Abstract
Neurons enzymatically isolated from the adult rat superior cervical ganglion (SCG) were investigated using the whole-cell variant of the patch-clamp technique. Currentclamp studies revealed the following mean passive and active membrane properties: resting membrane potential, −54.9 mV; input resistance, 349 MΩ; action potential (AP) threshold, −29.8 mV; AP overshoot, 53.3 mV; AP maximum rate of rise, 166.4 V/s; and AP duration, 3.2 ms. Chemosensitivity to acetylcholine remained intact following enzymatic dispersion. Voltage-clamp studies of a transient tetrodotoxin-sensitive Na+ current revealed activation and inactivation processes which could be fit to modified Boltzmann equations. Na+ current activation parameters for the half activation potential (Vh) and slope factor (K) were −23.3 mV and 5.3 mV, respectively. Inactivation parameters forVh andK were −59.3 mV and 7.6 mV, respectively. Voltage-clamp studies also revealed a high voltageactivated sustained inward current which was eliminated upon removal of external Ca2+, greatly reduced by 500 μM Cd2+, and supported by Ba2+ or Sr2+. Tail current analysis of this Ca2+ current revealed a sigmoidal activation. A low voltage-activated transient Ca2+ current was not observed. We conclude that isolated SCG neurons retain the properties of neurons in intact ganglia and provide several advantages over conventional preparations for the study of voltagegated membrane currents.
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Adams PR, Brown DA, Constanti A (1982) M-currents and other potassium currents in bullfrog sympathetic neurones. J Physiol (Lond) 330:537–572
Almers W, McCleskey EW (1984) Non-selective conductance in calcium channels of frog muscle: calcium selectivity in a singlefile pore. J Physiol (Lond) 353:585–608
Belluzzi O, Sacchi O (1986) A quantitative description of the sodium current in the rat sympathetic neurone. J Physiol (Lond) 380:275–291
Belluzzi O, Sacchi O, Wanke E (1985a) A fast transient outward current in the rat sympathetic neurone studied under voltageclamp conditions. J Physiol (Lond) 358:91–108
Belluzzi O, Sacchi O, Wanke E (1985b) Identification of delayed potassium and calcium currents in the rat sympathetic neurone under voltage-clamp. J Physiol (Lond) 358:109–129
Bossu JL, Feltz A (1984) Patch-clamp study of tetrodotoxin-resistant sodium currents in group C sensory neurones. Neurosci Lett 51:241–246
Bossu JL, Feltz A (1986) Inactivation of the low-threshold transient calcium current in rat sensory neurones: evidence for a dual process. J Physiol (Lond) 376:341–357
Bossu JL, Feltz A, Thomann JM (1985) Depolarization elicits two distinct calcium currents in vertebrate sensory neurons. Pflügers Arch 403:360–368
Brown DA, Selyanko AA (1985) Two components of muscarinesensitive membrane currents in rat sympathetic neurons. J Physiol (Lond) 358:335–363
Carbone E, Lux HD (1984a) A low-voltage-activated calcium conductance in embryonic chick sensory neurons. Biophys J 46:413–418
Carbone E, Lux HD (1984b) A low voltage-activated, fully inactivating Ca2+ channel in vertebrate sensory neurones. Nature (Lond) 310:501–502
Carbone E, Lux HD (1986) Sodium channels in cultured chick dorsal root ganglion neurons. Eur Biophys J 13:259–271
Carbone E, Lux HD (1987a) Kinetics and selectivity of a lowvoltage-activated calcium current in chick and rat sensory neurones. J Physiol (Lond) 386:547–570
Carbone E, Lux HD (1987b) Single low-voltage-activated calcium current in chick and rat sensory neurones. J Physiol (Lond) 386:571–601
Constanti A, Brown DA (1981) M-currents in voltage-clamped mammalian sympathetic neurones. Neurosci Lett 24:289–294
Cull-Candy SG, Mathie A (1986) Ion channels activated by acetylcholine and γ-aminobutyric acid in freshly dissociated sympathetic neurons of the rat. Neurosci Lett 66:275–280
Cull-Candy SG, Magnus CJ, Mathie A (1986) Freshly dissociated rat sympathetic neurones: a preparation suitable for patch-clamp recording. J Physiol (Lond) 377:2P
Derkach VA, North RA, Selyanko AA, Skok VI (1987) Single channels activated by acetylcholine in rat superior cervical ganglion. J Physiol (Lond) 388:141–151
Dolphin AC, Scott RH (1987) Calcium channel currents and their inhibition by (−)-baclofen in rat sensory neurones: modulation by guanine nucleotides. J Physiol (Lond) 386:1–17
Fedulova SA, Kostyuk PG, Veselovsky NS (1985) Two types of calcium channels in the somatic membrane of new-born rat dorsal root ganglion neurones. J Physiol (Lond) 359:431–446
Fenwick EM, Marty A, Neher E (1982a) A patch-clamp study of bovine chromaffin cells and their sensitivity to acetylcholine. J Physiol (Lond) 331:577–597
Fenwick EM, Marty A, Neher E (1982b) Sodium and calcium channels in bovine chromaffin cells. J Physiol (Lond) 331:599–635
Fernandez JM, Fox AP, Krasne S (1984) Membrane patches and whole-cell membranes: a comparison of electrical properties in rat clonal pituitary (GH3) cells. J Physiol (Lond) 356:565–585
Forscher P, Oxford GS (1985) Modulation of calcium channel by norepinephrine in internally dialyzed avian sensory neurons. J Gen Physiol 85:743–763
Freschi JE (1983) Membrane currents of cultured rat sympathetic neurons under voltage clamp. J Neurophysiol 50:1460–1478
Fukuda J, Kameyama M (1980) A tissue-culture of nerve cells from adult mammalian ganglia and some electrophysiological properties of the nerve cells in vitro. Brain Res 202:249–255
Galvan M (1982) A transient outward current in rat sympathetic neurones. Neurosci Lett 31:295–300
Galvan M, Adams PR (1982) Control of calcium current in rat sympathetic neurons by norepinephrine. Brain Res 244:135–144
Galvan M, Sedlmeir C (1983) An N-shaped current-voltage relationship in rat sympathetic neurones. Pflügers Arch 398:78–80
Galvan M, Sedlmeir C (1984) Outward currents in voltage-clamped rat sympathetic neurones. J Physiol (Lond) 356:115–133
Hagiwara S, Byerly L (1981) Calcium channels. Annu Rev Neurosci 4:69–125
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100
Hess P, Fox AP, Lansman JB, Nilius B, Nowycky MC, Tsien RW (1986a) Calcium channel types in cardiac, neuronal, and smooth muscle-derived cells: differences in gating, permeation, and pharmacology. In: Ritchie JM, Keynes RD, Bolis L (eds) Ion channels in neural membranes. Liss, New York, pp 227–252
Hess P, Tsien RW (1984) Mechanism of ion permeation through calcium channels. Nature (Lond) 309:453–456
Hess P, Lansman JB, Tsien RW (1986b) Calcium channel selectivity for divalent and monovalent cations. Voltage and concentration dependence of single channel current in ventricular heart cells. J Gen Physiol 88:293–320
Hille B (1971) The permeability of the sodium channel to organic cations in myelinated nerve. J Gen Physiol 58:599–619
Hodgkin AL, Huxley AF (1952) Currents carried by sodium and potassium ions through the membrane of the giant axon of Loligo. J Physiol (Lond) 116:449–472
Holtz GG, Rane SG, Dunlap K (1986) GTP-binding proteins mediate transmitter inhibition of voltage-dependent calcium channels. Nature (Lond) 319:670–672
Ikeda SR, Schofield GG (1987) Tetrodotoxin-resistant sodium current of rat nodose neurones: Monovalent cation selectivity and divalent cation block. J Physiol (Lond) 389:255–270
Ikeda SR, Schofield GG, Weight FF (1986) Na+ and Ca2+ currents of acutely isolated adult rat nodose ganglion cells. J Neurophysiol 55:527–539
Ikeda SR, Schofield GG, Weight FF (1987) Somatostatin blocks a calcium current in acutely isolated adult rat superior cervical ganglion neurons. Neurosci Lett 81:123–128
Kay AR, Wong RKS (1986) Isolation of neurons suitable for patch-clamping from adult mammalian central nervous systems. J Neurosci Meth 16:227–238
Kostyuk PG, Krishtal OA, Pidoplicko VI (1975) Effect of internal fluoride and phosphate on membrane currents during intracellular dialysis of nerve cells. Nature (Lond) 257:691–693
Kostyuk PG, Veselovsky NS, Tsyndrenko AY (1981a) Ionic currents in the somatic membrane of rat dorsal root ganglion neurons. I. Sodium currents. Neuroscience 6:2423–2430
Kostyuk PG, Veselovsky NS, Fedulova SA (1981b) Ionic currents in the somatic membrane of rat dorsal root ganglion neurons. II. Calcium currents. Neuroscience 6:2431–2437
Lee KS, Akaike N, Brown AM (1977) Trypsin inhibits the action of tetrodotoxin on neurones. Nature (Lond) 265:751–753
Marchetti C, Carbone E, Lux HD (1986) Effects of dopamine and noradrenaline on Ca2+ channels of cultured sensory and sympathetic neurons of chick. Pflügers Arch 406:104–111
Marrion NV, Smart TG, Brown DA (1987) Membrane currents in adult rat superior cervical ganglia in dissociated tissue culture. Neurosci Lett 77:55–60
Matteson DR, Armstrong CM (1986) Properties of two types of calcium channels in clonal pituitary cells. J Gen Physiol 87:161–182
McAfee DA, Yarowsky PJ (1979) Calcium-dependent potentials in the mammalian sympathetic neurone. J Physiol (Lond) 290:507–523
Miller RJ (1987) Multiple calcium channels and neuronal function. Science 235:46–52
Nowycky MC, Fox AP, Tsien RW (1985) Three types of neuronal calcium channel with different calcium agonist sensitivity. Nature (Lond) 316:440–443
Perri V, Sacchi O, Casella C (1970) Electrical properties and synaptic connections of the sympathetic neurons in the rat and guineapig superior cervical ganglion. Pflügers Arch 314:40–54
Pettman B, Louis JC, Sensenbrenner M (1979) Morphological and biochemical maturation of neurons cultured in the absence of glial cells. Nature (Lond) 281:378–380
Schofield GG, Ikeda SR, Weight FF (1986) Three potassium currents in acutely isolated adult rat superior cervical ganglion cells. Soc Neurosci Abstr 12:1344
Scott BS (1982) Adult neurons in cell culture: electrophysiology characterization and use in neurobiological research. Prog Neurobiol 19:187–211
Wakshull E, Johnson MI, Burton H (1979) Postnatal rat sympathetic neurons in culture, I. A comparison with embryonic neurons. J Neurophysiol 42:1410–1425
Wanke E, Ferroni A, Malgaroli A, Ambrosini A, Pozzan T, Meldolesi J (1987) Activation of a muscarinic receptor selectively inhibits a rapidly inactivated Ca2+ current in rat sympathetic neurons. Proc Natl Acad Sci USA 84:4313–4317
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Schofield, G.G., Ikeda, S.R. Sodium and calcium currents of acutely isolated adult rat superior cervical ganglion neurons. Pflugers Arch. 411, 481–490 (1988). https://doi.org/10.1007/BF00582368
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DOI: https://doi.org/10.1007/BF00582368