Oviductal protease and trypsin treatment enhance sperm-envelope interaction in Bufo arenarum coelomic eggs.
We describe the morphological and biochemical changes in Bufo arenarum coelomic egg envelopes (CE) following passage through the oviduct. In this species, the transformation of the CE into the vitelline envelope (VE) leads to the acquisition of fertilizability and involves the cleavage of a glycoprotein component. Electrophoretic patterns indicate that a pars recta oviductal protease selectively hydrolyzes in vitro the 84 and the 55 kDa glycoproteins of the CE. During the CE to VE transformation, the relative concentrations of gp48, 42 and 39 kDa also change. In in vitro tests, sperm binding to envelope glycoprotein occurs when they are exposed to VE but not when treated with CE, and VE labeled glycoproteins bind to the head and mid piece of the sperm. The gp39 VE component has 100% identity with internal domains of the sequence deduced from ovarian cDNA for the homologous zona pellucida glycoprotein type C (ZPC) protein precursor in B. arenarum. The effects of trypsin as a substitute for oviductal protease were also examined. Trypsin selectively attacks the 84 and the 55 kDa glycoproteins without hydrolyzing other components and renders coelomic eggs fertilizable in a jelly water preparation. Therefore, trypsin can mimic in vitro the biological action of the oviductal protease. However, it does not wholly mimic the biological action of the oviduct which, in B. arenarum at least, exceeds a mere proteolytic effect. This fact was verified by the lower fertility rates and the abnormal embryo development found when trypsin-treated coelomic eggs were fertilized in vitro.
Llanos
Ricardo J
RJ
Departamento de Biología del Desarrollo, Instituto Superior de Investigaciones Biológicas, T4000ILI San Miguel de Tucumán, República Argentina.
Barrera
Daniel
D
Valz-Gianinet
Jorge N
JN
Miceli
Dora C
DC
D000595:Amino Acid Sequence; D000818:Animals; D002022:Bufo arenarum / Q000502:physiology*; D005260:Female; D005298:Fertility / Q000187:drug effects; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism; D066298:In Vitro Techniques; D008297:Male; D008969:Molecular Sequence Data; D010057:Oviducts / Q000201:enzymology; D010063:Ovum / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D012697:Serine Endopeptidases / Q000494:pharmacology*; D013084:Sperm-Ovum Interactions / Q000187:drug effects* / Q000502:physiology; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology; D014357:Trypsin / Q000494:pharmacology*; D014817:Vitelline Membrane / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism
D006023:Glycoproteins; D012697:Serine Endopeptidases; C074599:oviductin; D014357:Trypsin