Unknown,Transcriptomics,Genomics,Proteomics

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3'-UTR-seq of Mus musculus female and male hypothalamus across the pubertal transition

ABSTRACT: The hypothalamus is a functionally and cellularly complex tissue controlling many developmental processes, including puberty. While key hormonal aspects of puberty regulation in the hypothalamus are well established, understanding the genes, cell-types, and epigenetic mechanisms underlying and regulating puberty is limited. Here, we performed 3’-UTR-seq on the hypothalamus from both sexes of C57BL/6J mice at 5 ages spanning pubertal transition (postnatal days 12, 22, 27, 32, 37) (4-5 replicates per sex at each age) to examine genome-wide age- and sex-biased trends in gene expression in a cell-type aware manner. Sample collection, RNA extraction, and sequencing was completed using the same protocols and on the same mice as PMID: 3622112 (E-MTAB-9459). QuantSeq 3’mRNA-seq libraries were constructed from total RNA using an automated method with Agilent NGS Workstation. The resulting single-end libraries were sequenced at SickKids TCAG core on the Illumina v4 flow cell with SR50 bp cycles extended to 68 bp. A customized pipeline was developed and used for the analysis of reads obtained (PMID: 36221127 for details). Processed reads were mapped to mouse genome (mm10).

INSTRUMENT(S): Illumina HiSeq 2500, Agilent NGS Workstation

ORGANISM(S): Mus musculus

SUBMITTER: Dustin Sokolowski

PROVIDER: E-MTAB-12340 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Puberty is a complex physiological event measured by various indicator traits in genetic improvement programs of beef cattle; thus, developing a more complete understanding of the genes and regulatory pathways and networks involved in puberty will provide knowledge to help improve genetic selection strategies. Herein, we characterized the transcriptome of five reproductive tissues (i.e. hypothalamus, pituitary gland, ovary, uterus, and endometrium) as well as tissues known to be relevant to growth and metabolism needed for cattle to achieve puberty (i.e., longissimus dorsi muscle, fat, and liver). These tissues were collected from pre (PRE)- and post (POST)-pubertal Brangus (3/8 Brahman; Bos indicus x 5/8 Angus; Bos taurus) heifers derived from a population of cattle used to identify QTL associated with fertility traits. In order to exploit the power of complementary omics analyses, PRE and POST puberty co-expression gene networks were constructed by combining the results from RNA-Seq, GWAS, and bovine transcription factors. RNA-Seq of 8 tissues among PRE and POST Brangus heifers revealed 1515 differentiallyexpressed and 943 tissue-specific genes within the 17,832 genes confirmed by metrics of RNA-Seq analysis. Combining the results from RNA-Seq and GWAS indentified a total of 25 QTL associated to heifer fertility. The hypothalamus experienced the most notable up-regulation of genes via puberty. Complementary omics procedures revealed 2,450 co-expressed genes across the 8 tissues relative to puberty. The PRE network had 372,861 connections whereas the POST network had 328,357 connections. A sub-network from this process revealed key transcriptional regulators (i.e., PITX2, FOXA1, TSG1D1, DACH2, LHX4, PROP1 and SIX6). Results from multiples sources of omics data will facilitate the design of breeding strategies to improve fertility in Bos indicus-influenced composite cattle. Sixty-one samples from PRE and POST pubertal composite beef heifers were analyzed with RNA-Seq. The transcriptome of five reproductive tissues (i.e. hypothalamus, pituitary gland, ovary, uterus, and endometrium) as well as tissues known to be relevant to metabolism andbody morphometrics needed for cattle to achieve puberty (i.e.,) was characterized. These tissues were collected from pre (PRE)- and post (POST)-pubertal Brangus (3/8 Brahman x 5/8 Angus) heifers derived from a population of cattle used to identify QTL associated with fertility. Total RNA was purified using a Trizol protocol (Invitrogen, Carlsbad, CA). Sequencing libraries were made using TruSeq RNA Sample Preparation kit of Illumina (San Diego, CA).

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3'-UTR-seq of Mus musculus female and male hypothalamus across the pubertal transition

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