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Application of ITGA2 gene in regulation and control of formation of rice-colored fat of pigs

A gene and fat technology, applied in the application field of ITGA2 gene in regulating the formation of beige fat in pigs

Active Publication Date: 2022-03-18
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on porcine beige fat marker genes, so it is urgent to establish a method for screening and functional verification of porcine beige fat marker genes, so as to provide new ideas for the treatment of diseases related to fat metabolism disorders

Method used

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  • Application of ITGA2 gene in regulation and control of formation of rice-colored fat of pigs
  • Application of ITGA2 gene in regulation and control of formation of rice-colored fat of pigs
  • Application of ITGA2 gene in regulation and control of formation of rice-colored fat of pigs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 1 Establishment of porcine beige adipocyte differentiation system in vitro

[0081] In this example, SVF cells (vascular stromal cells) were subcutaneously isolated from the groin of Tibetan pigs, and induced to differentiate into beige adipocytes. Through comparative genomics, the genes expressed more in beige adipocytes than white adipocytes were analyzed, and beige adipocytes were analyzed. Morphological and molecular characteristics of adipocytes ( figure 1 , A).

[0082] 1. Isolation, culture and differentiation induction of porcine SVF cells:

[0083] Take the subcutaneous white adipose tissue of the groin of 1-month-old male Tibetan piglets, cut it, wash it with DPBS containing 5% penicillin / streptomycin, shred it, and put it in Dulbecco Hanks balanced salt solution containing 2mg / ml type I collagenase at 37°C Digest in medium for 60min, filter the tissue suspension with a 70μm cell strainer, centrifuge at 1500r / min for 10min, resuspend the cell pellet ...

Embodiment 2

[0088] Example 2 The ITGA2 gene is a gene specifically highly expressed in pig beige fat

[0089] Previously, it was found that beige adipocytes appeared in the inguinal subcutaneous adipose tissue of Tibetan pigs after acute cold stimulation. In this example, a total of 95 genes ( image 3, A), the top 20 differentially expressed genes include the reported KCNK3 and ITGA2 ( image 3 , B), these genes are mainly enriched in lipid synthesis and transport, lipid metabolism and other pathways ( image 3 , C). The gene set of the reported human and mouse brown core screening genes contains the known beige fat marker genes KCNK3, EBF2 and CD36, but does not contain the ITGA2 gene, indicating that ITGA2 is a gene specifically expressed in pig beige fat ( image 3 , D~E).

Embodiment 3

[0090] Example 3 ITGA2 gene is an essential gene for porcine beige fat

[0091] In this example, RNAi technology was used to knock down the ITGA2 gene to verify its role in the differentiation process of beige fat. Experimental design such as Figure 4 As shown in middle A, transfection reagents were added at the -2d, 0d and 3d of the beige fat differentiation, and the differentiation was induced to the 6th day, and the expression of the ITGA2 gene was detected. The results showed that after inhibiting the expression of the ITGA2 gene ( Figure 4 , B), the differentiation efficiency of beige adipocytes was significantly reduced ( Figure 4 , C and D). By using the seahorse system to detect the OCR (oxygen consumption rate) of beige adipocytes, it was found that after inhibiting the expression of the ITGA2 gene, the basal respiration and heat production of the cells were significantly reduced. The experimental results show that the ITGA2 gene is necessary for the formation o...

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Abstract

The invention discloses an application of an ITGA2 gene in regulating and controlling the formation of rice-colored fat of a pig. According to the present invention, the RNA-seq technology is adopted, the gene expression difference of the mature beige fat cells and the white fat cells of the Tibetan pig is analyzed, the beige fat specific high expression gene is screened, and the pig specific beige fat regulation gene ITGA2 is successfully identified based on the human, mouse and pig comparative genomics technology; the expression of the ITGA2 gene is knocked down by further utilizing an RNAi technology, and it is proved through oil red O dyeing and a seharse experiment that ITGA2 is needed for the formation and function maintenance of the rice-colored fat of the pig. According to the invention, the effect of ITGA2 in the formation process of the rice-colored fat of the pig is discovered for the first time, and the discovery has important significance for regulating and controlling fat deposition of the pig by utilizing the rice-colored fat.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of ITGA2 gene in regulating the formation of pig beige fat. Background technique [0002] Mammalian adipose tissue is an important endocrine organ regulating energy balance. Classical adipose tissue can be divided into two functional groups: white adipose tissue stores excess energy in the form of triglycerides, and brown adipose tissue dissipates energy in the form of heat through adaptive thermogenesis. Under various stimuli such as cold exposure, drugs, and exercise, beige adipocytes appear in white adipose tissue, and they have similar characteristics to brown adipocytes, such as the presence of multilocular lipid droplets and uncoupling protein 1 (UCP1)-positive cells. Molecular features of human and mouse beige adipocytes have now been characterized, and similar to brown adipocytes, beige adipocytes' thermogenic activity is dependent on the activation of UCP1. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/113C12Q1/6888A61K48/00A61K38/17A61P3/04A61P3/00
CPCC07K14/70546C12N15/1138C12Q1/6888A61K48/005A61K38/1777A61P3/04A61P3/00C12N2310/14C12Q2600/124C12Q2600/158Y02P60/87
Inventor 陶聪张立兰王彦芳陈川河刘嘉莉
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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