[Calcium-dependent chloride channels in plasma membrane of oocytes from toad, Bufo bufo gargarizans]

Sheng Li Xue Bao. 2006 Oct 25;58(5):471-6.
[Article in Chinese]

Abstract

In this paper, membrane current properties of the fully-grown oocytes from toad, Bufo bufo gargarizans, were studied by using two-microelectrode voltage clamp technique. Axion of adult female toad was destroyed, and then ovarian lobes containing oocytes in stage I to VI were removed and incubated in Ca(2+)-free ND96 solution with collagenase (1.5 mg/ml) for 1 h. Subsequently, the oocytes were washed in Ca(2+)-free ND96 solution for 10 min to completely remove the follicular layer. For the experiments only the oocytes in stage V and VI were selected and used during 1 to 5 d. The membrane was depolarized from a holding potential of -80 mV to +60 mV in 10 mV step. It was found that a sustained outward current was elicited by depolarization. Potassium channel blockers (tetraethylammonium chloride, TEA, 10 mmol/L and 4-aminopyridine, 4-AP, 10 mmol/L) reduced the outward current to (23.4+/-0.72)% of the maximum. However, further addition of chloride channel blocker (5-nitro-2, 3-phenypropylamino benzoate, NPPB, 30 micromol/L) could almost completely block the outward current to (2.1+/-0.08)% of the maximum. In the presence of TEA and 4-AP, removal of extracellular Ca(2+) or adding verapamil (40 micromol/L), could also reduce the outward current to (2.2+/-0.04) % and (3.1+/-0.15) % of the maximum, respectively. It is concluded that calcium-dependent chloride channels exist in plasma membrane of Bufo bufo gargarizans oocytes, besides potassium channels.

Publication types

  • English Abstract

MeSH terms

  • 4-Aminopyridine / toxicity
  • Animals
  • Bufo bufo
  • Calcium / metabolism*
  • Cell Membrane / metabolism
  • Chloride Channels / drug effects
  • Chloride Channels / physiology*
  • Female
  • Nitrobenzoates / pharmacology
  • Oocytes / metabolism*
  • Tetraethylammonium Compounds / pharmacology
  • Verapamil / pharmacology

Substances

  • Chloride Channels
  • Nitrobenzoates
  • Tetraethylammonium Compounds
  • 5-nitro-2-(3-phenylpropylamino)benzoic acid
  • 4-Aminopyridine
  • Verapamil
  • Calcium