Localization of the Chaperone Proteins GRP78 and HSP60 on the Luminal Surface of Bovine Oviduct Epithelial Cells and Their Association with Spermatozoa
Upon their transit through the female genital tract, bovine spermatozoa bind to oviduct epithelial cells, where they are maintained alive for long periods of time until fertilization. Although carbohydrate components of the oviduct epithelial cell membrane are involved in these sperm/oviduct interac...
Gespeichert in:
Veröffentlicht in: | Biology of reproduction 2004-12, Vol.71 (6), p.1879-1889 |
---|---|
Hauptverfasser: | , , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
Zusammenfassung: | Upon their transit through the female genital tract, bovine spermatozoa bind to oviduct epithelial cells, where they are maintained
alive for long periods of time until fertilization. Although carbohydrate components of the oviduct epithelial cell membrane
are involved in these sperm/oviduct interactions, no protein candidate has been identified to play this role. To identify
the oviduct factors involved in their survival, sperm cells were preincubated for 30 min with apical membranes isolated from
oviduct epithelial cells, washed extensively, and further incubated for up to 12 h in the absence of apical membranes. During
this incubation, sperm viability, motility, and acrosomal integrity were improved compared with cells preincubated in the
absence of apical membranes. This suggests that, during the 30-min preincubation with apical membrane extracts, either an
oviductal factor triggered intracellular events resulting in positive effects on spermatozoa or that such a factor strongly
attached to sperm cells to promote a positive action. Similarly, spermatozoa were incubated with apical membranes isolated
from oviduct epithelial cells labeled with [ 35 S]-methionine and, upon extensive washes, proteins were separated by two-dimensional (2-D) gel electrophoresis to identify
the factors suspected to have beneficial effects on spermatozoa. The six major proteins, according to their signal intensity
on the autoradiographic film, were extracted from a 2-D gel of oviduct epithelial cell proteins run in parallel and processed
for N-terminal sequencing of the first 15 amino acids. Of these, one was identical to heat shock protein 60 (HSP60) and one
to the glucose-regulated protein 78 (GRP78). Their identities and association with spermatozoa were confirmed using an antibody
directed against these proteins. This paper reports the localization of both GRP78 and HSP60 on the luminal/apical surface
of oviduct epithelial cells, their binding to spermatozoa, and the presence of endogenous HSP60 in the sperm midpiece. |
---|---|
ISSN: | 0006-3363 1529-7268 |
DOI: | 10.1095/biolreprod.103.026849 |