Induction of the Secretion of LH and GH by Orexin A and Ghrelin is Controlled in Vivo by Leptin and Photoperiod in Sheep

Open access


The influence of leptin on orexin A and the interaction of leptin with ghrelin in regulating the gonadotropic and somatotropic axes in seasonally polyestrous animals are not well understood. This study examined the effects of these factors as well as the mediating roles of specific ovine leptin antagonist (SOLA; mutant D23L/L39A/D40A/F41A) and photoperiod on luteinizing hormone (LH) and growth hormone (GH) secretion. Twenty-four ovariectomized, estradiol-implanted ewes were used in a replicated switchback design. The ewes were assigned randomly to 1 of 6 treatments (infused into the third ventricle 3 times at 0 (dusk), 1, and 2 h) as follows: control, Ringer-Locke buffer; leptin, 0.5 μg/kg b.w.; orexin A, 0.3 μg/kg b.w.; ghrelin, 2.5 μg/kg b.w.; SOLA, 50 μg/kg b.w. + orexin A, 0.3 μg/kg b.w.; and SOLA, 50 μg/kg b.w. + ghrelin, 2.5 μg/kg b.w. Blood samples (5 ml) were collected at 15-min intervals for 4 h. SOLA + orexin A resulted in an increase (P<0.01) in the LH plasma concentration during short-day (SD) and long-day (LD) photoperiods. However, ghrelin and SOLA + ghrelin had the opposite effect. SOLA + orexin A resulted in an increase (P<0.001) in the GH concentration compared with leptin or orexin A during the LD season. Ghrelin and SOLA + ghrelin increased the GH concentration (P<0.01) regardless of the season. In summary, LH and GH secretion are seasonally dependent on relationships that are subject to photoperiodic regulation, and leptin is an important regulator of the effects of ghrelin and orexin A on the activities of the gonadotropic and somatotropic axes in sheep.

Accorsi P.A., Munno A., Gamberoni M., Viggiani R., De Ambrogi M., Tamanini C., Seren E. (2007). Role of leptin on growth hormone and prolactin secretion by bovine pituitary explants. J. Dairy Sci., 90: 1683-1691.

Amstalden M., Garcia M.R., Stanko R.L., Nizielski S.E., Morrison C.D., Keisler D.H. (2002). Central infusion of recombinant ovine leptin normalizes plasma insulin and stimulatesanovel hypersecretion of luteinizing hormone after short-term fasting in mature beef cows. Biol. Reprod., 66: 1555-1561.

Amstalden M., Zieba D.A., Edwards J.F., Harms P.G., Welsh T.H. Jr., Stanko R.L. (2003). Leptin acts at the bovine adenohypophysis to enhance basal and gonadotropin-releasing hormone-mediated release of luteinizing hormone: differential effects are dependent upon nutritional history. Biol. Reprod., 69: 1539-1544.

Anukulkitch C., Rao A., Pereira A., Mc Ewan J., Clarke I.J. (2010). Expression of genes for appetite-regulating peptides in the hypothalamus of genetically selected lean and fat sheep. Neuroendocrinology, 91: 223-238.

Archer Z.A., Findlay P.A., Rhind S.M., Mercer J.G., Adam C.L. (2002). Orexin gene expression and regulation by photoperiod in the sheep hypothalamus. Regul. Pept., 104: 41-45.

Backholer K., Smith J., Clarke I.J. (2009). Melanocortins may stimulate reproduction by activating orexin neurons in the dorsomedial hypothalamus and kisspeptin neurons in the preoptic area of the ewe. Endocrinology, 150: 5488-5497.

Barb C.R., Matteri R.L. (2005). Orexin-Bmodulates luteinizing hormone and growth hormone secretion from porcine pituitary cells in culture. Domest. Anim. Endocrinol., 28: 331-337.

Barenton B., Ravault J., Chabanet C., Daveau A., Pelletier J., Ortavant R. (1988). Photoperiodic control of growth hormone secretion and body weight in rams. Domest. Anim. Endocrinol., 5: 247-255.

Chen C., Xu R. (2003). The in vitro regulation of growth hormone secretion by orexins. Endocrine, 22: 57-66.

Clarke I.J. (2014). Interface between metabolic balance and reproduction in ruminants: focus on the hypothalamus and pituitary. Horm. Behav., 66: 15-40.

Cocchi D., De Gennaro Colonna V., Bagnasco M., Bonacci D., Müller E.E. (1999). Leptin regulates GHsecretion in the rat by acting on GHRHand somatostatinergic functions. J. Endocrinol., 162: 95-99.

Dovolou E., Chadio S., Messinis I.E., Rekkas C.A., Deligiannis C., Kalogiannis D., Amiridis G.S. (2013). Human ghrelin decreases pituitary response to Gn RHin superovulated ewes. Theriogenology, 80: 262-268.

Dvorak P., Becka S., Krejci P., Chrpova M. (1978). Radioimmunoassay of bovine growth hormone. Radiochem. Radioanal. Lett., 34: 160-166.

Grouselle D., Chaillou E., Caraty A., Bluet- Pajot M.T., Zizzari P., Tillet Y., Epelbaum J. (2008). Pulsatile cerebrospinal fluid and plasma ghrelin in relation to growth hormone secretion and food intake in the sheep. J. Neuroendocrinol., 20: 1138-1146.

Harrison J.L., Miller D.W., Findlay P.A., Adam C.L. (2008). Photoperiod influences the central effects of ghrelin on food intake, GHand LHsecretion in sheep. Neuroendocrinology, 87:182-192.

Hashizume T., Horiuchi M., Nonaka S., Kasuya E., Kojima M., Hosoda H., Kangawa K. (2005). Effects of ghrelin on growth hormone secretion in vivo in ruminants. Regul. Pept., 126: 61-65.

Iqbal J., Manley T.R., Ciofi P., Clarke I.J. (2005). Reduction in adiposity affects the extent of afferent projections to growth hormone-releasing hormone and somatostatin neurons and the degree of colocalization of neuropeptides in growth hormone-releasing hormone and somatostatin cells of the ovine hypothalamus. Endocrinology, 146: 4776-4785.

Iqbal J., Kurose Y., Canny B., Clarke I.J. (2006). Effects of central infusion of ghrelin on food intake and plasma levels of growth hormone, luteinizing hormone, prolactin, and cortisol secretion in sheep. Endocrinology, 147: 510-519.

Iqbal J., Pompolo S., Murakami T., Grouzmann E., Sakurai T., Meister B., Clarke I.J. (2001). Immunohistochemical characterization of localization of long-form leptin receptor (OB-Rb) in neurochemically defined cells in the ovine hypothalamus. Brain Res., 920: 55-64.

Kirsz K., Szczesna M., Molik E., Misztal T., Wojtowicz A.K., Zieba D.A. (2012). Seasonal changes in the interactions among leptin, ghrelin, and orexin in sheep. J. Anim. Sci., 90:2524-2531.

Kirsz K., Szczesna M., Dudek K., Bartlewski P.M., Zieba D.A. (2014). Influence of season and nutritional status on the direct effects of leptin, orexin-Aand ghrelin on luteinizing hormone and growth hormone secretion in the ovine pituitary explant model. Domest. Anim. Endocrinol., 48: 69-76.

Kohsaka A., Watanobe H., Kakizaki Y., Suda T., Schiöth H.B. (2001). Asignificant participation of orexin-A,apotent orexigenic peptide, in the preovulatory luteinizing hormone and prolactin surges in the rat. Brain Res., 898: 166-170.

Lebrethon M.C., Aganina A., Fournier M., G érard A., Parent A.S., Bourguignon J.P. (2007). Effects of in vivo and in vitro administration of ghrelin, leptin and neuropeptide mediators on pulsatile gonadotrophin-releasing hormone secretion from male rat hypothalamus before and after puberty. J. Neuroendocrinol., 19: 181-188.

López M., Seoane L.M., Tovar S., Nogueiras R., Diéguez C., Señarís R. (2004). Orexin-Aregulates growth hormone-releasing hormone m RNAcontent inanucleus-specific manner and somatostatin m RNAcontent inagrowth hormone-dependent fashion in the rat hypothalamus. Eur. J. Neurosci., 19: 2080-2088.

Luque R.M., Huang Z.H., Shah B., Mazzone T., Kineman R.D. (2007). Effects of leptin replacement on hypothalamic-pituitary growth hormone axis function and circulating ghrelin levels in ob/ob mice. Am. J. Physiol. - Endoc. M., 292: E891-E899.

Miller D., Findlay P.A., Morrison M.A., Raver N., Adam C.L. (2002). Seasonal and dosedependent effects of intracerebroventricular leptin on LHsecretion and appetite in sheep. J. Endocrinol., 175: 395-404.

Nagatani S., Zeng Y., Keisler D.H., Foster D.L., Jaffe C.A. (2000). Leptin regulates pulsatile luteinizing hormone and growth hormone secretion in the sheep. Endocrinology, 141:3965-3975.

Nonaka S., Hashizume T., Kasuya E. (2005). Effects of leptin on the release of luteinizing hormone, growth hormone and prolactin from cultured bovine anterior pituitary cells. Anim. Sci. J., 76: 435-440.

Polkowska J., Gajewska A., Wańkowska M., Misztal T., Wójcik - Gładysz A. (2012). The effect of intracerebroventricular infusions of ghrelin or short fasting on the gene expression and immunoreactivity of neuropeptide Yin the hypothalamic neurons in prepubertal female lambs:amorphofunctional study. J. Chem. Neuroanat., 46: 45-50.

Pu S., Jain M.R., Kalra P.S., Kalra S.P. (1998). Orexins,anovel family of hypothalamic neuropeptides, modulate pituitary luteinizing hormone secretion in an ovarian steroid-dependent manner. Regul. Pept., 78: 133-136.

Russel A.J.F., Doney J.M., Gunn R.G. (1969). Subjective assessment of body fat in live sheep. J. Agric. Sci., 72: 451-454.

Russell S.H., Small C.J., Kennedy A.R., Stanley S.A., Seth A., Murphy K.G., Taheri S., Ghatei M.A., Bloom S.R. (2001). Orexin Ainteractions in the hypothalamo-pituitary gonadal axis. Endocrinology, 142: 5294-5302.

Sartin J.L., Dyer C., Matteri R., Buxton D., Buonomo F., Shores M., Baker J., Osborne J.A., Braden T., Steele B. (2001). Effect of intracerebroventricular orexin-Bon food intake in sheep. J. Anim. Sci., 79: 1573-1577.

Seoane L., Tovar S., Perez D., Mallo F., Lopez M., Señaris R., Casanueva F., Dieguez C. (2004). Orexin Asuppresses in vivo GHsecretion. Eur. J. Endocrinol., 150: 731-736.

Sirotkin A.V., Grossmann R. (2007). The role of ghrelin and some intracellular mechanisms in controlling the secretory activity of chicken ovarian cells. Comp. Biochem. Phys. A, 147: 239-246.

Sirotkin A.V., Chrenková M., Nitrayová S., Patraš P., Darlak K., Valenzuela F., Pinilla L., Tena - Sempere M. (2008). Effects of chronic food restriction and treatments with leptin or ghrelin on different reproductive parameters of male rats. Peptides, 29:1362-1368.

Small C.J., Goubillon M.L., Murray J.F., Siddiqui A., Grimshaw S.E., Young H., Sivanesan V., Kalamatianos T., Kennedy A.R., Coen C.W., Bloom S.R., Wilson C.A. (2003). Central orexin Ahas site-specific effects on luteinizing hormone release in female rats. Endocrinology, 144: 3225-3236.

Spicer L.J., Francisco C.C. (1997). The adipose obese gene product, leptin: evidence ofadirect inhibitory role in ovarian function. Endocrinology, 138: 3374-3379.

Stupnicki R., Madej A. (1976). Radioimmunoassay of LHin blood plasma of farm animals. Endokrinologie, 68: 6-13.

Sun G., Tian Z., Yao Y., Li H., Higuchi T. (2006). Central and/or peripheral immunoreactivity of orexin-Ain pregnant rats and women. J. Mol. Endocrinol., 36: 131-138.

Szczesna M., Kirsz K., Kucharski M., Szymaszek P., Zieba D.A. (2013). Obesity and leptin resistance: the role of growth hormone. Health, 5: 29-39.

Tamura T., Irahara M., Tezuka M., Kiyokawa M., Aono T. (1999). Orexins, orexigenic hypothalamic neuropeptides, suppress the pulsatile secretion of luteinizing hormone in ovariectomized female rats. Biochem. Biophys. Res. Commun., 264: 759-762.

Towhidi A., Khazali H., Zhandi M. (2007). Leptin isametabolic signal for Gn RH-LH/FSH Axis in feed-restricted ewes. Asian Australas. J. Anim. Sci., 20: 1039-1048.

Traczyk W., Przekop F. (1963). Methods of investigation of the function of the hypothalamus and hypophysis in chronic experiment in sheep. Acta Physiol. Pol., 14: 217-226.

Tritos N.A., Mastaitis J.W., Kokkotou E., Maratos- Flier E. (2001). Characterization of melanin concentrating hormone and preproorexin expression in the murine hypothalamus. Brain Res., 895: 160-166.

Wójcik - Gładysz A., Wańkowska M., Misztal T., Romanowicz K., Polkowska J. (2009). Effect of intracerebroventricular infusion of leptin on the secretory activity of the Gn RH/LH axis in fasted prepubertal lambs. Anim. Reprod. Sci., 114: 370-383.

Zachow R.J., Magoffin D.A. (1997). Direct intraovarian effects of leptin: impairment of the synergistic action of insulin-like growth factor-Ion follicle-stimulating hormone-dependent estradiol- 17 beta production by rat ovarian granulosa cells. Endocrinology, 138: 847-850.

Zieba D., Amstalden M., Gallino J.L., Edwards J.F., Harms P.G., Williams G.L.(2003 a). Effects of leptin on basal and GHRH-stimulated GHsecretion from the bovine adenohypophysis are dependent upon nutritional status. J. Endocrinol., 178: 83-89.

Zieba D.A., Amstalden M., Maciel M.N., Keisler D.H., Raver N., Gertler A., Williams GL. (2003 b). Divergent effects of leptin on luteinizing hormone and insulin secretion are dose dependent. Exp. Biol. Med., 228: 325-330.

Zieba D.A., Szczesna M., Klocek - Gorka B., Molik E., Misztal T., Williams G.L., Romanowicz K., Stepien E., Keisler D.H., Murawski M. (2008 a). Seasonal effects of central leptin infusion on secretion of melatonin and prolactin and on SOCS-3 gene expression in ewes. J. Endocrinol., 198: 147-155.

Zieba D.A., Szczesna M., Klocek - Gorka B., Williams G.L. (2008 b). Leptin asanutritional signal regulating appetite and reproductive processes in seasonally-breeding ruminants. J. Physiol. Pharmacol., 59 (Suppl. 9): 7-18.

Zieba D.A., Kirsz K., Molik E., Romanowicz K., Wojtowicz A.K. (2011). Effects of orexigenic peptides and leptin on melatonin secretion during different photoperiods in seasonal breeding ewes: an in vitro study. Domest. Anim. Endocrinol., 40: 139-146.

Annals of Animal Science

The Journal of National Research Institute of Animal Production

Journal Information

IMPACT FACTOR 2017: 1.018
5-year IMPACT FACTOR: 0.959

CiteScore 2017: 1.01

SCImago Journal Rank (SJR) 2017: 0.413
Source Normalized Impact per Paper (SNIP) 2017: 0.822

Cited By


All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 214 214 22
PDF Downloads 60 60 8