Study of the mechanism of action for wich estradiol stimulates PGF2a endometrial synthesis in female bovine

• Grant number: 14/08815-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2014
• End date: June 30, 2015
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Claudia Maria Bertan Membrive
• Grantee: Bruna Folegatti Santana
• Host Institution: Universidade Estadual Paulista (UNESP). Campus Experimental de Dracena. Dracena , SP, Brazil

Abstract

In bovine females release of PGF2a can be induced in vivo by estradiol ( E2 ) , however, the role of E2 in the synthesis of PGF2a has not yet been clarified . Whereas the increase plasma concentrations of PGF2 3.5 hours after the application of E2 in vivo , it is believed that E2 activates not only enzymes, but also stimulates the synthesis of proteins essential for the production of PGF2a . It is known that bovine endometrial explants from animals injected with E2 , and cultivation of bovine endometrial cells ( BEND ) exposed to E2 , to be treated with calcium ionophore ( CI ) had a marked increase in the release of PGF2a compared with those who did not received E2 and E2 treated exclusively with or CI . The hypothesis is that E2 stimulates the expression and / or activity of PKC and PLA2 enzymes, both calcium -dependent activation, involved in the synthesis of endometrial PGF2a . The hypothesis is that E2 stimulates the expression and / or activity of PKC and PLA2 enzymes, both calcium -dependent activation, involved in the synthesis of endometrial PGF2a . The aim of this study is to identify groups of genes involved in the synthesis of endometrial PGF2a in bovine females , whose expression is modulated by E2 by RT - qPCR and further substantiate the different proteins identified as in experiment 2 by Western blotting . Cyclical Nellore females ( n = 60 ) , non-lactating , will be paired on the 17th of a synchronized estrous cycle , injected with 0 mg ( n = 30 ) or 17² - estradiol 3mg ( n = 30 ) and then be subdivided within each group to have a uterine biopsy of the uterine horn ipsilateral to the corpus luteum performed 2 hours ( n = 6 n = 6 0 mg to 3 mg de17² -estradiol ) or 3 hours (n = 6 n = 6 0 mg to 3 mg de17² - estradiol) or 4 hours (n = 6 n = 6 0 mg to 3 mg de17² - estradiol) or 5 hours (n = 6 n = 6 0 mg to 3 mg de17² -estradiol ) or 6 hours (n = 6 n = 6 0 mg to 3 mg de17² - estradiol) . Tissue samples will be evaluated for mRNA expression by RT- qPCR for specific proteins involved in the synthesis of endometrial PGF2 , among these aldo - keto redutase1B1 , Aldo - keto reductase 1C3 , oxytocin receptor, phospholipase A24A , PKC± , PKC² , PKC³ , cyclooxygenase 1 and cyclooxygenase 2 .