ACTIVATION OF BOVINE OOCYTES FOLLOWING IN1 ...

13 downloads 74 Views 3MB Size Report
sperrn injection (ICSI) in bovine oocytes. Oocytes recovered fiom abattoir-derived ovaries were centrifbged for 5 min at 6000xg to facilitate sperm injection.
ACTIVATION OF BOVINE OOCYTES FOLLOWING IN1RACYTOPLASMIC SPERM INJECTION (ICSI)

Jin-Tae Chung

A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfilment of the requiiements for the degiee of Master of Science

Department of Animal Science Macdonald Campus McGiII University Montreal, Quebec, Canada

@

February 1999

National Library

Bibliothèque nationale du Canada

Acquisitions and Bibliographie Services

Acquisitions et services bibliographiques

395 Wellington Street OttawaON K l A W canada

Ottawa ON K l A W canala

395. Ne WdbglOn

The author has granted a nonexclusive licence dowing the National Libra~yof Canada to reproduce, loan, distribute or sell copies of this thesis in microfom, paper or electronic formats.

L'auteur a accordé une Licence non exclusive permettant à la Bibliothèque nationale du Canada de reproduire, prêter, distribuer ou vendre des copies de cette thèse sous la forme de rnicrofiche/£ilm, de reproduction sur papier ou sur format électronique.

The author retains ownership of the copyright in this thesis. Neither the thesis nor substantial extracts fkom it may be printed or otherwise reproduced without the author's permission.

L'auteur conserve la propriété du droit d'auteur qui protège cette thèse. Ni la thèse ni des extraits substantiels de celle-ci ne doivent être imprimés ou autrement reproduits sans son autorisation.

Suggested short title:

lntracytoplasmic sperm injection of bovine oocytes

ACTIVATION OF BOVINE OOCYTES FOLLOWING INTRACYTOPLASMIC SPERM INJECTION (KSI) ABSTRACT

M.Sc. Jin-Tae Chung Animal Science

The objective of this study was to develop a reliable method for intracytoplasmic sperrn injection (ICSI) in bovine oocytes. Oocytes recovered fiom abattoir-derived ovaries were centrifbged for 5 min at 6000xg to facilitate sperm injection. S p e m were pre-treated in vilro with 5mM dithiothreitol @TT), and diluted (approximately 1 5 ) with 5% poIyvinylpyrrolidone (PVP)in 0.9% saline. After sperm injection, various activation

procedures were compared. Initially, 3 h after activation with 5pM Ionomycin (A23 187), oocytes with second polar bodies were selected and treated with 1.9mM 6dimethylarninopurine (DMAP). The cleavage rate of sperm-injected oocytes treated with Ionomycin and DMAP was higher than with Ionomycin alone (62.1 vs. 27.3%, p