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Effect of coculture with stallion spermatozoa on de novo protein synthesis and secretion by equine oviduct epithelial cells
1995
Thomas, P.G.A. | Ignotz, G.G. | Ball, B.A. | Brinsko, S.P. | Currie, W.B.
Adhesion of equine spermatozoa to homologous oviduct epithelial cells (OEC) in vitro results in specific changes in spermatozoa and OEC function. To test the hypothesis that adhesion of spermatozoa affects protein synthesis and secretion by OEC, the following treatment groups were established in culture: OEC with culture medium only; control spermatozoa in culture medium only; OEC in coculture with spermatozoa; and OEC and spermatozoa in coculture, but physically separated by a microporous membrane. The experiment was replicated within each of 4 ejaculates from 3 stallions. De novo protein secretion by OEC was measured and compared by incorporation of [35S]methionine, and evaluated, using two-dimensional polyacrylamide gel electrophoresis and fluorography. Monolayers of OEC secreted a large number of proteins of molecular mass ranging from 14 to 205 kd. Adhesion of spermatozoa consistently caused reduced synthesis of 2 OEC secretory proteins and new or increased synthesis of 6 proteins. When spermatozoa and OEC were separated by a microporous membrane, some but not all of these changes were duplicated. Synthesis of 3 OEC secretory proteins, unaffected by binding of spermatozoa, was reduced when spermatozoa were prevented from contact with OEC by a microporous membrane. Adhesion of equine spermatozoa to homologous OEC monolayers and presence of equine spermatozoa resulted in qualitative and quantitative changes in synthesis and secretion of proteins by OEC. These changes have implications for storage, longevity, and maturation of spermatozoa.
Mostrar más [+] Menos [-]Capacitation-like membrane changes and prolonged viability in vitro of equine spermatozoa cultured with uterine tube epithelial cells
1993
Ellington, J.E. | Ball, B.A. | Blue, B.J. | Wilker, C.E.
Reliable capacitation of equine spermatozoa has been a major obstacle in the development of equine in vitro fertilization. Experiments were done to compare in vitro capacitation of equine spermatozoa by use of heparin/caffeine, calcium ionophore, uterine tube epithelial cell (UTEC)-conditioned medium, and direct culturing of spermatozoa with UTEC (coculturing). Capacitation-like changes, as determined by chlortetracycline membrane staining patterns, developed with UTEC-conditioned medium and coculturing, equivalent to that with calcium ionophore. Both of these treatments induced more (P < 0.05) capacitation-like changes than did the control, a modified Tyrode's medium. More (P < 0.05) spermatozoa were viable after 24 hours of UTEC coculturing than in the control incubation.
Mostrar más [+] Menos [-]Scanning electron microscopy of pathologic changes in the epithelial surfaces of the uterus and uterine tubes of cows
1990
Bollo, E. | Biolatti, B. | Pau, S. | Galloni, M.
Genital tracts from 15 cows with catarrhal and purulent inflammation of the uterus and uterine tubes, cystic hyperplasia of the endometrium, or hydrosalpinx were evaluated by use of scanning electron microscopy to determine epithelial changes associated with these conditions. Uterine epithelium was revealed to be easily damaged, even in the course of mild inflammation, whereas epithelium of the uterine tube was more resistant.
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