Anti-Mullerian hormone, testosterone, and insulin-like peptide 3 as biomarkers of Sertoli and Leydig cell function during deslorelin-induced testicular downregulation in the dog
Anti-Mullerian hormone, testosterone, and insulin-like peptide 3 as biomarkers of Sertoli and Leydig cell function during deslorelin-induced testicular downregulation in the dog
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DOI：
10.1016/j.theriogenology.2021.08.017
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发表时间：
2021-09-14
期刊：
影响因子：
2.8
通讯作者：
Muller, Linda
中科院分区：
文献类型：
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作者：
Balogh, Orsolya;Somoskoi, Bence;Muller, Linda
抗苗勒管激素（AMH）和胰岛素样肽3（INSL 3）在男性不育中的作用尚未完全了解。我们使用下调的睾丸作为促性腺激素依赖性不育的模型。在植入4.7mg地洛瑞林（Suprelorin（R），Virbac）（DES组）（第0天）的5只成年雄性比格犬中研究血清睾酮和AMH浓度。在地洛瑞林处理后112天（16周），通过qPCR和免疫组织化学评价LH受体（LHR）和雄激素受体（AR）、AMH、2型AMH受体（AMHR 2）、INSL 3及其受体（RXFP 2）的睾丸表达，并与未处理的成年（CON，n = 6）和青春期前（PRE，n = 8）犬进行比较。在研究第14天（4只犬）或第21天（1只犬）发生无精症时，血清睾酮浓度显著降低，并在第105天（第15周）达到基线水平。相比之下，血清AMH仅在无精症发作后开始增加，并在个体犬中在第49-105天达到测定的最大可检测浓度。DES组睾丸LHR基因表达低于CON组和PRE组（P < 0.0001），而AR基因表达与CON组相似，显著高于PRE组（P < 0.0001）。DES组睾丸AMH表达水平居中，CON组最低，PRE组最高（P < 0.006）。AMHR 2基因表达在组间相似。AMH蛋白仅在支持细胞中检测到，而AMHR 2免疫反应主要在间质细胞中检测到，这似乎在DES中增加。INSL 3和RXFP 2基因表达显着下调DES睾丸沿着明显弱的间质细胞免疫信号相比CON。总之，地洛瑞林治疗引起睾丸LH不敏感，而不影响雄激素敏感性，去分化的支持细胞和间质细胞。在DES中，AMH-AMHR 2反馈环的上调和INSL 3-RXFP 2前馈环的下调是旁分泌-自分泌机制，其可以独立于促性腺激素额外调节睾酮产生。我们的研究结果支持AMH和INSL 3作为睾丸功能的独特生物标志物和旁分泌-自分泌调节因子参与支持细胞和间质细胞之间的密切相互作用。(c)2021作者爱思唯尔公司出版这是一个在CC BY-NC-ND许可证下的开放获取文章（http：//creativecommons.org/licenses/by-nc-nd/4.0/）。
The role of anti-Mullerian hormone (AMH) and insulin-like peptide 3 (INSL3) in male infertility is not fully understood. We used the downregulated testis as a model of gonadotropin-dependent infertility. Serum testosterone and AMH concentrations were studied in five adult male Beagles implanted (day 0) with 4.7 mg deslorelin (Suprelorin (R), Virbac) (DES group). Testicular expression of LH receptor (LHR) and androgen receptor (AR), AMH, type 2 AMH receptor (AMHR2), INSL3 and its receptor (RXFP2) was evaluated 112 days (16 weeks) after deslorelin treatment by qPCR and immunohistochemistry, and compared to untreated adult (CON, n = 6) and prepubertal (PRE, n = 8) dogs. Serum testosterone concentration decreased significantly by the onset of aspermia on study day 14 (four dogs) or day 21 (one dog), and was baseline on day 105 (week 15). In contrast, serum AMH started to increase only after the onset of aspermia and reached the maximum detectable concentration of the assay by day 49-105 in individual dogs. Testicular LHR gene expression in DES was lower than in CON and PRE (P < 0.0001), while AR gene expression in DES was similar to CON and significantly higher than PRE (P < 0.0001). Testicular AMH expression in DES was intermediate compared to the lowest mRNA levels found in CON and the highest in PRE (P < 0.006). AMHR2 gene expression was similar between groups. AMH protein was detected in Sertoli cells only, while AMHR2 immunoreactivity was principally detected in Leydig cells which appeared to be increased in DES. INSL3 and RXFP2 gene expression was significantly downregulated in the DES testis along with noticeably weak Leydig cell immunosignals compared to CON. In conclusion, deslorelin treatment caused testicular LH insensitivity without affecting androgen sensitivity, and de-differentiation of Sertoli and Leydig cells. In DES, upregulation of the AMH-AMHR2 feed-back loop and downregulation of the INSL3-RXFP2 feed-forward loop are paracrine-autocrine mechanisms that may additionally regulate testosterone production independent of gonadotropins. Our results support AMH and INSL3 as unique biomarkers and paracrine-autocrine regulators of testis function involved in the intimate interplay between Sertoli and Leydig cells. (c) 2021 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).