Functional characterization of a water channel of the nematode Caenorhabditis elegans.

A genome project for the species Caenorhabditis elegans has demonstrated the presence of eight cDNAs belonging to the major intrinsic protein (MIP) family. We previously characterized one of these cDNAs known as C01G6.1. C01G6.1 was confirmed to be a water channel and newly designated as AQP-CE1 [Am. J. Physiol. 275 (1998) C1459-C1464]. In this paper, we examined the function of another MIP protein encoded by F40F9.9. This cDNA encodes a 274-amino acid protein showing a high sequence identity with mammalian aquaporin-8 (AQP8) water channel (35%) and d-TIP (34%), an AQP of Arabidopsis. The expression of F40F9.9 in Xenopus oocytes increased the osmotic water permeability (P(f)) 10.4-fold, and the activation energy for P(f) from Arrhenius plot was 4.7 kcal/mol, suggesting that F40F9.9 is a water channel (AQP-CE2). AQP-CE2 was not permeable to glycerol or urea. Oocyte P(f) was reversibly inhibited by 58% after an incubation with 0.3 mM HgCl(2). To identify the mercury-sensitive site, four individual cysteine residues in AQP-CE2 (at positions 47, 132, 149, 259) were altered to serine by site-directed mutagenesis. Of these mutants, only C132S had a P(f) similar to that of the wild-type together with an acquired mercury resistance, suggesting that Cys-132 is the mercury-sensitive site. Similar results were obtained by the mutation of Cys-132 to alanine (C132A). Replacement of Cys-132 with tryptophan decreased P(f) by 64%, but P(f) was still 2.5 times higher than that of the control. Cys-132 is located in the transmembrane helix 3, close to the transition to the extracellular loop C. These results suggest that the transmembrane helix 3, including Cys-132, might participate in the aqueous pore formation, or, alternatively, that Cys-132 might contribute to the construction of the AQP protein.

Transcriptional regulation of the CLC-K1 promoter by myc-associated zinc finger protein and kidney-enriched Krüppel-like factor, a novel zinc finger repressor.

The expression of CLC-K1 and CLC-K2, two kidney-specific CLC chloride channels, is transcriptionally regulated on a tissue-specific basis. Previous studies have shown that a GA element near their transcriptional start sites is important for basal and cell-specific activities of the CLC-K1 and CLC-K2 gene promoters. To identify the GA-binding proteins, the human kidney cDNA library was screened by a yeast one-hybrid system. A novel member of the Cys2-His2 zinc finger gene designated KKLF (for "kidney-enriched Krüppel-like factor") and the previously isolated MAZ (for "myc-associated zinc finger protein") were cloned. KKLF was found to be abundantly expressed in the liver, kidneys, heart, and skeletal muscle, and immunohistochemistry revealed the nuclear localization of KKLF protein in interstitial cells in heart and skeletal muscle, stellate cells, and fibroblasts in the liver. In the kidneys, KKLF protein was localized in interstitial cells, mesangial cells, and nephron segments, where CLC-K1 and CLC-K2 were not expressed. A gel mobility shift assay revealed sequence-specific binding of recombinant KKLF and MAZ proteins to the CLC-K1 GA element, and the fine-mutation assay clarified that the consensus sequence for the KKLF binding site was GGGGNGGNG. In a transient-transfection experiment, MAZ had a strong activating effect on transcription of the CLC-K1-luciferase reporter gene. On the other hand, KKLF coexpression with MAZ appeared to block the activating effect of MAZ. These results suggest that a novel set of zinc finger proteins may help regulate the strict tissue- and nephron segment-specific expression of the CLC-K1 and CLC-K2 channel genes through their GA cis element.

Lack of a relation between serum potassium concentration and exercise hyperpnea in patients with chronic heart disease.

Exertional dyspnea, a major symptom of patients with chronic heart failure, mainly stems from an abnormally high ventilatory response to exercise. However, there has been considerable controversy surrounding the mechanisms of respiratory control during exercise, especially regarding the role of serum potassium. We investigated the relation between serum potassium concentration [K+] and ventilation (VE) during exercise before and after oral supplements of potassium chloride in cardiac patients. Thirteen patients with chronic heart disease performed a 6-min constant-work-rate exercise (65.8+/-11.1 W) with respiratory gas measurements before initiating oral supplements of potassium chloride, 4 weeks after continued supplements, and 4 weeks after discontinuing supplements. Blood was sampled from a forearm vein at rest before exercise and at the end of exercise for measurement of [K+] and blood gases. The [K+] at rest was 3.66+/-0.30 mmol/L before oral supplements of potassium and significantly increased to 4.08+/-0.31 mmol/L (p<0.01) after supplements. In spite of the significant increases in the [K+], resting VE was not changed. While serum [K+] during exercise was significantly higher after potassium supplements than before, exercise VE was not influenced by the changes in [K+] throughout the study period. The findings of the present study strongly suggest that the chronic increase in the serum [K+] has no influence on the resting or exercise VE in patients with heart disease.

Nonlinear ablation targeting an isthmus of critically slow conduction detected by high-density electroanatomical mapping for atypical atrial flutter.

Focused high-density atrial endocardial mapping was performed with a three-dimensional electroanatomical mapping system or a multielectrode basket catheter in six men and two women (mean age = 54 years) with atypical atrial flutter (AFL) to characterize its reentry circuit and identify its isthmus of critically slow conduction (ICSC). Activation mapping revealed figure-8 reentry with ICSC between a surgical atrial scars in three atypical AFLs following atriotomy, and between the crista terminalis (CT) and the inferior (IVC) or superior (SVC) vena cavae in atypical right atrial (RA) AFL in absence of prior atriotomy. Figure-8 double loop reentry was documented in one RA atypical AFL. ICSC was characterized by concealed entrainment with a post-pacing interval identical to the AFL cycle length, and a mid-diastolic fractionated electrogram, 129 +/- 23 ms in duration, spanning the isoelectric line between double potentials on adjacent area of conduction block. All AFLs were successfully ablated with 4.9 +/- 4.3 RF pulses applied at ICSC. A possible mechanism of atypical AFL consists of figure-8 reentry with ICSC between surgical scars in postoperative AFL, and between the CT and the IVC/SVC in RA AFL not preceded by cardiac surgery. Late and partial regeneration of conduction across the atriotomy scar can create an ICSC. Nonlinear ablation targeting ICSC can cure atypical AFL, whether it follows surgery or not.

Electrocardiographic characteristics of left ventricular outflow tract tachycardia.

Catheter ablation of idiopathic left ventricular outflow tract tachycardia (LVOT-VT) is rare because a safe ablation technique at this site has not been described, and serious complications may occur. This study compared the QRS morphology of LVOT-VT with that of idiopathic right ventricular outflow tract tachycardia. A comparison was made between the electrocardiographic characteristics of LVOT-VT originating from the supravalvular region of a coronary cusp (Supra-Ao group) with those of LVOT-VT originating from the infravalvular endocardial region of a coronary cusp of the aortic valve within the LV (Infra-Ao group). After precise mapping of the right ventricle, left ventricle, pulmonary artery, coronary cusps, and proximal portion of the anterior interventricular vein, there were 17 patients in whom VT was thought to be located at the LVOT by both activation and pace mapping. They were divided between a Supra-Ao group (n = 8), and an Infra-Ao group (n = 9). Analysis of the 12-lead electrocardiogram (ECG) revealed an S wave in lead I in all 17 patients. A precordial R wave transition was also observed at V1 or V2 in 16 patients (94%). In 7 of 8 patients (88%) with Supra-Ao LVOT-VT, no S wave was observed in either V5 or V6. In contrast, an Rs pattern was observed in both V5 and V6, or in V6 only, in 100% of the patients with Infra-Ao LVOT-VT. A LVOT-VT should be suspected when the ECG shows an S wave in lead I and an R/S ratio greater than 1 in lead V1 or V2, versus a coronary cusp location if there is no S wave in either lead V5 or V6.

Isolation and characterization of the human CLC-5 chloride channel gene promoter.

The human CLC-5 chloride channel is expressed mainly in the kidney and its mutations cause Dent's disease (a familial renal tubular syndrome with hypercalciuria, tubular proteinuria, rickets, nephrocalcinosis, and eventual renal failure). To gain insight into the regulatory mechanism of CLC-5 expression, a genomic clone that contains the 5'-flanking region of the human CLC-5 gene was isolated and characterized. Two types of 5'-ends of cDNA were isolated by 5'-rapid amplification of cDNA ends, and one of them, approximately 2.1 kbp upstream of ATG-containing exon II, was first identified in human. The major promoter activity was detected in the 5'-flanking region of this newly identified exon Ia. The sequence of the proximal 5'-flanking region contained an activator protein (AP)-1-like site and cAMP-responsive element, but it lacked a TATA box, a GC-rich element, and an SP-1 site. Deletion analysis of the 5'-flanking region showed that the fragments containing the AP-1-like element (TGACTCC) positioned at -38 exhibited high promoter activities in CLC-5 expressing LLC-PK1 cells, but that further deletions not containing this AP-1-like element resulted in a great loss of luciferase activities. Gel-retardation analysis demonstrated the existence of a specific protein binding to this AP-1-like element in LLC-PK1 cells, which seemed to differ from an authentic AP-1. This study clarified the key element of the human CLCN5 promoter, and the mutation in this region could be the cause of Dent's disease.

Identification of an acid-activated Cl(-) channel from human skeletal muscles.

ClC-4 gene was isolated as a putative Cl(-) channel. Due to a lack of functional expression of ClC-4, its physiological role remains unknown. We isolated a human ClC-4 clone (hClC-4sk) from human skeletal muscles and stably transfected it to Chinese hamster ovary cells. Whole cell patch-clamp studies showed that the hClC-4sk channel was activated by external acidic pH and inhibited by DIDS. It passed a strong outward Cl(-) current with a permeability sequence of I(-) > Cl(-) > F(-). The hClC-4sk has consensus sites for phosphorylation by protein kinase A (PKA); however, stimulation of PKA had no effect on the currents. hClC-4sk mRNA was expressed in excitable tissues, such as heart, brain, and skeletal muscle. These functional characteristics of hClC-4sk provide a clue to its physiological role in excitable cells.

Isolation and characterization of kidney-specific CLC-K2 chloride channel gene promoter.

CLC-K1 and CLC-K2 are highly homologous kidney-specific chloride channels, but they are expressed in the different nephron segments. To understand the molecular mechanisms of kidney-specific and nephron-segment-specific expression of CLC-K channel genes, the rat ClC-K2 gene promoter was cloned and compared with that of CLC-K1. In the 1.5-kb pair 5'-flanking region of the CLC-K2 gene, no TATA box was identified around the transcriptional start site, and the proximal region (-32 to -68) was characterized by a GA-rich motif that had a significant sequence similarity to that of the previously isolated CLC-K1 gene promoter. In contrast, the distal portion did not have significant sequence similarity to that of CLC-K1. Reporter gene assay and gel-retardation analysis revealed that the GA-rich motif and the binding of a specific protein(s) to this element were indispensable for the basal promoter activity of the CLC-K2 gene. These results suggest that the GA-rich element may have an important role in the promoter activities of the kidney-specific CLC-K1 and -K2 genes, but that the GA-element alone is not sufficient for the strict regulation of nephron-segment-specific expression of CLC-K1 and CLC-K2 genes.

Thermal preconditioning protects rat cardiac muscle cells from doxorubicin-induced apoptosis.

Doxorubicin (DOX=adriamycine), an effective chemotherapeutic agents for cancers, has severe cardiotoxicity. In the paresent study, we examined the protective effect of thermal preconditioning (TP) against apoptosis of rat cardiac muscle cells induced by DOX. Treatment with DOX (10 microM) for 24 hrs resulted in apoptosis of cardiac muscle cells, which was evaluated by examining "DNA ladder" formation and TUNEL staining. The number of TUNEL-positive cells was significantly decreased in cells subjected to TP by incubation at 42 degrees C for 30 min, 24 hrs prior to DOX-treatment. Antisense oligonucleotides of the heat shock protein (HSP) 70 blunted this effect. These results indicate that DOX-induced apoptosis in cardiac muscle cells is prevented by TP, at least in part, via a HSP70-mediated mechanism.

Acute lung injury after inhalation of water-proofing spray while smoking a cigarette.

A 34-year-old Japanese woman developed acute lung injury soon after inhaling a water-proofing spray which she applied onto her ski suit while smoking a cigarette at the same time. She initially demonstrated arterial hypoxemia (PaO2 = 59 mm Hg) and ground-glass opacities in both lung fields on the CT scan, which both returned to normal without any medication. Several water-proofing sprays, which are easily obtainable in Japan, contain 1,1,1-trichloroethane, liquefied petroleum gas and fluoride resin. Although these components have not been reported to be toxic to the lung yet, high concentrations of these components and/or the pyrolytic products of fluoride resin may have caused acute lung injury in this case.

Mercury-sensitive residues and pore site in AQP3 water channel.

Water channel function of all aquaporins (AQPs) but AQP4 can be inhibited by mercurial reagents. Mercurial reagents are believed to bind specifically to cysteine residues and block the aqueous pore of AQPs. Because of the low homology of AQP3 to other AQPs, it is not certain whether the pore structure of AQP3 is similar to that of the others. Determination of mercury-sensitive cysteine residues in AQP3 and comparison with those in other AQPs will help to resolve this question. When AQP3 was expressed in Xenopus oocytes, incubation with 0.3 mM HgCl2 decreased its osmotic water permability (Pf) by approximately 30%. To identify the mercury-sensitive site, six individual cysteine residues in human AQP3 (at positions 11, 29, 40, 91, 174, and 267) were altered by site-directed mutagenesis. Mutants of C11S and C11A had a similar basal Pf to wild-type but acquired mercury resistance. Replacement of Cys-11 with Trp, which possesses a large side chain, did not change Pf. Mercurial inhibition of Pf was still observed in five other Cys-to-Ser mutants. These results suggest that Cys-11 is the mercury-sensitive residue in AQP3 and that this residue might be independent of water channel function. Mutation of Tyr-212, a position corresponding to the mercury-sensitive residues in AQP1 and AQP2, to cysteine enhanced the mercurial inhibition of Pf. Y212W had no water channel activity. Expression of AQP3 increased glycerol permeability (Pgly) 3.1-fold, whereas Pgly of Y212W-expressing oocytes was similar to Pgly of control oocytes. Cysteine mutation at Tyr-212 increased the inhibitory effect of mercury on Pgly. These results suggest that the structure of the aqueous pore of AQP3 resembles those of AQP1 and AQP2 and support the hypothesis that water and small molecules share a common pore in AQP3.

Iron deposition at mineralization fronts and osteoid formation following chronic cadmium exposure in ovariectomized rats.

To investigate whether chronic exposure of cadmium (Cd) chloride induces osteomalacic lesions similar to Itai-itai disease (IID), ovariectomized rats were injected intravenously with the cadmium at doses of 0.05 and 0.5 mg/kg/day, 5 days per week, for 50 weeks. In six rats in the 0.5 mg/kg group, the administration was continued for up to 70 weeks. In the 0.5 mg/kg group, the plasma concentration of calcium was similar in the treatment and control groups throughout the treatment period. The urinary excretion of calcium increased from 20 weeks and the increase became marked from 40 weeks. Histopathologically, osteoid seams in the femur, tibia, and humerus were increased from 50 weeks, and these changes became prominent at 70 weeks. Hypertrophy and hyperplasia of chief cells in the parathyroid were also observed from 50 weeks. The osteoid morphometry of the trabecular bone of the femur and sternum revealed a dose-dependent increase in osteoid/bone volumes. Roentgenographs of the antebrachial and metacarpal bones taken at 70 weeks showed so-called paper bone. The bone Cd content markedly increased until 25 weeks, but thereafter decreased linearly for up to 70 weeks. In contrast to the Cd content, the iron content decreased until 25 weeks, but thereafter increased until 70 weeks. Undecalcified section of the humerus showed the deposition of iron and formation of osteoid at mineralization fronts. Our data suggest that osteomalacic lesions were caused by chronic Cd intoxication, and that iron, as well as Cd, was involved in osteoid formation.

Endothelin production in the rabbit collecting ducts of acute renal failure models. An immunohistochemical study.

Endothelin 1 (ET-1) production was examined in the rabbit nephron in acute renal failure (ARF) induced by uranyl acetate administration or by clamping the renal artery. Uranyl acetate dissolved in saline was injected intravenously at a dose of 0.8 mg/kg (n = 12). In the ischemic kidney experiment, 60 min of left renal artery clamping was carried out 1 week after removing the right kidney (n = 8). Plasma and urine concentrations of ET-1 were measured 0, 24, and 48 h after treatment, and immunohistochemical studies of renal tissues obtained 48 h after the experiments were carried out using ET-1 monoclonal antibody. The fractional excretion of ET-1 increased from 10 to 89% at 48 h in the uranyl acetate treated group and from 6 to 15% at 24 h in the renal artery clamping group, suggesting the existence of ET-1 secretion from the nephron in both types of ARF. By immunohistochemical examination, strong ET-1 expression was noted only in the collecting ducts. No staining was observed in other parts of the nephron in both types of ARF. The present study indicates that the increased expression of ET-1 probably reflects production by the collecting ducts in both rabbit ARF models.

Hyponatremia with consciousness disturbance caused by omeprazole administration. A case report and literature review.

A 68-year-old man developed severe consciousness disturbance after daily administration of 20 mg omeprazole for four days for the treatment of bleeding gastric ulcer. Systemic investigation revealed severe hyponatremia (111 meq/liter). Consciousness did not become clear until his sodium intake was increased to 480 meq/day, and his serum sodium concentrations reached 130 meq/liter. After the discontinuation of omeprazole, his serum sodium levels returned to the normal range with only minimum supplementation of sodium in the form of dietary sodium chloride intake of 10 g/day. Although the mechanism of hyponatremia induced by omeprazole is not clear, an excessive loss of urinary sodium appears to be more likely than water retention with an increase in fluid intake. The literature was also reviewed.

Cloning, tissue distribution, and intrarenal localization of ClC chloride channels in human kidney.

Two kidney-specific chloride channels, ClC-K1 and ClC-K2, have been isolated from rat kidney. In the present study, we sought to isolate human homologue of rat ClC-K2 chloride channel that was present in the thick ascending limb of Henle's loop and collecting ducts. Human kidney cDNA library was screened with the whole rat ClC-K2 cDNA probe. Two highly homologous but not identical cDNAs were isolated and sequenced. Northern analysis showed that both clones were expressed only in kidney among various human tissues, demonstrating that kidney-specific ClC family members were also present in human kidney. Because both clones had almost the same nucleotide identity (approximately 80%) with rat ClC-K2, we could not determine by sequence alone which human clone corresponded to rat ClC-K2. Accordingly, we performed reverse transcription PCR using dissected human nephron segments and identified the site of expression of each clone in human nephron segments. One clone was only expressed in the thin limb of Henle's loop and the other was expressed in glomeruli, proximal tubules, and collecting ducts. We identified the latter clone as human ClC-K2 based on the localization of rat ClC-K1 and ClC-K2. Identification of human ClC-K2 clone will be of help in understanding the genetic involvement of chloride channel in disorders of chloride transport such as Bartter's syndrome.

Two isoforms of a chloride channel predominantly expressed in thick ascending limb of Henle's loop and collecting ducts of rat kidney.

Complementary DNAs encoding rat kidney chloride channels (ClC-K2L and ClC-K2S) were isolated by a polymerase chain reaction cloning strategy. Degenerate primers were designed based on the significant amino acid identity of the previously cloned chloride channels (ClC-0, -1, -2, and -K1). The 687-amino acid protein encoded by ClC-K2L is about 80% identical to rat ClC-K1 and about 40% identical to ClC-0, -1, and -2. ClC-K2S encodes a 632-amino acid protein in which 55 amino acids containing the putative second membrane-spanning domain of ClC-K2L are deleted. Chloride currents induced by both clones were very similar in terms of inhibitor sensitivity and anion selectivity (Br- > I- > Cl- >> cyclamate-). Northern blot with total ClC-K2L as a probe under high stringency revealed its message predominantly in kidney, especially in the outer and inner medulla. Reverse transcription polymerase chain reaction technique using microdissected nephron segments revealed that the main site of expression of both clones in kidney was the thick ascending limb of Henle's loop and collecting ducts, where the existence of a variety of chloride channels and their importance for maintaining body fluid homeostasis have been demonstrated. These results suggest that ClC-K2L and -K2S are chloride channels in the thick ascending limb and collecting ducts and may be important routes for transcellular chloride transport like ClC-K1.

Induction of nitric oxide synthase gene by interleukin-1 beta in cultured rat cardiocytes.

BACKGROUND: Impaired myocardial contractility in septic shock is protracting, which may be caused by cytokine-induced nitric oxide (NO) synthesis in the heart. However, the cellular mechanism by which cytokines induce nitric oxide synthase (NOS) in cardiocytes remains obscure. METHODS AND RESULTS: We studied the effect of human recombinant interleukin-1 beta (IL-1 beta) on synthesis of NO2-/NO3- (NOx) and the expression of NOS mRNA and protein in cultured neonatal rat cardiocytes. IL-1 beta dose-dependently (0.1 to 10 ng/mL) stimulated NOx production as a function of time (6 to 48 hours). Northern blot analysis using complementary DNAs for rat brain-type constitutive (c) NOS and mouse macrophage-type inducible (i) NOS as probes showed that IL-1 beta induced expression of mRNA for iNOS but not for cNOS, starting after 6 hours and reaching a maximum after 48 hours in cardiocytes. IL-1 beta similarly induced iNOS mRNA expression in cultured adult rat cardiocytes in a time-dependent manner. Western blot analysis using specific antibody against the N-terminal fragment of mouse iNOS revealed the expression of 130-kD iNOS-like protein in IL-1 beta-treated cardiocytes. Northern blotting and immunocytochemical study revealed that IL-1 beta-induced iNOS mRNA and iNOS-like immunoreactivity were exclusively localized to cardiac myocytes but also to nonmyocytes, to a lesser extent. NG-mono-methyl-L-arginine, an NOS inhibitor, completely blocked the IL-1 beta-induced NOx production, whose effect was reversed by L-arginine but not by D-arginine. Dexamethasone inhibited the IL-1 beta-induced NOx production as well as iNOS mRNA expression. Cycloheximide and actinomycin D completely inhibited the IL-1 beta-induced NOx production and iNOS mRNA expression. Neither a calmodulin inhibitor (W-7), a protein kinase C inhibitor (calphostin C), nor a Ca2+ channel antagonist (nicardipine) showed any effect on the IL-1 beta-induced NOx production. CONCLUSIONS: These data demonstrate that IL-1 beta induces macrophage-type iNOS mRNA expression mainly by cardiac myocytes but also by nonmyocytes to a lesser extent, and subsequent de novo protein synthesis of iNOS leads to excessive local production of NO by cardiocytes.

Cadmium-induced osteomalacic and osteopetrotic lesions in ovariectomized rats.

The effects of long-term administration of cadmium (Cd) chloride on the bone were studied using ovariectomized rats. The rats were injected iv with the compound at doses of 1.0 and 2.0 mg/kg, 5 days a week, for 13 weeks. The serum concentrations of calcium and inorganic phosphorus were significantly increased from 8 weeks in the 2.0 mg/kg group. The bone Cd content was gradually increased for 13 weeks in a dose-dependent manner. Calcium and phosphorus contents in the bone, and serum levels of parathyroid hormone and osteocalcin, were not significantly different between Cd-treated and control rats. Histopathologically, chronic Cd nephropathy such as tubular atrophy and interstitial fibrosis was observed with clinical polyuria and increased enzymuria. The skeletal changes were detected mainly in the femur and tibia. In the metaphysis of Cd-treated rats, cancellous bone mass increased with time. This change was detected as an increased opacity by a roentgenogram. In the cortical bone of the midshaft haversian canals were dilated with clearly bordered osteoid seams and showed a motheaten pattern in rats in the 2.0 mg/kg group at 13 weeks. In the present study, we report Cd nephropathy and osteomalacic changes in ovariectomized rats with iv injection of CdCl2 for 13 weeks. Although an involvement of the indirect action of Cd through renal failure could not be ruled out in this experiment, our biochemical and pathological data suggested that osteomalacia was induced by a direct action of Cd on the bone through abnormal calcium homeostasis.

Cloning and expression of a protein kinase C-regulated chloride channel abundantly expressed in rat brain neuronal cells.

cDNA (CIC-3) encoding a protein kinase C-regulated chloride channel was cloned and characterized. The open reading frame encodes 760 amino acids, which possess significantly amino acid identity with previously cloned CIC chloride channels. The chloride currents expressed in Xenopus oocytes injected with CIC-3 cRNA were completely blocked by activation of protein kinase C by 12-O-tetradecanoylphorbol 13-acetate. Abundant expression of CIC-3 mRNA was observed in rat brain, especially in the olfactory bulb, hippocampus, and cerebellum. These findings suggest that CIC-3 may play an important role in neuronal cell function through regulation of membrane excitability by protein kinase C.