The endocannabinoid system in Leydig cells

BESIO MORENO MARCOS; PEREYRA, ELBA NORA; RAICESTRINIDAD; PIGNATARO OMAR P.

Buenos Aires

Simposio; rontiers in Biosciences-3; 2018

The endocannabinoid system in Leydig cellsRaices T1, Besio Moreno M1, Pereyra EN1, Pignataro OP1,2.1Institute of Biology and Experimental Medicine (IBYME-CONICET).2Department of Biological Chemistry, School of Sciences, University of Buenos Aires.The endocannabinoid system consists of a lipid signalling network, which comprises endogenous ligands, their cannabinoid receptors and the enzymes that catalyze the formation and degradation of endocannabinoids. The main ligands are anandamide (AEA) and 2-arachidonoyl glycerol. Endocannabinoid receptors are CB1, CB2 and the transient receptor potential vanilloid 1 (TRPV1). Research on this complex network is beginning to shift from the events in the nervous system to other physiopathological functions. However, its effects on the endocrine population of the testis, the Leydig cells (LCs), still need to be clarified. In the present study, we used the mice tumoral cell line MA-10 to unveil the role of the endocannabinoid system in LCs steroidogenesis. Firstly, we incubated the cells with AEA at concentrations ranging from 0,1nM to 1000nM. We found that anandamide (AEA) stimulates basal and db-cAMP stimulated steroidogenesis, having a maximum effect at 10nM concentration. At the concentration range tested, AEA has no effect on cell proliferation. With the aim of assessing the participation of cannabinoid receptors in AEA effect, we treated the cells with 10nM AM251 (CB1 receptor antagonist) and 100nM WIN 55,212-2 (WIN, CB1 and CB2 receptor agonist). Our results indicate that WIN concentration-dependently stimulates steroidogenesis. This effect is reverted by CB1 antagonist, AM215, which suggests that CB1 activation positively regulates LCs steroidogenesis. We also studied the effect of TRPV1 using capsazepine, a specific antagonist. We found that 100nM capsazepine stimulates steroidogenesis, meaning that TRPV1 activation has a negative effect. This comprises the first functional evidence of TRPV1 presence in LCs. In conclusion, the results presented herein provide an approximation on how the endocannabinoid system might be directly affecting LCs functions.