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1.
Jpn J Physiol ; 51(5): 569-76, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11734077

ABSTRACT

A cation channel from Tetrahymena cilia is permeable to both monovalent and divalent cations. A single-file two-site channel model was introduced for explaining the single channel currents of the channel in mixed solutions of K(+) and Ca(2+). In the model it was assumed that two potassium ions or one calcium ion can bind to the binding sites, and that the potassium ions between the binding sites are in a fast equilibrium condition. Single channel currents were calculated from the values of rate constants, ionic concentrations on both sides of the membrane, and the membrane voltages. This model could explain all the observed single channel currents of the channel in K(+) or Ca(2+) solution and in mixed solutions of K(+) and Ca(2+). The values of the reversal potential in the bi-ionic condition could distinguish this single-file two-site channel model from the single-site channel model or the model in which each ion permeates through the same channel independently (the Goldman-Hodgkin-Katz equation). Experimental data supported this model.


Subject(s)
Calcium Channels/physiology , Models, Biological , Potassium Channels/physiology , Animals , Calcium/pharmacokinetics , Cilia/physiology , Permeability , Potassium/pharmacokinetics , Tetrahymena/physiology
2.
J Clin Neurophysiol ; 18(2): 185-90, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11435811

ABSTRACT

The authors studied the origin of the scalp P13-like potential in median somatosensory evoked potentials, which have been reported to be preserved in patients with cervicomedullary lesions or in brain death. There were five patients with high to middle cervical lesions (C2/3 or C3/4 level). Small P13-like potentials after P11 were identified for all patients with a noncephalic reference but not with an ear reference. Their onset latencies were slightly earlier than the expected latency of the true P13/14 onset. In two patients, delayed true P13/14s followed by N18s were identified with both noncephalic and ear references. The authors argue that the P13-like potential observed in these patients is a different entity from scalp P13 in normal subjects. Because the C3/4 vertebral level corresponds to the C5 cord level, the origin of the P13-like potential must be below C5, contradicting the previous opinion that it is generated at the cervicomedullary junction or at the high cervical dorsal column. The authors named this potential lower cervical P13 (or lcP13), and present an opinion that it is generated by the beginning of the second spinal ascending volley, which has been described by direct-recording studies in humans.


Subject(s)
Evoked Potentials, Somatosensory , Multiple Sclerosis/physiopathology , Spinal Cord Diseases/physiopathology , Spinal Cord/physiopathology , Spinal Osteophytosis/physiopathology , Adult , Aged , Evoked Potentials, Somatosensory/physiology , Female , Humans , Magnetic Resonance Imaging , Male , Median Nerve , Middle Aged , Multiple Sclerosis/complications , Multiple Sclerosis/diagnosis , Neck/pathology , Reaction Time , Spinal Cord/pathology , Spinal Cord Diseases/complications , Spinal Cord Diseases/diagnosis , Spinal Osteophytosis/complications , Spinal Osteophytosis/diagnosis
3.
Intern Med ; 39(10): 788-93, 2000 Oct.
Article in English | MEDLINE | ID: mdl-11030201

ABSTRACT

OBJECTIVE: Interferon has been used as a new therapeutic agent for glomerulonephritis since a manifest relationship between membranoproliferative glomerulonephritis (MPGN) and hepatitis C virus (HCV) infection was documented. However, several side effects and rebound phenomenon have been significant problems. We retrospectively evaluated the therapeutic effect and safety of the standard treatment with steroids and/or immunosuppressive agents for MPGN patients with an HCV infection. METHODS: Remission and renal survival rates as well as clinical and histological data were compared between MPGN groups with or without an HCV infection. In addition, the hepatic function was followed-up after the treatment. PATIENTS: The subjects were 42 biopsy proven MPGN patients. Seven were positive for an HCV infection. Secondary causes of MPGN excluding an HCV infection were omitted. Most patients were treated with steroids and/or immunosuppressive agents. RESULTS: The mean age of the MPGN patients with an HCV infection was significantly higher than that of those without an HCV infection. The renal function and the interstitial change of the former group were significantly worse than those of the latter. Nevertheless, remission and renal survival rates were not significantly different between the two groups. None in the HCV positive MPGN group showed an impairment of hepatic function during the clinical course. However, 2 subjects died from severe pneumonia during the treatment. CONCLUSION: The standard treatment with steroids and/or immunosuppressive agents did not reveal a statistical difference in the therapeutic efficacy between MPGN patients with or without an HCV infection. However, some in the former group showed a poor prognosis.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Glomerulonephritis, Membranoproliferative/drug therapy , Glucocorticoids/therapeutic use , Hepatitis C/drug therapy , Immunosuppressive Agents/therapeutic use , Platelet Aggregation Inhibitors/therapeutic use , Adult , Aged , Biopsy , Drug Therapy, Combination , Female , Glomerulonephritis, Membranoproliferative/pathology , Glomerulonephritis, Membranoproliferative/virology , Hepatitis C/pathology , Hepatitis C/virology , Humans , Kidney Function Tests , Liver Function Tests , Male , Middle Aged , Prognosis , Retrospective Studies
4.
J Membr Biol ; 168(3): 283-7, 1999 Apr 01.
Article in English | MEDLINE | ID: mdl-10191362

ABSTRACT

A Paramecium cell responded to heat and cold stimuli, exhibiting increased frequency of directional changes in its swimming behavior. The increase in the frequency of directional changes was maintained during heating, but was transient during cooling. Although variations were large, as expected with this type of electrophysiological recording, results consistently showed a sustained depolarization of deciliated cells in response to heating. Depolarizations were also consistently observed upon cooling. However, these depolarizations were transient and not continuous throughout the cooling period. These depolarizations were lost or became small in Ca2+-free solutions. In a voltage-clamped cell, heating induced a continuous inward current and cooling induced a transient inward current under conditions where K+ currents were suppressed. The heat-induced inward current was not affected significantly by replacing extracellular Ca2+ with equimolar concentrations of Ba2+, Sr2+, Mg2+, or Mn2+, and was lost upon replacing with equimolar concentration of Ni2+. On the other hand, the cold-induced inward current was not affected significantly by Ba2+, or Sr2+, however the decay of the inward current was slowed and was lost or became small upon replacing with equimolar concentrations of Mg2+, Mn2+, or Ni2+. These results indicate that Paramecium cells have heat-activated Ca2+ channels and cold-activated Ca2+ channels and that the cold-activated Ca2+ channel is different from the heat-activated Ca2+ channel in the ion selectivity and the calcium-dependent inactivation.


Subject(s)
Calcium Channels/metabolism , Calcium/metabolism , Paramecium tetraurelia/metabolism , Protozoan Proteins/metabolism , Animals , Cations, Divalent/pharmacology , Cold Temperature , Hot Temperature , Ion Transport , Locomotion , Membrane Potentials , Paramecium tetraurelia/physiology , Patch-Clamp Techniques
5.
Hum Pathol ; 30(3): 331-8, 1999 Mar.
Article in English | MEDLINE | ID: mdl-10088553

ABSTRACT

To clarify the process and mechanisms of the development and progression of peripheral lung adenocarcinoma, we investigated the relationships among the patterns of basement membrane (BM), stromal fibrosis, and the expressions of gelatinase A and tissue inhibitor of metalloproteinases-2 (TIMP-2) in 33 lesions of atypical alveolar cell hyperplasia (AAH) and 48 lesions of lung adenocarcinoma, including 24 lesions of bronchioloalveolar carcinoma (BAC). We found that the architecture of alveolar BM was intact in all 33 AAH lesions and 11 nonsclerosing BAC lesions that formed no central scar, suggesting that these lesions are early-stage intraepithelial neoplasia. The preexistent BM of the lung was disrupted, and the BM components around the neoplastic glands were disrupted or absent in the area of the central scar of some sclerosing BAC lesions with collapse fibrosis alone (2 of 4) and in those of all of the adenocarcinoma lesions associated with desmoplastic stromal fibrosis (nine sclerosing BAC and 24 non-BAC tumors). These results suggested that, in lung adenocarcinomas, destruction of the BM was correlated with the formation of a central scar, particularly with desmoplasia. It is likely that adenocarcinomas with a central scar are advanced and invasive cancers potentially having metastatic activity. The expression of gelatinase A and TIMP-2 was associated with central scar formation as well as with destruction of the BM components. Both the neoplastic and stromal cells expressed gelatinase A and TIMP-2 and probably play a role in tumor cell invasion.


Subject(s)
Adenocarcinoma/pathology , Basement Membrane/pathology , Gelatinases/metabolism , Lung Neoplasms/pathology , Metalloendopeptidases/metabolism , Tissue Inhibitor of Metalloproteinase-2/metabolism , Adenocarcinoma/enzymology , Fibrosis , Humans , Hyperplasia , Lung Neoplasms/enzymology , Matrix Metalloproteinase 2 , Stromal Cells/pathology
6.
J Neurochem ; 69(1): 377-87, 1997 Jul.
Article in English | MEDLINE | ID: mdl-9202332

ABSTRACT

Hypoxia (5% O2) enhanced catecholamine release in cultured rat adrenal chromaffin cells. Also, the intracellular free Ca2+ concentration ([Ca2+]i) increased within 3 min in approximately 50% of the chromaffin cells under hypoxic stimulation. The increase depended on the presence of extracellular Ca2+. Nifedipine and omega-conotoxin decreased the population of the cells that showed the hypoxia-induced [Ca2+]i increase, showing that the Ca2+ influx was attributable to L- and N-type voltage-dependent Ca2+ channels. The membrane potential was depolarized during the perfusion with the hypoxic solution and returned to the basal level following the change to the normoxic solution (20% O2). Membrane resistance increased twofold under the hypoxic condition. The current-voltage relationship showed a hypoxia-induced decrease in the outward K+ current. Among the K+ channel openers tested, cromakalim and levcromakalim, both of which interact with ATP-sensitive K+ channels, inhibited the hypoxia-induced [Ca2+]i increase and catecholamine release. The inhibitory effects of cromakalim and levcromakalim were reversed by glibenclamide and tolbutamide, potent blockers of ATP-sensitive K+ channels. These results suggest that some fractions of adrenal chromaffin cells are reactive to hypoxia and that K+ channels sensitive to cromakalim and glibenclamide might have a crucial role in hypoxia-induced responses. Adrenal chromaffin cells could thus be a useful model for the study of oxygen-sensing mechanisms.


Subject(s)
Calcium/metabolism , Catecholamines/metabolism , Chromaffin Cells/metabolism , Potassium Channel Blockers , Animals , Benzopyrans/pharmacology , Buffers , CHO Cells , Carotid Body/chemistry , Carotid Body/metabolism , Cell Hypoxia/physiology , Chromaffin Cells/chemistry , Chromaffin Cells/cytology , Cricetinae , Cromakalim , Diazoxide/pharmacology , Glioma , Glyburide/pharmacology , Guanidines/pharmacology , Hybrid Cells , Hypoglycemic Agents/pharmacology , Male , Membrane Potentials/physiology , Minoxidil/pharmacology , Neuroblastoma , Parasympatholytics/pharmacology , Patch-Clamp Techniques , Pinacidil , Potassium/pharmacology , Potassium Channels/agonists , Potassium Channels/metabolism , Pyrroles/pharmacology , Rats , Rats, Wistar , Vasodilator Agents/pharmacology
7.
Biochim Biophys Acta ; 1327(1): 52-60, 1997 Jul 05.
Article in English | MEDLINE | ID: mdl-9247166

ABSTRACT

A non-selective cation channel was found in mutant Paramecium cells (K115). This cell had been selected as a resistant mutant in a high-K+ solution. In patch clamp studies of these cells in the inside-out configuration, this channel was activated by bath applications of elevated Ca2+ concentrations. The channels became very active when the Ca2+ concentration was above 3.2 microM. The channel was also activated by depolarization. The voltage dependency was steep upon depolarization, whereas upon hyperpolarization the channel activity barely changed. This channel had poor selectivity for monovalent alkali cations. Using the Goldman-Hodgkin-Katz equation for the reversal potential, the permeability ratios with respect to K+ for Na+, Rb+, Cs+ and Li+ were nearly 1. Although the permeability ratios were similar for each cation, the single channel conductances differed. The single channel conductances were 467 pS with K+ as the charge carrier, 406 pS with Na+, 397 pS with Rb+, 253 pS with Cs+ and 198 pS with Li+ upon depolarization in 100 mM cation solutions. A similar calcium-activated large conductance channel was observed in the wild-type (G3) Paramecium cells but was very rare.


Subject(s)
Calcium/pharmacology , Ion Channels/metabolism , Paramecium/metabolism , Animals , Cell Membrane/metabolism , Electric Conductivity , Ion Channels/antagonists & inhibitors , Ion Channels/drug effects , Patch-Clamp Techniques , Potassium Chloride/pharmacology
8.
Cell Biochem Funct ; 15(2): 113-7, 1997 Jun.
Article in English | MEDLINE | ID: mdl-9253163

ABSTRACT

Changes of intracellular calcium concentration ([Ca2+]i) induced by the extracellular application of ATP and bradykinin in mouse mammary tumour cells (MMT060562) were investigated by image analysis of fluo-3 fluorescence at 24 degrees C and 35 degrees C. ATP (0.1-100 microM) and bradykinin (0.1 nM-1 microM) induced the increase of [Ca2+]i at both temperatures and Ca(2+)-depletion did not affect these [Ca2+]i responses. Both [Ca2+]i responses became more sensitive at 35 degrees C than at 24 degrees C. A clear latency of [Ca2+]i increase after the application of the agonists was observed, and it changed with the concentration of the agonist. As concentrations of ATP or bradykinin became lower, the latency and rise time became longer. At higher concentrations, the latency and rise time approached a constant value. The latency shortened remarkably at 35 degrees C. These results suggested the involvement of a regenerative or threshold process in the [Ca2+]i responses in mammary tumour cells.


Subject(s)
Calcium/metabolism , Mammary Neoplasms, Experimental/pathology , Temperature , Adenosine Triphosphate/pharmacology , Animals , Bradykinin/pharmacology , Dose-Response Relationship, Drug , Image Processing, Computer-Assisted , Mice , Tumor Cells, Cultured
9.
Cell Struct Funct ; 22(6): 579-83, 1997 Dec.
Article in English | MEDLINE | ID: mdl-9591049

ABSTRACT

Under voltage clamp, Paramecium tetraurelia was used to examine the cold-induced inward current and its inhibition by a calmodulin antagonist, W-7 [N-(6 aminohexyl)-5-chloro-1-naphthalenesulphonamide]. Cooling of the cell caused an inward current. The amplitude of the current was increased as the membrane potential was made more positive than the resting potential, and it was significantly blocked by using CsCl-filled electrodes and tetraethylammonium in the bath solution, suggesting that the current was accompanied mainly by a decrease in K+ conductance. The cold-induced inward current was reversibly inhibited by the external application of W-7 in a concentration-dependent manner. EGTA-microinjection into the cell also reduced the current. These results indicate that the decrease in K+ conductance induced by cooling is Ca(2+)-dependent and is inhibited by W-7.


Subject(s)
Calmodulin/antagonists & inhibitors , Cold Temperature , Paramecium tetraurelia/physiology , Potassium/metabolism , Sulfonamides/pharmacology , Animals , Biological Transport/drug effects , Biological Transport/physiology , Calcium/metabolism , Calcium Channels/physiology , Chelating Agents/pharmacology , Egtazic Acid/pharmacology , Paramecium tetraurelia/chemistry , Patch-Clamp Techniques , Potassium Channels/physiology
10.
Cell Struct Funct ; 22(5): 493-500, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9431453

ABSTRACT

Wild type and calmodulin mutants (cam) of Paramecium tetraurelia were examined for cold-sensitive responses. Among mutants tested, cam12 and cam13 mutants, which have substitutions in N-terminal lobe of calmodulin molecule, reduced both responses in the swimming and the membrane potential. Under voltage clamp conditions, the cooling stimulus to the wild type cell induced a transient inward current whose amplitude increased with the rate of temperature drop. The cam12 cell did not induce inward currents in response to cooling with a rate slower than -0.4 degree C/s. The reduced current response of cam12 mutant was restored by an external application of a phosphodiesterase inhibitor, theophylline. Also, an intracellular injection of hydrolysis-resistant cyclic nucleotides, either 8-bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP) or 8-bromoguanosine 3,5'-cyclic monophosphate (8-Br-cGMP), restored the current response. Such restoration was accompanied by shifts of the resting potential to hyperpolarized levels and by an increase in the membrane conductance. The results suggest the possibility that calmodulin and cyclic nucleotide regulate K+ channels responsive to the cooling stimulus.


Subject(s)
Calmodulin/physiology , Nucleotides, Cyclic/pharmacology , Paramecium tetraurelia/metabolism , Thermosensing/physiology , Animals , Calmodulin/drug effects , Cell Movement/physiology , Cells, Cultured , Cold Temperature , Cyclic AMP/analysis , Cyclic AMP/pharmacology , Cyclic GMP/analysis , Cyclic GMP/pharmacology , Membrane Potentials/drug effects , Membrane Potentials/physiology , Mutation , Nucleotides, Cyclic/administration & dosage , Paramecium tetraurelia/drug effects , Patch-Clamp Techniques , Radioimmunoassay , Theophylline/pharmacology , Thermosensing/drug effects
11.
J Membr Biol ; 154(2): 163-7, 1996 Nov.
Article in English | MEDLINE | ID: mdl-8929290

ABSTRACT

The concentration of intracellular calcium, [Ca2+]i, in Paramecium was imaged during cold-sensitive response by monitoring fluorescence of two calcium-sensitive dyes, Fluo-3 and Fura-Red. Cooling of a deciliated Paramecium caused a transient increase in [Ca2+]i at the anterior region of the cell. Increase in [Ca2+]i was not observed at any region in Ca(2+)-free solution. Under the electrophysiological recording, a transient depolarization of the cell was observed in response to cooling. On the voltage-clamped cell, cooling induced a transient inward current under conditions where K+ currents were suppressed. These membrane depolarizations and inward currents in response to cooling were lost upon removing extracellular Ca2+. The cold-induced inward current was lost upon replacing extracellular Ca2+ with equimolar concentration of Co2+, Mg2+ or Mn2+, but it was not affected significantly by replacing with equimolar concentration of Ba2+ or Sr2+. These results indicate that Paramecium cells have Ca2+ channels that are permeable to Ca2+, Ba2+ and Sr2+ in the anterior soma membrane and the channels are opened by cooling.


Subject(s)
Calcium/metabolism , Cold Temperature , Image Processing, Computer-Assisted/methods , Paramecium/metabolism , Animals , Calcium/analysis , Calcium Channels/metabolism , Cell Membrane/metabolism , Humans , Membrane Potentials , Paramecium/chemistry , Patch-Clamp Techniques , Potassium Channels/metabolism
12.
Nihon Ronen Igakkai Zasshi ; 30(1): 65-9, 1993 Jan.
Article in Japanese | MEDLINE | ID: mdl-8474230

ABSTRACT

A 78-year-old man was admitted to our hospital with anemia and jaundice. Hematological studies revealed hyperchromic macrocytic anemia, and biochemical studies revealed findings of hemolysis. The folic acid level was low and megaloblasts were observed in the bone marrow. From these findings, the patient was diagnosed as having megaloblastic anemia due to folic acid deficiency. This patient had been a heavy alcohol drinker in the habit of drinking alcohol without meals. He began to eat regular meals in the hospital, and the anemia and jaundice improved gradually. Since liver cirrhosis was suspected, amino acid analysis of the urine was performed, and abnormal excretion of beta-amino-isobutyric acid (BAIB) was found. According to the amount of BAIB excreted, the Japanese population can be divided into low and high BAIB excretors comprising 65% and 36%, respectively. BAIB is also considered to reflect dissimilation of thymine. The present patient was included in the high excretion group because of the abnormally high urine level of BAIB, which was considered to be caused by ineffective hematopoiesis in the bone marrow as a result of his megaloblastic anemia. For this reason, dissimilation of thymine was considered to have been active in this patient.


Subject(s)
Aminoisobutyric Acids/urine , Anemia, Megaloblastic/urine , Aged , Humans , Male
13.
Biochem Biophys Res Commun ; 186(2): 1037-41, 1992 Jul 31.
Article in English | MEDLINE | ID: mdl-1497637

ABSTRACT

Ion channels were incorporated into planar lipid bilayers following fusion of vesicles from the membrane of an insulin-secreting beta-cell line, HIT T15. The channel was completely blocked by 0.5 mM ATP. The channel retained the same ATP-dependence, voltage-sensitivity and single channel conductance as the ATP-regulated K+ channel that found in isolated membrane patches.


Subject(s)
Adenosine Triphosphate/pharmacology , Islets of Langerhans/physiology , Lipid Bilayers , Potassium Channels/physiology , Animals , Cell Line , Cell Membrane/physiology , Cricetinae , Electric Conductivity , Insulin/metabolism , Insulin Secretion , Ion Channel Gating/drug effects , Membrane Potentials/drug effects , Mesocricetus , Potassium Channels/drug effects , Probability
14.
Diabetologia ; 35(7): 619-23, 1992 Jul.
Article in English | MEDLINE | ID: mdl-1379561

ABSTRACT

This study evaluates the use of the planar lipid bilayer as a functional assay of Ca(2+)-activated K+ channel activity for use in purification of the channel protein. Ca(2+)-activated K+ channels from the plasma membrane of an insulin-secreting hamster Beta-cell line (HIT T15) were incorporated into planar lipid bilayers. The single channel conductance was 233 picoSiemens (pS) in symmetrical 140 mmol/l KCl and the channel was strongly K(+)-selective (PCl/PK = 0.046; PNa/PK = 0.027). Channels incorporated into the bilayer with two orientations. In 65% of cases, the probability of the channel being open was increased by raising calcium on the cis side of the bilayer (to which the membrane vesicles were added) or by making the cis side potential more positive. At a membrane potential of + 20 mV, which is close to the peak of the Beta-cell action potential, channel activity was half-maximal at a Ca2+ concentration of about 15 mumol/l. Charybdotoxin greatly reduced the probability of the channel being open when added to the side opposite to that at which Ca2+ activated the channel. These results resemble those found for Ca(2+)-activated K+ channels in native Beta cell membranes and indicate that the channel properties are not significantly altered by incorporation in a planar lipid bilayer.


Subject(s)
Calcium/pharmacology , Islets of Langerhans/physiology , Lipid Bilayers , Potassium Channels/physiology , Animals , Cell Line, Transformed , Cell Membrane/physiology , Charybdotoxin , Egtazic Acid/pharmacology , Electric Conductivity , Kinetics , Mathematics , Membrane Potentials/drug effects , Potassium Channels/drug effects , Scorpion Venoms/pharmacology
15.
Biophys J ; 56(6): 1217-23, 1989 Dec.
Article in English | MEDLINE | ID: mdl-2482084

ABSTRACT

The cation-selective channel from Tetrahymena cilia is permeable to both monovalent and divalent cations. The single channel conductance in mixed solutions of K+ and Ca2+ was determined by the Gibbs-Donnan ratio of K+ and Ca2+, and the binding sites of this channel were considered to be always occupied by two potassium ions or by one calcium ion under the experimental conditions: 5-90 mM K+ and 0.5-35 mM Ca2+ (Oosawa and Kasai, 1988). A two-barrier model for the channel was introduced and the values of Michaelis-Menten constants and maximum currents carried by K+ and Ca2+ were calculated using this model. Single channel current amplitudes and reversal potentials were calculated from these values. The calculated single-channel currents were compared with those obtained experimentally. The calculated reversal potentials were compared with the resting potentials of Tetrahymena measured in various concentrations of extracellular K+ and Ca2+. The method of calculation of ionic currents and reversal potentials presented here is helpful for understanding the properties of the channels permeable to both monovalent and divalent cations.


Subject(s)
Calcium/metabolism , Ion Channels/metabolism , Models, Biological , Potassium/metabolism , Animals , Cations, Divalent , Cations, Monovalent , Cilia/metabolism , Kinetics , Mathematics , Membrane Potentials , Tetrahymena/metabolism
16.
J Physiol ; 408: 223-32, 1989 Jan.
Article in English | MEDLINE | ID: mdl-2476556

ABSTRACT

1. Ionic currents were studied in Xenopus laevis oocytes using the patch-clamp and the whole-cell voltage-clamp techniques. 2. Single-channel currents were recorded from the cell-attached patches in oocytes injected with rat brain mRNA when glutamate was applied locally outside the patch. The single-channel conductance was 3.66 pS, and the extrapolated equilibrium potential was -23.0 mV, indicating that the channels were chloride selective. 3. Single-channel currents with similar characteristics were observed in cell-attached patches in native oocytes in response to injection of inositol 1,4,5-trisphosphate (IP3) or Ca2+. 4. Whole-cell currents were evoked by glutamate in oocytes injected with rat brain mRNA. They usually showed an oscillatory component, and reversed direction at about the chloride equilibrium potential. Injection of IP3 or Ca2+ into a native oocyte evoked a transient whole-cell current. The reversal potential was near the chloride equilibrium potential, and it changed from negative to positive in low-chloride solution. 5. The results suggest that the glutamate receptors are not directly coupled with the endogenous chloride channels but indirectly activate these via the messenger system IP3-Ca2+.


Subject(s)
Chlorides/metabolism , Ion Channels/metabolism , Oocytes/metabolism , Receptors, Neurotransmitter/physiology , Animals , Brain , Calcium/metabolism , Female , Glutamates/pharmacology , Inositol 1,4,5-Trisphosphate , Inositol Phosphates/pharmacology , Membrane Potentials/drug effects , RNA, Messenger/pharmacology , Rats , Receptors, Glutamate , Xenopus laevis
17.
Biophys J ; 54(3): 407-10, 1988 Sep.
Article in English | MEDLINE | ID: mdl-2462927

ABSTRACT

A single cation-channel from Tetrahymena cilia was incorporated into planar lipid bilayers. This channel was voltage-independent and is permeable to K+ and Ca2+. In the experiments with mixed solutions where the concentrations of K+ and Ca2+ were varied, the single-channel conductance was found to be influenced by the Gibbs-Donnan ratio. The data are explained by assuming that the binding sites of this channel were always occupied by two potassium ions or one calcium ion under the present experimental conditions (5 mM-90 mM K+ and 0.5 mM-35 mM Ca2+) and these bound cations determined the channel conductivity.


Subject(s)
Calcium/metabolism , Cilia/physiology , Ion Channels/physiology , Potassium/metabolism , Tetrahymena/physiology , Animals , Electric Conductivity , Mathematics , Membrane Potentials , Models, Theoretical
18.
Cell Struct Funct ; 13(1): 51-60, 1988 Feb.
Article in English | MEDLINE | ID: mdl-2453296

ABSTRACT

The single-channel properties for monovalent and divalent cations of a voltage-independent cation channel from Tetrahymena cilia were studied in planar lipid bilayers. The single-channel conductance reached a maximum value as the K+ concentration was increased in symmetrical solutions of K+. The concentration dependence of the conductance was approximated to a simple saturation curve (a single-ion channel model) with an apparent Michaelis constant of 16.3 mM and a maximum conductance of 354 pS. Divalent cations (Ca2+, Ba2+, Sr2+, and Mg2+) also permeated this channel. The sequence of permeability determined by zero current potentials at high ionic concentrations was Ba2+ greater than or equal to K+ greater than or equal to Sr2+ greater than Mg2+ greater than Ca2+. Single-channel conductances for Ca2+ were nearly constant (13.9 pS-20.5 pS) in the concentrations between 0.5 mM and 50 mM Ca-gluconate. In the experiments with mixed solutions of K+ and Ca2+, a maximum conductance of Ca2+ (gamma Camax) and an apparent Michaelis constant of Ca2+ (K Cam) were obtained by assuming a simple competitive relation between the cations. Gamma Camax and K Cam were 14.0 pS and 0.160 mM, respectively. Single-channel conductances in mixed solutions were well-fitted to this competitive model supporting that this cation channel behaves as a single-ion channel. This channel had relatively high-affinity Ca2+-binding sites.


Subject(s)
Calcium/metabolism , Ion Channels/metabolism , Lipid Bilayers/analysis , Potassium/metabolism , Tetrahymena/metabolism , Animals , Cations, Divalent/metabolism , Cations, Monovalent/metabolism , Cell Membrane , Cilia/analysis , Tetrahymena/analysis
19.
Am J Physiol ; 250(3 Pt 1): C361-4, 1986 Mar.
Article in English | MEDLINE | ID: mdl-2420192

ABSTRACT

Single-channel conductance of the K+ channel from sarcoplasmic reticulum (SR) was reduced by aminoglycoside antibiotics such as neomycin and ribostamycin and also by n-hexylamine from either side of the membrane in a dose- and voltage-dependent manner. K+ channels were incorporated into an artificial phospholipid bilayer. This inhibition follows a single-site titration curve. The voltage dependence of the inhibition is explained by assuming that these drugs bind to the open state of a single channel on one site located approximately 40% of the way through the membrane from the cis side (the side to which SR vesicles are added) when drugs are added to the cis side and bind on another site located approximately 40% of the way through the membrane from the trans side (the opposite side to the cis side) when drugs are added to the trans side.


Subject(s)
Anti-Bacterial Agents/pharmacology , Ion Channels/drug effects , Potassium/metabolism , Sarcoplasmic Reticulum/drug effects , Amines/pharmacology , Aminoglycosides/pharmacology , Animals , Binding Sites/drug effects , Biological Transport, Active/drug effects , In Vitro Techniques , Ion Channels/metabolism , Neomycin/pharmacology , Rabbits , Ribostamycin/pharmacology , Sarcoplasmic Reticulum/metabolism
20.
Am J Physiol ; 249(1 Pt 1): C177-9, 1985 Jul.
Article in English | MEDLINE | ID: mdl-2409811

ABSTRACT

A single cation channel from Tetrahymena cilia was incorporated into planar lipid bilayers. This channel selected for K+, Na+, and Li+ over Cl- and gluconate-, and its single channel conductance (at +25 mV) was 211 +/- 8 pS (mean +/- SE) in 100 mM K+-gluconate. The channel was not voltage dependent and may contribute to the resting K+ conductance of ciliary membrane.


Subject(s)
Cations/metabolism , Ion Channels/physiology , Lipid Bilayers , Tetrahymena/physiology , Animals , Cilia/physiology , Electrophysiology , Intracellular Membranes/physiology , Membrane Potentials , Potassium/metabolism
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