{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:30:41Z","timestamp":1791419441656,"version":"4.3.3"},"reference-count":37,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2024,3,16]],"date-time":"2024-03-16T00:00:00Z","timestamp":1710547200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Italian Ministry of University and Research","award":["20204YRYS5"],"award-info":[{"award-number":["20204YRYS5"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Mitochondria-Associated Endoplasmic Reticulum Membranes (MAMs) mediate the communication between the Endoplasmic Reticulum (ER) and the mitochondria, playing a fundamental role in steroidogenesis. This study aimed to understand how D-aspartate (D-Asp), a well-known stimulator of testosterone biosynthesis and spermatogenesis, affects the mechanism of steroidogenesis in rat testes. Our results suggested that D-Asp exerts this function through MAMs, affecting lipid trafficking, calcium signaling, ER stress, and mitochondrial dynamics. After 15 days of oral administration of D-Asp to rats, there was an increase in both antioxidant enzymes (SOD and Catalase) and in the protein expression levels of ATAD3A, FACL4, and SOAT1, which are markers of lipid transfer, as well as VDAC and GRP75, which are markers of calcium signaling. Additionally, there was a decrease in protein expression levels of GRP78, a marker of aging that counteracts ER stress. The effects of D-Asp on mitochondrial dynamics strongly suggested its active role as well. It induced the expression levels of proteins involved in fusion (MFN1, MFN2, and OPA1) and in biogenesis (NRF1 and TFAM), as well as in mitochondrial mass (TOMM20), and decreased the expression level of DRP1, a crucial mitochondrial fission marker. These findings suggested D-Asp involvement in the functional improvement of mitochondria during steroidogenesis. Immunofluorescent signals of ATAD3A, MFN1\/2, TFAM, and TOMM20 confirmed their localization in Leydig cells showing an intensity upgrade in D-Asp-treated rat testes. Taken together, our results demonstrate the involvement of D-Asp in the steroidogenesis of rat testes, acting at multiple stages of both MAMs and mitochondrial dynamics, opening new opportunities for future investigation in other steroidogenic tissues.<\/jats:p>","DOI":"10.3390\/cells13060523","type":"journal-article","created":{"date-parts":[[2024,3,18]],"date-time":"2024-03-18T06:58:39Z","timestamp":1710745119000},"page":"523","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":31,"title":["Steroidogenesis Upregulation through Mitochondria-Associated Endoplasmic Reticulum Membranes and Mitochondrial Dynamics in Rat Testes: The Role of D-Aspartate"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0757-8430","authenticated-orcid":false,"given":"Debora","family":"Latino","sequence":"first","affiliation":[{"name":"Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania \u2018Luigi Vanvitelli\u2019, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7357-5353","authenticated-orcid":false,"given":"Massimo","family":"Venditti","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine, Section Human Physiology and Integrated Biological Functions, University of Campania \u2018Luigi Vanvitelli\u2019, 80138 Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Falvo","sequence":"additional","affiliation":[{"name":"Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania \u2018Luigi Vanvitelli\u2019, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giulia","family":"Grillo","sequence":"additional","affiliation":[{"name":"Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania \u2018Luigi Vanvitelli\u2019, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandra","family":"Santillo","sequence":"additional","affiliation":[{"name":"Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania \u2018Luigi Vanvitelli\u2019, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4025-8765","authenticated-orcid":false,"given":"Imed","family":"Messaoudi","sequence":"additional","affiliation":[{"name":"LR11ES41: G\u00e9netique, Biodiversit\u00e9 et Valorisation des Bioressources, Institut Sup\u00e9rieur de Biotechnologie, Universit\u00e9 de Monastir, Monastir 5000, Tunisia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariem","family":"Ben Rhouma","sequence":"additional","affiliation":[{"name":"LR11ES41: G\u00e9netique, Biodiversit\u00e9 et Valorisation des Bioressources, Institut Sup\u00e9rieur de Biotechnologie, Universit\u00e9 de Monastir, Monastir 5000, Tunisia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1007-7840","authenticated-orcid":false,"given":"Sergio","family":"Minucci","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine, Section Human Physiology and Integrated Biological Functions, University of Campania \u2018Luigi Vanvitelli\u2019, 80138 Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriella","family":"Chieffi Baccari","sequence":"additional","affiliation":[{"name":"Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania \u2018Luigi Vanvitelli\u2019, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4673-6729","authenticated-orcid":false,"given":"Maria Maddalena","family":"Di Fiore","sequence":"additional","affiliation":[{"name":"Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania \u2018Luigi Vanvitelli\u2019, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,3,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"102343","DOI":"10.1016\/j.ceca.2020.102343","article-title":"Mitochondria Associated Membranes (MAMs): Architecture and physiopathological role","volume":"94","author":"Barazzuol","year":"2021","journal-title":"Cell Calcium"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1016\/j.mce.2016.08.050","article-title":"The ER-mitochondria couple: In life and death from steroidogenesis to tumorigenesis","volume":"441","author":"Lalli","year":"2017","journal-title":"Mol. 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