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Arginine is a conditionally essential amino acid involved in metabolic regulation; however, its protective effects on the male reproductive system under heat stress remain unclear. Using a heat-stressed Rongchang boar model, this study aimed to explore the possible mechanism of dietary arginine supplementation on sperm quality and testicular function. Twenty-four Rongchang boars were assigned to three treatments: thermoneutral control (CON), heat-stressed control (HS), or heat-stressed treatment supplemented with 0.8% dietary arginine (HA). Results showed that heat stress reduced feed intake, body weight, sperm viability, sperm concentration, and sperm motion parameters (p &lt; 0.05), accompanied by testicular histological damage. However, arginine supplementation improved sperm viability and straight-line velocity (p &lt; 0.05), with partial preservation of testicular morphology. Furthermore, single-cell RNA sequencing revealed heat stress-associated changes in germ-cell composition and enrichment of autophagy-related pathways. Transmission electron microscopy and qRT-PCR showed endoplasmic reticulum swelling, autophagic vesicle accumulation, and increased expression of GABARAPL1, ULK1, BECN1, and CALM3 in heat-stressed testes (p &lt; 0.05); these alterations were attenuated by arginine supplementation, as indicated by reduced ULK1 and BECN1 expression (p &lt; 0.05). Metabolomic and targeted analyses indicated improved circulating arginine availability and reshaped testicular arginine-related amino acid\/polyamine metabolism after arginine supplementation (p &lt; 0.05). Overall, dietary arginine supplementation alleviated heat stress-induced testicular dysfunction, which was accompanied by alterations in arginine-related metabolic regulation and autophagy-associated cellular stress responses.<\/jats:p>","DOI":"10.3390\/ani16172682","type":"journal-article","created":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T08:02:27Z","timestamp":1787817747000},"page":"2682","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Dietary Arginine Supplementation Mitigates Heat Stress-Induced Testicular Dysfunction Through Arginine-Related Metabolic Remodeling and Autophagy-Associated Stress Adaptation in a Rongchang Boar Model"],"prefix":"10.3390","volume":"16","author":[{"given":"Xiao","family":"Lin","sequence":"first","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kun","family":"Wu","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingchang","family":"Ren","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8348-4199","authenticated-orcid":false,"given":"Yong","family":"Zhuo","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6258-2729","authenticated-orcid":false,"given":"Bin","family":"Feng","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lianqiang","family":"Che","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengfeng","family":"Fang","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"},{"name":"College of Food Science, Sichuan Agricultural University, Ya\u2019an 625014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4415-8542","authenticated-orcid":false,"given":"Shengyu","family":"Xu","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lun","family":"Hua","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2385-7528","authenticated-orcid":false,"given":"Guangmang","family":"Liu","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaojun","family":"Jiang","sequence":"additional","affiliation":[{"name":"Sichuan DaBeiNong Agriculture and Animal Husbandry Technology Co., Ltd., Chengdu 611430, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Li","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"De","family":"Wu","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Lin","sequence":"additional","affiliation":[{"name":"Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"\u021aogoe, D., and Minc\u0103, N.A. 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