Volume: 49 Issue: 2
Year: 2018, Page: 63-66,
Received: Aug. 10, 2017 Accepted: Aug. 25, 2017 Published: July 1, 2018
The study was carried out in 12 normal and 12 repeat breeder crossbred cows. These cows were further divided into four groups of six animals in each group. The animals were subjected to transvaginal oocyte recovery (TVOR) for a period of twomonths without any treatment at once weekly and twice weekly interval. Culture grade oocytes retrieved were subjected to in vitro maturation for 24hat 38.5o C and 5 per cent CO2 . Nuclear maturation was assessed with Hoechst 33342 stain and oocyte viability was assessed with fluorescein diacetate (FDA) stain. Results revealed that nuclear maturation based on Hoechst 33342 as 80.00 and 76.19 per cent respectively for normal and repeat breeders. Assessment ofviability of oocytes based on fluorescein diacetate (FDA)stain revealed that viability of oocytes in normal breeders (83.33 per cent) was higher than repeat breeiders (75 per cent). The present study revealed that nuclear maturation rate and oocyte viability is less in repeat breeders when compared to normal breeders
Keywords: Transvaginal oocyte recovery, Oocyte viability nuclear maturation, Hoechst 33342, Fluorescein diacetate (FDA)
Arndt-Jovin, D.J. and Jovin, T.M. 1977Analysis and sorting of living cells according to deoxyribonucleic acid content.J.Histochem.Cytochem.25: 585–589.
Chazotte, B. 2011.Labeling Nuclear DNA withHoechst 33342.Cold Spring HarbProtoc; 2011; doi:10.1101/pdb.prot5557: 83-85.
Gall, L., De Smedt, V., Crozet, N., Ruffini, S. andSévellee, C.1996.Meiotically incompetent and competent goatoocytes: timing of nuclear events and protein phosphorylation.Theriogenology.46: 825–35.
Katska, L. and Smorag, Z. 1985. The influence of culture temperature on in vitro maturation of bovine oocytes.Anim. Reprod. Sci.9: 205-212.
Lelande, M.E., Ling, V. & Miller, R.G. 1981. Hoechst 33342dye uptake as a probe ofmembrane permeability changesin mammalian cells. Proc. Natl. Acad. Sci. USA 7: 363–367.
Liang, Y., Rakwongrit, D., Phermthai, T., Somfai, T., Nagai, T. and Parnpai, R. 2012. Cryopreservation of immature buffalo oocytes: Effects of cytochalasin B pretreatment on the efficiency of cryotop and solid surface vitrification methods. Anim. Sci. J.83: 630-638
Magnus, P.K. 2005.Effect of ovum retrieval methods and cumulus ooocyte complex morphology on in vitro maturation of bovine oocytes.M.V.Sc thesis, Kerala Agricultural University, Thrissur. 125p
Mohr, L.R.and Trounson, A.O.1980. The use of fluorescein diacetate to assess embryo viability in the mouse.J. Reprod. Fertil. 58:189-96
Rotman, B. and Papermaster, B. W. 1966 Membrane properties of living mammalian cells as studied by enzymatic hydrolysis of fluorogenic esters. Proc. Nat. Acad. Sd. USA. 55: 134-41.
Pateria, A.K. and Rawal, C.V.S. 1990. White side test for subclinical metritis in Buffaloes. Indian J. of Anim. Reprod., 11, 142-144.
Sakhong, D., Vongpralub, T., Katawatin, S. and Sirisathien, S. 2012. Ultrasound – guided trans-vaginal follicular aspiration and development of vitrified-thawed indigenous beef cattle (bosindicus) oocytes after in vitro fertilization. Thai J. Vet. Med. 42: 509-516.
Smith, L.C. 1993. Membrane and intracellular effects of ulteaviolet irradiation with Hoechst 33342on bovine secondary oocytes matured in vitro. J. Reprod. Fertil.99: 39-4.
Zuccotti, M., Piccinelli, A., Rossi, P.G., Garagna, S. &Redi, C.A. 1995. Chromatin organization during mouse oocytegrowth.Mol. Reprod.Dev. 41, 479–85
© 2018 Abhilash et al. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.