Polyadenylated tail length variation pattern in ultra-rapid vitrified bovine oocytes

Research (Published online: 14-10-2016)

6. Polyadenylated tail length variation pattern in ultra-rapid vitrified bovine oocytes - D. J. Dutta, Himangshu Raj and Hiramoni Dev

Veterinary World, 9(10): 1070-1074

   doi: 10.14202/vetworld.2016.1070-1074

Abstract

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Aim: The current study aims at investigating the polyadenylated (poly[A]) tail length of morphologically high and low competent oocytes at different developmental stages. Furthermore, effect of ultra-rapid vitrification on the poly(A) tail length was studied.

Materials and Methods: Fresh bovine cumulus oocyte complexes from abattoir originated ovaries were graded based on morphological characters and matured in vitro. Cryopreservation was done by ultra-rapid vitrification method. mRNA was isolated from different categories of oocyte and subjected to ligation-mediated poly(A) test followed by polymerase chain reaction for determining the poly(A) tail length of β actin, gap junction protein alpha 1 (GJA1), poly(A) polymerase alpha (PAPOLA), and heat shock 70 kDa protein (HSP70) transcripts.

Results: GJA1, PAPOLA, and HSP70 showed significantly higher poly(A) in immature oocytes of higher competence irrespective of vitrification effects as compared to mature oocytes of higher competence.

Conclusion: mRNA poly(A) tail size increases in developmentally high competent immature bovine oocytes. There was limited effect of ultra-rapid vitrification of bovine oocytes on poly(A).

Keywords: ligation-mediated poly(A) test, oocyte, poly-adenylated tail, vitrification.