Application of a zona pellucida binding assay (ZBA) in the domestic cat benefits from the use of in vitro matured oocytes.
Acta Vet Scand
; 49: 28, 2007 Oct 01.
Article
em En
| MEDLINE
| ID: mdl-17908298
ABSTRACT
BACKGROUND:
Zona pellucida binding assays (ZBAs) have proven useful in determining the fertilising ability of spermatozoa in several species. Most ZBAs use fresh or salt-stored oocytes collected from fresh ovaries but because ovaries are not easy to obtain on a regular basis, chilled and frozen-thawed ovaries have been tested, with varying results. The present study tested the hypothesis that cat spermatozoa, either fresh or frozen-thawed, can bind to homologous zona pellucida of oocytes retrieved from frozen-thawed queen ovaries to a similar extent as they can bind to the zona pellucida of fresh, in vitro matured oocytes.METHODS:
Ovaries were collected from queens after routine ovario-hysterectomy and either stored in NaCl at -20 degrees C until use (treatment animals), or used fresh (controls). Cumulus-oocyte complexes (COCs) were retrieved by ovarian slicing from either source and used directly (immature oocytes from frozen-thawed ovaries; treatment animals) or after in vitro maturation (IVM) (fresh ovaries; controls) for 24 hours in TCM 199, supplemented with 1 IU hCG/mL and 0.5 IU eCG/mL and 0.5% bovine serum albumin (BSA). The oocytes were incubated for 4 hours in 5% CO2 in air at 38 degrees C and 100% humidity in the presence of 5 x 106 fresh or frozen-thawed spermatozoa/mL. Representative samples of oocytes were processed for scanning electron microscopy (SEM).RESULTS:
Both fresh and frozen-thawed spermatozoa bound to the in vitro matured zona pellucida but significantly fewer, or no, spermatozoa bound to frozen-thawed, immature zona pellucida (P < 0.001). Also, more fresh spermatozoa than frozen-thawed spermatozoa bound to the zona pellucida (P < 0.001). The zona pellucida surface differed in morphology (SEM), with in vitro matured oocytes showing a dense surface with few fenestrations in contrast to their frozen-thawed, immature counterparts, where fenestrations were conspicuously larger.CONCLUSION:
In conclusion, under the conditions of the present study, immature oocytes recovered from ovaries frozen immersed in NaCl at -20 degrees C are less suitable for use in feline ZBA.
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Base de dados:
MEDLINE
Assunto principal:
Oócitos
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Preservação do Sêmen
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Interações Espermatozoide-Óvulo
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Espermatozoides
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Criopreservação
Idioma:
En
Ano de publicação:
2007
Tipo de documento:
Article