{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:36:02Z","timestamp":1760146562251,"version":"build-2065373602"},"reference-count":54,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2024,11,18]],"date-time":"2024-11-18T00:00:00Z","timestamp":1731888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>The grey squirrel, an invasive species, threatens the Eurasian red squirrel\u2019s conservation, particularly in Umbria, Italy. Understanding its reproductive biology is essential to limiting its reproductive success. This study investigates the NGF system and its receptors (NTRK1 and p75NTR) in the testes of male grey squirrels, following prior research on female reproductive biology. NGF plays a role in testicular morphogenesis and spermiogenesis in animals and humans. As part of the LIFE Project U-SAVEREDS, eighteen squirrels were captured and classified into three morphotypes (immature, pubertal, and active spermatogenesis). NGF and its receptors were analyzed using real-time PCR, western blotting, immunohistochemistry, and plasma levels measured via ELISA. NGF qPCR expression levels were significantly higher during puberty compared to the immature and spermatogenesis stages (p &lt; 0.01). Immunohistochemistry revealed NGF in Leydig cells, with stronger staining in pubertal and mature squirrels, while NTRK1 was found in Leydig cells in immature squirrels and germ cells in pubertal and mature ones. NGF receptors were observed in Sertoli cells in pubertal and mature squirrels. Plasma NGF levels showed a significant upregulation in pubertal squirrels (135.80 \u00b1 12 pg\/mL) compared to those in the immature (25.60 \u00b1 9.32 pg\/mL) and spermatogenesis stages (34.20 \u00b1 6.06 pg\/mL), with a p value &lt; 0.01. The co-localization of NGF and its receptors suggests that NGF, produced by Leydig cells, regulates testis development and reproductive success through autocrine or paracrine mechanisms, potentially involving an unidentified pathway.<\/jats:p>","DOI":"10.3390\/ani14223318","type":"journal-article","created":{"date-parts":[[2024,11,18]],"date-time":"2024-11-18T13:03:14Z","timestamp":1731934994000},"page":"3318","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["From Gene to Protein: Unraveling the Reproductive Blueprint of Male Grey Squirrels via Nerve Growth Factor (NGF) and Cognate Receptors"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8799-1200","authenticated-orcid":false,"given":"Francesca","family":"Mercati","sequence":"first","affiliation":[{"name":"Department of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3695-9275","authenticated-orcid":false,"given":"Gabriella","family":"Guelfi","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5073-6540","authenticated-orcid":false,"given":"Antonello","family":"Bufalari","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6451-3751","authenticated-orcid":false,"given":"Cecilia","family":"Dall\u2019Aglio","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0634-0386","authenticated-orcid":false,"given":"Chiara","family":"Suvieri","sequence":"additional","affiliation":[{"name":"Department of Medicine and Surgery, University of Perugia, Piazzale Settimio Gambuli, 1, 06129 Perugia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paolo","family":"Cocci","sequence":"additional","affiliation":[{"name":"School of Bioscience and Veterinary Medicine, University of Camerino, Via Gentile III Da Varano, 62032 Camerino, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesco Alessandro","family":"Palermo","sequence":"additional","affiliation":[{"name":"School of Bioscience and Veterinary Medicine, University of Camerino, Via Gentile III Da Varano, 62032 Camerino, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8871-550X","authenticated-orcid":false,"given":"Polina","family":"Anipchenko","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1040-6245","authenticated-orcid":false,"given":"Camilla","family":"Capaccia","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3993-6258","authenticated-orcid":false,"given":"Beniamino","family":"Cenci-Goga","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Massimo","family":"Zerani","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1727-4118","authenticated-orcid":false,"given":"Margherita","family":"Maranesi","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,11,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1284","DOI":"10.1152\/physrev.1980.60.4.1284","article-title":"Physiology of Nerve Growth Factor","volume":"60","author":"Thoenen","year":"1980","journal-title":"Physiol. 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