Biophysical and pharmacological characterization of spermatogenic T-type calcium current in mice lacking the CaV3.1 (alpha1G) calcium channel: CaV3.2 (alpha1H) is the main functional calcium channel in wild-type spermatogenic cells

J Cell Physiol. 2004 Jul;200(1):116-24. doi: 10.1002/jcp.10480.

Abstract

Mammalian acrosome reaction (AR) requires successive activation of three different types of calcium channels (T-type channels, Inositol-3-phosphate (InsP3) receptors, and TRPC2 channels). All the calcium signaling is under the control of the activation of the first-one, a T-type calcium channel. The molecular characterization of the T-type calcium channel is still a matter of debate, previous reports showing the presence of transcripts for Ca(V)3.1 and Ca(V)3.2 subunits. Using mice deficient for Ca(V)3.1 subunit, we show that the T-type current density in spermatogenic cells is not reduced in deficient mice versus control mice. We characterized the biophysical and pharmacological properties of T-type current in spermatogenic cells from Ca(V)3.1 deficient mice. Biophysical and pharmacological properties of spermatogenic T-type current from wild-type and Ca(V)3.1 deficient mice demonstrate that Ca(V)3.3 does not contribute to T-type current. Moreover, nickel and amiloride inhibit T-type currents in deficient and wild-type mice with similar potencies. These results demonstrate that T-type currents in spermatogenic cells is due to Ca(V)3.2 subunit and that Ca(V)3.1 contributes to a very negligible extent to the T-type currents. Thus, the deficient Ca(V)3.1 mouse model allows the characterization of native Ca(V)3.2 currents in spermatogenic cells. Spermatogenic Ca(V)3.2 currents present specific feature in comparison to the cloned Ca(V)3.2 current so far. More particularly, the time-dependence of recovery from short-term inactivation of native spermatogenic Ca(V)3.2 is close to 100 millisecond, a value expected for Ca(V)3.1 current.

Publication types

  • Comparative Study

MeSH terms

  • Amiloride / pharmacology
  • Animals
  • Biophysical Phenomena
  • Biophysics*
  • Calcium / metabolism
  • Calcium Channel Blockers / pharmacology*
  • Calcium Channels / drug effects*
  • Calcium Channels / genetics
  • Calcium Channels / metabolism
  • Calcium Channels, T-Type / deficiency
  • Calcium Channels, T-Type / genetics
  • Calcium Channels, T-Type / physiology*
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Inhibitory Concentration 50
  • Male
  • Mice
  • Mice, Knockout
  • Nickel / pharmacology
  • Patch-Clamp Techniques
  • Spermatozoa / cytology
  • Spermatozoa / drug effects
  • Spermatozoa / metabolism*
  • Time Factors

Substances

  • Calcium Channel Blockers
  • Calcium Channels
  • Calcium Channels, T-Type
  • Amiloride
  • Nickel
  • Calcium