AUTHOR=Zhao Le , Yang Haili , Li Xingchun , Zhou Yumei , Liu Taolu , Zhao Yongju 

TITLE=Transcriptome-based selection and validation of optimal reference genes in perirenal adipose developing of goat (Capra hircus)

JOURNAL=Frontiers in Veterinary Science

VOLUME=9

YEAR=2022

URL=https://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2022.1055866

DOI=10.3389/fvets.2022.1055866

ISSN=2297-1769

ABSTRACT=<p>Brown adipose tissue (BAT) is mainly present in young mammals and is important for maintaining body temperature in neonatal mammals because of its ability to produce non-shivering thermogenesis. There is usually a large amount of BAT around the kidneys of newborn kids, but the BAT gradually “whiting” after birth. Screening and validating appropriate reference genes is a prerequisite for further studying the mechanism of goat brown adipose tissue “whiting” during the early stages. In this study, the expression stability of 17 candidate reference genes: 12 <italic>COPS8, SAP18, IGF2R, PARL, SNRNP200, ACTG1, CLTA, GANAB, GABARAP, PCBP2, CTSB</italic>, and <italic>CD151</italic>) selected based on previous transcriptome data as new candidate reference genes, 3 (<italic>PFDN5, CTNNB1</italic>, and <italic>EIF3M</italic>) recommended in previous studies, and 2 traditional reference genes (<italic>ACTB</italic> and <italic>GAPDH</italic>) was evaluated. Real-time quantitative PCR (RT-qPCR) technology was used to detect the expression level of candidate reference genes during goat BAT “whiting”. Four algorithms: Normfinder, geNorm, ΔCt method, and BestKeeper, and two comprehensive algorithms: ComprFinder and RefFinder, were used to analyze the stability of each candidate reference genes. <italic>GABARAP, CLTA, GAPDH</italic>, and <italic>ACTB</italic> were identified as the most stable reference genes, while <italic>CTNNB1, CTSB</italic>, and <italic>EIF3M</italic> were the least stable. Moreover, two randomly selected target genes <italic>IDH2</italic> and <italic>RBP4</italic>, were effectively normalized using the selected most stable reference genes. These findings collectively suggest that <italic>GABARAP, CLTA, GAPDH</italic>, and <italic>ACTB</italic> are relatively stable reference genes that can potentially be used for the development of perirenal fat in goats.</p>