Author:
Sidhu K. S.,Mate K. E.,Molinia F. C.,Glazier A. M.,Rodger J. C.
Abstract
Previous studies have demonstrated that
co-culture of brushtail possum epididymal spermatozoa with oviduct epithelial
cell monolayers prolongs sperm survival and results in the re-orientation of
the sperm head and tail to the T-shape (thumbtack) configuration.
Transformation of sperm to thumbtack orientation is believed to be associated
with marsupial sperm capacitation. Here we report that incubation in
oviduct-conditioned media also significantly prolongs sperm survival and
results in the transformation of sperm to the thumbtack orientation. The major
objective of the current study was to examine the proteins present in the
conditioned media, to determine whether any of these proteins specifically
bound to sperm and the relationship between these proteins and sperm survival
and thumbtack orientation. Co-culturing brushtail possum sperm with
biotin-labeled proteins in conditioned media (CM) from ampulla, isthmus and
uterine explants demonstrated strong binding of two proteins of molecular mass
230 and 61 kD and weak binding of nine proteins of molecular mass 200, 180,
120, 140, 55, 52, 48, 34, 30 kD to sperm within 30 min. The binding of the
61-kD protein from the conditioned media appeared specific as increasing
concentrations of non-labeled oviduct proteins, but not serum proteins,
inhibited the binding of labeled proteins. The binding of oviduct and uterine
proteins in the conditioned media significantly prolonged sperm survival and
percentage motility and also transformed a large number of sperm to a
thumbtack orientation. The implication of binding of these proteins is
discussed in the context of sperm survival and capacitation in this species.
Subject
Developmental Biology,Endocrinology,Genetics,Molecular Biology,Animal Science and Zoology,Reproductive Medicine,Biotechnology
Cited by
8 articles.
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