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The effect of kinetic heat shock on bovine oocyte maturation and subsequent gene expression of targeted genes

Published online by Cambridge University Press:  08 June 2017

Krishna C. Pavani
Affiliation:
Department of Agrarian Sciences, CITA-A (Research Centre for Agricultural and Environmental Sciences and Technology of the Azores), Animal Reproduction, University of the Azores, Angra do Heroísmo 9700-042, Portugal Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, University of Ghent, Salisburylaan 133, B-9820 Merelbeke, Belgium
António Rocha
Affiliation:
ICBAS, Abel Salazar Biomedical Institute, University of Porto, Campus Agrário de Vairão, Rua Padre Armando Quintas, 4485–661 Vairão, Porto, Portugal
Erica Baron
Affiliation:
Centro Universitário Monte Serrat, Rua Comendador Martins, 52 - Vila Matias, Santos – SP, 11015–530, Brazil
Joana Lourenço
Affiliation:
Department of Agrarian Sciences, CITA-A (Research Centre for Agricultural and Environmental Sciences and Technology of the Azores), Animal Reproduction, University of the Azores, Angra do Heroísmo 9700-042, Portugal
Marwa Faheem
Affiliation:
Department of Animal Production Science, Faculty of Agriculture, Cairo University, 12613 Giza, Egypt
Fernando Moreira da Silva*
Affiliation:
Departamento de Ciências Agrárias, Secção Reprodução, Campus de Angra do Heroísmo, Universidade dos Açores, Rua Capitão João D'Avila, 9700-042 Angra do Heroísmo, Portugal
*
All correspondence to: Fernando Moreira da Silva. Departamento de Ciências Agrárias, Secção Reprodução, Campus de Angra do Heroísmo, Universidade dos Açores, Rua Capitão João D'Avila, 9700-042 Angra do Heroísmo, Portugal, Tel.: +351 295 402200, Fax: +351 295 402209, Email: [email protected]

Summary

The exposure of oocytes to heat stress during the maturation process results in harmful effects to their internal organelles, low fertilization capability and higher embryonic losses. In the present experiment the effect of heat shock (HS) during the maturation process was assessed. In Assay 1, oocytes from winter (December–March; n = 100) and summer (June–September; n = 100) months were collected and matured to analyse their HS tolerance. Total RNA was extracted from matured oocytes and cDNA synthesis was performed, followed by qPCR for selected genes (Cx43, CDH1, DNMT1, HSPA14), compared with two reference genes (GAPDH and SDHA). In Assay 2, oocytes collected during the winter were subjected to kinetic HS by stressing them at 39.5°C for 6, 12, 18 or 24 h and were afterwards matured at control temperature (38.5°C), and subsequently subjected to the previously described gene analysis procedure. Results of Assay 1 show that summer-collected oocytes exhibited lower maturation rate than winter-collected oocytes, which may be due to the down-regulation of the HSPA 14 gene. Assay 2 showed that 6 h of HS had no effect on gene regulation. CDH1 and DNMT1 up-regulation was observed starting at 12 h, which may represent the effect of heat shock on oocyte development.

Type
Short Communication
Copyright
Copyright © Cambridge University Press 2017 

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