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  • Author or Editor: Philip G. A. Thomas x
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Abstract

Objective

To examine glycoconjugates in the isthmic and ampullar regions of the uterine tube (oviduct) of horses during estrus, diestrus, and pregnancy.

Sample Population

Oviductal samples from 17 mares.

Procedure

Oviducts were collected during estrus (n = 3), diestrus (n = 3), or pregnancy (n = 3), embedded, and snap frozen in liquid nitrogen. Frozen sections (5 to 6 μm in thickness) were stained with 100 μg/ml of fluorescein-isothiocyanate-conjugated lectin (30 min at 38.5 C) and were evaluated by use of epifluorescence microscopy and video image analysis. Specificity of lectins was established by blocking with the corresponding carbohydrate. Dolichos biflorus agglutinin (DBA)-affinity studies on western blots of oviductal lavage fluid, oviductal explant conditioned media, and apical membrane proteins from isthmic and ampullar regions of oviducts were used to identify glycoproteins with galactosyl residues.

Results

Use of 4 lectins resulted in differential labeling of the luminal surface of the oviductal epithelium. Both DBA and soybean agglutinin labeled the apical epithelium of the isthmus, but not the ampullar oviduct. Soybean agglutinin resulted in more-intensely labeled epithelium in the isthmic region of oviducts during estrus and pregnancy than during diestrus. The DBA labeled a number of glycoproteins in conditioned media from both regions of the oviduct. These glycoproteins ranged from 14 to 200 kd, with major glycoproteins identified at 31 and 57 kd.

Conclusions

The predominant glycoconjugates in the oviduct of horses are galactosyl residues. There are regional differences in the distribution of these galactosyl glycoconjugates in the isthmic and ampullar oviduct. (Am J Vet Res 1997;58:816–822)

Free access
in American Journal of Veterinary Research

SUMMARY

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.

Free access
in American Journal of Veterinary Research

Summary

Development of 1- to 2-cell in vivo fertilized equine embryos cultured with or without uterine tubal epithelial cells (utec) was studied. One- to 2-cell embryos (n = 26) were collected surgically from the uterine tubes of pony mares 1 day after ovulation. Four- to 8-cell embryos (n = 9) were collected 2 days after ovulation. Presumptive zygotes and 2-cell embryos were cultured with (n = 17) or without (n = 9) utec, and all 4- to 8-cell embryos were cocultured with utec as positive controls. Uterine tubal epithelial cells were used as cell suspensions within 2 weeks after initiation of cultures. Embryos were cultured to blastocysts or until the embryo had morphologic degeneration. Six presumptive zygotes failed to cleave in vitro. Development to blastocysts of 1-cell (4 of 11) and 2-cell (2 of 6) embryos cocultured with utec was similar. Coculture of 1- to 2-cell embryos with utec significantly (P = 0.05) improved development to blastocysts, compared with culture in medium alone (35 vs 0%, respectively); however, development to blastocysts of 1- to 2-cell embryos cocultured with utec was less (P < 0.025) than that of 4- to 8-cell embryos cocultured with utec (35 vs 89%, respectively).

Free access
in American Journal of Veterinary Research