title: Kisspeptin drives germ cell progression in the anuran amphibian Pelophylax esculentus: a study carried out in ex vivo testes.
abstract: Kisspeptin, via Gpr54 receptor, regulates puberty onset in most vertebrates. Thus, the direct involvement of kisspeptin activity in testis physiology was investigated in the anuran amphibian, Pelophylax esculentus. In this vertebrate gpr54 mRNA has been localized in both interstitial compartment and spermatogonia (SPG), whereas SPG proliferation requires the cooperation between estradiol and testicular Gonadotropin releasing hormone (Gnrh). In the pre-reproductive period, dose response curve to assess the effects of Kisspeptin-10 (Kp-10) was carried out in vitro (dose range: 10(-9)-10(-6)M; incubation times: 1 and 4h); proliferative activity and germ cell progression were evaluated by expression analysis of proliferating cell nuclear antigen (pcna), estrogen receptor beta (erβ), Gnrh system (gnrh1, gnrh2, gnrhr1, r2, r3) and by the count of empty, mitotic and meiotic tubules. All selected markers were up regulated at 4h Kp-10 incubation. Histological analysis also proved the increase of mitotic activity and the progression of spermatogenesis. Besides Kp-10 modulation of testicular Gnrh system, in vitro treatment with 17β-estradiol (10(-6)M) ± the antagonist ICI182-780 (10(-5)M) revealed gnrh2 and gnrhr3 estrogen dependent expression. In the reproductive period, testes were incubated for 1 and 4h with Kp-10 (10(-7)M) or Kp-10 (10(-7)M)+kisspeptin antagonist [Kp-234 (10(-6)M)]. Results obtained in the pre-reproductive period were confirmed and Kp-234 completely counteracted Kp-10 effects. In conclusion, Kp-10 modulated the expression of pcna, erβ, gnrhs and gnrhrs, inducing the progression of the spermatogenesis.
authors.affiliation: Dipartimento di Medicina Sperimentale Sezione "F. Bottazzi", Seconda Università di Napoli, Via Costantinopoli 16, 80138 Napoli, Italy. Electronic address: rosanna.chianese@unina2.it.
authors.forename: Rosanna
authors.initials: R
authors.lastname: Chianese
authors.affiliation: Dipartimento di Medicina Sperimentale Sezione "F. Bottazzi", Seconda Università di Napoli, Via Costantinopoli 16, 80138 Napoli, Italy. Electronic address: enza.ciaramella@unina2.it.
authors.forename: Vincenza
authors.initials: V
authors.lastname: Ciaramella
authors.affiliation: Dipartimento di Medicina Sperimentale Sezione "F. Bottazzi", Seconda Università di Napoli, Via Costantinopoli 16, 80138 Napoli, Italy. Electronic address: silvia.fasano@unina2.it.
authors.forename: Silvia
authors.initials: S
authors.lastname: Fasano
authors.affiliation: Dipartimento di Medicina Sperimentale Sezione "F. Bottazzi", Seconda Università di Napoli, Via Costantinopoli 16, 80138 Napoli, Italy. Electronic address: riccardo.pierantoni@unina2.it.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Pierantoni
authors.affiliation: Dipartimento di Scienze Motorie e del Benessere, Università di Napoli Parthenope, Via Medina 40, 80133 Napoli, Italy. Electronic address: rosaria.meccariello@uniparthenope.it.
authors.forename: Rosaria
authors.initials: R
authors.lastname: Meccariello
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D004958:Estradiol / Q000494:pharmacology; D047629:Estrogen Receptor beta / Q000378:metabolism; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism; D066298:In Vitro Techniques; D059648:Kisspeptins / Q000494:pharmacology*; D008297:Male; D008540:Meiosis / Q000187:drug effects; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011893:Rana esculenta / Q000235:genetics / Q000378:metabolism*; D011966:Receptors, LHRH / Q000378:metabolism; D012098:Reproduction / Q000187:drug effects; D013094:Spermatozoa / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013737:Testis / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
chemical_list: D047629:Estrogen Receptor beta; C106628:KISS1 protein, human; D059648:Kisspeptins; D018809:Proliferating Cell Nuclear Antigen; D012333:RNA, Messenger; D011966:Receptors, LHRH; D007987:Gonadotropin-Releasing Hormone; D004958:Estradiol
keywords: ERβ; Gnrh; Kisspeptin; Pcna; Spermatogonial proliferation