Animal Reproduction Science 170 (2016) 157–169 Contents lists available at ScienceDirect Animal Reproduction Science jou rn al hom epage : w ww.elsevier.com/locate/anir eprosci Tissue localization of GM-CSF receptor in bovine ovarian follicles and its role on glucose uptake by mural granulosa cells O.A. Peralta a,1 , D. Bucher b,1 , C. Angulo b , M.A. Castro b , M.H. Ratto c,d,∗ , Il. Concha b,∗ a Departamento de Fomento de la Producción Animal, Facultad de Ciencias Veterinarias y Pecuarias, Universidad de Chile, Santiago, Chile b Instituto de Bioquímica y Microbiología, Universidad Austral de Chile, Valdivia, Chile c Instituto de Ciencia Animal, Universidad Austral de Chile, Valdivia, Chile d Ross University School of Veterinary Medicine, St. Kitts, Basseterre, West Indies a r t i c l e i n f o Article history: Received 27 January 2016 Received in revised form 25 April 2016 Accepted 28 April 2016 Available online 7 May 2016 Keywords: Bovine follicle development Granulosa cells Oocyte Glucose transporters Cattle a b s t r a c t The granulocyte-macrophage colony stimulating factor (GM-CSF) is a multifunctional cytokine implicated in proliferation, differentiation, and activation of several cell types including those involved in hematopoiesis and reproduction. In the present study, the expression of the - and -subunit genes of GM-CSF receptor during follicular develop- ment in cattle was assessed. The spatial association of - and -subunits of GM-CSF with follicle stimulating hormone receptor (FSHR) and 3-hydroxysteroid dehydrogenase (3- HSD), and the temporal associations with gene expression of hexose transporters (GLUTs) in granulosa cells of cattle were also evaluated. The effect of GM-CSF on the functional- ity of hexose transporters was also determined in an in vitro primary culture of granulosa cells. The spatial association of subunits of the GM-CSF receptor with 3-HSD and FSHR sug- gests a potential steroidogenic regulation of GM-CSF in granulosa cells. Immunodetection of GLUTs and uptake kinetic assays confirmed expression and functionality of these genes for hexose transporters in granulosa cells of cattle. Treatment of granulosa cells with GM-CSF, FSH or insulin- like growth factor-I (IGF-I) alone increased 2-deoxyglucose (DOG) or 3-0- methylglucose (OMG) uptake; however, when cells were treated with various combination of these factors there were no additive effect. Unexpectedly, the combination of GM-CSF and FSH decreased DOG uptake compared to FSH treatment alone. Thus, the expression pattern of GM-CSF receptor subunit genes during follicle development in cattle and pro- motion of DOG and OMG uptake in granulosa cells indicate a role for GM-CSF, FSH and/or IGF-I alone in regulating granulosa cell metabolic activity, specifically by promoting glucose uptake. © 2016 Elsevier B.V. All rights reserved. ∗ Corresponding authors E-mail addresses: mratto@rossvet.edu.kn (M.H. Ratto), conchagrabinger@uach.cl (Il. Concha). 1 Equal contributors. 1. Introduction The granulocyte-macrophage colony stimulating factor (GM-CSF) is a multifunctional cytokine initially identified as a regulator of proliferation, differentiation, and activa- tion of myeloid hematopoietic cells (Ruef and Coleman, http://dx.doi.org/10.1016/j.anireprosci.2016.04.014 0378-4320/© 2016 Elsevier B.V. All rights reserved.