Ultrastructural Changes in Bovine Oocytes Induced During In Vitro Production of Embryos

Despite substantial progress in the development of procedures for in vitro maturation, in vitro fertilisation and in vitro culture, the production of viable embryos in most laboratories is around 20-30%. The objective of this study was to examine ultrastructural changes induced in bovine oocytes dur...

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Main Author:	Kanwal, Kiron Deep Singh
Format:	Thesis
Language:	English English
Published:	1999
Subjects:	Cattle - Growth Embryology - Cultures and culture media Reproductive technology
Online Access:	http://psasir.upm.edu.my/id/eprint/12173/1/FPV_1999_4_A.pdf
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id	my-upm-ir.12173
record_format	uketd_dc
spelling	my-upm-ir.121732013-05-27T07:51:14Z Ultrastructural Changes in Bovine Oocytes Induced During In Vitro Production of Embryos 1999-03 Kanwal, Kiron Deep Singh Despite substantial progress in the development of procedures for in vitro maturation, in vitro fertilisation and in vitro culture, the production of viable embryos in most laboratories is around 20-30%. The objective of this study was to examine ultrastructural changes induced in bovine oocytes during various stages of in vitro embryo production as these changes could contribute towards the low viability of the embryos. The oocytes recovered from slaughterhouse ovanes were transported in modified Dulbecco's phosphate buffered saline for various periods of time at two different temperatures, in vitro matured (WM), in vitro fertilised (WF) and in vitro cultured (WC) to morulae stages. After submitting to various stages mentioned above the oocytes were processed for transmission electron microscopy and examined in Hitachi 7100 or Phillips CM 12 electron microscopes at 75 or 80 kV respectively. Results exhibited that holding of oocytes in commonly used transport medium induced changes in the organelles such as rough endoplasmic reticulum, cortical granules, mitochondria and Golgi complexes both in the cumulus cells and those present in the oocytes. The severity of these changes was higher in oocytes transported at 2-4 C than those transported at 35-370 C the effect of which was reflected by the viability test. None of the cleaved oocytes (0/49) in the former while an average of 18.2% (10/55) of the oocytes in the latter developed to morulae. In the second experiment, the oocytes which were submitted to IVM, exhibited ultrastructural changes such as incomplete cumulus expansion, swelling of mitochondria, reduced incidence of cortical granules and accumulation of lipid droplets which were probably the factors affecting the viability of oocytes. In the third experiment, examination of in vitro produced morulae (35/121, 28.9%) revealed the presence of a large number of lipid droplets, vacuoles and numerous mitochondria undergoing the process of degeneration which ultimately may affect the viability of embryos. It was concluded that the ultrastructural changes induced during various stages of in vitro embryo production contribute towards low viability of the in vitro produced embryos. Cattle - Growth Embryology - Cultures and culture media Reproductive technology 1999-03 Thesis http://psasir.upm.edu.my/id/eprint/12173/ http://psasir.upm.edu.my/id/eprint/12173/1/FPV_1999_4_A.pdf application/pdf en public phd doctoral Universiti Putra Malaysia Cattle - Growth Embryology - Cultures and culture media Reproductive technology Faculty of Veterinary Medicine English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Cattle - Growth Embryology - Cultures and culture media Reproductive technology
spellingShingle	Cattle - Growth Embryology - Cultures and culture media Reproductive technology Kanwal, Kiron Deep Singh Ultrastructural Changes in Bovine Oocytes Induced During In Vitro Production of Embryos
description	Despite substantial progress in the development of procedures for in vitro maturation, in vitro fertilisation and in vitro culture, the production of viable embryos in most laboratories is around 20-30%. The objective of this study was to examine ultrastructural changes induced in bovine oocytes during various stages of in vitro embryo production as these changes could contribute towards the low viability of the embryos. The oocytes recovered from slaughterhouse ovanes were transported in modified Dulbecco's phosphate buffered saline for various periods of time at two different temperatures, in vitro matured (WM), in vitro fertilised (WF) and in vitro cultured (WC) to morulae stages. After submitting to various stages mentioned above the oocytes were processed for transmission electron microscopy and examined in Hitachi 7100 or Phillips CM 12 electron microscopes at 75 or 80 kV respectively. Results exhibited that holding of oocytes in commonly used transport medium induced changes in the organelles such as rough endoplasmic reticulum, cortical granules, mitochondria and Golgi complexes both in the cumulus cells and those present in the oocytes. The severity of these changes was higher in oocytes transported at 2-4 C than those transported at 35-370 C the effect of which was reflected by the viability test. None of the cleaved oocytes (0/49) in the former while an average of 18.2% (10/55) of the oocytes in the latter developed to morulae. In the second experiment, the oocytes which were submitted to IVM, exhibited ultrastructural changes such as incomplete cumulus expansion, swelling of mitochondria, reduced incidence of cortical granules and accumulation of lipid droplets which were probably the factors affecting the viability of oocytes. In the third experiment, examination of in vitro produced morulae (35/121, 28.9%) revealed the presence of a large number of lipid droplets, vacuoles and numerous mitochondria undergoing the process of degeneration which ultimately may affect the viability of embryos. It was concluded that the ultrastructural changes induced during various stages of in vitro embryo production contribute towards low viability of the in vitro produced embryos.
format	Thesis
qualification_name	Doctor of Philosophy (PhD.)
qualification_level	Doctorate
author	Kanwal, Kiron Deep Singh
author_facet	Kanwal, Kiron Deep Singh
author_sort	Kanwal, Kiron Deep Singh
title	Ultrastructural Changes in Bovine Oocytes Induced During In Vitro Production of Embryos
title_short	Ultrastructural Changes in Bovine Oocytes Induced During In Vitro Production of Embryos
title_full	Ultrastructural Changes in Bovine Oocytes Induced During In Vitro Production of Embryos
title_fullStr	Ultrastructural Changes in Bovine Oocytes Induced During In Vitro Production of Embryos
title_full_unstemmed	Ultrastructural Changes in Bovine Oocytes Induced During In Vitro Production of Embryos
title_sort	ultrastructural changes in bovine oocytes induced during in vitro production of embryos
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Veterinary Medicine
publishDate	1999
url	http://psasir.upm.edu.my/id/eprint/12173/1/FPV_1999_4_A.pdf
_version_	1747811320843993088

Ultrastructural Changes in Bovine Oocytes Induced During In Vitro Production of Embryos

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