Optical recording of electrical activity from parallel fibres and other cell types in skate cerebellar slices in vitro.
1. A reliable and simple fish brain slice preparation was obtained from the cerebellum of the skate, and its properties were described. 2. A potentiometric oxonol dye, RH-482, and multiple site optical recording of transmembrane voltage (MSORTV) were used to reveal the electrophysiological properties of the parallel fibre action potential and to measure its conduction (0.13 m/s). The parallel fibre action potential was blocked in the presence of tetrodotoxin (TTX) and prolonged by tetraethylammonium (TEA), suggesting that the upstroke depends upon sodium entry and the repolarization upon potassium efflux. An after-hyperpolarization results from a calcium-dependent potassium conductance. 3. A second potentiometric dye, RH-155, differing only slightly from RH-482, exhibited a high affinity for glial cell membrane, and could be used to monitor changes in extracellular potassium concentration by detecting changes in glial membrane potential. 4. Calcium channel blockers such as cadmium ions blocked the optical signal that reflected the extracellular accumulation of potassium. 5. Interventions that modified the extracellular volume, and thereby affected the accumulation of potassium, produced large changes in the optical signal that monitored glial depolarization. Hypertonic and hypotonic bathing solutions resulted in decreases and increases, respectively, in the magnitude of the extrinsic absorption change that tracked potassium accumulation. 6. Blocking sodium-potassium pump activity by means of ouabain prolonged the time course of the optical signal that was related to potassium accumulation in the extracellular space. 7. Extracellular potassium accumulation was revealed to be critically dependent upon intracellular calcium ions.
Konnerth
A
A
Department of Physiology, University of Pennsylvania, Philadelphia 19104.
Obaid
A L
AL
Salzberg
B M
BM
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D002104:Cadmium / Q000494:pharmacology; D002531:Cerebellum / Q000502:physiology*; D004555:Electric Fish / Q000502:physiology*; D005456:Fluorescent Dyes; D007555:Isoxazoles; D009412:Nerve Fibers / Q000502:physiology*; D009431:Neural Conduction / Q000187:drug effects; D009457:Neuroglia / Q000502:physiology; D011188:Potassium / Q000502:physiology; D012860:Skates, Fish / Q000502:physiology*; D019789:Tetraethylammonium; D013757:Tetraethylammonium Compounds / Q000494:pharmacology; D013779:Tetrodotoxin / Q000494:pharmacology; D013997:Time Factors
D005456:Fluorescent Dyes; D007555:Isoxazoles; D013757:Tetraethylammonium Compounds; D002104:Cadmium; D013779:Tetrodotoxin; D019789:Tetraethylammonium; D011188:Potassium