Biomedical subjects

C K Ifune

Publications and source records attributed to C K Ifune.

6 recordsLinked to original sources

PubMed1993

Modulation of acetylcholine-elicited currents in clonal rat phaeochromocytoma (PC12) cells by internal polyphosphates.

1. Whole-cell voltage clamp techniques were used to examine acetylcholine (ACh)-elicited currents in differentiated cells of the rat phaeochromocytoma cell line, PC12. 2. In the absence of intracellular Mg2+, the whole-cell current-voltage relationship for the ACh-elicited current displayed inward rectification which was reduced in part by the presence of 5 mM internal adenosine 5'-triphosphate (ATP). 3. The reduction in the rectification attributed to ATP developed over the first 15-20 min of whole-cell recording. Similar results were obtained with a non-hydrolysable ATP analogue, adenosine-5'-O-3-thiotriphosphate (ATP gamma S), or cytosine 5'-triphosphate (CTP) in the internal solution, but not with adenosine 5'-diphosphate (ADP) or pyrophosphate. 4. The magnitude of the ACh-elicited current was also dependent on recording time and the composition of the internal pipette solution. The magnitude of the peak ACh-elicited current increased over time when the cell was internally perfused with the control solution or a pipette solution containing pyrophosphate, ATP gamma S, or ADP. The largest sustained increases in ACh-elicited current were observed in the presence of internal pyrophosphate or ATP gamma S. In contrast, with internal ATP or CTP, the whole-cell current initially increased, then steadily decreased with recording time. 5. The desensitization rate of the ACh-elicited current increased with recording time irrespective of the composition of the intracellular solution. 6. The actions of the compounds tested make it likely that the changes in the whole-cell current-voltage relationship, peak current, and desensitization are produced by separate mechanisms. The mechanisms underlying these changes are unknown, but the ability of the compounds to chelate divalent cations is unlikely to be the explanation. Other unlikely explanations include phosphorylation of the ACh receptor or regulation by GTP-binding proteins.

Acetylcholine↗

PubMed1992

Inward rectification of acetylcholine-elicited currents in rat phaeochromocytoma cells.

1. Currents elicited by acetylcholine (ACh) were studied in the rat phaeochromocytoma cell line, PC12, using patch-clamp techniques. 2. Whole-cell ACh-elicited currents are inwardly rectifying and intracellular Mg2+ can play a role in determining the extent of whole-cell current rectification. Increasing the intracellular Mg2+ concentration, [Mg2+]i, augmented the rectification. The effects of increased [Mg2+]i on the whole-cell current can be explained by the block of receptor channels by Mg2+. 3. In the nominal absence of internal divalent cations, however, a substantial degree of rectification remains. This rectification is probably not due to divalent cations, as buffering the external Mg2+ concentration to 50 microM and the internal concentration to nominally 0 Mg2+ did not reduce the rectification. The remanent rectification was not due to block by the main permeant cation, Na+. Using K+ or Cs+ as the main monovalent cation inside the cell did not diminish the rectification. Neither replacing the pH buffer, HEPES, with phosphate buffer nor increasing the intracellular pH removed the rectification. 4. For ACh receptor channels in excised patches, the voltage dependence of the probability of being open (Popen) stemmed mainly from the voltage dependence of the channel burst duration. The channel opening rate was relatively voltage independent. The weak voltage dependence displayed by the channel burst duration was insufficient to account for the reduced whole-cell outward current at positive potentials. The mean burst duration of the channel did not have a simple logarithmic relationship with voltage. 5. In the absence of intracellular Mg2+, the instantaneous current-voltage relationship for whole-cell currents was linear suggesting that the I-V relationship of single channels in perfused cells is linear and does not contribute to the rectification of the whole-cell current. 6. In perfused cells, receptor channels had a low steady-state probability of being open at positive potentials compared to channels in excised patches. Voltage jumps to positive potentials revealed a process in perfused cells which could account for the low Popen. Relaxations of agonist-induced current at +40 mV had a large, exponentially decaying component that quickly closed channels (rate constant, tau, approximately 400 microseconds). The mechanism responsible for this decay could explain the rectification that remains in the absence of intracellular divalent cations.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholine↗

PubMed1991

Voltage-dependent block by magnesium of neuronal nicotinic acetylcholine receptor channels in rat phaeochromocytoma cells.

1. The effects of Mg2+ on the single-channel conductance of neuronal nicotinic acetylcholine receptors were examined using receptors expressed by the rat phaeochromocytoma cell line, PC12. PC12 cells express at least three conductance classes of channels that are activated by acetylcholine, the largest conductance class being the most prevalent. This receptor channel is blocked by intracellular and extracellular Mg2+. 2. The effects of Mg2+ are asymmetrical; at a given concentration, internal Mg2+ is more effective at blocking outward currents than external Mg2+ is at blocking inward currents. Receptor channels are blocked at concentrations of Mg2+ that are low compared to the concentration of the main permeant cation, Na+, and the block is voltage dependent. 3. The block by Mg2+ is not complete as Mg2+ can permeate the channel. With 80 mM-extracellular Mg2+ (no extracellular Na+), the channel has an inward slope conductance of 2.9 pS. 4. The block by extracellular Mg2+ can be described by a one site, two barrier model for the channel which includes a negative surface charge on the external surface of the membrane. The parameters of the model place the binding site for Mg2+ at 52% of the membrane field from the outside with an apparent dissociation constant of 14 mM. However, the same parameters cannot describe the block by intracellular Mg2+. The deviations from the model suggest that the receptor channel may have more than one binding site for Mg2+.

Acetylcholine↗

PubMed1990

Regulation of sodium currents and acetylcholine responses in PC12 cells.

Voltage-gated sodium currents and acetylcholine-elicited currents in clonal rat pheochromocytoma cells (PC12) were studied using the whole-cell patch-clamp technique. After treatment of cultures with nerve growth factor (NGF, 2-4 nM) for 5 or more days, both Na currents and ACh responses increased by 5-7-fold. We tested the ability of a number of treatments reported to induce physiological differentiation in neuroblastoma or neuroblastoma-glioma hybrid cells. We found that no treatment was as effective as NGF, and mitotic inhibitors and 8-bromocyclic AMP reduced the efficacy of NGF at increasing both sodium currents and ACh responses. Some treatments were able to selectively reduce or enhance the ability of NGF to induce ACh responses or sodium currents. Dexamethasone, in particular, completely blocked the NGF-induced increase in ACh response, while leaving Na currents unaffected. Furthermore, in individual cells the Na current density and ACh current density are uncorrelated. These observations indicate that physiological differentiation in PC12 cells is regulated differently than in neuroblastoma cells and, further, in PC12 cells sodium currents and ACh responses are independently regulated.

Acetylcholine↗

PubMed1990

Rectification of acetylcholine-elicited currents in PC12 pheochromocytoma cells.

The current-voltage (I-V) relationship for acetylcholine-elicited currents in the rat pheochromocytoma cell line PC12 is nonlinear. Two voltage-dependent processes that could account for the whole-cell current rectification were examined, receptor channel gating and single receptor channel permeation. We found that both factors are involved in the rectification of the whole-cell currents. The voltage dependence of channel gating determines the shape of the I-V curve at negative potentials. The single-channel I-V relationship is inwardly rectifying and largely responsible for the characteristic shape of the whole-cell I-V curve at positive potentials. The rectification of the single-channel currents is produced by the voltage-dependent block of outward currents by intracellular Mg2+ ions.

Acetylcholine↗

PubMed1984

Hemispheric differences in split-brain monkeys viewing and responding to videotape recordings.

Eight split-brain monkeys were tested for hemispheric differences in their viewing of and responses to colored videotape recordings of monkeys, people, animals, and scenery. The number of facial expressions elicited from the right hemisphere was significantly greater than the number made when using the left hemisphere. Monkeys also tended to look longer when viewing with their right hemispheres than when viewing with the left.

Animals↗