Glucose-regulated protein 78 (Grp78/BiP) is secreted by human oviduct epithelial cells and the recombinant protein modulates sperm-zona pellucida binding.
OBJECTIVE
To determine the secretion of Grp78 by human oviduct epithelial cells, its association to spermatozoa, and its involvement in gamete interaction.


METHODS
Prospective study.


METHODS
Basic research laboratory.


METHODS
Semen samples obtained from normozoospermic volunteers. Tubal tissue provided by patients undergoing hysterectomies. Oocytes collected from women undergoing IVF-ET.


METHODS
Analysis of Grp78 expression and secretion by oviductal tissue. Gamete incubation with recombinant Grp78 (rec-Grp78).


METHODS
Assessment of protein expression and secretion by immunohistochemistry and Western immunoblotting, respectively. Evaluation of rec-Grp78 binding to human spermatozoa by immunocytochemistry, and analysis of its effect upon gamete interaction using the hemizona assay.


RESULTS
Grp78 was found in the surface of oviduct epithelial cells. Soluble Grp78 was detected in oviductal fluids from women in the periovulatory period and in oviductal tissue conditioned medium. Rec-Grp78 was able to bind to the sperm acrosomal cap, and its presence during gamete interaction led to a decrease in the number of spermatozoa bound to the zona pellucida (ZP). When calcium ions from the incubation medium were replaced by strontium, rec-Grp78 enhanced sperm-ZP interaction.


CONCLUSIONS
Grp78 is expressed and secreted by oviduct epithelial cells. The protein would bind to the gametes and may modulate their interaction in a calcium-dependent manner.
Marín-Briggiler
Clara I
CI
Instituto de Biología y Medicina Experimental, National Research Council of Argentina (CONICET), University of Buenos Aires, 1428ADN Buenos Aires, Argentina.
González-Echeverría
María F
MF
Munuce
María J
MJ
Ghersevich
Sergio
S
Caille
Adriana M
AM
Hellman
Ulf
U
Corrigall
Valerie M
VM
Vazquez-Levin
Mónica H
MH
D000328:Adult; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D000091342:Endoplasmic Reticulum Chaperone BiP; D004847:Epithelial Cells / Q000378:metabolism*; D005187:Fallopian Tubes / Q000378:metabolism*; D005260:Female; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008597:Menstrual Cycle; D008875:Middle Aged; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000378:metabolism*; D046509:Tissue Culture Techniques; D015044:Zona Pellucida / Q000378:metabolism*
D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D002118:Calcium