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. 1991 May;436:739–767. doi: 10.1113/jphysiol.1991.sp018577

Single calcium channels in rat and guinea-pig hippocampal neurons.

T J O'Dell 1, B E Alger 1
PMCID: PMC1181532  PMID: 1648133

Abstract

1. Calcium (Ca2+) channels were studied under whole-cell clamp and in cell-attached patch recordings from acutely isolated and tissue-cultured rat hippocampal neurons. Under whole-cell voltage clamp of tissue-cultured neurons the current-voltage plot for Ca2+ channel current was biphasic, with a 'hump' on the descending phase of the plot when the cell was held at -80 mV and stepped to various command potentials. When determined from a holding potential of -40 mV the plot was no longer biphasic. 2. In cell-attached patch experiments extracellular isotonic potassium gluconate was used to zero the cell membrane potential. When the patch electrode solution contained monovalent cations (150 mM-Li+ in most experiments) and no Ca2+, a small channel (approximately 13 pA) could be clearly distinguished in tissue-cultured neurons from a large dihydropyridine-sensitive channel (approximately 36 pS). With Ba2+ as the charge carrier it was more difficult to resolve small channel openings. The small channel was activated during voltage steps from negative holding potentials (-80 to -100 mV) over a range of potentials (-70 mV and less negative). The channels inactivated rapidly (time constants of 20-40 ms) during the voltage step. Steady-state inactivation and activation functions were well fitted by single Boltzmann equations of the form y = 1/[1 + exp [V-V0.5)/k)] and y = 1/[1 + exp [V0.5-V)/k)], where V0.5 = -82.9 +/- 2.4 and -55.2 +/- 1.5 mV and k = 4.5 +/- 0.6 and 4.9 +/- 0.6 mV, respectively. These small channels were not found in acutely isolated adult cells. 3. Ensemble averages of small-channel activity in numerous sweeps were very similar in time course to the T currents recorded in the whole-cell mode. Often small channels occurred in clusters, with many channels in a single patch. In these multichannel patches the voltage dependence of the kinetics of the channel was clearly revealed. 4. Small channels were insensitive to the dihydropyridine nifedipine (20 microM) and to TTX (1-5 microM), but the Li+ current through this channel was readily blocked by including 2 mM-Ca2+ in the recording pipette. Small-channel activity persisted for minutes in off-cell patches, and in cell-attached patches was not affected by phorbol esters. 5. The large channel was studied with electrodes filled with 110 mM-Ca2+ or Ba2+ (single-channel conductance approximately 24 pS in Ba2+). Activity of this channel was dramatically increased by the dihydropyridine Bay K 8644 and suppressed by nifedipine.(ABSTRACT TRUNCATED AT 400 WORDS)

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Selected References

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  1. Akaike N., Kostyuk P. G., Osipchuk Y. V. Dihydropyridine-sensitive low-threshold calcium channels in isolated rat hypothalamic neurones. J Physiol. 1989 May;412:181–195. doi: 10.1113/jphysiol.1989.sp017610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aosaki T., Kasai H. Characterization of two kinds of high-voltage-activated Ca-channel currents in chick sensory neurons. Differential sensitivity to dihydropyridines and omega-conotoxin GVIA. Pflugers Arch. 1989 Jun;414(2):150–156. doi: 10.1007/BF00580957. [DOI] [PubMed] [Google Scholar]
  3. Armstrong C. M., Matteson D. R. Two distinct populations of calcium channels in a clonal line of pituitary cells. Science. 1985 Jan 4;227(4682):65–67. doi: 10.1126/science.2578071. [DOI] [PubMed] [Google Scholar]
  4. Armstrong D., Eckert R. Voltage-activated calcium channels that must be phosphorylated to respond to membrane depolarization. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2518–2522. doi: 10.1073/pnas.84.8.2518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baraban J. M., Snyder S. H., Alger B. E. Protein kinase C regulates ionic conductance in hippocampal pyramidal neurons: electrophysiological effects of phorbol esters. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2538–2542. doi: 10.1073/pnas.82.8.2538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bazzi M. D., Nelsestuen G. L. Association of protein kinase C with phospholipid monolayers: two-stage irreversible binding. Biochemistry. 1988 Sep 6;27(18):6776–6783. doi: 10.1021/bi00418a020. [DOI] [PubMed] [Google Scholar]
  7. Blaxter T. J., Carlen P. L., Niesen C. Pharmacological and anatomical separation of calcium currents in rat dentate granule neurones in vitro. J Physiol. 1989 May;412:93–112. doi: 10.1113/jphysiol.1989.sp017605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brown D. A., Griffith W. H. Persistent slow inward calcium current in voltage-clamped hippocampal neurones of the guinea-pig. J Physiol. 1983 Apr;337:303–320. doi: 10.1113/jphysiol.1983.sp014625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Carbone E., Lux H. D. A low voltage-activated, fully inactivating Ca channel in vertebrate sensory neurones. Nature. 1984 Aug 9;310(5977):501–502. doi: 10.1038/310501a0. [DOI] [PubMed] [Google Scholar]
  10. Carbone E., Lux H. D. Kinetics and selectivity of a low-voltage-activated calcium current in chick and rat sensory neurones. J Physiol. 1987 May;386:547–570. doi: 10.1113/jphysiol.1987.sp016551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Carbone E., Lux H. D. Omega-conotoxin blockade distinguishes Ca from Na permeable states in neuronal calcium channels. Pflugers Arch. 1988 Nov;413(1):14–22. doi: 10.1007/BF00581223. [DOI] [PubMed] [Google Scholar]
  12. Catterall W. A. The molecular basis of neuronal excitability. Science. 1984 Feb 17;223(4637):653–661. doi: 10.1126/science.6320365. [DOI] [PubMed] [Google Scholar]
  13. Chad J. E., Eckert R. An enzymatic mechanism for calcium current inactivation in dialysed Helix neurones. J Physiol. 1986 Sep;378:31–51. doi: 10.1113/jphysiol.1986.sp016206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Coulter D. A., Huguenard J. R., Prince D. A. Calcium currents in rat thalamocortical relay neurones: kinetic properties of the transient, low-threshold current. J Physiol. 1989 Jul;414:587–604. doi: 10.1113/jphysiol.1989.sp017705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Crunelli V., Lightowler S., Pollard C. E. A T-type Ca2+ current underlies low-threshold Ca2+ potentials in cells of the cat and rat lateral geniculate nucleus. J Physiol. 1989 Jun;413:543–561. doi: 10.1113/jphysiol.1989.sp017668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. DeRiemer S. A., Strong J. A., Albert K. A., Greengard P., Kaczmarek L. K. Enhancement of calcium current in Aplysia neurones by phorbol ester and protein kinase C. Nature. 1985 Jan 24;313(6000):313–316. doi: 10.1038/313313a0. [DOI] [PubMed] [Google Scholar]
  17. Docherty R. J., Brown D. A. Interaction of 1,4-dihydropyridines with somatic Ca currents in hippocampal CA1 neurones of the guinea pig in vitro. Neurosci Lett. 1986 Sep 25;70(1):110–115. doi: 10.1016/0304-3940(86)90447-7. [DOI] [PubMed] [Google Scholar]
  18. Doerner D., Abdel-Latif M., Rogers T. B., Alger B. E. Protein kinase C-dependent and -independent effects of phorbol esters on hippocampal calcium channel current. J Neurosci. 1990 May;10(5):1699–1706. doi: 10.1523/JNEUROSCI.10-05-01699.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Doerner D., Pitler T. A., Alger B. E. Protein kinase C activators block specific calcium and potassium current components in isolated hippocampal neurons. J Neurosci. 1988 Nov;8(11):4069–4078. doi: 10.1523/JNEUROSCI.08-11-04069.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Dolphin A. C., Forda S. R., Scott R. H. Calcium-dependent currents in cultured rat dorsal root ganglion neurones are inhibited by an adenosine analogue. J Physiol. 1986 Apr;373:47–61. doi: 10.1113/jphysiol.1986.sp016034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Dolphin A. C., Scott R. H. Calcium channel currents and their inhibition by (-)-baclofen in rat sensory neurones: modulation by guanine nucleotides. J Physiol. 1987 May;386:1–17. doi: 10.1113/jphysiol.1987.sp016518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ewald D. A., Pang I. H., Sternweis P. C., Miller R. J. Differential G protein-mediated coupling of neurotransmitter receptors to Ca2+ channels in rat dorsal root ganglion neurons in vitro. Neuron. 1989 Feb;2(2):1185–1193. doi: 10.1016/0896-6273(89)90185-2. [DOI] [PubMed] [Google Scholar]
  23. Fisher R. E., Gray R., Johnston D. Properties and distribution of single voltage-gated calcium channels in adult hippocampal neurons. J Neurophysiol. 1990 Jul;64(1):91–104. doi: 10.1152/jn.1990.64.1.91. [DOI] [PubMed] [Google Scholar]
  24. Fox A. P., Nowycky M. C., Tsien R. W. Kinetic and pharmacological properties distinguishing three types of calcium currents in chick sensory neurones. J Physiol. 1987 Dec;394:149–172. doi: 10.1113/jphysiol.1987.sp016864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Fox A. P., Nowycky M. C., Tsien R. W. Single-channel recordings of three types of calcium channels in chick sensory neurones. J Physiol. 1987 Dec;394:173–200. doi: 10.1113/jphysiol.1987.sp016865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Gray R., Johnston D. Noradrenaline and beta-adrenoceptor agonists increase activity of voltage-dependent calcium channels in hippocampal neurons. Nature. 1987 Jun 18;327(6123):620–622. doi: 10.1038/327620a0. [DOI] [PubMed] [Google Scholar]
  27. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  28. Hess P., Prod'Hom B., Pietrobon D. Mechanisms of interaction of permeant ions and protons with dihydropyridine-sensitive calcium channels. Ann N Y Acad Sci. 1989;560:80–93. doi: 10.1111/j.1749-6632.1989.tb24082.x. [DOI] [PubMed] [Google Scholar]
  29. Hirning L. D., Fox A. P., McCleskey E. W., Olivera B. M., Thayer S. A., Miller R. J., Tsien R. W. Dominant role of N-type Ca2+ channels in evoked release of norepinephrine from sympathetic neurons. Science. 1988 Jan 1;239(4835):57–61. doi: 10.1126/science.2447647. [DOI] [PubMed] [Google Scholar]
  30. Horn R., Marty A. Muscarinic activation of ionic currents measured by a new whole-cell recording method. J Gen Physiol. 1988 Aug;92(2):145–159. doi: 10.1085/jgp.92.2.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Huang F. L., Yoshida Y., Cunha-Melo J. R., Beaven M. A., Huang K. P. Differential down-regulation of protein kinase C isozymes. J Biol Chem. 1989 Mar 5;264(7):4238–4243. [PubMed] [Google Scholar]
  32. Johnston D., Hablitz J. J., Wilson W. A. Voltage clamp discloses slow inward current in hippocampal burst-firing neurones. Nature. 1980 Jul 24;286(5771):391–393. doi: 10.1038/286391a0. [DOI] [PubMed] [Google Scholar]
  33. Kay A. R., Miles R., Wong R. K. Intracellular fluoride alters the kinetic properties of calcium currents facilitating the investigation of synaptic events in hippocampal neurons. J Neurosci. 1986 Oct;6(10):2915–2920. doi: 10.1523/JNEUROSCI.06-10-02915.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Kay A. R., Wong R. K. Calcium current activation kinetics in isolated pyramidal neurones of the Ca1 region of the mature guinea-pig hippocampus. J Physiol. 1987 Nov;392:603–616. doi: 10.1113/jphysiol.1987.sp016799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Kay A. R., Wong R. K. Isolation of neurons suitable for patch-clamping from adult mammalian central nervous systems. J Neurosci Methods. 1986 May;16(3):227–238. doi: 10.1016/0165-0270(86)90040-3. [DOI] [PubMed] [Google Scholar]
  36. Kostyuk P. G., Shuba YaM, Savchenko A. N. Three types of calcium channels in the membrane of mouse sensory neurons. Pflugers Arch. 1988 Jun;411(6):661–669. doi: 10.1007/BF00580863. [DOI] [PubMed] [Google Scholar]
  37. Kostyuk P., Akaike N., Osipchuk Y. u., Savchenko A., Shuba Y. a. Gating and permeation of different types of Ca channels. Ann N Y Acad Sci. 1989;560:63–79. doi: 10.1111/j.1749-6632.1989.tb24081.x. [DOI] [PubMed] [Google Scholar]
  38. Lacerda A. E., Rampe D., Brown A. M. Effects of protein kinase C activators on cardiac Ca2+ channels. Nature. 1988 Sep 15;335(6187):249–251. doi: 10.1038/335249a0. [DOI] [PubMed] [Google Scholar]
  39. Lansman J. B., Hess P., Tsien R. W. Blockade of current through single calcium channels by Cd2+, Mg2+, and Ca2+. Voltage and concentration dependence of calcium entry into the pore. J Gen Physiol. 1986 Sep;88(3):321–347. doi: 10.1085/jgp.88.3.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Lipscombe D., Kongsamut S., Tsien R. W. Alpha-adrenergic inhibition of sympathetic neurotransmitter release mediated by modulation of N-type calcium-channel gating. Nature. 1989 Aug 24;340(6235):639–642. doi: 10.1038/340639a0. [DOI] [PubMed] [Google Scholar]
  41. Lux H. D., Carbone E., Zucker H. Block of Na+ ion permeation and selectivity of Ca channels. Ann N Y Acad Sci. 1989;560:94–102. doi: 10.1111/j.1749-6632.1989.tb24083.x. [DOI] [PubMed] [Google Scholar]
  42. MacDonald R. L., Skerritt J. H., Werz M. A. Adenosine agonists reduce voltage-dependent calcium conductance of mouse sensory neurones in cell culture. J Physiol. 1986 Jan;370:75–90. doi: 10.1113/jphysiol.1986.sp015923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Macdonald R. L., Werz M. A. Dynorphin A decreases voltage-dependent calcium conductance of mouse dorsal root ganglion neurones. J Physiol. 1986 Aug;377:237–249. doi: 10.1113/jphysiol.1986.sp016184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Meyers D. E., Barker J. L. Whole-cell patch-clamp analysis of voltage-dependent calcium conductances in cultured embryonic rat hippocampal neurons. J Neurophysiol. 1989 Mar;61(3):467–477. doi: 10.1152/jn.1989.61.3.467. [DOI] [PubMed] [Google Scholar]
  45. Nagle D. S., Blumberg P. M. Regional localization by light microscopic autoradiography of receptors in mouse brain for phorbol ester tumor promoters. Cancer Lett. 1983 Feb;18(1):35–40. doi: 10.1016/0304-3835(83)90115-5. [DOI] [PubMed] [Google Scholar]
  46. Nishizuka Y. The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature. 1988 Aug 25;334(6184):661–665. doi: 10.1038/334661a0. [DOI] [PubMed] [Google Scholar]
  47. Nowycky M. C., Fox A. P., Tsien R. W. Long-opening mode of gating of neuronal calcium channels and its promotion by the dihydropyridine calcium agonist Bay K 8644. Proc Natl Acad Sci U S A. 1985 Apr;82(7):2178–2182. doi: 10.1073/pnas.82.7.2178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Nowycky M. C., Fox A. P., Tsien R. W. Three types of neuronal calcium channel with different calcium agonist sensitivity. Nature. 1985 Aug 1;316(6027):440–443. doi: 10.1038/316440a0. [DOI] [PubMed] [Google Scholar]
  49. Plummer M. R., Logothetis D. E., Hess P. Elementary properties and pharmacological sensitivities of calcium channels in mammalian peripheral neurons. Neuron. 1989 May;2(5):1453–1463. doi: 10.1016/0896-6273(89)90191-8. [DOI] [PubMed] [Google Scholar]
  50. Prod'hom B., Pietrobon D., Hess P. Direct measurement of proton transfer rates to a group controlling the dihydropyridine-sensitive Ca2+ channel. Nature. 1987 Sep 17;329(6136):243–246. doi: 10.1038/329243a0. [DOI] [PubMed] [Google Scholar]
  51. Rane S. G., Dunlap K. Kinase C activator 1,2-oleoylacetylglycerol attenuates voltage-dependent calcium current in sensory neurons. Proc Natl Acad Sci U S A. 1986 Jan;83(1):184–188. doi: 10.1073/pnas.83.1.184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Sanguinetti M. C., Kass R. S. Voltage-dependent block of calcium channel current in the calf cardiac Purkinje fiber by dihydropyridine calcium channel antagonists. Circ Res. 1984 Sep;55(3):336–348. doi: 10.1161/01.res.55.3.336. [DOI] [PubMed] [Google Scholar]
  53. Segal M., Barker J. L. Rat hippocampal neurons in culture: Ca2+ and Ca2+-dependent K+ conductances. J Neurophysiol. 1986 Apr;55(4):751–766. doi: 10.1152/jn.1986.55.4.751. [DOI] [PubMed] [Google Scholar]
  54. Segal M. Rat hippocampal neurons in culture: responses to electrical and chemical stimuli. J Neurophysiol. 1983 Dec;50(6):1249–1264. doi: 10.1152/jn.1983.50.6.1249. [DOI] [PubMed] [Google Scholar]
  55. Snutch T. P., Leonard J. P., Gilbert M. M., Lester H. A., Davidson N. Rat brain expresses a heterogeneous family of calcium channels. Proc Natl Acad Sci U S A. 1990 May;87(9):3391–3395. doi: 10.1073/pnas.87.9.3391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Sperelakis N. Regulation of calcium slow channels of cardiac muscle by cyclic nucleotides and phosphorylation. J Mol Cell Cardiol. 1988 Mar;20 (Suppl 2):75–105. doi: 10.1016/0022-2828(88)90334-3. [DOI] [PubMed] [Google Scholar]
  57. Strong J. A., Fox A. P., Tsien R. W., Kaczmarek L. K. Stimulation of protein kinase C recruits covert calcium channels in Aplysia bag cell neurons. Nature. 1987 Feb 19;325(6106):714–717. doi: 10.1038/325714a0. [DOI] [PubMed] [Google Scholar]
  58. Swandulla D., Armstrong C. M. Calcium channel block by cadmium in chicken sensory neurons. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1736–1740. doi: 10.1073/pnas.86.5.1736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Takahashi K., Tateishi N., Kaneda M., Akaike N. Comparison of low-threshold Ca2+ currents in the hippocampal CA1 neurons among the newborn, adult and aged rats. Neurosci Lett. 1989 Aug 14;103(1):29–33. doi: 10.1016/0304-3940(89)90480-1. [DOI] [PubMed] [Google Scholar]
  60. Thompson S., Coombs J. Spatial distribution of Ca currents in molluscan neuron cell bodies and regional differences in the strength of inactivation. J Neurosci. 1988 Jun;8(6):1929–1939. doi: 10.1523/JNEUROSCI.08-06-01929.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Yaari Y., Hamon B., Lux H. D. Development of two types of calcium channels in cultured mammalian hippocampal neurons. Science. 1987 Feb 6;235(4789):680–682. doi: 10.1126/science.2433765. [DOI] [PubMed] [Google Scholar]
  62. Yatani A., Codina J., Imoto Y., Reeves J. P., Birnbaumer L., Brown A. M. A G protein directly regulates mammalian cardiac calcium channels. Science. 1987 Nov 27;238(4831):1288–1292. doi: 10.1126/science.2446390. [DOI] [PubMed] [Google Scholar]

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