Relationship between kisspeptin-10, neurokinin B and dynorphin A in the course of normal and delayed puberty in ewes

Natalia Szysiak; Urszula Kosior-Korzecka; Monika Greguła-Kania; Krzysztof Patkowski; Mateusz Fila; Andrzej Junkuszew

doi:10.2478/jvetres-2026-0010

Abstract

Introduction

Kisspeptin (KiSS), neurokinin B (NKB) and dynorphin A (Dyn A) participate in the neuroendocrine regulation of reproductive system development and functioning; however, their roles in the onset of sexual maturity and pathomechanism of delayed puberty have not yet been fully elucidated. The aim of the study was to determine changes in the kisspeptin-10 (KiSS-10), NKB and Dyn A concentration in blood plasma collected from ewe lambs during puberty in relation to the first ovulation time, and to examine the relationship between these neuropeptides.

Material and Methods

Twenty-four ewe lambs were divided into two groups: from single (S) pregnancies and from twin (T) pregnancies. To determine the KiSS-10, NKB and Dyn A concentration with ELISA, blood was collected from all sheep at 4, 5, 6, 7, 8, 9 and 10 months of age. Singletons and twins had their first ovulations at 8 and 10 months of age, respectively.

Results

Changes in the KiSS-10 and NKB concentrations were positively correlated with age in both S (r = 0.86) and T ewes (r = 0.89). A negative correlation was found between KiSS-10 and Dyn A (r = –0.55 and –0.98, for S and T, respectively) and between NKB and Dyn A (r = –0.89 and –0.94, for S and T, respectively).

Conclusion

The initiation of ovarian activity is associated with concurrent increases in plasma KiSS-10 and NKB concentrations with age and reduced Dyn A concentrations in both groups. Our results show that these neuropeptides may regulate the timing of reproductive activity onset in sheep.

References

Abadjieva D., Shumkov K., Kistanova E., Kacheva D., Georgiev B.: Opportunities for the improvement of the reproductive performances in female animals. Bio Anim Husb 2011, 27, 365–372, doi: 10.2298/BAH1103365A.
Open DOI Search in Google Scholar Back to article
Ahima R.S.: No Kiss1ng by leptin during puberty? J Clin Invest 2011, 121, 34–36, doi: 10.1172/JCI45813.
Open DOI Search in Google Scholar Back to article
Amstalden M., Coolen L.M., Hemmerle A.M., Billings H.J., Connors J.M., Goodman R.L., Lehman M.N.: Neurokinin 3 receptor immunoreactivity in the septal region, preoptic area and hypothalamus of the female sheep: colocalisation in neurokinin B cells of the arcuate nucleus but not in gonadotrophin-releasing hormone neurones. J Neuroendocrinol 2010, 22, 1–12, doi: 10.1111/j.1365-2826.2009.01930.x.
Open DOI Search in Google Scholar Back to article
Billings H.J., Connors J.M., Altman S.N., Hileman S.M., Holaskova I., Lehman M.N., McManus C.J., Nestor C.C., Jacobs B.H., Goodman R.L.: Neurokinin B acts via the neurokinin-3 receptor in the retrochiasmatic area to stimulate luteinizing hormone secretion in sheep. Endocrinology 2010, 151, 3836–3846, doi: 10.1210/en.2010-0174.
Open DOI Search in Google Scholar Back to article
Castellano J.M., Navarro V.M., Fernández-Fernández R., Nogueiras R., Tovar S., Roa J., Vazquez M.J., Vigo E., Casanueva F.F., Aguilar E., Pinilla L., Dieguez C., Tena-Sempere M.: Changes in hypothalamic KiSS-1 system and restoration of pubertal activation of the reproductive axis by kisspeptin in undernutrition. Endocrinology 2005, 146, 3917–3925, doi: 10.1210/en.2005-0337.
Open DOI Search in Google Scholar Back to article
De Roux N., Genin E., Carel J.-C., Matsuda F., Chaussain J.-L., Milgrom E.: Hypogonadotropic hypogonadism due to loss of function of the KiSS1-derived peptide receptor GPR54. Proc Natl Acad Sci U S A 2003, 100, 10972–10976, doi: 10.1073/pnas.1834399100.
Open DOI Search in Google Scholar Back to article
Harlow K., Renwick A.N., Shuping S.L., Sommer J.R., Lents C.A., Knauer M.T., Nestor C.C.: Evidence that pubertal status impacts kisspeptin/neurokinin B/dynorphin neurons in the gilt. Biol Reprod 2021, 105, 1533–1544, doi: 10.1093/biolre/ioab189.
Open DOI Search in Google Scholar Back to article
Hausman G.J., Barb C.R., Lents C.A.: Leptin and reproductive function. Biochimie 2012, 94, 2075–2081, doi: 10.1016/j.biochi.2012.02.022.
Open DOI Search in Google Scholar Back to article
Kosior-Korzecka U., Patkowski K., Bobowiec R., Wójcik M., Tusińska E.: Sustained opposite relationships between anabolic hormones in preweaning triplet lambs born to obese mothers. Bull Vet Inst Pulawy 2012, 56, 109–114, doi: 10.2478/v10213-012-0020-6.
Open DOI Search in Google Scholar Back to article
Lapatto R., Pallais J.C., Zhang D., Chan Y.-M., Mahan A., Cerrato F., Le W.W., Hoffman G.E., Seminara S.B.: Kiss1 –/– mice exhibit more variable hypogonadism than Gpr54–/–mice. Endocrinology 2007, 148, 4927–4936, doi: 10.1210/en.2007-0078.
Open DOI Search in Google Scholar Back to article
Li Q., Smith J.T., Henry B., Rao A., Pereira A., Clarke I.J.: Expression of genes for kisspeptin (KISS1), neurokinin B (TAC3), prodynorphin (PDYN), and gonadotropin inhibitory hormone (RFRP) across natural puberty in ewes. Physiol Rep 2020, 8, e14399, doi: 10.14814/phy2.14399.
Open DOI Search in Google Scholar Back to article
Lopez J.A., Bedenbaugh M.N., McCosh R.B., Weems P.W., Meadows L.J., Wisman B., Coolen L.M., Goodman R.L., Hileman S.M.: Does dynorphin play a role in the onset of puberty in female sheep? J Neuroendocrinol 2016, 28, e12445, doi: 10.1111/jne.12445.
Open DOI Search in Google Scholar Back to article
Nakahara T., Uenoyama Y., Iwase A., Oishi S., Nakamura S., Minabe S., Watanabe Y., Deura C., Noguchi T., Fujii N., Kikkawa F., Maeda K., Tsukamura H.: Chronic peripheral administration of kappa-opioid receptor antagonist advances puberty onset associated with acceleration of pulsatile luteinizing hormone secretion in female rats. J Reprod Dev 2013, 59, 479–484, doi: 10.1262/jrd.2013-046.
Open DOI Search in Google Scholar Back to article
Nestor C.C., Briscoe A.M.S., Davis S.M., Valent M., Goodman R.L., Hileman S.M.: Evidence of a role for kisspeptin and neurokinin B in puberty of female sheep. Endocrinology 2012, 153, 2756–2765, doi: 10.1210/en.2011-2009.
Open DOI Search in Google Scholar Back to article
Pinilla L., Aguilar E., Dieguez C., Millar R.P., Tena-Sempere M.: Kisspeptins and reproduction: physiological roles and regulatory mechanisms. Physiol Rev 2012, 92, 1235–1316, doi: 10.1152/physrev.00037.2010.
Open DOI Search in Google Scholar Back to article
Plant T.M., Ramaswamy S., DiPietro M.J.: Repetitive activation of hypothalamic G protein-coupled receptor 54 with intravenous pulses of kisspeptin in the juvenile monkey (Macaca mulatta,) elicits a sustained train of gonadotropin-releasing hormone discharges. Endocrinology 2006, 147, 1007–1013, doi: 10.1210/en.2005-1261.
Open DOI Search in Google Scholar Back to article
Qiu J., Fang Y., Bosch M.A., Rønnekleiv O.K., Kelly M.J.: Guinea pig kisspeptin neurons are depolarized by leptin via activation of TRPC channels. Endocrinology 2011, 152, 1503–1514, doi: 10.1210/en.2010-1285.
Open DOI Search in Google Scholar Back to article
Radwańska P., Kosior-Korzecka U.: Relationships between leptin, the KiSS-1/GPR54 system and thyrotropic axis activity in ewe lambs predisposed to the delayed puberty. Small Rumin Res 2016, 144, 6–16, doi: 10.1016/j.smallrumres.2016.07.019.
Open DOI Search in Google Scholar Back to article
Ramaswamy S., Seminara S.B., Ali B., Ciofi P., Amin N.A., Plant T.M.: Neurokinin B stimulates GnRH release in the male monkey (Macaca mulatta) and is colocalized with kisspeptin in the arcuate nucleus. Endocrinology 2010, 151, 4494–4503, doi: 10.1210/en.2010-0223.
Open DOI Search in Google Scholar Back to article
Ramaswamy S., Seminara S.B., Plant T.M.: Evidence from the agonadal juvenile male rhesus monkey (Macaca mulatta) for the view that the action of neurokinin B to trigger gonadotropin-releasing hormone release is upstream from the kisspeptin receptor. Neuroendocrinology 2011, 94, 237–245, doi: 10.1159/000329045.
Open DOI Search in Google Scholar Back to article
Redmond J.S., Macedo G.G., Velez I.C., Caraty A., Williams G.L., Amstalden M.: Kisspeptin activates the hypothalamic–adenohypophyseal–gonadal axis in prepubertal ewe lambs. Reproduction 2011, 141, 541–548, doi: 10.1530/REP-10-0467.
Open DOI Search in Google Scholar Back to article
Rosales Nieto C.A., Ferguson M.B., Macleay C.A., Briegel J.R., Martin G.B., Thompson A.N.: Selection for superior growth advances the onset of puberty and increases reproductive performance in ewe lambs. Animal 2013, 7, 990–997, doi: 10.1017/S1751731113000074.
Open DOI Search in Google Scholar Back to article
Rosales Nieto C.A., Thompson A.N., Macleay C.A., Briegel J.R., Hedger M.P., Ferguson M.B., Martin G.B.: Relationships among body composition, circulating concentrations of leptin and follistatin, and the onset of puberty and fertility in young female sheep. Anim Reprod Sci 2014, 151, 148–156, doi: 10.1016/j.anireprosci.2014.10.008.
Open DOI Search in Google Scholar Back to article
Seminara S.B., Messager S., Chatzidaki E.E., Thresher R.R., Acierno J.S., Shagoury J.K., Bo-Abbas Y., Kuohung W., Schwinof K.M., Hendrick A.G., Zahn D., Dixon J., Kaiser U.B., Slaugenhaupt S.A., Gusella J.F., O’Rahilly S., Carlton M.B.L., Crowley W.F., Aparicio S.A.J.R., Colledge W.H.: The GPR54 gene as a regulator of puberty. N Engl J Med 2003, 349, 1614–1627, doi: 10.1056/NEJMoa035322.
Open DOI Search in Google Scholar Back to article
Smith J.T., Acohido B.V., Clifton D.K., Steiner R.A.: KiSS-1 neurones are direct targets for leptin in the ob/ob mouse. J Neuroendocrinol 2006, 18, 298–303, doi: 10.1111/j.1365-2826.2006.01417.x.
Open DOI Search in Google Scholar Back to article
Szysiak N., Kosior-Korzecka U., Greguła-Kania M., Patkowski K., Fila M., Junkuszew A.: Effect of neurokinin B and dynorphin A on kisspeptin-10 secretion from the anterior pituitary cells of pubescent ewes in vitro. J Vet Res 2025, 69, 299–304, doi: 10.2478/jvetres-2025-0026.
Open DOI Search in Google Scholar Back to article
Szysiak N., Kosior-Korzecka U., Longo V., Patkowski K., Greguła-Kania M., Nowakiewicz A., Bochniarz M., Junkuszew A.: Influence of neurokinin B, dynorphin A and kisspeptin-10 on in vitro gonadotropin secretion by anterior pituitary cells isolated from pubescent ewes. J Vet Res 2025, 69, 121–129, doi: 10.2478/jvetres-2025-0003.
Open DOI Search in Google Scholar Back to article
Tolson K.P., Chappell P.E.: The changes they are a-timed: metabolism, endogenous clocks, and the timing of puberty. Front Endocrinol 2012, 3, 45, doi: 10.3389/fendo.2012.00045.
Open DOI Search in Google Scholar Back to article
Topaloglu A.K., Reimann F., Guclu M., Yalin A.S., Kotan L.D., Porter K.M., Serin A., Mungan N.O., Cook J.R., Ozbek M.N., Imamoglu S., Akalin N.S., Yuksel B., O’Rahilly S., Semple R.K.: TAC3 and TACR3 mutations in familial hypogonadotropic hypogonadism reveal a key role for neurokinin B in the central control of reproduction. Nat Genet 2009, 41, 354–358, doi: 10.1038/ng.306.
Open DOI Search in Google Scholar Back to article
Uenoyama Y., Nagae M., Tsuchida H., Inoue N., Tsukamura H.: Role of KNDy neurons expressing kisspeptin, neurokinin B, and dynorphin A as a GnRH pulse generator controlling mammalian reproduction. Front Endocrinol 2021, 12, 724632, doi: 10.3389/fendo.2021.724632.
Open DOI Search in Google Scholar Back to article
Wang J., Sun L., Zhang T., Zhou H., Lou Y.: Effect of peripheral administration of kisspeptin-10 on dynamic LH secretion in prepubertal ewes. Asian Australas J Anim Sci 2012, 25, 785–788, doi: 10.5713/ajas.2011.11390.
Open DOI Search in Google Scholar Back to article
Weems P.W., Coolen L.M., Hileman S.M., Hardy S., McCosh R.B., Goodman R.L., Lehman M.N.: Evidence that dynorphin acts upon KNDy and GnRH neurons during GnRH pulse termination in the ewe. Endocrinology 2018, 159, 3187–3199, doi: 10.1210/en.2018-00435.
Open DOI Search in Google Scholar Back to article
Weems P.W., Witty C.F., Amstalden M., Coolen L.M., Goodman R.L., Lehman M.N.: κ-opioid receptor is colocalized in GnRH and KNDy cells in the female ovine and rat brain. Endocrinology 2016, 157, 2367–2379, doi: 10.1210/en.2015-1763.
Open DOI Search in Google Scholar Back to article
Woller M., Tessmer S., Neff D., Nguema A.A., Roo B.V., Waechter-Brulla D.: Leptin stimulates gonadotropin releasing hormone release from cultured intact hemihypothalami and enzymatically dispersed neurons. Exp Biol Med 2001, 226, 591–596, doi: 10.1177/153537020122600613.
Open DOI Search in Google Scholar Back to article
Yang Y., Xiong X.-Y., Yang L., Xie L., Huang H.: Testing of kisspeptin levels in girls with idiopathic central precocious puberty and its significance. Exp Ther Med 2015, 9, 2369–2373, doi: 10.3892/etm.2015.2398.
Open DOI Search in Google Scholar Back to article