Evaluation of Incubation Parameters in a TUNEL assay for Detecting Apoptotic cells in Cumulus Oocyte Complexes and in vitro produced early Embryos of Buffalo

Order of Publishing in Issue: 
7
Volume :10
Issue :3
July, 2016
Page No: 
274-279
Authors: 
Asit Jain[1], Tripti Jain[2], G K Sachdeva, Sachinandan De and Tirtha Kumar Datta*
Address: 
Animal Genomics Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal-132001, Haryana
Address: 
[1]Currently at Department of Animal Genetics and Breeding, College of Veterinary Science and Animal Husbandry, CGKV, Anjora, Durg, Chhattisgarh, India
Address: 
[2]Currently at Animal Biotechnology Centre, CGKV, Durg, Chhattisgarh, India
Email-ID: 
tirthadatta@gmail.com

Terminal deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL) is used widely for detecting the apoptotic status in biological cells of diverse origin. Reproducing the TUNEL procedure with specialized cells like cumulus oocyte complexes (COCs) poses technical challenge in view of mammalian oocytes which undergo very dynamic transitions in the course of oocyte maturation within a short time. The current work describes a standardized TUNEL protocol where the process of fixation and permeabilization were optimized using different concentrations of paraformaldehyde and Triton X-100 with sodium citrate, respectively. Effectiveness of the procedure was validated at different time intervals of oocyte culture using known oocyte stimulators. The procedure was further validated in the in vitro produced early embryos. Consistent and specific signal could be obtained using the procedure depicting the apoptotic status of cumulus cells and early embryos at different cell stages in buffalo.

Keywords: 
Apoptosis, Cumulus Cells, Buffalo, TUNEL, Fixative
AttachmentSize
PDF1.45 MB

 
To download full text articles and issues in PDF at no cost.All you have to do is fill out a brief registration form. It's easy and it's free. At any time, you may request removal of your contact information from our database.
Click here to Register | Click here if you have already registered

2018 ABAP Awards

Invitation for Nominations to 2019 ABAP Awards
The Association of Biotechnology and Pharmacy invites nominations for the following awards from the eligible talented people with proven track record and achievements on or before August 31, 2019 in the prescribed format.

DOWNLOAD CIRCULAR | APPLICATION FORM

AWARDS

Upcoming 0 events

  • No upcoming events available

User login