共查询到20条相似文献,搜索用时 31 毫秒
1.
Laverty G Anttila A Carty J Reddy V Yum J Arnason SS 《Comparative biochemistry and physiology. Part A, Molecular & integrative physiology》2012,161(1):53-60
In primary cell cultures of the avian (Gallus gallus) renal proximal tubule parathyroid hormone and cAMP activation generate a Cl−-dependent short circuit current (ISC) response, consistent with net transepithelial Cl− secretion. In this study we investigated the expression and physiological function of the Na-K-2Cl (NKCC) transporter and CFTR chloride channel, both associated with Cl− secretion in a variety of tissues, in these proximal tubule cells. Using both RT-PCR and immunoblotting approaches, we showed that NKCC and CFTR are expressed, both in proximal tubule primary cultures and in a proximal tubule fraction of non-cultured (native tissue) fragments. We also used electrophysiological methods to assess the functional contribution of NKCC and CFTR to forskolin-activated ISC responses in filter grown cultured monolayers. Bumetanide (10 μM), a specific blocker of NKCC, inhibited forskolin activated ISC by about 40%, suggesting that basolateral uptake of Cl− is partially mediated by NKCC transport. In monolayers permeabilized on the basolateral side with nystatin, forskolin activated an apical Cl− conductance, manifested as bidirectional diffusion currents in the presence of oppositely directed Cl− gradients. Under these conditions the apical conductance appeared to show some bias towards apical-to-basolateral Cl− current. Two selective CFTR blockers, CFTR Inhibitor 172 and GlyH-101 (both at 20 μM) inhibited the forskolin activated diffusion currents by 38-68%, with GlyH-101 having a greater effect. These data support the conclusion that avian renal proximal tubules utilize an apical CFTR Cl− channel to mediate cAMP-activated Cl− secretion. 相似文献
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We observed intermediate conductance channels in approximately 20% of successful patch-clamp seals made on collecting tubules
dissected from Ambystoma adapted to 50 mm potassium. These channels were rarely observed in collecting tubules taken from animals which were maintained in tap water.
Potassium-adaptation either leads to an increase in the number of channels present or activates quiescent channels.
In cell-attached patches the conductance averaged 30.3 ± 2.4 (9) pS. Since replacement of the chloride in the patch pipette
with gluconate did not change the conductance, the channel carries cations, not anions. Notably, channel activity was observed
at both positive and negative pipette voltages. When the pipette was voltage clamped at 0 mV or positive voltages, the current
was directed inward, consistent with the movement of sodium into the cell. The pipette voltage at which the polarity of the
current reversed (movement of potassium into the pipette) was −29.6 ± 6.5(9) mV.
Open probability at 0 mV pipette voltage was 0.08 ± 0.03 and was unaffected when the apical membrane was exposed to either
2 × 10−6 or 2 × 10−5
m of amiloride. Exposure of the basolateral surface of the tubule to a saline containing 15 mm potassium caused a significant increase (P less than 0.001) in the open probability of these channels to 0.139 ± 0.002 without affecting the conductance of the apical
channel.
These data illustrate the presence of an intermediate conductance, poorly selective, amiloride-insensitive cation channel
in native vertebrate collecting tubule. We postulate that, at least in amphibia, this channel may be used to secrete potassium.
Received: 14 January 2000/Revised: 16 June 2000 相似文献
4.
Zuzana Tomaskova 《FEBS letters》2010,584(10):2085-2092
This minireview focuses on observation of the properties, functional significance, and modulation of single chloride channels in the mitochondrial inner membrane using two electrophysiological methods - the patch-clamp and bilayer lipid membrane methods. Measurements of parameters such as conductance, Cl−/K+ selectivity, voltage or pH dependence as well as their modulation by endogenous and exogenous compounds using individual mitochondrial chloride channels result in an unexpectedly wide range of values. This paper discusses the origin of this wide variety of channel parameters and the possible involvement of these channels in mitochondrial membrane potential oscillations, apoptosis, carrier function, and mitochondrial fusion and fission. 相似文献
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A. Huanosta-Gutiérrez A.E. Espino-SaldañaJ.P. Reyes A. PétrizR. Miledi A. Martínez-Torres 《Biochemical and biophysical research communications》2014
Oocytes of Xenopus tropicalis elicit a Ca2+-dependent outwardly rectifying, low-activating current (ICl,Ca) that is inhibited by Cl− channel blockers. When inactivated, ICl,Ca shows an exponentially decaying tail current that is related to currents generated by TMEM16A ion channels. Accordingly, RT-PCR revealed the expression of five alternatively spliced isoforms of TMEM16A in oocytes, which, after expression in HEK-293 cells, gave rise to fully functional Cl− channels. Upon hyperpolarization to −80 mV a transient current was observed only in isoforms that carry the exon 1d, coding for two potentially phosphorylatable Threonine residues. The identified isoforms are differentially expressed in several tissues of the frog. Thus, it appears that X. tropicalis oocytes express TMEM16A that gives rise to a Ca2+-dependent Cl− current, which is different from the previously reported voltage-dependent outwardly rectifying Cl− current. 相似文献
7.
Ugarte G Delgado R O'Day PM Farjah F Cid LP Vergara C Bacigalupo J 《The Journal of membrane biology》2005,207(3):151-160
We report that Drosophila retinal photoreceptors express inwardly rectifying chloride channels that seem to be orthologous to mammalian ClC-2 inward
rectifier channels. We measured inwardly rectifying Cl− currents in photoreceptor plasma membranes: Hyperpolarization under whole-cell tight-seal voltage clamp induced inward Cl− currents; and hyperpolarization of voltage-clamped inside-out patches excised from plasma membrane induced Cl− currents that have a unitary channel conductance of ∼3.7 pS. The channel was inhibited by 1 mM Zn2+ and by 1 mM 9-anthracene, but was insensitive to DIDS. Its anion permeability sequence is Cl− = SCN−> Br−>> I−, characteristic of ClC-2 channels. Exogenous polyunsaturated fatty acid, linolenic acid, enhanced or activated the inward
rectifier Cl− currents in both whole-cell and excised patch-clamp recordings. Using RT-PCR, we found expression in Drosophila retina of a ClC-2 gene orthologous to mammalian ClC-2 channels. Antibodies to rat ClC-2 channels labeled Drosophila photoreceptor plasma membranes and synaptic regions. Our results provide evidence that the inward rectification in Drosophila retinal photoreceptors is mediated by ClC-2-like channels in the non-transducing (extra-rhabdomeral) plasma membrane, and
that this inward rectification can be modulated by polyunsaturated fatty acid.
G. Ugarte and R. Delgado contributed equally to this work. 相似文献
8.
Summary K+, Cl–, and Ca2+ channels in the vacuolar membrane of tobacco cell suspension cultures have been investigated using the patch-clamp technique. In symmetrical 100mM K+, K+ channels opened at positive vacuolar membrane potentials (cytoplasmic side as reference) had different conductances of 57 pS and 24 pS. K+ channel opened at negative vacuolar membrane potentials had a conductance of 43 pS. The K+ channels showed a significant discrimination against Na+ and Cl–. The Cl– channel opened at positive vacuolar membrane potentials for cytoplasmic Cl– influx had a high conductance of 110pS in symmetrical 100mM Cl–. When K+ and Cl– channels were excluded from opening, no traces were found of Ca2+ channel activity for vacuolar Ca2+ release induced by inositol 1,4,5-trisphosphate or other events. However, we found a 19pS Ca2+ channel which allowed influx of cytoplasmic Ca2+ into the vacuole when the Ca2+ concentration on the cytoplasmic side was high. When Ca2+ was substituted by Ba2+, the conductance of the 19 pS channel became 30 pS and the channel showed a selectivity sequence of Ba2+Sr2+Ca2+Mg2+=10.60.60.21. The reversal potentials of the channel shifted with the change in Ca2+ concentration on the vacuolar side. The channel could be efficiently blocked from the cytoplasmic side by Cd2+, but was insensitive to La3+, Gd3+, Ni2+, verapamil, and nifedipine. The related ion channels in freshly isolated vacuoles from red beet root cells were also recorded. The coexistence of the K+, Cl–, and Ca2+ channels in the vacuolar membrane of tobacco cells might imply a precise classification and cooperation of the channels in the physiological process of plant cells. 相似文献
9.
Epithelial cells of toad (Bufo bufo) skin were isolated by treatments of the epidermis with collagenase and trypsin. Cl- channels in the basolateral membrane from soma or neck of mitochondria-rich cells were studied in cell-attached and excised
inside-out configurations. Of a total of 87 sealed patches only 28 (32%) were electrically active, and in these we identified
four different types of Cl- channels. The two major populations constituted Ohmic Cl- channels with limiting conductance (γ125/125) of 10 pS and 30 pS, respectively. A much rarer 150 pS Ohmic Cl- channel was also characterized. From i/V relationships of individual channels the following Goldman-Hodgkin-Katz permeabilities were calculated, 2.2 (±0.1) × 10-14, 5.7 (±0.7) × 10-14, and 32 (±2) × 10-14 cm3/sec, for the 10, 30 and 150 pS Cl- channels, respectively. The 30 pS channel was activated by hyperpolarization. The gating kinetics of the 150 pS channel was
complex with burstlike closures within openings of long duration. The fourth type of Cl- channel was studied in patches generating `noisy currents' with no discrete single-channel events, but with vanishing fluctuations
at pipette potentials near E
Cl. Noise analysis revealed a power spectrum with cutoff frequencies of 1.2 and 13 Hz, indicating that resolution of kinetic
steps was limited by small channel currents rather than fast channel gating. From the background noise level we estimated
the channel conductance to be less than 1.7 pS. Despite the fact that the majority of patches did not contain electrically
active Cl- channels, patches being active, generally, contained more than a single active channel. Thus, for the above three types of
resolvable channels, the mean number of active channels per patch amounted to 2.1, 1.4, and 2.0, respectively. This observation,
like the finding of few patches with several unresolvable channels, indicates that electrically active Cl- channels are organized in clusters.
Received: 10 October 1996/Revised: 8 January 1997 相似文献
10.
Several RNA-cleaving deoxyribozymes (DNAzymes) have been reported for efficient cleavage of purine-containing junctions, but none is able to efficiently cleave pyrimidine-pyrimidine (Pyr-Pyr) junctions. We hypothesize that a stronger Pyr-Pyr cleavage activity requires larger DNAzymes with complex structures that are difficult to isolate directly from a DNA library; one possible way to obtain such DNAzymes is to optimize DNA sequences with weak activities. To test this, we carried out an in vitro selection study to derive DNAzymes capable of cleaving an rC-T junction in a chimeric DNA/RNA substrate from DNA libraries constructed through chemical mutagenesis of five previous DNAzymes with a kobs of ∼ 0.001 min− 1 for the rC-T junction. After several rounds of selective amplification, DNAzyme descendants with a kobs of ∼ 0.1 min− 1 were obtained from a DNAzyme pool. The most efficient motif, denoted “CT10-3.29,” was found to have a catalytic core of ∼ 50 nt, larger than other known RNA-cleaving DNAzymes, and its secondary structure contains five short duplexes confined by a four-way junction. Several variants of CT10-3.29 exhibit a kobs of 0.3-1.4 min− 1 against the rC-T junction. CT10-3.29 also shows strong activity (kobs > 0.1 min− 1) for rU-A and rU-T junctions, medium activity (> 0.01 min− 1) for rC-A and rA-T junctions, and weak activity (> 0.001 min− 1) for rA-A, rG-T, and rG-A junctions. Interestingly, a single-point mutation within the catalytic core of CT10-3.29 altered the pattern of junction specificity with a significantly decreased ability to cleave rC-T and rC-A junctions and a substantially increased ability to cleave rA-A, rA-T, rG-A, rG-T, rU-A, and rU-T junctions. This observation illustrates the intricacy and plasticity of this RNA-cleaving DNAzyme in dinucleotide junction selectivity. The current study shows that it is feasible to derive efficient DNAzymes for a difficult chemical task and reveals that DNAzymes require more complex structural solutions for such a task. 相似文献
11.
Victor Sabanov 《生物化学与生物物理学报:生物膜》2007,1768(11):2714-2725
The Cl− channels of brown adipocytes electrophysiologically resemble outwardly rectifying Cl− channels (ORCC). To study tentative Ca2+ regulation of these channels, we attempted to control Ca2+ levels at the cytoplasmic side of the inside-out membrane patches with Ca2+-chelating agents. However, we found that the commonly used Ca2+-chelators EGTA and BAPTA by themselves influenced the Cl− channel currents, unrelated to their calcium chelating effects. Consequently, in this report we delineate effects of Ca2+-chelators (acting from the cytoplasmic side) on the single Cl− channel currents in patch-clamp experiments. Using fixed (1-2 mM) concentrations of chelators, two types of Cl− channels were identified, as discriminated by their reaction to the Ca2+-chelators and by their conductance: true-blockage channels (31 pS) and quasi-blockage channels (52 pS). In true-blockage channels, EGTA and BAPTA inhibited channel activity in a classical flickery type manner. In quasi-blockage channels, chelators significantly shortened the duration of individual openings, as in a flickering block, but the overall channel activity tended to increase. This dual effect of mean open time decrease accompanied by a tendency of open probability to increase we termed a quasi-blockage. Despite the complications due to the chelators as such, we could detect a moderate inhibitory effect of Ca2+. The anionic classical Cl− channel blockers DIDS and SITS could mimic the true/quasi blockage of EGTA and BAPTA. It was concluded that at least in this experimental system, standard techniques for Ca2+ level control in themselves could fundamentally affect the behaviour of Cl− channels. 相似文献
12.
Sitsapesan R 《The Journal of membrane biology》1999,168(2):159-168
Tonoplast K+ channels of Chara corallina are well characterized but only a few reports mention anion channels, which are likely to play an important role in the tonoplast
action potential and osmoregulation of this plant. For experiments internodal cells were isolated. Cytoplasmic droplets were
formed in an iso-osmotic bath solution according to a modified procedure. Ion channels with conductances of 48 pS and 170
pS were detected by the patch-clamp technique. In the absence of K+ in the bath solution the 170 pS channel was not observed at negative pipette potential values. When Cl− on either the vacuolar side or the cytoplasmic side was partly replaced with F−, the reversal potential of the 48 pS channel shifted conform to the Cl− equilibrium potential with similar behavior in droplet-attached and excised patch mode. These results showed that the 48
pS channel was a Cl− channel. In droplet-attached mode the channel rectified outward current flow, and the slope conductance was smaller. When
Chara droplets were formed in a bath solution containing low (10−8
m) Ca2+, then no Cl− channels could be detected either in droplet-attached or in inside-out patch mode. Channel activity was restored if Ca2+ was applied to the cytoplasmic side of inside-out patches. Rectification properties in the inside-out patch configuration
could be controlled by the holding pipette potential. Holding potential values negative or positive to the calculated reversal
potential for Cl− ions induced opposite rectification properties. Our results show Ca2+-activated Cl− channels in the tonoplast of Chara with holding potential dependent rectification.
Received: 30 March 1999/Revised: 10 August 1999 相似文献
13.
Summary Patch-clamp methods were used to study single-channel events in isolated oxyntic cells and gastric glands fromNecturus maculosa. Cell-attached, excised inside-out and outside-out patches from the basolateral membrane frequently contained channels which had conductances of 67±21 pS in 24% of the patches and channels of smaller conductance, 33±6 pS in 56% of the patches. Channels in both classes were highly selective for K+ over Na+ and Cl–, and shared linear current-voltage relations. The 67-pS channel was activated by membrane depolarization, whereas the activity of the 33-pS channel was relatively voltage independent. The larger conductance channels were activated by intracellular Ca2+ in the range between 5 and 500nm, but unaffected by cAMP. The smaller conductance channels were activated by cAMP, but not Ca2+. The presence of K+ channels in the basolateral membrane which are regulated by these known second messengers can account for the increase in conductance and the hyperpolarization of the membrane observed upon secretagogue stimulation. 相似文献
14.
Molecular-dynamics simulations were carried out to ascertain which of the potential multimeric forms of the transmembrane peptaibol channel, antiamoebin, is consistent with its measured conductance. Estimates of the conductance obtained through counting ions that cross the channel and by solving the Nernst-Planck equation yield consistent results, indicating that the motion of ions inside the channel can be satisfactorily described as diffusive. The calculated conductance of octameric channels is markedly higher than the conductance measured in single channel recordings, whereas the tetramer appears to be nonconducting. The conductance of the hexamer was estimated to be 115 ± 34 pS and 74 ± 20 pS, at 150 mV and 75 mV, respectively, in satisfactory agreement with the value of 90 pS measured at 75 mV. On this basis, we propose that the antiamoebin channel consists of six monomers. Its pore is large enough to accommodate K+ and Cl− with their first solvation shells intact. The free energy barrier encountered by K+ is only 2.2 kcal/mol whereas Cl− encounters a substantially higher barrier of nearly 5 kcal/mol. This difference makes the channel selective for cations. Ion crossing events are shown to be uncorrelated and follow Poisson statistics. 相似文献
15.
Antonella Gradogna Paola Imbrici Giovanni Zifarelli Antonella Liantonio Diana Conte Camerino Michael Pusch 《生物化学与生物物理学报:生物膜》2014
CLC-K chloride channels and their subunit, barttin, are crucial for renal NaCl reabsorption and for inner ear endolymph production. Mutations in CLC-Kb and barttin cause Bartter syndrome. Here, we identified two adjacent residues, F256 and N257, that when mutated hugely alter in Xenopus oocytes CLC-Ka's biphasic response to niflumic acid, a drug belonging to the fenamate class, with F256A being potentiated 37-fold and N257A being potently blocked with a KD ~ 1 μM. These residues are localized in the same extracellular I–J loop which harbors a regulatory Ca2 + binding site. This loop thus can represent an ideal and CLC-K specific target for extracellular ligands able to modulate channel activity. Furthermore, we demonstrated the involvement of the barttin subunit in the NFA potentiation. Indeed the F256A mutation confers onto CLC-K1 a transient potentiation induced by NFA which is found only when CLC-K1/F256A is co-expressed with barttin. Thus, in addition to the role of barttin in targeting and gating, the subunit participates in the pharmacological modulation of CLC-K channels and thus represents a further target for potential drugs. 相似文献
16.
Hideaki Ando Katsuhiro KawaaiKatsuhiko Mikoshiba 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》2014
IRBIT (also called AHCYL1) was originally identified as a binding protein of the intracellular Ca2 + channel inositol 1,4,5-trisphosphate (IP3) receptor and functions as an inhibitory regulator of this receptor. Unexpectedly, many functions have subsequently been identified for IRBIT including the activation of multiple ion channels and ion transporters, such as the Na+/HCO3− co-transporter NBCe1-B, the Na+/H+ exchanger NHE3, the Cl− channel cystic fibrosis transmembrane conductance regulator (CFTR), and the Cl−/HCO3− exchanger Slc26a6. The characteristic serine-rich region in IRBIT plays a critical role in the functions of this protein. In this review, we describe the evolution, domain structure, expression pattern, and physiological roles of IRBIT and discuss the potential molecular mechanisms underlying the coordinated regulation of these diverse ion channels/transporters through IRBIT. This article is part of a Special Issue entitled: Calcium signaling in health and disease. Guest Editors: Geert Bultynck, Jacques Haiech, Claus W. Heizmann, Joachim Krebs, and Marc Moreau. 相似文献
17.
Priti KatroliaPeng Zhou Pan ZhangQiaojuan Yan Yinan LiZhengqiang Jiang Haibo Xu 《Carbohydrate polymers》2012,87(1):480-490
A novel β-mannanase gene (CsMan5A) was cloned from Chaetomium sp. CQ31 and expressed in Pichia pastoris. It had an open reading frame of 1251 bp encoding 416 amino acids and contained two introns. The deduced amino acid sequence shared the highest similarity (73%) with the β-mannanase from Emericella nidulans and belongs to glycosyl hydrolase family 5. The recombinant β-mannanase (CsMan5A) was secreted at extremely high levels of 50,030 U mL−1 and 6.1 mg mL−1 in high cell density fermentor. The purified enzyme was optimally active at pH 5.0 and 65 °C and displayed broad pH stability (pH 5.0-11.0) and exhibited specificity towards locust bean gum (Km = 3.1 mg mL−1), guar gum (Km = 9.3 mg mL−1) and konjac powder (Km = 10.5 mg mL−1). It efficiently degraded mannan polysaccharides into mannose and mannooligosacccharides, and also hydrolyzed mannotriose and mannotetraose. These properties make CsMan5A highly useful in food, feed and paper/pulp industries. 相似文献
18.
Depletion of intracellular Ca2 + stores in mammalian cells results in Ca2 + entry across the plasma membrane mediated primarily by Ca2 + release-activated Ca2 + (CRAC) channels. Ca2 + influx through these channels is required for the maintenance of homeostasis and Ca2 + signaling in most cell types. One of the main features of native CRAC channels is fast Ca2 +-dependent inactivation (FCDI), where Ca2 + entering through the channel binds to a site near its intracellular mouth and causes a conformational change, closing the channel and limiting further Ca2 + entry. Early studies suggested that FCDI of CRAC channels was mediated by calmodulin. However, since the discovery of STIM1 and Orai1 proteins as the basic molecular components of the CRAC channel, it has become apparent that FCDI is a more complex phenomenon. Data obtained using heterologous overexpression of STIM1 and Orai1 suggest that, in addition to calmodulin, several cytoplasmic domains of STIM1 and Orai1 and the selectivity filter within the channel pore are required for FCDI. The stoichiometry of STIM1 binding to Orai1 also has emerged as an important determinant of FCDI. Consequently, STIM1 protein expression levels have the potential to be an endogenous regulator of CRAC channel Ca2 + influx. This review discusses the current understanding of the molecular mechanisms governing the FCDI of CRAC channels, including an evaluation of further experiments that may delineate whether STIM1 and/or Orai1 protein expression is endogenously regulated to modulate CRAC channel function, or may be dysregulated in some pathophysiological states. 相似文献
19.
The plasma membrane of Chara corallina was made accessible for patch pipettes by cutting a small window through the cell wall of plasmolyzed internodal cells. With pipettes containing Cl– as Ca2+ or Ba2+ (50 or 100 mm), but not as Mg2+ or K+ salt, it was possible to record in the cell-attached mode for long periods with little channel activity, randomly interspersed with intervals of transient activation of two Cl– channel types (cord conductance at +50 mV: 52 and 16 pS, respectively). During these periods of transient channel activity, variable numbers (up to some 10) of the two Cl– channel types activated and again inactivated over several 100 msec in a coordinated fashion. Transient Cl channel activity was favored by voltages positive of the free running membrane voltage (> –45 mV); but positive voltage alone was neither a sufficient nor a necessary condition for activtion of these channels. Neither type of Cl– channel was markedly voltage dependent. A third, nonselective 4 pS channel is a candidate for Ca2+ translocation. The activity of this channel does not correlate in time with the transient activity of the Cl– channels. The entire set of results is consistent with the following microscopic mechanism of action potentials in Chara, concerning the role of Ca2+ and Cl– for triggering and time course: Ca2+ uptake does not activate Cl– channels directly but first supplies a membrane-associated population of Ca2+ storage sites. Depolarization enhances discharge of Ca2+ from these elements (none or few under the patch pipette) resulting in a local and transient increase of free Ca2+ concentration ([Ca2+]cyt) at the inner side of the membrane before being scavenged by the cytoplasmic Ca2+ buffer system. In turn, the transient rise in [Ca2+]cyt causes the transient activity of those Cl– channels, which are more likely to open at an elevated Ca2+ concentration.The financial support by the Deutsche Forschungsgemeinschaft is gratefully acknowledged. 相似文献
20.
L. C. Stoner B. G. Engbretson S. C. Viggiano D. J. Benos P. R. Smith 《The Journal of membrane biology》1995,144(2):147-156
Patch clamp methods were used to characterize sodium channels on the apical membrane of Ambystoma distal nephron. The apical membranes were exposed by everting and perfusing initial collecting tubules in vitro. In cell-attached patches, we observed channels whose mean inward unitary current averaged 0.39±0.05 pA (9 patches). The conductance of these channels was 4.3±0.2 pS. The unitary current approached zero at a pipette voltage of –92 mV. When clamped at the membrane potential the channel expressed a relatively high open probability (0.46). These characteristics, together with observation that doses of 0.5 to 2 m amiloride reversibly inhibited the channel activity, are consistent with the presence of the high amiloride affinity, high sodium selectivity channel reported for rat cortical collecting tubule and cultured epithelial cell lines.We used antisodium channel antibodies to identify biochemically the epithelial sodium channels in the distal nephron of Ambystoma. Polyclonal antisodium channel antibodies generated against purified bovine renal, high amiloride affinity epithelial sodium channel specifically recognized 110, 57, and 55 kDa polypeptides in Ambystoma and localized the channels to the apical membrane of the distal nephron. A polyclonal antibody generated against a synthetic peptide corresponding to the C-terminus of Apx, a protein associated with the high amiloride affinity epithelial sodium channel expressed in A6 cells, specifically recognized a 170 kDa polypeptide. These data corroborate that the apically restricted sodium channels in Ambystoma are similar to the high amiloride affinity, sodium selective channels expressed in both A6 cells and the mammalian kidney.This work was supported by American Heart Association, New York Affiliate Grant 91007G (LCS) and National Institute of Diabetes and Digestive and Kidney Disease Grants DK-37206 (DJB) and DK46705 (PRS). 相似文献