Fate of lactadherin P47 during post-testicular maturation and capacitation of boar spermatozoa.
Polyclonal avian antibody was used partially to characterize the pig sperm lactadherin P47. P47 is a mosaic protein, composed of two epidermal growth factor (EGF)-like domains and two C1/C2 domains. P47 is homologous to the bovine mammary gland protein MGP 53/57 and mouse milk fat globule protein. Expression of P47 along the male genital tract and its localization on spermatozoa during post-testicular maturation and capacitation were studied. P47 was detected in the testis and in all parts of the epididymis by immunohistochemistry and by western blots of tissue extracts. By indirect immunocytochemistry, P47 was localized at the apical ridge of the sperm head in testicular, epididymal and ejaculated spermatozoa. The fluorescence intensity progressed during sperm transit from caput to cauda epididymis, probably caused by the ongoing expression and subsequent accumulation of P47 on the sperm surface. During the time course of capacitation, P47 appears to be unmasked by the release of coating proteins and appears to migrate from the apical ridge onto the entire acrosomal region, showing an intensive fluorescence pattern after 3 h capacitation in vitro. The kinetics of signal changes during in vitro capacitation were different in epididymal and ejaculated spermatozoa, indicating accelerated capacitational plasma membrane destabilization in epididymal spermatozoa.
Petrunkina
A M
AM
Institute for Reproductive Medicine, School of Veterinary Medicine Hannover, Germany.
Läkamp
A
A
Gentzel
M
M
Ekhlasi-Hundrieser
M
M
Töpfer-Petersen
E
E
D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002352:Carrier Proteins / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D004822:Epididymis / Q000737:chemistry; D005837:Genitalia, Male / Q000737:chemistry*; D007150:Immunohistochemistry / Q000379:methods; D036341:Intercellular Signaling Peptides and Proteins; D008297:Male; D008856:Microscopy, Fluorescence; D013075:Sperm Capacitation / Q000502:physiology*; D013079:Sperm Maturation / Q000502:physiology*; D013094:Spermatozoa / Q000737:chemistry*; D013552:Swine / Q000378:metabolism*; D013737:Testis / Q000737:chemistry
D002352:Carrier Proteins; D036341:Intercellular Signaling Peptides and Proteins; C112214:P47 sperm protein, Sus scrofa