Testosterone exerts profound effects on reproduction and energy homeostasis. Like other orexigenic hormones, it increases endocannabinoid tone within the hypothalamic feeding circuitry. Therefore, we tested the hypothesis that testosterone upregulates the expression of diacylglycerol lipase (DAGL)α in the hypothalamic arcuate nucleus (ARC) to increase energy intake via enhanced endocannabinoid-mediated retrograde inhibition of anorexigenic proopiomelanocortin (POMC) neurons. Energy intake, meal patterns, and energy expenditure were evaluated in orchidectomized, male guinea pigs treated subcutaneously with testosterone propionate (TP; 400 μg) or its sesame oil vehicle (0.1 mL). TP rapidly increased energy intake, meal size, O2 consumption, CO2 production, and metabolic heat production, all of which were antagonized by prior administration of the DAGL inhibitor orlistat (3 μg) into the third ventricle. These orlistat-sensitive, TP-induced increases in energy intake and expenditure were temporally associated with a significant elevation in ARC DAGLα expression. Electrophysiological recordings in hypothalamic slices revealed that TP potentiated depolarization-induced suppression of excitatory glutamatergic input onto identified ARC POMC neurons, which was also abolished by orlistat (3 μM), the CB1 receptor antagonist AM251 (1 μM), and the AMP-activated protein kinase inhibitor compound C (30 μM) and simulated by transient bath application of the dihydrotestosterone mimetic Cl-4AS-1 (100 nM) and testosterone-conjugated bovine serum albumin (100 nM). Thus, testosterone boosts DAGLα expression to augment retrograde, presynaptic inhibition of glutamate release onto ARC POMC neurons that, in turn, increases energy intake and expenditure. These studies advance our understanding of how androgens work within the hypothalamic feeding circuitry to affect changes in energy balance.

Keywords:
Testosterone, Proopiomelanocortin, Diacylglycerol lipase-α, Depolarization-induced suppression of excitation, 2-Arachidonoylglycerol, Estradiol
1.
Davis ME, Potter EL: The response of the human fetal reproductive system to the administration of diethylstilbestrol and testosterone propionate during early pregnancy. Endocrinology 1948;42:370-378.
2.
Phoenix CH, Goy RW, Gerall AA, Young WC: Organizing action of prenatally administered testosterone propionate on the tissues mediating mating behavior in the female guinea pig. Endocrinology 1959;65:369-382.
3.
van Wagenen G: Maturity induced by testosterone in the young male monkey. Fed Proc 1947;6(pt 2):219.
4.
Cheng P, Casida LE: Effects of testosterone propionate upon sexual libido and the production of semen and sperm in the rabbit. Endocrinology 1949;44:38-48.
5.
Bardin CW: The anabolic action of testosterone. N Engl J Med 1996;335:52-53.
6.
Bhasin S, Storer TW, Berman N, Yarasheski KE, Clevenger B, Phillips J, Lee WP, Bunnell TJ, Casaburi R: Testosterone replacement increases fat-free mass and muscle size in hypogonadal men. J Clin Endocrinol Metab 1997;82:407-413.
7.
Sinchak K, Wagner EJ: Estradiol signaling in the regulation of reproduction and energy balance. Front Neuroendocrinol 2012;33:342-363.
8.
Nohara K, Zhang Y, Waraich RS, Laque A, Tiano JP, Tong J, Münzberg H, Mauvais-Jarvis F: Early-life exposure to testosterone programs the hypothalamic melanocortin system. Endocrinology 2011;152:1661-1669.
9.
Rana K, Fam BC, Clarke MV, Pang TPS, Zajac JD, MacLean HE: Increased adiposity in DNA binding-dependent androgen receptor knockout male mice associated with decreased voluntary activity and not insulin resistance. Am J Physiol Endocrinol Metab 2011;301:E767-E778.
10.
Borgquist A, Meza C, Wagner EJ: The role of AMP-activated protein kinase in the androgenic potentiation of cannabinoid-induced changes in energy homeostasis. Am J Physiol Endocrinol Metab 2015;308:E482-E495.
11.
Anukulkitch C, Rao A, Dunshea FR, Blache D, Lincoln GA, Clarke IJ: Influence of photoperiod and gonadal status on food intake, adiposity, and gene expression of hypothalamic appetite regulators in a seasonal mammal. Am J Physiol Regul Integr Comp Physiol 2007;292:R242-R252.
12.
Fan W, Yanase T, Nomura M, Okabe T, Goto K, Sato T, Kawano H, Kato S, Nawata H: Androgen receptor null male mice develop late-onset obesity caused by decreased energy expenditure and lipolytic activity but show normal insulin sensitivity with high adiponectin secretion. Diabetes 2005;54:1000-1008.
13.
Rodríguez AM, Monjo M, Roca P, Palou A: Opposite actions of testosterone and progesterone on UCP1 mRNA expression in cultured brown adipocytes. Cell Mol Life Sci 2002;59:1714-1723.
14.
Lin H-Y, Xu Q, Yeh S, Wang R-S, Sparks JD, Chang C: Insulin and leptin resistance with hyperleptinemia in mice lacking androgen receptor. Diabetes 2005;54:1717-1725.
15.
Sheppard KM, Padmanabhan V, Coolen LM, Lehman MN: Prenatal programming by testosterone of hypothalamic metabolic control neurones in the ewe. J Neuroendocrinol 2011;23:401-411.
16.
Borgquist A, Wagner EJ: On the cannabinoid regulation of energy homeostasis: past, present and future; in Murillo-Rodríguez E, Onaivi ES, Darmani NA, Wagner EJ (eds): Endocannabinoids: Molecular, Pharmacological, Behavioral and Clinical Features. Oak Park, Bentham Science, 2013, pp 60-91.
17.
Di Marzo V, Goparahu SK, Wang L, Liu J, Bátkai S, Járai Z, Fezza F, Miura GI, Palmiter RD, Sugiura T, Kunos G: Leptin-regulated endocannabinoids are involved in maintaining food intake. Nature 2001;410:822-825.
18.
Malcher-Lopes R, Di S, Marcheselli VS, Weng F-J, Stuart CT, Bazan NG, Tasker JG: Opposing crosstalk between leptin and glucocorticoids rapidly modulates synaptic excitation via endocannabinoid release. J Neurosci 2006;26:6643-6650.
19.
Kola B, Farkas I, Christ-Crain M, Wittmann G, Lolli F, Amin F, Harvey-White J, Liposits Z, Kunos G, Grossman AB, Fekete C, Korbonits M: The orexigenic effect of ghrelin is mediated through central activation of the endogenous cannabinoid system. PLoS One 2008;3:e1797.
20.
Kirkham TC, Williams CM, Fezza F, Di Marzo V: Endocannabinoid levels in rat limbic forebrain and hypothalamus in relation to fasting, feeding and satiation: stimulation of eating by 2-arachidonyl glycerol. Br J Pharmacol 2002;136:550-557.
21.
Yoshida T, Fukaya M, Uchigashima M, Miura E, Kamiya H, Kano M, Watanabe M: Localization of diacylglycerol lipase-α around postsynaptic spine suggests close proximity between production site of an endocannabinoid, 2-arachidonyl-glycerol, and presynaptic cannabinoid CB1 receptor. J Neurosci 2006;26:4740-4751.
22.
Di Marzo V, Ligresti A, Cristino L: The endocannabinoid system as a link between homeostatic and hedonic pathways involved in energy balance regulation. Int J Obes (Lond) 2009;33(suppl 2):S18-S24.
23.
Pérez-Morales M, López-Colomé AM, Méndez-Díaz M, Ruiz-Contreras AE, Prospéro-García O: Inhibition of diacylglycerol lipase (DAGL) in the lateral hypothalamus of rats prevents the increase in REMS and food ingestion induced by PAR1 stimulation. Neurosci Lett 2014;578:117-121.
24.
Borgquist A, Meza C, Wagner EJ: Role of neuronal nitric oxide synthase in the estrogenic attenuation of cannabinoid-induced changes in energy homeostasis. J Neurophysiol 2015;113:904-914.
25.
Farhang B, Pietruszewski L, Lutfy K, Wagner EJ: The role of the NOP receptor in regulating food intake, meal pattern, and the excitability of proopiomelanocortin neurons. Neuropharmacology 2010;59:190-200.
26.
Hashimotodani Y, Ohno-Shosaku T, Yamazaki M, Sakimura K, Kano M: Neuronal protease-activated receptor 1 drives synaptic retrograde signaling mediated by the endocannabinoid 2-arachidonoylglycerol. J Neurosci 2011;31:3104-3109.
27.
Schmidt A, Harada S-I, Kimmel DB, Bai C, Chen F, Rutledge SJ, Vogel RL, Scafonas A, Gentile MA, Nantermet PV, McElwee-Witmer S, Pennypacker B, Masarachia P, Sahoo SP, Kim Y, Meissner RS, Hartman GD, Duggan ME, Rodan GA, Towler DA, Ray WJ: Identification of anabolic selective androgen receptor modulators with reduced activities in reproductive tissues and sebaceous glands. J Biol Chem 2009;284:36367-36376.
28.
Ho J, Cox JM, Wagner EJ: Cannabinoid-induced hyperphagia: correlation with inhibition of proopiomelanocortin neurons? Physiol Behav 2007;92:507-519.
29.
Borgquist A, Rivas VM, Kachani M, Sinchak K, Wagner EJ: Gonadal steroids differentially modulate the actions of orphanin FQ/nociceptin at a physiologically relevant circuit controlling female sexual receptivity. J Neuroendocrinol 2014;26:329-340.
30.
Kim KW, Jo Y-H, Zhao L, Stallings NR, Chua SC, Parker KL: Steroidogenic factor 1 regulates expression of the cannabinoid receptor 1 in the ventromedial hypothalamic nucleus. Mol Endocrinol 2008;22:1950-1961.
31.
Lindberg D, Chen P, Li C: Conditional viral tracing reveals that steroidogenic factor 1-positive neurons of the dorsomedial subdivision of the ventromedial hypothalamus project to the autonomic centers of the hypothalamus and hindbrain. J Comp Neurol 2013;521:3167-3190.
32.
Cardinal P, André C, Quarta C, Bellocchio L, Clark S, Elie M, Leste-Lasserre T, Maitre M, Gonzales D, Cannich A, Pagotto U, Marsicano G, Cota D: CB1 cannabinoid receptor in SF1-expressing neurons of the ventromedial hypothalamus determines metabolic responses to diet and leptin. Mol Metab 2014;3:705-716.
33.
Hirsch E, Collier G: The ecological determinants of reinforcement in the guinea pig. Physiol Behav 1974;12:239-249.
34.
Horton BJ, West CE, Turley SD: Diurnal variation in the feeding pattern of guinea pigs. Nutr Metabol 1975;18:294-301.
35.
Chai J-K, Blaha V, Meguid MM, Laviano A, Yang Z-J, Varma M: Use of orchidectomy and testosterone replacement to explore meal number-to-meal size relationship in male rats. Am J Physiol Regul Integr Comp Physiol 1999;45:R1366-R1373.
36.
Batterham MJ, Garsia R: A comparison of megestrol acetate, nandrolone decanoate and dietary counselling for HIV associated weight loss. Int J Androl 2001;24:232-240.
37.
Storer TW, Woodhouse LJ, Sattler F, Singh AB, Schroeder ET, Beck K, Padero M, Mac P, Yarasheski KE, Geurts P, Willemsen A, Harms MK, Bhasin S: A randomized, placebo-controlled trial of nandrolone decanoate in human immunodeficiency virus-infected men with mild to moderate weight loss with recombinant human growth hormone as active reference treatment. J Clin Endocrinol Metab 2005;90:4474-4482.
38.
Wagner EJ: Sex differences in cannabinoid-regulated biology: a focus on energy homeostasis. Front Neuroendocrinol 2016;40:101-109.
39.
Conde K, Meza C, Kelly MJ, Sinchak K, Wagner EJ: Estradiol rapidly attenuates ORL-1 receptor-mediated inhibition of proopiomelanocortin neurons via Gq-coupled, membrane-initiated signaling. Neuroendocrinology 2016;103:787-805.
40.
Badawy ZS, Chohan KR, Whyte DA, Penefsky HS, Brown OM, Souid A-K: Cannabinoids inhibit the respiration of human sperm. Fertil Steril 2009;91:2471-2476.
41.
Whyte DA, Al-Hammadi S, Balhaj G, Brown OM, Penefsky HS, Souid A-K: Cannabinoids inhibit cellular respiration of human oral cancer cells. Pharmacology 2010;85:328-335.
42.
Perwitz N, Fasshauer M, Klein J: Cannabinoid receptor signaling directly inhibits thermogenesis and alters expression of adiponectin and visfatin. Horm Metab Res 2006;38:356-358.
43.
Washburn N, Borgquist A, Wang K, Jeffery GS, Kelly MJ, Wagner EJ: Receptor subtypes and signal transduction mechanisms contributing to the estrogenic attenuation of cannabinoid-induced changes in energy homeostasis. Neuroendocrinology 2013;97:160-175.
44.
Bénard G, Massa F, Puente N, Lourenço J, Bellocchio L, Soria-Gómez E, Matias I, Delamarre A, Metna-Laurent M, Cannich A, Hebert-Chatelain E, Mulle C, Ortega-Gutiérrez S, Martín-Fontecha M, Klugmann M, Guggenhuber S, Lutz B, Gertsch J, Chaouloff F, López-Rodríguez ML, Grandes P, Rossignol R, Marsicano G: Mitochondrial CB1 receptors regulate neuronal energy metabolism. Nat Neurosci 2012;15:558-564.
45.
Koch M, Varela L, Kim JG, Kim JD, Hernández-Nuño F, Simonds SE, Castorena CM, Vianna CR, Elmquist JK, Morozov YM, Rakic P, Bechmann I, Cowley MA, Szigeti-Buck K, Dietrich MO, Gao X-B, Diano S, Horvath TL: Hypothalamic POMC neurons promote cannabinoid-induced feeding. Nature 2015;519:45-50.
46.
Kola B, Hubina E, Tucci SA, Kirkham TC, Garcia EA, Mitchell SE, Williams LM, Hawley SA, Hardie DG, Grossman AB, Korbonits M: Cannabinoids and ghrelin have both central and peripheral metabolic and cardiac effects via AMP-activated protein kinase. J Biol Chem 2005;280:25196-25201.
47.
Estrada M, Espinosa A, Müller M, Jaimovich E: Testosterone stimulates intracellular calcium release and mitogen-activated protein kinases via a G protein-coupled receptor in skeletal muscle cells. Endocrinology 2003;144:3586-3597.
48.
Vicencio JM, Ibarra C, Estrada M, Chiong M, Soto D, Parra V, Diaz-Araya G, Jaimovich E, Lavandero S: Testosterone induces an intracellular calcium increase by a nongenomic mechanism in cultured rat cardiac myocytes. Endocrinology 2006;147:1386-1395.
49.
Wilson C, Contreras-Ferrat A, Venegas N, Osorio-Fuentealba C, Pávez M, Montoya K, Durán J, Maass R, Lavandero S, Estrada M: Testosterone increases GLUT4-dependent glucose uptake in cardiomyocytes. J Cell Physiol 2013;228:2399-2407.
50.
Loss ES, Jacobus AP, Wassermann GF: Rapid signaling responses in Sertoli cell membranes induced by follicle stimulating hormone and testosterone: calcium inflow and electrophysiological changes. Life Sci 2011;89:577-583.
51.
Gu S, Honisch S, Kounenidakis M, Alkahtani S, Alarifi S, Alevizopoulos K, Stournaras C, Lang F: Membrane androgen receptor down-regulates c-Src-activity and beta-catenin transcription and triggers GSK-3beta-phosphorylation in colon tumor cells. Cell Physiol Biochem 2014;34:1402-1412.
52.
Kreitzer AC, Regehr WG: Retrograde inhibition of presynaptic calcium influx by endogenous cannabinoids at excitatory synapses onto Purkinje cells. Neuron 2001;29:717-727.
53.
Rancz EA, Häusser M: Dendritic calcium spikes are tunable triggers of cannabinoid release and short-term synaptic plasticity in cerebellar Purkinje neurons. J Neurosci 2006;26:5428-5437.
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2016
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