Functional expression and intracellular signaling of UTP-sensitive P2Y receptors in theca-interstitial cells.

Reproductive Biology and Endocrinology : RB&E
Francisco G Vázquez-CuevasRogelio O Arellano

Abstract

Purinergic receptors are expressed in the ovary of different species; their physiological roles remain to be elucidated. UTP-sensitive P2Y receptor activity may regulate cell proliferation. The aim of the present work was to study the functional expression of these receptors in theca/interstitial cells (TIC). TIC were isolated by centrifugation in a Percoll gradient. P2Y receptors and cellular markers in TIC were detected by RT-PCR and Western blot. Intracellular calcium mobilization induced by purinergic drugs was evaluated by fluorescence microscopy, phosphorylation of MAPK p44/p42 and of cAMP response element binding protein (CREB) was determined by Western blot and proliferation was quantified by [3H]-thymidine incorporation into DNA. RT-PCR showed expression of p2y2r and p2y6r transcripts, expression of the corresponding proteins was confirmed. UTP and UDP, agonists for P2Y2 and P2Y6 receptors, induced an intracellular calcium increase with a maximum of more than 400% and 200% of basal level, respectively. The response elicited by UTP had an EC50 of 3.5 +/- 1.01 microM, while that for UDP was 3.24 +/- 0.82 microM. To explore components of the pathway activated by these receptors, we evaluated the phosphorylation induced by...Continue Reading

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Oct 7, 2009·Proceedings of the National Academy of Sciences of the United States of America·Rogelio O ArellanoFrancisco Vázquez-Cuevas

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Citations

Oct 12, 2012·Reproduction, Fertility, and Development·F G Vázquez-CuevasR O Arellano
Nov 23, 2013·Purinergic Signalling·Geoffrey Burnstock
Nov 26, 2013·Purinergic Signalling·Geoffrey Burnstock
Aug 15, 2015·Molecular Reproduction and Development·Angélica S Martínez-Ramírez, Francisco G Vázquez-Cuevas
May 12, 2015·Autonomic Neuroscience : Basic & Clinical·Ivana BjelobabaStanko S Stojilkovic
May 20, 2015·Autonomic Neuroscience : Basic & Clinical·George I Gorodeski
Jan 10, 2019·Molecular Reproduction and Development·Anaí Del Rocío Campos-ContrerasFrancisco Gabriel Vázquez-Cuevas
Apr 1, 2020·International Journal of Molecular Sciences·Erandi Velázquez-MirandaFrancisco G Vázquez-Cuevas
Sep 25, 2016·Reproductive Sciences·Vaibhave UbbaRajesh Kumar Jha
May 1, 2017·Reproductive Sciences·Vaibhave UbbaRajesh Kumar Jha
Oct 26, 2021·Molecular Reproduction and Development·Ana Patricia Juárez-MercadoFrancisco G Vázquez-Cuevas

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Methods Mentioned

BETA
PCR
immunoprecipitation
fluorescence microscopy
PMA

Software Mentioned

Pro Plus
ImageJ
GraphPad Prism
Image
Imagenes
BLAST

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