and secrete a bicarbonate-rich
solution and form a HCO3-rich mucus layer that coats the mucosa and
protects the epithelial cells. Bicarbonate is alkaline (a base), and it neutralizes
the acid secreted by the parietal cells, producing water in the process. This
continuous supply of bicarbonate is the main way that the stomach protects
itself from digesting itself and the overall acidic environment.
Peristalsis, a rhythmic contraction of muscles of the alimentary tract,
moves the partially digested food from the stomach into the small intestine.
The small intestine is made up of three partsthe duodenum, jejunum,
 THE DIGESTIVE SYSTEM BREAKS DOWN FOOD 11

absorptive cells capillary

artery
vessel vein
carrying
blood
villi

lumen

muscle
layers lymphatic
villi vessel

lumen of small intestine

Na+ glucose 2K+ H2O

osmosis
Na+/K+-
ATPase

GLUT-2
transport
protein

glucose blood 3Na+ H2O osmosis

Figure 5 Small intestine epithelial cells and glucose transport.

and ileum, in that order. Digestion is completed, and assimilation of the

components of food (sugars, amino acids, and fats) occurs in the small
intestine. As with the stomach, the small intestine has populations of spe-
cialized cells that absorb nutrients. Most enzymes that break down macro-
molecules in the lumen of the small intestine are secreted by the pancreas.
As with the stomach and most other organs, the small intestine is
lined with epithelial cells of various types that play specialized roles in the
digestive process (Figure 5). Many of these cells play multiple roles and
can take up a variety of nutrients. The cells involved in glucose transport
will now be examined.
The three major classes of nutrientsproteins, lipids (fats), and
carbohydratesall undergo digestion in the lumen of the small intes-
tine. Proteins are degraded into peptides and amino acids before absorp-
tion. Lipids (fats) are degraded into fatty acids and glycerol. Some
12 CELLS IN TISSUES

carbohydrates are degraded into simple sugars or monosaccharides, such

as glucose. Enzymes break down starch to shorter oligosaccharides, and
then enzymes located on the lumen end of epithelial cells break these
short chains into monosaccharides.
The inner wall of the small intestine is also called the mucosa and is
lined with epithelial cells. Structurally, the mucosa is highly wrinkled.
Microscopic villi project from these folds, and capillaries project into the
villi from outside the small intestine. Capillaries are small blood vessels
that form a network between arterioles and venules in the circulatory sys-
tem. Nutrients, vitamins, and even toxins, pass from the lumen through
epithelial cells, into the inside of the villi, and then into the capillaries
or lymphatic vessels where they are transported to the rest of the body.
Lymphatic vessels are thin walled, valved structures that carry lymph,
a colorless fluid that contains white blood cells. This movement occurs
through diffusion or active transport. Individual epithelial cells also have
finger-like projections known as microvilli. These adaptations increase the
surface area of the small intestine. Undigested and unabsorbed material
passes into the large intestine.
Pia Rder and her colleagues studied glucose absorption and trans-
port in small intestine epithelial cells in mice. They took advantage of
mutant strains of mice that lacked the ability to produce two trans-
membrane proteins known to transport glucose: the sodium-dependent
glucose cotransporter (SGLT1) and glucose transporter 2 (GLUT2). The
researchers bred sglt1 wild-type (sglt1+/+), sglt1 mutant (sglt1/), glut2
wild-type (glut2+/+), and glut2 mutant (glut2/) mice.
Mice were fed a solution with radioactive 14C glucose. The research-
ers standardized the amount given to mice based on body mass. After
15 minutes the animals were euthanized, blood was collected, and
the small intestine was dissected out, everted, and washed thoroughly
for analysis. Blood was centrifuged and analyzed for radioactive 14C
(Figure 6). Sections of small intestine were used to measure incorpo-
rated radioactivity, which was used to determine glucose retention
(see Figure 6).
Sections of the epithelium of the jejunum were incubated with anti-
bodies for SGLT1 or GLUT2 and stained for cell nuclei. This allowed
the researchers to visualize localization of SGLT1 and GLUT2 along the
 THE DIGESTIVE SYSTEM BREAKS DOWN FOOD 13

tracer in plasma (dpm/10 ml)

350 ** 900 **

over 15 min. (nmol/cm)

glucose accumulation
300 750
250
600
200
450
150
300
100
50 150

0 0
A sglt1+/+ sglt1/ B sglt1+/+ sglt1/

tracer in plasma (dpm/10 ml)

750 * 1400 **
over 15 min. (nmol/cm)
glucose accumulation

1200
600
1000
450 800

300 600
400
150
200
0 0
C glut2+/+ glut2/ D glut2+/+ glut2/

Figure 6 Effects of SGLT1 and GLUT2 on glucose accumulation

and blood plasma glucose, as measured by amount of radioactive
tracer. Bars on left are wild-type and bars on right are mutant mice.
Error bars represent 1 standard error (SE). Statistical analyses
were performed using a t-test. *p-value 0.05. **p-value 0.01.
n = 4 to 6 mice per group. A, Mean accumulation of glucose in
intestinal tissue samples for sglt1 wild-type and mutant mice. B,Mean
amount of glucose in blood plasma for sglt1 wild-type and mutant
mice. C, Mean accumulation of glucose in intestinal tissue samples
for glut2 wild-type and mutant mice. D, Mean amount of glucose in
blood plasma for glut2 wild-type and mutant mice.
Source: Rder et al. 2014, Figures 1 and 3. PLoS One 9(2): e89977. doi:10.1371/journal.pone.
0089977. 2014 Rder et al.

epithelial cell membranes (Figure 7). In the images, rows of epithelial cells
that line up to form the villi of the small intestine can be seen (see also
Figure 5). Note where SGLT1 localizes relative to the cell nuclei and how
that differs from the localization of GLUT2.
By knocking out the sglt1 gene in mice and then comparing
responses to wild-type mice, scientists can determine the effect of the
SGLT1 transmembrane protein. This is also true for the glut2 mutants.
14 CELLS IN TISSUES

A B

C D

Figure 7 Immunostained cells from wild-type and mutant mice

jejunum epithelial cells showing localization of transmembrane glucose
transport proteins (gray shading) and cell nuclei (white/light gray
shading; used for reference). A, Localization of SGLT1 from sglt1+/+
wild-type mice. Two villi are shown with rows of epithelial cells.
Bright blue nuclei are cells within the villi, not the lumen. Note
that SGLT1 localizes away from the nucleus along the membrane
in contact with the lumen. B, Localization of SGLT1 from sglt1/
mutant mice. One villi is shown. C, Localization of GLUT2
from wild-type glut2+/+ mice. A portion of one villi is shown. D,
Localization of GLUT2 from mutant glut2/ mice. Portions of three
villi are shown.
Source: Rder et al., 2014, Figures 5 and 6. PLoS One 9(2): e89977. doi:10.1371/journal.pone.
0089977. 2014 Rder et al.

Mutantsglt1/ mice take up very little glucose into epithelial cells and
show very little glucose in the blood relative to wild-type mice. The scien-
tists concluded that the function of SGLT1 was to transport glucose into
the epithelial cell. If glucose is not getting into the epithelial cell, then it
cannot find its way into the blood. The localization of SGLT1 near the
lumen further confirms this. The lack of SGLT1 in the mutants confirms
that the mutant really is deficient in producing that protein.
 THE DIGESTIVE SYSTEM BREAKS DOWN FOOD 15

Glucose accumulation in epithelial cells is higher in mutant glut2/

mice than wild-type mice. But glucose concentrations are lower in the
blood of those mutant mice than in the wild-type mice. This suggests
that glucose can enter the epithelial cell but cannot get out. So it enters
but doesnt leave the epithelial cells in mutant glut2/ mice. It enters and
leaves normally in wild-type mice, and the response is similar to the
sglt1+/+ mice. GLUT2 localizes in membranes on the sides and away
from the lumen, appearing to surround the nucleus of epithelial cells (see
Figure 7). Again, the mutant is deficient in GLUT2.
In summary, epithelial cells of the small intestine possess two trans-
membrane proteins: one that is sodium-dependent and brings glucose
into the cell from the lumen, and one that transports it into the villi to
be picked up by the blood capillaries. These epithelial cells possess many
other specialized proteins that are used to take up other sugars, amino
acids, fatty acids, and other nutrients. This is convincing evidence for the
existence and function of two of these transmembrane proteins.
To complete this short tour of the digestive system, material that has
not been digested and absorbed by the small intestine enters the large
intestine. Here fermentation of any remaining digestible matter by the
gut flora occurs. Any remaining semisolid waste (termed feces) is removed
by the coordinated contractions of the intestinal walls (peristalsis), which
propels it forward to reach the rectum and exit via defecation from the
anus. Specialized cells occur in this organ too, although they will not be
examined here. Data on only a small part of the mammalian digestive sys-
tem has been explored here. Specialized populations of cells exist in each
organ and function to break down and absorb various nutrients into the
body. In the next chapter, populations of cells of another organ system,
the endocrine system, and how they also affect the whole organism, will
be examined.

Bibliography
Baron JH: The discovery of gastric acid, Gastroenterology 76:10561064,
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Baumont W: Experiments and observations on the gastric juice and physiol-
ogy of digestion, Cambridge, MA, 1929, Harvard University Press.
16 CELLS IN TISSUES

Rder PV, Geillinger KE, Zietek TS, et al.: The role of SGLT1 and GLUT2
in intestinal glucose transport and sensing, PLoS One 9(2):e89977,
2014.
Kousoulis AA, Tsoucalas G, Armenis I, et al.: From the hungry acid to
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 Index
Accessory glands, 3 Fats. See Lipids
Alimentary canal, 1, 2 Feces, 15
mammalian digestive 5-bromodeoxyuridine (BrdU), 30
system, 23
Antagonist, 8 Gallbladder, 3
Assimilation, 1, 2 Ganglia, 24
Gastric pits, 56
Betticher, Daniel, 34, 37 Gastrin, 8
Bicarbonate, 10 Glucose absorption and transport, 12
Bmi-1, 3032 Glucose transporter 2 (GLUT2),
effects on neural stem cells, 3334 1215
loss effect on self-renewal of CNS/ Gut. See Alimentary canal
PNS stem cells, 31
Hematopoietic stem cells (HSCs), 27
Cancer cells, 27, 38 Hemolymph, 19
ethical, legal, social implications of, Histamine, 78
3841 Homeostasis, 2
versus stem cells, 2741 Hormones, 17
Capillaries, 12 juvenile, 22, 23
Carbohydrates, 12 molting, 20, 22, 24, 2526
Carcinoma, 34 Human digestive system, anatomy
CDKN2, 32, 33, 35, 36, 38 of, 3
Central nervous system (CNS), 30, 31
Connective tissue, 5 Information, 1
Corpus allatum, 21, 23 Ingestion, 1, 2
Critical period, 19
Cyclic adenosine monophosphate Juvenile hormone, 2224
(cAMP), 89
Large intestine, 2
da Vinci, Leonardo, 2 Lipids, 11
Differentiated cells, 27 Liver, 3
Digestion, 1, 2 Lumen, 5
Luminescence detection system, 35
Endocrine system, 17, 25 Lung cancer, 34, 35, 36, 37
Epidermis, 18 Lymphatic vessels, 12
Epithelial cells, 5
small intestine, 11, 15 Medical intervention decision-
Epithelium, 5 making, ethical, legal, social
Esophagus, 2 implications of, 3841
Exoskeleton, 17 Metamorphosis, 21
molting of, 17, 18, 20 Microvilli, 12
50 INDEX

Mitogen, 29 Reverse transcriptase polymerase

Molting, 20, 22, 24, 2526 chain reaction (RT-PCR), 36
Morrison, Sean, 30, 32, 33 Rhodnius prolixus (R. prolixus), 18,
mRNA, 36 21, 23
Mucosa, 5, 12 brain transplant experiments, 24
Mucus, 10 decapitation experiments, 20, 23
Multicellular organisms, 1 molting of, 20
Multipotent, 28 Rder, Pia, 12
Muscle cells, 1
smooth, 5 Salivary glands, 3
Muscularis externa, 5 Schiavo, Terri, 3941
Schwann, Theodor, 10
Neurons, 1 Second messenger, 8
Neurosecretory cells, 25 Small cell lung carcinoma
Non-small cell lung carcinoma (SCLC), 34
(NSCLC), 34, 38 Small intestine, 2
epithelial cells and glucose
Organ transplantation, 40 transport, 11
Smooth muscle, 5
Pancreas, 3 Sodium-dependent glucose
Parietal cells cotransporter (SGLT1),
activation of, 6 1214
pH changes on, 67 Stem cells, 27, 38
representation at rest and under versus cancer cells, 2741
stimulation, 10 cell cycle regulation in, 2829
Pepsin, 10 multipotent, 28
Pepsinogen, 10 population of, 27
Peripheral nervous system (PNS), tissue-specific, 28
30, 31 Stomach, 2
Peristalsis, 10 layers and cell types of, 5
Persistent vegetative state (PVS), 39 lumen of, 5
Phosphorylate retinoblastoma
(Rb), 33 Tissues, 1
Population, 1 connective, 5
Proteins, 11 Tissue-specific stem cell, 28
Prothoracic gland, 25 Tubulovesicles, 9
Protocerebrum, 25 Tumors, 27
Proton pump inhibitor (PPI), 8 Tumor suppressors, 29, 34
Proto-oncogenes, 29, 34
Prout, William, 4 Wigglesworth, Vincent, 1726
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