Contents The proliferation and differentiation ability of testicular Sertoli cells directly affects spermatogenesis and male reproductive development. WNT proteins are involved in the regulation of cell proliferation, differentiation and spermatogenesis. Therefore, to study whether lncRNAs, which regulate the expression of WNT proteins during cell proliferation and differentiation, are worthwhile. In this study, testicular tissue from the Dazu black goat (Capra, goat, Chongqing, China) at neonatal time (less than 7 days old), early puberty time (45 days old) and sexual maturity time (90 days old) at three ages was subjected to high‐throughput sequencing to predict testicular growth and development associated with WNT lncRNA. The final screening of lncWNT3‐IT may be targeted to regulate the expression of WNT3. At the same time, the expression of WNT3 was verified by lncWNT3‐IT by paraffin sectioning, fluorescence in situ hybridization, interference, overexpression, cytotoxicity assay, Western blotting and qPCR. The following results were obtained: lncWNT3‐IT was expressed in the testicular Sertoli cells and played a role in the Sertoli cell cytoplasm. Fluorescence in situ hybridization localization analysis showed that lncWNT3‐IT positively regulated the expression of WNT3, and through cell viability and cell proliferation experiments, it was found that the expression of lncWNT3‐IT assisted in Sertoli cell proliferation. In summary, lncWNT3‐IT can influence the proliferation of Sertoli cells by positively regulating the expression of WNT3.

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