Comprehensive transcriptome analysis of hypothalamus reveals genes associated with disorders of sex development in pigs

Abstract

BackgroundDisorders of sex development (DSD) is a chronic autoimmune disease characterized by systemic in ammation, pathological osteogenesis, and endocrine abnormality. However, its genetic etiology remains mostly unknown. In addition, little research focuses on the regulation mechanism from the view of transcriptomics in the hypothalamic-pituitary-gonadal axis (HPGA). The hypothalamus is the integrated center of the HPGA mediating neural, hormonal, and environmental stimulus to sex development. MethodsThree XX-DSD (SRY-negative) pig (DSD) and three NF pigs ( ve months old, 40 kg ± 5 kg) were selected by external genitalia observation and sex determining region Y gene (SRY) detection. The hypothalamus were sampled for RNA isolation, and the mRNA, lncRNA and miRNA expression pro les were analyzed by sequencing. ResultsA total of 1,258 lncRNAs, 1,086 mRNAs, and 61 microRNAs were found to differentially express in XX-DSD pigs compared with normal female pigs. Many genes in hormone biosynthesis and secretion pathway are signi cantly up-regulated, and the upregulation of GNRH1, KISS1 and AVP may be the candidate genes leading the abnormal secretion of GnRH. Next, we predicted the lncRNA-miRNA-mRNA co-expression triplets and constructed three competing endogenous RNA (ceRNA) potentially associated with DSD. Functional enrichment suggested TCONS_00340886, TCONS_00000204 and miR-181a were related to GnRH secretion. ConclusionsOur research revealed the rst transcriptomic pro le in the hypothalamus of XX-DSD pigs and provided new insight in coding and non-coding RNAs that may be associated with DSD in pigs.

Whole genome sequencing identifies a cryptic SOX9 regulatory element duplication underlying a case of 46,XX ovotesticular difference of sexual development

“…Although no genes were present within this region, the duplication involved a known regulatory element named RevSex that modulates SOX9 expression during development. Similar duplications encompassing the RevSex region have been previously reported in 46,XX OT‐DSD individuals (Figure 3 ) (Benko et al, 2011 ; Cox et al, 2011 ; Croft, Ohnesorg, Hewitt, Bowles, Quinn, Tan, Corbin, Pelosi, Van Den Bergen, & Sreenivasan, 2018 ; Croft, Ohnesorg, Hewitt, Bowles, Quinn, Tan, Corbin, Pelosi, van den Bergen, Sreenivasan, Knarston, et al, 2018 ; Croft, Ohnesorg, & Sinclair, 2018 ; Hyon et al, 2015 ; Kim et al, 2015 ; Ohnesorg et al, 2017 ; Tan et al, 2020 ; Xiao et al, 2013a ; Xiao et al, 2013b ). A re‐analysis of the proband's original chromosomal SNP microarray data was requested.…”

“…Although no genes were present within this region, the duplication involved a known regulatory element named RevSex that modulates SOX9 expression during development. Similar duplications encompassing the RevSex region have been previously reported in 46,XX OT-DSD individuals (Figure 3) (Benko et al, 2011;Cox et al, 2011;Croft, Ohnesorg, Hewitt, Bowles, Quinn, Tan, Corbin, Pelosi, van den Bergen, Sreenivasan, Knarston, et al, 2018;Hyon et al, 2015;Kim et al, 2015;Ohnesorg et al, 2017;Tan et al, 2020;Xiao et al, 2013a;Xiao et al, 2013b). A re-analysis of the F I G U R E 1 A 281 kilobase (kb) pathogenic copy number gain was detected at chromosome band 17q24.3, located 411 to 692 kb upstream of the SOX9 gene (arrow).…”

“…SRY translocation from Y to X chromosome and aberrations of downstream SOX9 are the most common causes (Ono & Harley, 2013 ). Emerging data also showed pathogenic variants of key genes involved in physiological sexual development such as NR5A , WNT4 , SOX3 , SOX5 , RSPO1 , or WT1 can also lead to 46,XX OT‐DSD via either promoting testiculogenesis or inhibiting ovogenesis (Domenice et al, 2016 ; Jurayyan & Nasir, 2011 ; Tan et al, 2020 ).…”

“…Previous reported patients with a RevSex duplication were detected by comparative genomic hybridization microarray (Croft, Ohnesorg, Hewitt, Bowles, Quinn, Tan, Corbin, Pelosi, van den Bergen, Sreenivasan, Knarston, et al, 2018 ; Kim et al, 2015 ; Tan et al, 2020 ; Xiao et al, 2013a ). Here we report the first patient with a RevSex duplication detected via whole genome sequencing.…”

Analyses of Molecular Characteristics and Enzymatic Activities of Ovine HSD17B3

“…Mutations of HSD17B3 genes cause a disorder of sexual development (DSD) and undermasculinizaion as a result of low T production and an abnormal A4/T ratio in multiple species, such as human, dog, and mouse [6,[8][9][10][11]. In livestock, DSD represents one of the major causes of reduced productivity [12][13][14][15]. Therefore, an understanding of sex differentiation-related genes, including HSD17B3, is necessary to explore new mutations/variants and to develop diagnostic tools to identify them.…”