KATP channels in mouse spermatogenic cells and sperm, and their role in capacitation.
Mammalian sperm must undergo a series of physiological changes after leaving the testis to become competent for fertilization. These changes, collectively known as capacitation, occur in the female reproductive tract where the sperm plasma membrane is modified in terms of its components and ionic permeability. Among other events, mouse sperm capacitation leads to an increase in the intracellular Ca(2+) and pH as well as to a hyperpolarization of the membrane potential. It is well known that ion channels play a crucial role in these events, though the molecular identity of the particular channels involved in capacitation is poorly defined. In the present work, we report the identification and potential functional role of K(ATP) channels in mouse spermatogenic cells and sperm. By using whole-cell patch clamp recordings in mouse spermatogenic cells, we found K(+) inwardly rectifying (K(ir)) currents that are sensitive to Ba(2+), glucose and the sulfonylureas (tolbutamide and glibenclamide) that block K(ATP) channels. The presence of these channels was confirmed using inhibitors of the ATP synthesis and K(ATP) channel activators. Furthermore, RT-PCR assays allowed us to detect transcripts for the K(ATP) subunits SUR1, SUR2, K(ir)6.1 and K(ir)6.2 in total RNA from elongated spermatids. In addition, immunoconfocal microscopy revealed the presence of these K(ATP) subunits in mouse spermatogenic cells and sperm. Notably, incubation of sperm with tolbutamide during capacitation abolished hyperpolarization and significantly decreased the percentage of AR in a dose-dependent fashion. Together, our results provide evidence for the presence of K(ATP) channels in mouse spermatogenic cells and sperm and disclose the contribution of these channels to the capacitation-associated hyperpolarization.
Acevedo
Juan José
JJ
Department of Developmental Genetics and Molecular Physiology, Institute of Biotechnology, UNAM, Cuernavaca, Mexico.
Mendoza-Lujambio
Irene
I
de la Vega-Beltrán
José Luis
JL
Treviño
Claudia L
CL
Felix
Ricardo
R
Darszon
Alberto
A
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001464:Barium / Q000378:metabolism / Q000494:pharmacology; D003981:Diazoxide / Q000378:metabolism / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005905:Glyburide / Q000378:metabolism / Q000494:pharmacology; D054086:KATP Channels; D008297:Male; D008564:Membrane Potentials; D051379:Mice; D018613:Microscopy, Confocal; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000378:metabolism; D020110:Pinacidil / Q000378:metabolism / Q000494:pharmacology; D024661:Potassium Channels, Inwardly Rectifying / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D011955:Receptors, Drug; D013075:Sperm Capacitation / Q000502:physiology*; D013091:Spermatogenesis / Q000502:physiology*; D013094:Spermatozoa / Q000166:cytology / Q000378:metabolism*; D064233:Sulfonylurea Receptors; D013997:Time Factors; D014044:Tolbutamide / Q000378:metabolism / Q000494:pharmacology; D000099161:Kcnj11 Channel
D018528:ATP-Binding Cassette Transporters; D000255:Adenosine Triphosphate; D001464:Barium; D003981:Diazoxide; D005905:Glyburide; D054086:KATP Channels; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D020110:Pinacidil; D024661:Potassium Channels, Inwardly Rectifying; D012333:RNA, Messenger; D011955:Receptors, Drug; D064233:Sulfonylurea Receptors; D014044:Tolbutamide; D000099161:Kcnj11 Channel; C577508:Abcc8 protein, mouse; C092520:uK-ATP-1 potassium channel