Evidence suggesting that the mouse sperm acrosome reaction initiated by the zona pellucida involves an alpha7 nicotinic acetylcholine receptor.
The mammalian sperm acrosome reaction (AR) is essential to fertilization and is believed to be initiated in vivo by ZP3, a glycoprotein component of the egg zona pellucida (ZP). Recently, we reported the results of antagonist studies suggesting that a nicotinic acetylcholine receptor (nAChR) containing an alpha7 subunit (alpha7nAChR) plays a role in the human sperm AR initiated by recombinant human ZP3 or by acetylcholine (ACh). Here, we show that ACh can initiate the mouse sperm AR and that antagonists of the nAChR inhibit the AR initiated by ACh or by ZP obtained from ovarian oocytes (isolated heat-solubilized mouse ZP). Preincubation with three antagonists of the nAChR, alpha-bungarotoxin (100 nM), alpha-conotoxin IMI (100 nM), and methyllycaconitine (100 nM), significantly blocked AR initiation by ACh or by isolated heat-solubilized mouse ZP (P </= 0.002). Because the only nAChR subunit known to bind all three antagonists is the alpha7, an alpha7nAChR appears to be involved in the mouse sperm AR initiated by mouse ZP or by ACh. The nAChR antagonists did not inhibit the AR initiated by calcium ionophore A23187, suggesting that the role of alpha7nAChR is upstream from Ca2+ influx. Pertussis toxin (PTX, 100 ng/ml) did not inhibit the AR initiated by ACh, suggesting that the alpha7nAChR might be a candidate for the PTX-insensitive, poorly selective cation channel shown previously to play a role in ZP-initiated mouse sperm AR. These studies with mouse sperm and ovary-derived ZP strongly support our previous conclusion that activation of an alpha7nAChR is important to the mammalian AR initiated by the egg ZP.
Son
Jung-Ho
JH
Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616-8643, USA.
Meizel
Stanley
S
D000109:Acetylcholine / Q000494:pharmacology; D000157:Aconitine / Q000031:analogs & derivatives* / Q000494:pharmacology; D020101:Acrosome Reaction / Q000502:physiology*; D000818:Animals; D002038:Bungarotoxins / Q000494:pharmacology; D018680:Cholinergic Antagonists / Q000494:pharmacology; D020916:Conotoxins / Q000494:pharmacology; D006358:Hot Temperature; D008297:Male; D051379:Mice; D018733:Nicotinic Antagonists / Q000494:pharmacology; D011978:Receptors, Nicotinic / Q000502:physiology*; D012995:Solubility; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000502:physiology*; D015044:Zona Pellucida / Q000502:physiology*; D064569:alpha7 Nicotinic Acetylcholine Receptor
D002038:Bungarotoxins; D018680:Cholinergic Antagonists; C579167:Chrna7 protein, human; C579166:Chrna7 protein, mouse; D020916:Conotoxins; D018733:Nicotinic Antagonists; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; C087686:alpha-conotoxin ImI; C054634:methyllycaconitine; D000109:Acetylcholine; D000157:Aconitine