Zebrafish are resilient to the loss of major diacylglycerol acyltransferase enzymes

Meredith H Wilson; Monica R Hensley; Meng-Chieh Shen; Hsiu-Yi Lu; Vanessa H Quinlivan; Elisabeth M Busch-Nentwich; John F Rawls; Steven A Farber

doi:10.1016/j.jbc.2024.107973

Zebrafish are resilient to the loss of major diacylglycerol acyltransferase enzymes

J Biol Chem. 2024 Nov 6;300(12):107973. doi: 10.1016/j.jbc.2024.107973. Online ahead of print.

Authors

Meredith H Wilson¹, Monica R Hensley¹, Meng-Chieh Shen², Hsiu-Yi Lu³, Vanessa H Quinlivan², Elisabeth M Busch-Nentwich⁴, John F Rawls³, Steven A Farber⁵

Affiliations

¹ Department of Biology, Johns Hopkins University, Baltimore, Maryland, USA; Department of Embryology, Carnegie Institution for Science, Baltimore, Maryland, USA.
² Department of Embryology, Carnegie Institution for Science, Baltimore, Maryland, USA.
³ Department of Molecular Genetics and Microbiology, Duke Microbiome Center, Duke University, Durham, North Carolina, USA.
⁴ School of Biological and Behavioural Sciences, Queen Mary University of London, London, UK.
⁵ Department of Biology, Johns Hopkins University, Baltimore, Maryland, USA; Department of Embryology, Carnegie Institution for Science, Baltimore, Maryland, USA. Electronic address: sfarber3@jh.edu.

Abstract

In zebrafish, maternally deposited yolk is the source of nutrients for embryogenesis prior to digestive system maturation. Yolk nutrients are processed and secreted to the growing organism by an extra-embryonic tissue, the yolk syncytial layer (YSL). The export of lipids from the YSL occurs through the production of triacylglycerol-rich lipoproteins. Here we report that mutations in the triacylglycerol synthesis enzyme, diacylglycerol acyltransferase-2 (Dgat2), cause yolk sac opacity due to aberrant accumulation of cytoplasmic lipid droplets in the YSL. Although triacylglycerol synthesis continues, it is not properly coupled to lipoprotein production as dgat2 mutants produce fewer, smaller, ApoB-containing lipoproteins. Unlike DGAT2-null mice, which are lipopenic and die soon after birth, zebrafish dgat2 mutants are viable, fertile, and exhibit normal mass and adiposity. Residual Dgat activity cannot be explained by the activity of other known Dgat isoenzymes, as dgat1a;dgat1b;dgat2 triple mutants continue to produce YSL lipid droplets and remain viable as adults. Further, the newly identified diacylglycerol acyltransferase, Tmem68, is also not responsible for the residual triacylglycerol synthesis activity. Unlike overexpression of Dgat1a and Dgat1b, monoacylglycerol acyltransferase-3 (Mogat3b) overexpression does not rescue yolk opacity, suggesting it does not possess Dgat activity in the YSL. However, mogat3b;dgat2 double mutants exhibit increased yolk opacity and often have structural alterations of the yolk extension. Quadruple mogat3b;dgat1a;dgat1b;dgat2 mutants either have severely reduced viability and stunted growth or do not survive past 3 days post fertilization, depending on the dgat2 mutant allele present. Our study highlights the remarkable ability of vertebrates to synthesize triacylglycerol through multiple biosynthetic pathways.

Keywords: acyltransferase; dgat; lipid droplet; lipoprotein secretion; membrane enzyme; mogat3; mutant; yolk syncytial layer; zebrafish.