Professional Documents
Culture Documents
www.sin-italy.org/jnonline www.jnephrol.com
Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, Ohio - USA
Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, Ohio - USA
Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, Ohio - USA
Abstract
[HCO-3]
pH = pK + log
[1]
s.PCO
The lungs control plasma PCO by exhaling the CO2 pro2
duced during aerobic respiration (1). The kidneys (2) regulate plasma [HCO3] by (i) reabsorbing the HCO3 filtered
in the glomeruli (~4,300 mmol/day) and (ii) transporting
into the plasma new HCO3 that neutralizes the H+ arising from sources such as the metabolic production nonvolatile acids (~70 mmol/day).
The renal proximal tubule (PT) is the major site of HCO3
reabsorption, reclaiming ~80% of the HCO3 filtered by
the glomerulus. Nearly all of the remaining 20% is reclaimed along the distal nephron segments (3). As shown
in Figure 1A, the PT cell secretes H+ from the cytosol
across the microvillus apical membrane into the lumen
via the Na-H exchanger (NHE), mainly NHE3 (4, 5), and
the vacuolar-type H+-ATPase (6). Carbonic anhydrase IV
is a membrane-associated carbonic anhydrase (CA) tethered by a glycosylphosphatidylinositol (GPI) anchor to
the outer leaflet of the apical membrane along the tubule
lumen, where it converts secreted H+ and luminal HCO3
to CO2 and H2O (7). The CO2 and H2O rapidly reenter the
cell across the apical membrane, facilitated, as we will
describe later, by the aquaporin 1 (AQP1) channel. In the
cytosol, CO2 and H2O are converted back into HCO3 and
H+ by CA II, the HCO3 exiting with Na+ across the basolateral membrane via the renal electrogenic Na+/HCO3
cotransporter (NBCe1-A) at a stoichiometry of 3:1 into
the interstitial space and ultimately the blood (8). Other
solutes (e.g., glucose, lactate, glutamine) move from the
lumen into the PT cell via a variety of Na+-coupled trans-
Introduction
The maintenance of a physiological ratio of the major bloodplasma buffer parameters, [HCO3] and PCO , is at the root of
2
S4
Fig. 1 - Model of acidbase metabolism by the proximal tubule (PT). A) Mechanism of HCO3 reabsorption in the early PT. The H+
pump (i.e., V-ATPase) and Na-K pump are unlabeled. B) Formation of new HCO3. AQP1 = aquaporin 1; CA II and CA IV = carbonic anhydrases 2 and 4; GDH = glutamate dehydrogenase; Gln = glutamine; Glu = glutamate; -KG2- = -ketoglutarate; NBCe1-A =
electrogenic Na/HCO3 cotransporter (1 variant A); NHE3 = Na-H exchanger 3; OAA = oxaloacetate; PEP = phosphoenolpyruvate;
PEPCK = phosphoenolpyruvate carboxykinase; SLC6A19 = system B neutral amino acid transporters; SNAT3 (aka SLC38A3) =
system N amino acid transporter. *Chronic metabolic acidosis up-regulates NHE3, GA, GDH, PEPCK and SNAT3.
(c) The final component of new HCO3 is coupled to ammonium secretion. The PT is the principal site of renal
ammonium synthesis (12), and the excretion of this NH4+
is the major route for excreting H+ equivalents in the urine,
in the following sequence of events. First, Na / amino acid
cotransporters mediate the uptake of glutamine (Gln)
across both the apical and basolateral membranes. The
system B neutral amino acid transporters (SLC6A19 and
SLC6A20) mediate the constitutive uptake of Gln across
the apical membrane (13), whereas the system N amino
acid transporter 3 (SNAT3 or SLC38A3) up-regulated
by acidosis mediates Gln uptake across the basolateral
membrane (13). Once inside the PT cell, the Gln enters
the mitochondrion and undergoes hydrolysis via glutaminase to form NH4+ plus glutamate, which then undergoes
oxidative deamination via glutamate dehydrogenase to
produce NH4+ plus -ketoglutarate (-KG2-). The newly
formed NH4+ dissociates to form intracellular H+ and NH3.
The PT cell extrudes the H+ across the apical membrane
via NHE3 and H+ pumps. The NH3 exits in parallel, probably both through the membrane lipid and via AQP1 (14).
Finally, the luminal H+ titrates the luminal NH3 to form
NH4+, much of which ultimately appears in the urine. The
metabolism of -KG2- produces CO2 and glucose (gluconeogenesis). In response to chronic metabolic acidosis, the PT adaptively up-regulates ammonium synthesis
to promote H+ excretion (12, 13, 15, 16).
In addition to responding to chronic acidbase disturbances, the PT also responds to acute disturbances. We
S5
Apical H+ extrusion
NHE3
The Na-H exchanger 3 (NHE3) is the most important H+-extruder along the PT apical membrane (17, 18). NHE3 utilizes
the inward Na+ gradient created by the basolateral Na-K
pump to exchange 1 H+ for 1 Na+, thereby extruding H+
against its electrochemical gradient (Fig. 1A). Adaptive responses to chronic respiratory or metabolic acidosis involve
increasing the abundance and activity of NHE3 protein in
the apical membrane (19).
The acute regulation of NHE3 involves the trafficking and
recycling of NHE3 along the microvilli of the apical membrane via interaction between the C-terminal PDZ motif of
NHE3 with the PDZ binding domain of the NHE regulatory
factor-1 (NHERF-1). In turn NHERF-1 links NHE3 to the actin
cytoskeleton through association with ezrin (20). Ezrin also
serves as a low-affinity cAMP kinase (PKA) anchoring protein (AKAP), enabling PKA to phosphorylate NHE3 (21). The
actin-based motor myosin VI is involved in trafficking NHE3
from the tip to the base of the microvilli thereby acutely suppressing H+ extrusion (20). A complex combination of phospho-serine modifications at the NHE3 carboxy terminus between amino acid residues 455 and 832 mediated both by
PKA and protein kinase C (PKC) are involved in controlling the majority of these acute regulatory stimuli. Generally,
NHE3 activity is inhibited in response to hormonal or other
stimuli that, via cAMP formation, enhance PKA. Conversely,
NHE3 activity is stimulated in response to PKC activation,
with serine modification being necessary (but not sufficient)
for increased NHE3 activity (21).
membrane VO ring-like structure that mediates H+ movement (subunits a-e) and a peripheral cytoplasmic V1 ATPase
(subunits A-H). Mutations in the kidney-specific subunit B1
encoded by ATP6V1B1 are found in individuals with autosomal-recessive distal renal tubular acidosis (dRTA) with
deafness (type 1b). Mutations occurring in another kidney
specific subunit a4, encoded by ATP6V0A4 cause dRTA
with preserved hearing (22). The trafficking of the a4 subunit
to the apical membrane along distal segments appears to
be the major regulatory mechanism for V-ATPase activity in
response to acid or NaHCO3 loading (23).
basolateral membranes of PT cells. Moreover, the movement of CO2 through apical AQP1 ought to contribute to
the reabsorption of HCO3. Preliminary work on isolated,
perfused mouse PTs (41) suggests that the knockout of
AQP1 reduces maximal HCO3 reabsorption by ~50%
CO2. Moreover, whereas the knockout of AQP1 has no effect on the transepithelial flux of HCO3 from the basolateral solution (bath) to the lumen, it reduces the transepithelial flux of CO2 by ~60%. This work would be the first
evidence that a channel plays a physiologically important
role in a mammalian tissue.
AE2 (SLC4A2)
Using microelectrodes on isolated PTs, Kondo and Frmter
demonstrated Cl-HCO3 exchange at the basolateral membrane of the S3 segment. Under physiological conditions,
this transporter would couple the exit of 1 HCO3 into the
interstitium to the uptake of 1 Cl (63). By semiquantitative
polymerase chain reaction (PCR), Brosius et al detected
AE2 in the convoluted and straight PT as well as more distal segments of rat kidney (64). Thus, AE2 may account for
the anion-exchange activity in the S3 segment.
Carbonic anhydrases
The carbonic anhydrase enzymes effectively bypass the
slow reaction in the sequence CO2 + H2O Slow H2CO3 Fast
HCO3 + H+, and thus are critical for HCO3 reabsorption
and the creation of new HCO3. Figure 1A summarizes the
disposition of CAs in the human PT.
CA II is the archetypal mammalian-class CA. It is a 29kDa cytosolic protein that is ubiquitously expressed, and
is among the fastest of CAs, achieving a turnover rate for
CO2 hydration of 1 106/s. Except for the thin ascending
limb, CA II is present throughout the nephron, accounting
for ~95% of the CA activity in the kidney (7). As illustrated
in Figure 1A, cytosolic CA II plays a central role by converting the CO2 that enters across the apical membrane
into H+ for secretion into the lumen plus HCO3 for export
across the basolateral membrane. The importance of CA
II is illustrated by the effect of inherited CA II deficiency,
which causes mixed proximal and distal (type 3) RTA (22)
accompanied by osteopetrosis (because of impaired osteoclast function) and cerebral calcification (65).
In humans, the 5% of renal CA activity not due to CA II
is accounted for by 2 integral membrane proteins: CA IV
and CA XII. The GPI-linked CA IV is localized both apically and basolaterally along the PT and TAL, and apically
in -intercalated cells of the cortical collecting duct and
cells of the medullary collecting duct (7). As shown in Figure 1A, apical CA IV catalyses HCO3 dehydration, thereby
consuming secreted H+ as well as generating membranepermeant CO2 and H2O. Consistent with this role, CA IV
has a higher HCO3 affinity and a greater HCO3 dehydratase activity than CA II (66).
The role of basolateral CA IV is unclear. Preliminary work
suggests that CA IV minimizes the changes in surface
pH caused by CO3= transport mediated by NBCe1-A, but
does not substantially change the activity of the cotransporter (52). Thus, CA IV may protect nearby proteins from
extreme pH fluctuations caused by NBCe1-A.
S7
Nitric oxide
Nitric oxide (NO) is produced in renal tubular epithelium by
the inducible isoform of nitric oxide synthase (iNOS). In situ
microperfusion studies on mouse PTs show that the knockout of iNOS reduces JV and JHCO . Nevertheless, the mouse
3
maintains normal acidbase status via unknown compensatory mechanisms (84).
Regulation by hormones
Angiotensin II
Angiotensin II (ANG II) is perhaps the most powerful hormonal modulator of Na+, fluid, and HCO3 reabsorption by
renal PTs. As we will see below (see the section Involvement of apical AT1 receptors, below), basolateral CO2
and HCO3 are equally powerful. Burg and Orloff were the
first to examine the effect of ANG II on fluid reabsorption
in isolated perfused PTs (68). Since then, many studies
involving isolated tubules (69-74) and micropuncture (7577) have reported that luminal or basolateral ANG II has
biphasic concentration-dependent effects, increasing the
fluid reabsorption rate (JV) and the HCO3 reabsorption
rate (JHCO ) at low ANG II concentrations, but decreasing
3
JV and JHCO at higher concentrations. ANG II acts via an3
giotensin II receptors type 1 (AT1) (78, 79), which are G
proteincoupled receptors (GPCRs), for both its stimulatory and inhibitory effects (80, 81).
Dopamine
Acting via autocrine and paracrine effects, dopamine is a
potent natriuretic hormone, acting in part by reducing apical NHE3 protein levels and thereby decreasing Na+ and
volume reabsorption by the PT (82).
Endothelin
Endothelins are small peptides that induce vasoconstriction but also have other important physiological roles. In
the renal PT, chronic metabolic acidosis increases endothelin-1 (ET-1) expression, enhancing its autocrine action on the apical endothelin-B receptor, which in turn
stimulates NHE3 (83).
S8
Basolateral pH
In the first series of experiments (Fig. 2A, B), we increased
pHBL from 6.8 to 8.0 while holding [HCO3]BL at 22 mM and
[CO2]BL at 5%. The middle symbol in Figure 2A (triangle) represents the JHCO under standard, equilibrated conditions: a
3
pHBL of 7.40, a [HCO3]BL of 22 mM and a [CO2]BL of 5%. The
triangle in Figure 2B represents the corresponding intracellular pH (pHi). Using OOE technology to lower pHBL to 6.8 or
raise it to 8.0 always at a [HCO3]BL of 22 mM and a [CO2]
of 5% produces a surprising result: no change in JHCO
BL
3
(Fig. 2A). Nevertheless, the isolated changes in pHBL produce sizeable changes in pHi (Fig. 2B). Thus, the PT cell can
not acutely regulate JHCO in response to changes in either
3
pHBL or pHi per se.
Basolateral [HCO3]
In the second series of experiments (Fig. 2C, D), we increased basolateral [HCO3] from 0 to 22 to 44 mM while
holding pHBL at 7.40 and [CO2]BL at 5%. The middle symbol
S9
Basolateral [CO2 ]
In the final series of these experiments (Fig. 3A, B), we increased [CO2]BL from 0 to 20% while holding pHBL at 7.40
and [HCO3]BL at either 22 mM (solid symbols) or 0 mM (open
symbols). The triangle in Figure 3A represents the same
standard, equilibrated conditions as in Figure 2A and C. Figure 3A shows that isolated increases in [CO2]BL from 0% to
20% cause graded increases in JHCO . As shown in Figure
3
S10
Fig. 4 - Replots of data from Figures 2 and 3. A) Dependence of JHCO3 on pHi for various out-of-equilibrium (OOE) protocols.
Here we plot the JHCO3 data from Figure 2A versus the pHi data from Figure 2B, and do the same for Figure 2C versus Figure 2D,
and for Figure 3A versus Figure 3B, respecting the symbols and colors in Figures 2 and 3. Data represented by solid symbols
from ref. (102); reproduced with permission in accordance with terms of original publication, Copyright 2005 by the National
Academy of Sciences. B) Dependence of JHCO3 on pHBL for various OOE protocols. As in panel A, we again replot the data from
Figures 2 and 3, but here as a function of pHBL. C) Dependence of JHCO3 on the [CO2]BL/[HCO3]BL ratio for various OOE protocols.
As in panels A and B, we replot the data from Figures 2 and 3, but here as a function of the ratio [CO2]BL/[HCO3]BL. D) Dependence of JHCO3 on the [HCO3]BL/[CO2]BL ratio for various OOE protocols. As in panel C, we replot the data from Figures 2 and 3,
but here as a function of the ratio [HCO3]BL/[CO2]BL. None of the 4 parameters on the x-axes can uniquely predict JHCO3. BL =
basolateral; JHCO3 = HCO3 reabsorption rate.
signal for regulating pHBL is that different acidbase disturbances can generate the same pHBL but different pHi values.
In other words, the pHi signal is degenerate (i.e., knowledge of pHi cannot inform the cell about the status of pHBL
or the identity of the acidbase disturbance). Furthermore,
if pHi were the critical signal, the dynamics of pHi regulation (e.g., the recovery of pHi from an acute intracellular acid
load; see (106-108)) would make JHCO quite unstable. The
3
PT cell faced with the dilemma of selfishly regulating its
own pHi while yet regulating blood pH as part of its raison
dtre seems to have evolved the only way it could have:
uncoupling pHi regulation from JHCO .
3
Figure 4B is similar to Figure 4A but a comparable plot with
pHBL on the abscissa, shows that pHBL is also not the unique
determinant of JHCO (see the vertical spread of JHCO data at
3
3
pHBL = 7.4).
Figure 4C is yet a third replot of the JHCO data in Figures
3
Involvement of EGFR
In the wake of the above work with OOE solutions (102), a
critical question is, how does the PT sense acute changes
of [CO2]BL and/or [HCO3]BL and transduce the signal(s)
in the PT cell to regulate bicarbonate reabsorption? In
studying the literature on gas-sensing by other organisms,
Patrice Bouyer (then in our group) learned that GillesGonzales et al (109) had demonstrated that the bacterium
Sinorhizobium meliloti (formerly Rhizobium meliloti) senses low O2 levels using a 2-component system comprising the regulatory proteins, FixL and FixJ. Low O2 levels
activate the His-kinase activity of the heme protein FixL,
which activates FixJ, which in turn activates genes encoding enzymes for nitrogen fixation (see (110)). In 1993,
Chang et al found that the ability of the plant Arabidopsis
to respond to ethylene, which acts as a hormone (111),
depends on the protein ETR1 (112), the C-terminal portion
of which is remarkably similar to both FixL and FixJ of the
bacterial 2-component system. Because the mammalian
cells do not have histidine kinases, Bouyer hypothesized
that the CO2-sensing mechanism of renal PTs requires a
receptor tyrosine kinase (RTK) or a receptor-associated
(i.e., soluble) tyrosine kinase (sTK) that would interact with
a membrane-bound CO2 sensor.
Zhou and Bouyer began to test a variety of tyrosine-kinase
S12
Involvement of RPTP
We became interested in receptor protein tyrosine phosphatase (RPTP) because it is a receptor protein tyrosine
phosphatase (see (117)) that has an extracellular ligandbinding domain that is homologous to the canonical CAs.
Joseph Schlessingers group cloned the cDNA encoding
RPTP (118) and created a RPTP-knockout mouse (119).
Barnea et al pointed out that the CA-like domain (CALD)
of RPTP lacks 2 of the 3 His residues needed for coordinating Zn2+, and thus suggested that the CALD would be
catalytically inactive (118). Preliminary work by Skelton et
al (120) indeed suggests that RPTP lacks CA activity, but
that a combination of 4 mutations (which render the CALD
more like CA II) engenders CA activity. Moreover, preliminary work by Zhou suggests that PTs from the RPTP-null
mouse cannot respond to alterations in either [CO2]BL (121)
or [HCO3]BL (122). RPTP mRNA is present in kidney (123),
and preliminary work that exploits a newly developed antibody suggests that RPTP is expressed at the basal but
not the lateral membrane of the PT (124). We hypothesize
that the CALD of RPTP senses CO2 and/or HCO3 and that
the phosphatase domain of RPTP then remodels ErbB1,
ErbB2 and/or other proteins responsible for transmitting the
CO2/HCO3 signal.
activity (138, 139) and enhanced HCO3 and water reabsorption (136). Evidence from rat PTs points to PKC- as
being the critical isoform (139). In preliminary work on isolated perfused rabbit S2 PTs, a PKC inhibitor eliminates
the CO2-evoked increase in JHCO (140). This result is con3
sistent with the hypothesis that PKC plays an important
role in the CO2 signal-transduction pathway.
Concluding remarks
Every cellular and bodily function depends on pH, everything from control of the cell cycle at one extreme
to the muscle contraction that underlies exercise at the
S13
References
1. Boron WF. Ventilation and perfusion of the lungs. In: Boron
WF, Boulpaep EL, eds. Medical physiology: a cellular and
molecular approach. 2nd ed. Philadelphia, PA: Saunders Elsevier; 2009:700-724.
2. Giebisch G, Windhager E. Transport of acids and bases. In:
Boron WF, Boulpaep EL, eds. Medical physiology: a cellular and molecular approach. Updated ed. Philadelphia, PA:
Saunders Elsevier; 2009:851-865.
3. Boron WF. Acid-base transport by the renal proximal tubule.
J Am Soc Nephrol. 2006;17:2368-2382.
4. Murer H, Hopfer U, Kinne R. Sodium/proton antiport in brushborder-membrane vesicles isolated from rat small intestine
and kidney. Biochem J. 1976;154:597-604.
5. Wang T, Hropot M, Aronson PS, Giebisch G. Role of NHE isoforms in mediating bicarbonate reabsorption along the nephron.
Am J Physiol Renal Physiol. 2001;281:F1117-F1122.
6. Gluck SL, Underhill DM, Iyori M, Holliday LS, Kostrominova
TY, Lee BS. Physiology and biochemistry of the kidney vacuolar H+-ATPase. Annu Rev Physiol. 1996;58:427-445.
S14
7. Purkerson JM, Schwartz GJ. The role of carbonic anhydrases in renal physiology. Kidney Int. 2007;71:103-115.
8. Soleimani M, Grassl SM, Aronson PS. Stoichiometry of Na+HCO3- cotransport in basolateral membrane vesicles isolated
from rabbit renal cortex. J Clin Invest. 1987;79:1276-1280.
9. Preston GM, Carroll TP, Guggino WB, Agre P. Appearance
of water channels in Xenopus oocytes expressing red cell
CHIP28 protein. Science. 1992;256:385-387.
10. Nielsen S, Smith BL, Christensen EI, Knepper MA, Agre
P. CHIP28 water channels are localized in constitutively
water-permeable segments of the nephron. J Cell Biol.
1993;120:371-383.
11. Schnermann J, Chou CL, Ma T, Traynor T, Knepper MA,
Verkman AS. Defective proximal tubular fluid reabsorption in
transgenic aquaporin-1 null mice. Proc Natl Acad Sci U S A.
1998;95:9660-9664.
12. Good DW, Burg M. Ammonia production by individual segments of the rat nephron. J Clin Invest. 1984;73:602-610.
13. Moret C, Dave MH, Schulz N, Jiang JX, Verrey F, Wagner CA.
Regulation of renal amino acid transporters during metabolic
acidosis. Am J Physiol Renal Physiol. 2007;292:F555-F566.
14. Musa-Aziz R, Jiang L, Chen LM, Behar KL, Boron WF. Con-
S15
48. Sassani P, Pushkin A, Gross E, et al. Functional characterization of NBC4: a new electrogenic sodium- bicarbonate cotransporter. Am J Physiol Cell Physiol. 2002;282:C408-C416.
49. Choi I, Aalkjr C, Boulpaep EL, Boron WF. An electroneutral
sodium/bicarbonate cotransporter NBCn1 and associated
sodium channel. Nature. 2000;405:571-575.
50. Wang CZ, Yano H, Nagashima K, Seino S. The Na+-driven Cl-/
HCO3- exchanger: cloning, tissue distribution, and functional
characterization. J Biol Chem. 2000;275:35486-35490.
51. Grichtchenko II, Choi I, Zhong X, Bray-Ward P, Russell JM,
Boron WF. Cloning, characterization, and chromosomal
mapping of a human electroneutral Na+-driven Cl-HCO3 exchanger. J Biol Chem. 2001;276:8358-8363.
52. Grichtchenko II, Boron WF. Surface-pH measurements in
voltage-clamped Xenopus oocytes co-expressing NBCe1
and CAIV: evidence for CO3= transport [abstract]. FASEB J.
2002;16:A795.
53. Igarashi T, Inatomi J, Sekine T, et al. Mutations in SLC4A4
cause permanent isolated proximal renal tubular acidosis
with ocular abnormalities. Nat Genet. 1999;23:264-266.
54. Horita S, Yamada H, Inatomi J, et al. Functional analysis of NBC1 mutants associated with proximal renal tubular acidosis and ocular abnormalities. J Am Soc Nephrol.
2005;16:2270-2278.
55. Suzuki M, Vaisbich MH, Yamada H, et al. Functional analysis of a novel missense NBC1 mutation and of other mutations causing proximal renal tubular acidosis. Pflgers Arch.
2008;455:583-593.
56. Dinour D, Chang MH, Satoh J, et al. A novel missense
mutation in the sodium bicarbonate cotransporter
(NBCe1/SLC4A4) causes proximal tubular acidosis and
glaucoma through ion transport defects. J Biol Chem.
2004;279:52238-52246.
57. Igarashi T, Inatomi J, Sekine T, et al. Novel nonsense mutation
in the Na+/HCO3- cotransporter gene (SLC4A4) in a patient
with permanent isolated proximal renal tubular acidosis and
bilateral glaucoma. J Am Soc Nephrol. 2001;12:713-718.
58. Demirci FY, Chang MH, Mah TS, Romero MF, Gorin MB. Proximal renal tubular acidosis and ocular pathology: a novel missense mutation in the gene (SLC4A4) for sodium bicarbonate
cotransporter protein (NBCe1). Mol Vis. 2006;12:324-330.
59. Lin SHP, Lo YF, Yang SS, Seki G. Severe metabolic acidosis
causes early lethality in NBC1 W516X knock-in mice [abstract]. J Am Soc Nephrol. 2009;20:33A.
60. Inatomi J, Horita S, Braverman N, et al. Mutational and functional analysis of SLC4A4 in a patient with proximal renal tubular acidosis. Pflgers Arch. 2004;448:438-444.
61. Igarashi T, Inatomi J, Sekine T, et al. Mutational and functional
analysis of the Na+/HCO3- cotransporter gene (SLC4AC) in
patients with permanent isolated proximal renal tubular acidosis and ocular abnormalities [abstract]. J Am Soc Nephrol.
2003;14:302A.
62. Boron WF, Chen L, Parker MD. Modular structure of sodium-coupled bicarbonate transporters. J Exp Biol. 2009;212:1697-1706.
S16
79. Sasaki K, Yamano Y, Bardhan S, et al. Cloning and expression of a complementary DNA encoding a bovine adrenal
angiotensin II type-1 receptor. Nature. 1991;351:230-233.
80. Baum M, Quigley R, Quan A. Effect of luminal angiotensin
II on rabbit proximal convoluted tubule bicarbonate absorption. Am J Physiol. 1997;273:F595-F600.
81. Zheng Y, Horita S, Hara C, et al. Biphasic regulation of renal
proximal bicarbonate absorption by luminal AT(1A) receptor.
J Am Soc Nephrol. 2003;14:1116-1122.
82. Bacic D, Kaissling B, McLeroy P, Zou L, Baum M, Moe OW.
Dopamine acutely decreases apical membrane Na/H exchanger NHE3 protein in mouse renal proximal tubule. Kidney Int. 2003;64:2133-2141.
83. Licht C, Laghmani K, Yanagisawa M, Preisig PA, Alpern RJ.
An autocrine role for endothelin-1 in the regulation of proximal tubule NHE3. Kidney Int. 2004;65:1320-1326.
84. Wang T. Role of iNOS and eNOS in modulating proximal
tubule transport and acid-base balance. Am J Physiol
Renal Physiol. 2002;283:F658-F662.
85. Boron WF. Acid-base physiology. In: Boron WF, Boulpaep EL,
eds. Medical physiology: a cellular and molecular approach.
2nd ed. Philadelphia, PA: Saunders Elsevier; 2009:652-671.
86. Wu MS, Biemesderfer D, Giebisch G, Aronson PS. Role
of NHE3 in mediating renal brush border Na+-H+ exchange: adaptation to metabolic acidosis. J Biol Chem.
1996;271:32749-32752.
87. Nowik M, Lecca MR, Velic A, Rehrauer H, Brandli AW, Wagner CA. Genome-wide gene expression profiling reveals renal
genes regulated during metabolic acidosis. Physiol Genomics. 2008;32:322-334.
88. Soleimani M, Bizal GL, McKinney TD, Hattabaugh YJ. Effect
of in vitro metabolic acidosis on luminal Na+/H+ exchange
and basolateral Na+:HCO3- cotransport in rabbit kidney proximal tubules. J Clin Invest. 1992;90:211-218.
89. Sasaki S, Berry CA, Rector FC Jr. Effect of luminal and peritubular HCO3- concentrations and PCO2 on HCO3- reabsorption in rabbit proximal convoluted tubules perfused in vitro. J
Clin Invest. 1982;70:639-649.
90. Mello-Aires M, MacLaughlin MA, Malnic G. Proximal tubular acidification in metabolic alkalosis. Braz J Med Biol Res.
1983;16:365-374.
91. Cogan MG. Effects of acute alterations in pCO2 on proximal HCO3-, Cl-, and H2O reabsorption. Am J Physiol.
1984;246:F21-F26.
92. Brazeau P, Gilman A. Effect of plasma CO2 tension on renal tubular reabsorption of bicarbonate. Am J Physiol. 1953;175:33-38.
93. Dorman PJ, Sullivan WJ, Pitts RF. The renal response to
acute respiratory acidosis. J Clin Invest. 1954;33:82-90.
94. Relman AS, Etsten B, Schwartz WB. The regulation of renal
bicarbonate reabsorption by plasma carbon dioxide tension.
J Clin Invest. 1953;32:972-978.
95. Chan YL, Giebisch G. Relationship between sodium and bicarbonate transport in the rat proximal convoluted tubule.
Am J Physiol. 1981;240:F222-F230.
96. Horie S, Moe O, Tejedor A, Alpern RJ. Preincubation in acid medium increases Na/H antiporter activity in cultured renal proximal
tubule cells. Proc Natl Acad Sci U S A. 1990;87:4742-4745.
97. Laghmani K, Preisig PA, Moe OW, Yanagisawa M, Alpern RJ.
Endothelin-1/endothelin-B receptor-mediated increases in
NHE3 activity in chronic metabolic acidosis. J Clin Invest.
2001;107:1563-1569.
98. Yamaji Y, Amemiya M, Cano A, et al. Overexpression of csk
inhibits acid-induced activation of NHE-3. Proc Natl Acad
Sci USA. 1995;92:6274-6278.
99. Yamaji Y, Tsuganezawa H, Moe OW, Alpern RJ. Intracellular acidosis activates c-Src. Am J Physiol. 1997;272:C886-C893.
100. Zhao J, Hogan EM, Bevensee MO, Boron WF. Out-of-equilibrium CO2/HCO3- solutions and their use in characterizing a
new K/HCO3 cotransporter. Nature. 1995;374:636-639.
101. Zhao J, Zhou Y, Boron WF. Effect of isolated removal of either
basolateral HCO3- or basolateral CO2 on HCO3- reabsorption
by rabbit S2 proximal tubule. Am J Physiol Renal Physiol.
2003;285:F359-F369.
102. Zhou Y, Zhao J, Bouyer P, Boron WF. Evidence from renal
proximal tubules that HCO3- and solute reabsorption are
acutely regulated not by pH but by basolateral HCO3- and
CO2. Proc Natl Acad Sci U S A. 2005;102:3875-3880.
103. Gluck S, Cannon C, Al-Awqati Q. Exocytosis regulates urinary acidification in turtle bladder by rapid insertion of H+
pumps into the luminal membrane. Proc Natl Acad Sci U S
A. 1982;79:4327-4331.
104. Schwartz GJ, Al-Awqati Q. Carbon dioxide causes exocytosis of vesicles containing H+ pumps in isolated perfused proximal and collecting tubules. J Clin Invest.
1985;75:1638-1644.
105. Bouyer P, Zhou Y, Boron WF. An increase in intracellular
calcium concentration that is induced by basolateral CO2
in rabbit renal proximal tubule. Am J Physiol Renal Physiol.
2003;285:F674-F687.
106. Roos A, Boron WF. Intracellular pH. Physiol Rev.
1981;61:296-434.
107. Bevensee MO, Boron WF. Regulation of intracellular pH. In:
Alpern RJ, Hebert SC, eds. Seldin and Giebischs the kidney:
physiology and pathophysiology. 4th ed. Burlington, MA:
Academic Press; 2008:1429-1480.
108. Boron WF. Regulation of intracellular pH. Adv Physiol Educ.
2004;28:160-179.
109. Gilles-Gonzalez MA, Ditta GS, Helinski DR. A hmoprotein
with kinase activity encoded by the oxygen sensor of Rhizobium meliloti. Nature. 1991;350:170-172.
110. Rodgers KR. Heme-based sensors in biological systems.
Curr Opin Chem Biol. 1999;3:158-167.
111. Ecker JR. The ethylene signal transduction pathway in plants.
Science. 1995;268:667-675.
112. Chang C, Kwok SF, Bleecker AB, Meyerowitz EM. Arabidopsis ethylene-response gene ETR1: similarity of product to
two-component regulators. Science. 1993;262:539-544.
113. Fry DW, Bridges AJ, Denny WA, et al. Specific, irreversible
S17
S18
CO2-induced stimulation of HCO3 reabsorption by renal proximal tubules. Am J Physiol Renal Physiol. 2007;293:F110-F120.
130. Burnier M. Angiotensin II type 1 receptor blockers. Circulation. 2001;103:904-912.
131. Fabiani ME, Dinh DT, Nassis L, Casley DJ, Johnston CI. In
vivo inhibition of angiotensin receptors in the rat kidney by
candesartan cilexetil: a comparison with losartan. Am J Hypertens. 2000;13:1005-1013.
132. Ito M, Oliverio MI, Mannon PJ, et al. Regulation of blood
pressure by the type 1A angiotensin II receptor gene. Proc
Natl Acad Sci U S A. 1995;92:3521-3525.
133. Sugaya T, Nishimatsu S, Tanimoto K, et al. Angiotensin II
type 1a receptor-deficient mice with hypotension and hyperreninemia. J Biol Chem. 1995;270:18719-18722.
134. Zhou Y, Boron WF. Effect of basolateral CO2 on the luminal
ANG II sensitivity of HCO3 reabsorption by rabbit S2 proximal
tubules [abstract]. FASEB J. 2008;22:760.2.
135. Weinman EJ, Shenolikar S. Protein kinase C activates the
renal apical membrane Na+/H+ exchanger. J Membr Biol.
1986;93:133-139.
136. Liu FY, Cogan MG. Role of protein kinase C in proximal bicarbonate absorption and angiotensin signaling. Am J Physiol.
1990;258:F927-F933.
137. Carraro-Lacroix LR, Malnic G. Signaling pathways involved with
the stimulatory effect of angiotensin II on vacuolar H+-ATPase in
proximal tubule cells. Pflgers Arch. 2006;452:728-736.
138. Houillier P, Chambrey R, Achard JM, Froissart M, Poggioli
J, Paillard M. Signaling pathways in the biphasic effect of
angiotensin II on apical Na/H antiport activity in proximal tubule. Kidney Int. 1996;50:1496-1505.
139. Karim ZG, Chambrey R, Chalumeau C, et al. Regulation by PKC
isoforms of Na+/H+ exchanger in luminal membrane vesicles isolated from cortical tubules. Am J Physiol. 1999;277:F773-F778.
140. Zhou Y, Bouyer P, Boron WF. CO2-evoked HCO3 reabsorption
in the rabbit S2 proximal tubule: Blockade by PKC inhibitor
RO-31-8220 [abstract]. FASEB J. 2005;19:A156.
141. Chen Y, Cann MJ, Litvin TN, et al. Soluble adenylyl cyclase
as an evolutionarily conserved bicarbonate sensor. Science.
2000;289:625-628.
142. Pastor-Soler N, Beaulieu V, Litvin TN, et al. Bicarbonate-regulated adenylyl cyclase (sAC) is a sensor that regulates pH-dependent V-ATPase recycling. J Biol Chem. 2003;278:49523-49529.
143. Hallows KR, Wang H, Edinger RS, et al. Regulation of epithelial Na+ transport by soluble adenylyl cyclase in kidney
collecting duct cells. J Biol Chem. 2009;284:5774-5783.
144. Tresguerres M, Parks SK, Salazar E, Levin LR, Goss GG,
Buck J. Bicarbonate-sensing soluble adenylyl cyclase is an
essential sensor for acid/base homeostasis. Proc Natl Acad
Sci U S A. 2010;107:442-447.
145. Ludwig MG, Vanek M, Guerini D, et al. Proton-sensing Gprotein-coupled receptors. Nature. 2003;425:93-98.
146. Jones WD, Cayirlioglu P, Kadow IG, Vosshall LB. Two
chemosensory receptors together mediate carbon dioxide
detection in Drosophila. Nature. 2007;445:86-90.