Ontogenetic development of adipose tissue in grass carp (Ctenopharyngodon idellus)

To investigate the adipose tissue development process during the early stages of grass carp (Ctenopharyngodon idellus) development, samples were collected from fertilized eggs to 30 days post-fertilization (dpf) of fish. Paraffin and frozen sections were taken to observe the characteristics of adipocytes in vivo by different staining methods, including hematoxylin and eosin (H&E), Oil red O, and BODIPY. The expression of lipogenesis-related genes of the samples at different time points was detected by real-time qPCR. In addition, protein expression level of peroxisome proliferator-activated receptors gamma (PPAR gamma) was detected by immunohistochemistry. The results showed that the neutral lipid droplets accumulated first in the hepatocytes of 14-dpf fish larvae, and visceral adipocytes appeared around the hepatopancreas on 16 dpf. As grass carp grew, the adipocytes increased in number and spread to other tissues. In 20-dpf fish larvae, the intestine was observed to be covered by adipose tissue. However, there was no significant change in the average size (30.40-40.01 mu m) of adipocytes during this period. Accordingly, the gene expression level of PPAR gamma and CCAAT/enhancer-binding proteins alpha (C/EBP alpha) was significantly elevated after fertilization for 12 days (p < 0.05), but C/EBP alpha declined at 20 dpf. Expression of lipoprotein lipase (LPL) increased from 2 to 16 dpf and then declined. In addition, immunoreaction of PPAR gamma was positive on hepatocytes after fertilization for 15 days. These results implied that the early developmental stage of adipose tissue is caused by active recruitment of adipocytes as opposed to hypertrophy of the cell. In addition, our study indicated that lipogenesis-related genes might regulate the ongoing development of adipose tissue.