scRNA of preadipocytes and mature adipocytes from mice exposed to a mouse model of early life stress

Background: Adipose tissue homeostasis is a complex process influenced by many mechanisms that become dysregulated with obesity. We have previously shown that Maternal Separation and Early Weaning (MSEW), a mouse model of early life stress, exacerbates high-fat diet (HF)-induced fat deposition in female mice, but not male mice. In addition, we have shown that this exacerbated adiposity is due to an increase in visceral adipose tissue, and fat depot known to be linked with cardio-metabolic dysfunction. Purpose: The aim of this study was to determine differential gene expression patterns in gonadal white adipose tissue (gWAT), a visceral fat, in female mice exposed to MSEW compared to controls. Adipose tissue is composed of many different stromal cell types including preadipocytes, mature adipocytes, and immune cells. Preadipocytes are the precursor stem cells that differentiate into mature adipocytes that can uptake and store lipids and have a more predominte endocrine function. Methods: Control and MSEW mice were weaned onto either a low-fat diet (LF) or a high-fat diet (HF) for 20 weeks. Fresh gonadal white adipose tissue was harvested after euthanasia and subjected to collagenase digestion. Tissue was centrifuged and filtered to separate the mature fraction from the stromal vascular fraction. Magnetic Activated Cell Sorting (MACs) was used to further isolate adipocytes from other cell types. Pre-and mature adipocytes were collected and saved in Purezol at -80oC for later RNA extraction using the Promega ReliaPrep RNA miniprep systems. Total RNA was sent to Novogene for scRNAseq analysis. Overall design: Groups (6 total): C_HF_P; MS_HF_P; C_HF_A; MS_HF_A; C_LF_P; MS_LF_P. C=control; MS=maternal separation and early weaning; HF=high-fat diet; LF= low-fat diet; P=preadipocyte; A=mature adipocyte.