Thymosins β-4 and β-10 are expressed in bovine ovarian follicles and upregulated in cumulus cells during meiotic maturation
Mohamad Salhab A , Pascal Papillier A , Christine Perreau A , Catherine Guyader-Joly B , Joelle Dupont A , Pascal Mermillod A and Svetlana Uzbekova A C
+ Author Affiliations
- Author Affiliations
A INRA, UMR85 Physiologie de la Reproduction et des Comportements, CNRS,UMR6175, Université de Tours; F-37380 Nouzilly, France.
B UNCEIA Département Recherche et Développement, 13 rue Jouët,94704 Maisons-Alfort, France.
C Corresponding author. Email: suzbekova@tours.inra.fr
Reproduction, Fertility and Development 22(8) 1206-1221 https://doi.org/10.1071/RD10015
Submitted: 28 January 2010 Accepted: 19 May 2010 Published: 1 October 2010
Abstract
β-Thymosins are small proteins that regulate the actin cytoskeleton and are involved in cell motility, differentiation, the induction of metalloproteinases, in anti-inflammatory processes and tumourigenesis. However, their roles in the ovary have not yet been elucidated. Using transcriptomics and real time reverse transcription–polymerase chain reaction validation, the present study demonstrates that thymosin β-4 (TMSB4) and thymosin β-10 (TMSB10) are upregulated in bovine cumulus cells (CCs) during in vitro maturation of cumulus–oocyte complexes (COCs) in parallel with an increase in mRNA expression of HAS2, COX2 and PGR genes. Using immunocytochemistry, both proteins were found to be localised mainly in granulosa cells, CCs and oocytes, in both the cytoplasm and nucleus, as well as being colocalised with F-actin stress fibres in CCs. Using different maturation mediums, we showed that the expression of TMSB10, but not TMSB4, was positively correlated with COC expansion and progesterone secretion and negatively correlated with apoptosis. Immunofluorescence, coupled with terminal deoxyribonucleotidyl transferase-mediated dUTP–digoxigenin nick end-labelling (TUNEL), demonstrated the absence of TMSB4 and/or TMSB10 in apoptotic cells. TMSB10 expression was higher in COCs matured in vivo than in vitro, and differences related to the age of the animal were observed. TMSB4 and/or TMSB10 expression was unchanged, whereas HAS2 overexpressed in CCs from oocytes that developed to the blastocyst stage in vitro compared with those that did not. Thus, TMSB4 and/or TMSB10 ovarian expression patterns suggest that these two thymosins may be involved in cumulus modifications during maturation.
Additional keywords: cow, ovary.
Acknowledgements
The authors thank Gael Ramé and Thierry Delpuech for technical assistance in the slaughterhouse and Virginie Mallard for help with the progesterone measurements. The authors are very grateful to Dr Hannah Brown for her help with English expression. This work was part of the program ANR-08-GENM-033 (OSCILE), sponsored by a grant from the French National Research Agency.
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