Department of Ecology, Tibetan Centre for Ecology and Conservation at WHU-TU, Hubei Key Laboratory of Cell Homeostasis, College of Life Science, Wuhan University, Wuhan 430072, China; Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, School of Life Science and Engineering, Foshan University, Foshan 528225, China.
College of Basic Medicine, Zunyi Medical University, Zunyi 563006, China.
J Steroid Biochem Mol Biol. 2021 Jun;210:105875. doi: 10.1016/j.jsbmb.2021.105875. Epub 2021 Mar 18.
XX sex reversal, also called XX disorders of sex development (XX-DSD), is a condition affecting the development of the gonads or genitalia, and is relatively common in pigs. However, its genetic etiology and transcriptional regulation mechanism in the hypothalamic-pituitary-gonadal axis (HPGA) remain mostly unknown. XX-DSD (SRY-negative) pigs and normal sows were selected by external genitalia observation. The hypothalamus, which is the integrated center of the HPGA was sampled for whole-transcriptome RNA-seq. The role of DEmiRNA was validated by its overexpression and knockdown in vitro. A total of 1,258 lncRNAs, 1,086 mRNAs, and 61 microRNAs differentially expressed in XX-DSD pigs compared with normal female pigs. Genes in the hormone biosynthesis and secretion pathway significantly up-regulated, and the up-regulation of GNRH1, KISS1 and AVP may associate with the abnormal secretion of GnRH. We also predicted the lncRNA-miRNA-mRNA co-expression triplets and constructed three competing endogenous RNA (ceRNA) potentially associated with XX-DSD. Functional enrichment studies suggested that TCONS_00340886, TCONS_00000204 and miR-181a related to GnRH secretion. Further, miR-181a inhibitor up-regulated GNRH1, PAK6, and CAMK4 in the GT1-7 cells. Conversely, transfection of miR-181a mimics obtained the opposite trends. The expression levels of FSHR, LHR, ESR1 and ESR2 were significantly higher in XX-DSD gondas than those in normal sows. Taken together, we proposed that the balance of endocrine had broken in XX-DSD pigs. The current study is the first to examine the transcriptomic profile in the hypothalamus of XX-DSD pigs. It provides new insight into coding and non-coding RNAs that may be associated with DSD in pigs.
XX 性反转，也称为 XX 性发育障碍（XX-DSD），是一种影响性腺或生殖器发育的疾病，在猪中较为常见。然而，其在下丘脑-垂体-性腺轴（HPGA）中的遗传病因和转录调控机制在很大程度上仍不清楚。我们通过外部生殖器观察选择了 XX-DSD（SRY 阴性）猪和正常母猪，然后对下丘脑（HPGA 的整合中心）进行了全转录组 RNA-seq 采样。通过体外过表达和敲低验证了差异表达 miRNA（DEmiRNA）的作用。与正常雌性猪相比，XX-DSD 猪的 1258 个长非编码 RNA（lncRNA）、1086 个 mRNA 和 61 个 microRNA 表达差异。激素生物合成和分泌途径中的基因显著上调，GNRH1、KISS1 和 AVP 的上调可能与 GnRH 的异常分泌有关。我们还预测了 lncRNA-miRNA-mRNA 共表达三元组，并构建了三个与 XX-DSD 相关的潜在竞争性内源 RNA（ceRNA）。功能富集研究表明，TCONS_00340886、TCONS_00000204 和与 GnRH 分泌相关的 miR-181a。此外，miR-181a 抑制剂在 GT1-7 细胞中上调了 GNRH1、PAK6 和 CAMK4。相反，转染 miR-181a 模拟物则获得了相反的趋势。XX-DSD 性腺中 FSHR、LHR、ESR1 和 ESR2 的表达水平明显高于正常母猪。总之，我们提出 XX-DSD 猪的内分泌平衡已经打破。本研究首次检测了 XX-DSD 猪下丘脑的转录组谱，为编码和非编码 RNA 与猪 DSD 相关提供了新的见解。
