Impact of PAMM gene knockout on gene expression in white and brown adipose tissues of mice
AIM:The protein peroxiredoxin-like 2 activated in M-CSF stimulated monocytes(PAMM)pos-sesses anti-inflammatory and antioxidant attributes and exhibits high expression in adipose tissue.To contribute new per-spectives to the functional research on PAMM,the objective of this study is to examine the influence of PAMM on the mor-phology and gene expression profiles of white and brown adipose tissues by using mice with PAMM gene knockout.METHODS:Homozygous knockout(KO)mice for the PAMM gene were modeled using techniques such as CRISPR-Cas9,with wild-type(WT)mice serving as controls.Subsequently,subcutaneous inguinal adipose tissue(IAT)and dor-sal brown adipose tissue(BAT)were procured,and tissue morphology was assessed through hematoxylin and eosin(HE)staining.Gene expression patterns were comprehensively analyzed using RNA sequencing(RNA-seq)and further con-firmed with quantitative reverse transcription-polymerase chain reaction(RT-qPCR).RESULTS:Genomic PCR and the product sequencing were used to validate gene knockout in mice.Notably,in PAMM knockout mice,there were no signifi-cant changes in the morphology of subcutaneous IAT and dorsal BAT.However,enrichment of gene sets associated with E2F targets and unfolded protein response was evident in either IAT or BAT in KO mice.A comparative analysis between WT and KO tissues revealed 33 genes exhibiting consistent changes in both IAT and BAT.Genes manifesting changes ex-clusively in IAT were predominantly located in extracellular and plasma membrane domains,contributing to immune and chemotactic responses,as well as calcium signaling pathways.Conversely,genes displaying changes solely in BAT were primarily distributed in extracellular matrix and nucleus regions,involved in glucose homeostasis and biological rhythms.RT-qPCR analysis of calcium signaling and biological rhythm-related genes corroborated trends observed in RNA-seq.The influence of PAMM knockout on differentially expressed genes between IAT and BAT was also evaluated in this study.While the majority of differential genes and pathways of BAT/IAT demonstrated consistency between WT and KO mice,noteworthy expression changes were observed in specific genes or pathways.CONCLUSION:PAMM knockout demon-strates no notable influence on the morphological characteristics of IAT and BAT.However,it exerts a significant effect on gene expression profiles and associated signaling pathways in both tissues,further indicating a potential impact on the crosstalk between IAT and BAT.
white adipose tissuebrown adipose tissuePAMM genegene knockout
国家科技期刊平台
NSTL
万方数据
维普
