Identification of differentially expressed genes associated with precocious puberty by suppression subtractive hybridization in goat pituitary tissues

Yufang Liu,Guiling Cao,Yujing Xie,Mingxing Chu

doi:10.1080/10495398.2021.1990940

Abstract

The aim of this study was to identify genes related to precocious puberty expressed in the pituitary of goats at different growth stages by suppression subtractive hybridization (SSH). The pituitary glands from Jining Gray (JG) goats (early puberty) and Liaoning Cashmere (LC) goats (late puberty) at 30, 90, and 180 days were used in this study. To identify differentially expressed genes (DEGs) in the pituitary glands, mRNA was extracted from these tissues, and SSH libraries were constructed and divided into the following groups: juvenile group (30-JG vs. 30-LC, API), puberty group (90-JG vs. 180-LC, BPI), and control group (90-JG vs. 90-LC, EPI). A total of 60, 49, and 58 DEGs were annotated by 222 Gene Ontology (GO) terms and 75 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Most of the DEGs were significantly enriched in GO terms related to ‘structural constituent of ribosome’, ‘translation’ and ‘GTP binding’, and numerous DEGs were also significantly enriched in KEGG terms related to the Jak-STAT signaling and oocyte meiosis pathways. Candidate genes associated with precocious puberty and sexual development were screened from the SSH libraries. These genes were analyzed to determine if they were expressed in the pituitary tissues of the goats at different growth stages and to identify genes that may influence the hypothalamic-pituitary-gonadal (HPG) axis. In this study, we found precocious puberty-related genes (such as PRLP0, EIF5A, and YWHAH) that may be interesting from an evolutionary perspective and that could be investigated for use in future goat breeding programs. Our results provide a valuable dataset that will facilitate further research into the reproductive biology of goats.

Highlights

Our study aimed to identify genes related to precocious puberty expressed in the pituitary of different growth stages goats using suppression subtractive hybridization (SSH) screening
222 Gene Ontology (GO) terms and 75 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were matched to those genes
Candidate genes to be involved in precocious puberty and sexual development were retrieved from the SSH libraries

Summary

Methods

All experimental procedures mentioned in the present study were approved by the Science Research Department (in charge of animal welfare issue) of the Institute of Animal Sciences, Chinese Academy of Agricultural Sciences (IAS-CAAS) (Beijing, China). Ethical approval was provided by the animal ethics committee of IASCAAS (No IAS2019-63). The pituitary tissues were collected from each goat, sacrificed after anesthesia with 3% pentobarbital sodium salt injection (20mg/kg body weight) (Merck, Darmstadt, Germany), and preserved in RNA later RNA Stabilization reagent (Qiagen, Hilden, Germany) and kept at -20°C until RNA isolation. According to the instruction of Dynabeads mRNA DIRECTTM Kit (Invitrogen, Inc. USA), the poly A+ RNA was isolated from pituitary. After mixed in equal quantity from three goats in each group, the poly A+RNA concentrated using RNA clean & concentration-5 mRNA (Zymo Research, Orange, CA, USA) to a suitable concentration

Results

Discussion

Conclusion