Cloning Of Duck C/EBPα, C/EBPβ, C/EBPδGenes And Developmental Expression In Duck Fat Tissues

The development of animal adipose is completed by the increase in the number of adipose cells and increases of the volume, development of adipose cell by a variety of genes regulation. C/EBPa,-β and-δ are three members of the CCAAT/enhancer binding proteins (C/EBPs) family, which is an important transcription factors in adipocyte differentiation. C/EBPs are more in-depth studies in mammalian fat development and differentiation of adipocyte in vitro, but there are no studies about roles of C/EBPs during development of duck fat development.In this research, the coding domain sequence (CDS) of C/EBPa,-β,-δ are cloned, then the relative expression levels of the three genes in28days duck different12tissues are detected by qRT-PCR method, and the developmental expression pattern of three genes are also researched by detecting the relative expression of subcutaneous adipose, abdominal adipose and liver in1-8weeks after the birth of the duck. The results showed as follows:1The CDS of duck C/EBPa is cloned for the first time, which is939bp length and encoding for313amino acids, the partial CDS sequences of C/EBPβ and C/EBPδ contain945bp and736bp, encoding315and242amino acids respectively.The homologies by comparison of the three genes among different species were highly conserved.2The C/EBPα, C/EBPβ and C/EBPδ abundance exhibit the significant difference among the various tissues. C/EBPa is most highly rich in (P<0.05) in the coeliac adipose the following expression rank is lung, subcutaneous adipose, liver, heart, kidney, intestinum tenue. C/EBPβ exhibits highest expression (P<0.05) in the lung, then in coeliac adipose, liver and subcutaneous adipose. C/EBPδ is the most highly abundant (P<0.05), then in coeliac adipose, liver, heart, muscle.3The mRNA expressions of C/EBPα,-β,-8were determined by real-time fluorescent quantitative PCR (q-PCR) method in postnatal duck (From week1to week8).The results showed that mRNAs (including C/EBPa, C/EBPβ and C/EBPδ) were expressed in seven tissues, including neck subcutaneous adipose, breast subcutaneous adipose, leg subcutaneous adipose, liver, abdominal subcutaneous adipose, back subcutaneous adipose and the abdominal subcutaneous adipose.4The expression pattern of C/EBPα shows that there is huge difference between the adipose tissues. The back subcutaneous adipose and the abdominal subcutaneous adipose get the peak expression at1w. The neck subcutaneous adipose, breast subcutaneous adipose, leg subcutaneous adipose and liver exhibit the highest abundance at4w, while the abundance of abdominal subcutaneous adipose peaks at6w.5The distinct expression pattern of C/EBPB in adipose tissues demonstrates that the neck subcutaneous adipose, abdominal subcutaneous adipose, back subcutaneous adipose and the coeliac adipose exhibits the highest transcript levels atl w, the leg subcutaneous adipose and reach highest level at3w, however the liver and breast subcutaneous adipose peak at4w.6The sharp expression pattern of C/EBPδ in adipose tissues reveals that expression levels of the neck subcutaneous adipose, back subcutaneous adipose and the coeliac adipose peak at1w, the leg subcutaneous adipose and reach at2w, however the liver and breast subcutaneous adipose peak at4w and the abdominal subcutaneous adipose get the highest abundance at6w.7There is a time sequence on C/EBPa,-β and-δ about regulating the adipose development of1-8w after the birth of duck, the expression of C/EBPa just after C/EBPβ and-δ. And the same time,C/EBPβ and-δ have Synergies in adipose development.