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

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Abstract

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 γ (PPAR γ) 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 μm) of adipocytes during this period. Accordingly, the gene expression level of PPAR γ and CCAAT/enhancer-binding proteins α (C/EBP α) was significantly elevated after fertilization for 12 days (p < 0.05), but C/EBP α declined at 20 dpf. Expression of lipoprotein lipase (LPL) increased from 2 to 16 dpf and then declined. In addition, immunoreaction of PPAR γ 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.

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Abbreviations

BAT:

Brown adipose tissue

WAT:

White adipose tissue

CVD:

Cardiovascular disease

PPAR γ:

Peroxisome proliferator-activated receptors

LPL:

Lipoprotein lipase

dpf:

Days post-fertilization

H&E:

Hematoxylin and eosin

PBS:

Phosphate-buffered saline

APES:

3-Aminopropyl-triethoxysilane

TBS:

Tris buffer solution

BSA:

Bovine serum albumin

C/EBP α:

CCAAT/enhancer-binding proteins α

FAS:

Fatty acid synthase

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Acknowledgments

The authors would thank Li Chao, Wang Yifei, and Yu Ping for their assistance in the study. This study was supported by the National Nature Science Foundation of China (NSFC, Grant Number 31072223).

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Authors and Affiliations

  1. College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, People’s Republic of China

    Pin Liu, Hong Ji, Chao Li, Jingjing Tian, Yifei Wang & Ping Yu

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  1. Pin Liu
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  2. Hong Ji
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  3. Chao Li
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Correspondence to Hong Ji.

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Liu, P., Ji, H., Li, C. et al. Ontogenetic development of adipose tissue in grass carp (Ctenopharyngodon idellus). Fish Physiol Biochem 41, 867–878 (2015). https://doi.org/10.1007/s10695-015-0053-x

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