A patch clamp study on reconstituted calcium permeable channels of human sperm plasma membranes.
Ionic flux is thought to be important in the initiating process of gamete interaction such as acrosome reaction. However, modern electrophysiological methods, intracellular recording and patch-clamping, are difficult to approach the ion channels in mammal sperm membrane of an intact sperm due to its small size. In this work, by reconstituting the channel protein into lipid bilayer, Ca2+ channels in human spermatozoa were investigated with voltage clamp technique. Membrane proteins isolated from human sperm of 12 healthy donors were incorporated into lipid bilayer via fusion. In a cis 50//trans 10 mmol/L CaCl2 solution system, two types of channel events with similar reversal potential near the value of a perfect Ca2+ electrode, and sensitive to nifedipine and verapamil, were observed. Their unit conductance was 40 and 25 pS respectively. Percentage of channel open time was not dependent to holding potential for the former. However, for the channels of 25 pS, the percentage increased when the holding potential was changed from -20 to 100 mV. Ca(2+)-permeable channels were also detected from the spermatozoon samples of two infertile donors. Abnormal open time of these channels indicates that there are some defects in the conformation of the channel protein of infertile sperm membrane.
Ma
X H
XH
Shanghai Institute of Physiology, Chinese Academy of Sciences, Shanghai 200031.
Shi
Y L
YL
D000328:Adult; D015220:Calcium Channels / Q000378:metabolism / Q000502:physiology*; D004594:Electrophysiology; D006801:Humans; D007248:Infertility, Male / Q000378:metabolism* / Q000503:physiopathology; D008051:Lipid Bilayers / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D008875:Middle Aged; D018408:Patch-Clamp Techniques; D011994:Recombinant Proteins / Q000378:metabolism; D013094:Spermatozoa / Q000378:metabolism* / Q000502:physiology
D015220:Calcium Channels; D008051:Lipid Bilayers; D008565:Membrane Proteins; D011994:Recombinant Proteins