Functional expression of a Ca2+-activated K+ channel in Xenopus oocytes injected with RNAs from the rat testis.
The present study investigated the feasibility of using Xenopus oocytes to express sperm ion channel by injection of RNAs extracted from the rat testis. The RNA-injected oocytes expressed an outwardly rectifying current which was dependent on K+ concentration and inhibitable by K+ channel blockers, charybdotoxin (CTX) and tetraethylammonium (TEA). The Ca2+ ionophore, ionomycin, could also stimulate current activation with similar current characteristics in the RNA-injected oocytes, suggesting the expression of a Ca2+-activated K+ channel. Immunolocalization indicated predominant Ca2+-activated K+ channel immunoreactivity associated with spermatogenic cells. Reverse transcriptase-polymerase chain reaction studies confirmed the expression of the Ca2+-activated K+ channel mRNA in isolated spermatogenic cells. Our results suggest that ion channels and/or receptors of spermatogenic cells could be investigated using the Xenopus oocyte as an expression system. The present study also suggests that sperm may possess a Ca2+-activated K+ channel which has been implicated in the process of sperm activation and gamete interaction.
Wu
W L
WL
Department of Physiology, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
So
S C
SC
Sun
Y P
YP
Zhou
T S
TS
Yu
Y
Y
Chung
Y W
YW
Wang
X F
XF
Bao
Y D
YD
Yan
Y C
YC
Chan
H C
HC
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D018999:Charybdotoxin / Q000494:pharmacology; D004594:Electrophysiology; D015870:Gene Expression / Q000235:genetics; D007150:Immunohistochemistry; D015759:Ionomycin / Q000494:pharmacology; D051036:Large-Conductance Calcium-Activated Potassium Channels; D008297:Male; D008845:Microinjections; D009865:Oocytes / Q000378:metabolism*; D010446:Peptide Fragments / Q000276:immunology; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000737:chemistry / Q000378:metabolism*; D024681:Potassium Channels, Calcium-Activated; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D013094:Spermatozoa / Q000378:metabolism; D013737:Testis / Q000737:chemistry*; D019789:Tetraethylammonium / Q000494:pharmacology; D014981:Xenopus
D051036:Large-Conductance Calcium-Activated Potassium Channels; D010446:Peptide Fragments; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D024681:Potassium Channels, Calcium-Activated; D012333:RNA, Messenger; D018999:Charybdotoxin; D015759:Ionomycin; D019789:Tetraethylammonium; D002118:Calcium