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detached oviduct magnum (A), and the adult female zebra finch with a history of infertility (B). A—Detached oviduct magnum. The tissue was 3 X 1.5 X 1 cm, 2.84 g, and covered by seeds (bird feed). It was red, wet, and spongy in texture. There was an

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in Journal of the American Veterinary Medical Association

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)

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in American Journal of Veterinary Research

Abstract

Objective

To compare the electrophoretic patterns of proteins synthesized and secreted by oviductal epithelial cell (OEC) explants obtained from young, fertile and aged, subfertile mares.

Animals

Young, fertile (n = 5; 2 to 7 years old) and aged, subfertile (n = 5; 17 to 24 years old) mares.

Procedure

2-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and computerized densitometry.

Results

Variation in the synthesis and secretion of poly-peptides from young, fertile mare OEC (YOEC) and aged, subfertile mare OEC (AOEC) was evidenced by differences in the intensity of radiolabeled polypeptides on fluorograms. Fluorograms for 9 acidic (isoelectric point, 5.09 to 6.50) proteins of low to medium molecular mass (18.2 to 85.0 kd) from YOEC had greater intensity than did those from AOEC. Fluorograms for 7 proteins (10.5 to 45.0 kd; isoelectric point, 5.80 to 6.92) from AOEC had greater intensity.

Conclusion

The differences detected in the fluorographic intensities of secreted proteins from YOEC and AOEC may be related to the disparity in embryo development observed between young, fertile and aged, subfertile mares.

Clinical Relevance

Failure to maintain pregnancy in aged, subfertile mares may be a result of a suboptimal oviductal environment exerting its effects on the conceptus during early cleavage. (Am J Vet Res 1996;57:1346-1353)

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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

Abstract

Objective

To test the hypotheses that the attachment of equine spermatozoa to uterine tubal (oviductal) epithelial cells (OEC) in vitro is mediated by glycoproteins, and that proteins with carbohydrate-binding properties are present in the periacrosomal plasma membrane of equine spermatozoa.

Animals

4 reproductively sound stallions, and 1 mare in estrus.

Procedures

In experiment 1a, fluorescent-labeled spermatozoa were cocultured with monolayers of OEC in the presence of 50 mM glucose, fructose, galactose, mannose, N-acetyl glucosamine, N-acetyl galactosamine, or N-acetyl neuraminic acid, or 10 mg of fetuin or asialofetuin/ml in modified Tyrode's solution (TALP), or in TALP alone. After 2 hours of coculture, numbers of attached spermatozoa were counted by fluorescence microscopy and analysis of digitized images. In experiment 1b, progressive motility, viability, acrosomal integrity, and capacitation status were determined in spermatozoa incubated for 2 hours in the presence of the respective monosaccharides and glycoproteins or in TALP alone. In experiment 2, proteins isolated from the periacrosomal plasma membrane of equine spermatozoa were subjected to galactose affinity chromatography and subsequent one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining.

Results

Numbers of spermatozoa attached to OEC were reduced (P < 0.05) after all treatments except N-acetyl glucosamine, compared with incubation in TALP alone. The lowest numbers of spermatozoa were bound in cultures incubated in the presence of galactose and asialofetuin. Spermatozoal motility was lower (P < 0.05) after incubation for 2 hours in the presence of fetuin, compared with control, and incubation in the presence of fetuin or asialofetuin caused a significant (P < 0.05) increase in the percentage of capacitated spermatozoa, compared with control. Affinity chromatography of periacrosomal plasma membrane proteins revealed a galactose-binding protein of about 66 kd.

Conclusion

Recognition of glycoconjugates with exposed galactosyl residues on OEC by galactose-binding proteins on the periacrosomal plasma membrane of equine spermatozoa could mediate the attachment of equine spermatozoa to OEC in vitro. (Am J Vet Res 1996;57:1635–1639)

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in American Journal of Veterinary Research

SUMMARY

Specimens of the uterine tube epithelium (ampulla) were obtained from 20 healthy, prepubertal, ovariohysterectomized gilts. A 2 to 3% proportion of atypical cilia was observed. Of 6,600 transversely sectioned cilia, 122 (1.8%) had microtubular disorganization, whereas 444 of 48,080 totally examined cilia (0.9%) were compound, 104 (0.2%) were swollen, and 44 (0.09%) were intracytoplasmic.

Free access
in American Journal of Veterinary Research
in Journal of the American Veterinary Medical Association

the oviduct, was identified left of midline and was not associated with the mineralized egg. Surgical exploration and removal of the suspected ectopic egg was recommended. The bird received an injection of leuprolide acetate f (800 μg/kg [364 μg

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in Journal of the American Veterinary Medical Association

clinical patients via in vitro maturation and transfer (surgical transfer to the oviduct of inseminated recipient mares) of oocytes collected after natural death or euthanasia of the donor mare. In that report, 6 oocyte transfer resulted in 8 foals or

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in Journal of the American Veterinary Medical Association

were within expected ranges for a female broiler breeder of this age, and there was minimal postmortem autolysis. The magnum of the oviduct was approximately 30 cm in length, and its wall was expanded by pinpoint to 5-mm-diameter, pale yellow

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in Journal of the American Veterinary Medical Association