Knockout of Butyrophilin Subfamily 1 Member A1 () Alters Lipid Droplet Formation and Phospholipid Composition in Bovine Mammary Epithelial Cells

Overview

Journal J Anim Sci Biotechnol

Publisher Biomed Central

Date 2020 Jul 9

PMID 32637097

Citations 6

Authors

Liqiang Han

Menglu Zhang

Zhiyang Xing

Danielle N Coleman

Yusheng Liang

Juan J Loor

Guoyu Yang

Affiliations

Soon will be listed here.

Abstract

Background: Milk lipids originate from cytoplasmic lipid droplets (LD) that are synthesized and secreted from mammary epithelial cells by a unique membrane-envelopment process. Butyrophilin 1A1 (BTN1A1) is one of the membrane proteins that surrounds LD, but its role in bovine mammary lipid droplet synthesis and secretion is not well known.

Methods: The objective was to knockout BTN1A1 in bovine mammary epithelial cells (BMEC) via the CRISPR/Cas9 system and evaluate LD formation, abundance of lipogenic enzymes, and content of cell membrane phospholipid (PL) species. Average LD diameter was determined via Oil Red O staining, and profiling of cell membrane phospholipid species via liquid chromatography-tandem mass spectrometry (LC-MS/MS).

Results: Lentivirus-mediated infection of the Cas9/sgRNA expression vector into BMEC resulted in production of a homozygous clone . The LD size and content decreased following gene knockout. The mRNA abundance of fatty acid synthase () and peroxisome proliferator-activated receptor-gamma () was downregulated in the clone. Subcellular analyses indicated that BTN1A1 and LD were co-localized in the cytoplasm gene knockout increased the percentage of phosphatidylethanolamine (PE) and decreased phosphatidylcholine (PC), which resulted in a lower PC/PE ratio.

Conclusions: Results suggest that plays an important role in regulating LD synthesis via a mechanism involving membrane phospholipid composition.

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