It is well documented that four gene families, including the glycerophosphate acyltransferases (GPATs), acylglycerophosphate acyltransferases (AGPATs), lipid phosphate phosphohydrolases (LPINs) and diacylglycerol acyltransferases (DGATs), are involved in the glycerophosphate pathway of
de novo
triglyceride (TG) biosynthesis in mammals. However, no systematic analysis has been conducted to characterize the gene families in poultry. In this study, the sequences of gene family members in the glycerophosphate pathway were obtained by screening the public databases. The phylogenetic tree, gene structures and conserved motifs of the corresponding proteins were evaluated. Dynamic expression changes of the genes at different developmental stages were analyzed by qRT-PCR. The regulatory characteristics of the genes were analyzed by
in vivo
experiments. The results showed that the
GPAT
,
AGPAT
and
LPIN
gene families have 2, 7 and 2 members, respectively, and they were classified into 2, 4 and 2 cluster respectively based on phylogenetic analysis. All of the genes except
AGPAT1
were extensively expressed in various tissues. Estrogen induction upregulated the expression of
GPAM
and
AGPAT2
, downregulated the expression of
AGPAT3
,
AGPAT9
,
LPIN1
and
LPIN2
, and had no effect on the expression of the other genes. These findings provide a valuable resource for further investigation of lipid metabolism in liver of chicken.