Targeted disruption of fad24, a regulator of adipogenesis, causes pre-implantation embryonic lethality due to the growth defect at the blastocyst stage

Abstract

In previous studies, we identified a novel gene, factor for adipocyte differentiation 24 (fad24), which plays an important role during the early stages of adipogenesis in mouse 3T3-L1 cells. Moreover, overexpression of fad24 increased the number of smaller adipocytes in white adipose tissue and improved glucose metabolic activity in mice, thus indicating that fad24 functions as a regulator of adipogenesis in vivo. However, the physiological roles of fad24 in vivo are largely unknown. In this study, we attempted to generate fad24-deficient mice by gene targeting. No fad24-null mutants were recovered after embryonic day 9.5 (E9.5). Although fad24-null embryos were detected in an expected Mendelian ratio of genotypes at E3.5, none of the homozygous mutants developed into blastocysts. In vitro culture experiments revealed that fad24-null embryos develop normally to the morula stage but acquire growth defects during subsequent stages. The number of nuclei decreased in fad24-deficient morulae compared with that in wild-type ones. These results strongly suggested that fad24 is essential for pre-implantation in embryonic development, particularly for the progression to the blastocyst stage.

Keywords: 4′,6-diamidino-2-phenylindole; Adipogenesis; Blastocyst; DAPI; E; Embryonic development; Embryonic lethality; HBO1; IVF; Morula; Neo; Noc; PBS; PCR; TK; embryonic day; factor for adipocyte differentiation; fad; fad24; histone acetyltransferase binding to ORC1; in vitro fertilization; neomycin resistance; nucleolar complex-associated protein; phosphate buffered saline; polymerase chain reaction; thymidine kinase.