Skip to main content
Log in

Voltage dependent calcium channels in cerebellar granule cell primary cultures

  • Published:
European Biophysics Journal Aims and scope Submit manuscript

Abstract

Voltage activated calcium channels were studied in rat cerebellar granule cells in primary culture. Macroscopic currents, carried by 20 mM Ba2+, were measured in the whole-cell configuration. Slowly inactivating macroscopic currents, with a maximum value at a membrane potential around 5 mV were recorded between the 1st and the 4th day in culture. These currents were completely blocked by 5 mM Co2+ , partially blocked by 10 μM nifedipine, and increased by 2 to 5 μM BAY K-8644. Two types of channels, in the presence of 80 mM Ba2+, were identified by single channel recording in cell-attached patches. The first type, which was dihydropyridine agonist sensitive, had a conductance of 18 pS, a half activation potential of more than 10 mV and did not inactivate. This type of channel was the only type found during the first four days in culture, although it was also present up to the 11th day. The second type of channel was dihydropyridine insensitive, had a conductance of 10 pS, a half activation potential less than −15 mV and displayed voltage dependent inactivation. This second type of channel was found in cells for more than four days in culture.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Armstrong CM, Bezanilla F (1977) Inactivation of the sodium channels. II Gating current experiments. J Gen Physiol 70:567–590

    Google Scholar 

  • Bean BP (1989a) Classes of calcium channels in vertebrate cells. Ann Rev Physiol 51:367–384

    Google Scholar 

  • Bean BP (1989b) More than one Ca2+ channel? TINS 12:128–130

    Google Scholar 

  • Carbone E, Lux HD (1984) Low voltage-activated calcium conductance in embrionic chick sensory neurons. Biophys J 46:413–418

    Google Scholar 

  • Carbone E, Lux HD (1987) Single low-voltage-activated calcium channels in chick and rat sensory neurones. J Physiol 386:571–601

    Google Scholar 

  • Connor JA, Tseng H-Y, Hockberger PE (1987) Depolarization- and transmitter-induced changes in intracellular Ca2+ of rat cerebellar granule cells in explant cultures. J Neurosci 7:1384–1399

    Google Scholar 

  • Cull-Candy SG, Howe JR, Odgen DC (1988) Noise and single channels activated by excitatory amino acids in rat cerebellar granule neurons. J Physiol 400:189–222

    Google Scholar 

  • Cull-Candy SG, Marshal CG, Odgen D (1989) Voltage-activated membrane currents in rat cerebellar granule neurones. J Physiol 414:179–199

    Google Scholar 

  • Droogmans G, Nilius B (1989) Kinetic properties of the cardiac T-type calcium channel in the guinea pig. J Physiol 414:627–650

    Google Scholar 

  • Fox AP, Nowycky MC, Tsien RW (1987) Kinetic and pharmacological properties distinguishing three types of calcium currents in chick sensory neurons. J Physiol 394:149–172

    Google Scholar 

  • Galdzicki Z, Lin F, Moran O, Novelli A, Puia G, Sciancalepore M (1990) Development of voltage activated potassium currents in cerebellar granule cells in culture. Int J Neurosci 56:193–200

    Google Scholar 

  • Hamill O, Marty A, Neher E, Sakmann B, Sigworth F (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100

    Google Scholar 

  • Hockberger PE, Tseng H-Y, Connor JA (1987) Immunochemical and electrophysiological differentiation of rat cerebellar granule cells explant cultures. J Neurosci 7:1370–1383

    Google Scholar 

  • Hodgkin AL, Huxley AF (1952) A quantitative description of membrane current and its application to conduction and excitation in nerve. J Physiol 117:500–544

    Google Scholar 

  • Jalonen T, Johansson S, Holopainen I, Oja AA, Arhem P (1990) Single-channel and whole-cell currents in rat cerebellar granule cells. Brain Res 535:33–38

    Google Scholar 

  • Kokubun S, Reuter H (1984) Dihydropyridine derivatives prolong the open state of Ca channels in cultured cardiac cells. Proc Natl Acad Sci 81:4824–4827

    Google Scholar 

  • Kingsbury A, Balazs R (1987) Effect of calcium agonists and antagonist on cerebellar granule cells. Eur J Pharmacol 140:275–283

    Google Scholar 

  • Levi G, Aloisi M, Ciotti M, Gallo V (1984) Autoradiographic localization and depolarization-induced release of acidic amino acids in differentiating cerebellar granule cells cultures. Brain Res 290:77–86

    Google Scholar 

  • Lin F, Moran O (1990) Voltage dependent sodium currents in cultured cerebellar granules. Biosci Rep 10:445–453

    Google Scholar 

  • Lin F, Conti F, Moran O (1991) Competitive blockage of the sodium channel by intracellular magnesium ions in central mammalian neurones. Eur Biophys J 19:109–118

    Google Scholar 

  • Marchetti C, Carignani C, Robello M (1990) A voltage dependent calcium current in cultured rat cerebellar granule cells. Proceedings of the IX Congresso della Society Italiana di Biofisica, Pura ed Applicata, p 185

  • Mori Y, Friedrich T, Kim MS, Mikami A, Nakai J, Ruth P, Bosse E, Hofmann F, Flockerzi V, Furiuchi T, Mikoshiba K, Imoto K, Tanabe T, Numa S (1991) Primary structure and functional expression from complementary DNA of a brain calcium channel. Nature 350:398–401

    Google Scholar 

  • Nowycky MC, Fox AP, Tsien RW (1985) Three types of neuronal calcium channel with different calcium agonist sensitivity. Nature 361:440–443

    Google Scholar 

  • Pusch M, Neher E (1988) Rates of diffusional exchange between small cells and a measuring patch pipette. Pflügers Arch 411: 204–211

    Google Scholar 

  • Reuter H, Porzing H, Kokubun S, Prod'hom B (1985) 1,4-dihydropyridines as tools in the study of Ca2+ channels. TINS 8:396–400

    Google Scholar 

  • Robello M, Carignani C, Marchetti C (1989) A transient voltage-dependent outward current in cultured cerebellar granules. Biosci Rep 9:451–457

    Google Scholar 

  • Sanguinetti MC, Kaas RS (1986) Volgate-dependent modulation of Ca channel current in heart cells by BAY K-8644. J Gen Physiol 88:369–392

    Google Scholar 

  • Sciancalepore M, Forti L, Moran O (1989) Changes of N-methyl-d-aspartate-activated channels of cerebellar granule cells with days in culture. Biochem Biophys Res Commun 165:481–487

    Google Scholar 

  • Sciancalepore M, Galdzicki Z, Zheng X, Moran O (1990) Kainate activated single channel currents as revealed by domoic acid. Eur Biophys J 19:63–68

    Google Scholar 

  • Sigworth FJ, Sine SM (1987) Data transformations for improved display and fitting of single-channel dwell time histograms. Biophys J 52:1047–1054

    Google Scholar 

  • Slesinger PA, Lansman JB (1990) Just one type of Ca2+ channel underlies the macroscopic Ca2+ current in cerebellar granule cells. Biophys J 57:524a

    Google Scholar 

  • Swandulla D, Carbone E, Lux HD (1991) Do calcium channel classification account for neuronal calcium channel activity? TINS 14:46–51

    Google Scholar 

  • Tsien RW (1983) Calcium channels in excitable cell membranes. Ann Rev Physiol 45:341–358

    Google Scholar 

  • Tsien RW, Lipscombe D, Madison DV, Bley KR, Fox AP (1988) Multiple types of calcium channels and their selective modulation. Trends Neurosci 11:431–443

    Google Scholar 

  • Wojcik WJ, Travagli RA, Costa E, Bertolino M (1990) Baclophen inhibits with high affinity an L-type — like voltage-dependent calcium channel in cerebellar granule cell cultures. Neuropharmacology 29:969–972

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Moran, O., Lin, F., Zegarra-Moran, O. et al. Voltage dependent calcium channels in cerebellar granule cell primary cultures. Eur Biophys J 20, 157–164 (1991). https://doi.org/10.1007/BF01561138

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01561138

Key words

Navigation