Differences in Early Fat Development and Gene Transcription Expression in the Adipose Tissues of Piglets with and Without Gut Microbiota

doi:10.13560/j.cnki.biotech.bull.1985.2020-0919

Abstract

Abstract:

We compared the difference of fat deposition in the early growth stage and the different transcriptional profiles of adipose tissue in sterile piglets and normal piglets,and evaluated the direct effects of intestinal microbial colonization on the early development of pig adipose tissue. The cervical subcutaneous fat of 25-day sterile piglets（3 in GF piglet group）and normal pigs（3 in normal piglet group）was collected. HE staining was used to observe the morphological differences of fat cells,Western Blot to detect the protein expression differences of fat synthesis regulators,ELISA to detect the content of cytokines secreted in fat tissues,and RNA-seq to analyze the gene expression profile differences in fat tissues and identify the key differential genes and relevant signal networks. Compared with the normal piglets,the fat deposition in the same age of the sterile piglets was less,and the thickness of back fat and the sizes of fat cells significantly reduced（P<0.01）. The expression of fatty acid binding protein 4（FABP4）,peroxisome proliferators activated receptor γ（PPARγ）,acetyl CoA carboxylase（ACC）and fatty acid synthase（FAS）in the fat decreased significantly（P<0.05）. Adiponectin（P=0.100）and leptin（P=0.095）also decreased in varied degrees. The results of transcriptome sequencing showed that there were 695 genes in the fat of sterile piglets,including 338 genes up-regulated and 357 genes down-regulated（P<0.05,fold change>2）. Accuracy of RNA-sequencing data was verified by the qRT-PCR results. KEGG analysis showed that the differentially expressed genes were mainly related to lipid synthesis metabolism,catabolism,lipid oxidation and energy homeostasis. The colonization and status of intestinal microorganisms significantly affect the development,metabolism and function of animal adipose tissue.

Key words: gut microorganism, fat, gene, transcriptome, pig

QIU Xiao-yu, LIU Zuo-hua, QI Ren-li. Differences in Early Fat Development and Gene Transcription Expression in the Adipose Tissues of Piglets with and Without Gut Microbiota[J]. Biotechnology Bulletin, 2021, 37(5): 56-66.

Figures/Tables 9

References 43

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样品名 Sample name	总序列数 Raw reads	干净序列数 Clean reads	总对比到的序列数 Total mapped reads	多重对比的序列数 Multiple mapped reads	唯一对比的序列数 Uniquely mapped reads
Normal A1	60 328 616	59 694 488	50 892 462（85.25%）	4 145 489（6.94%）	46 746 973（78.31%）
Normal A2	53 878 956	53 400 340	45 855 826（85.87%）	4 209 183（7.88%）	41 646 643（77.99%）
Normal A3	65 393 020	64 761 370	55 361 124（85.48%）	5 082 167（7.85%）	50 278 957（77.64%）
GF A1	48 721 914	48 260 760	41 315 919（85.61%）	2 945 880（6.10%）	38 370 039（79.51%）
GF A2	54 262 278	53 749 144	46 093 576（85.76%）	3 467 323（6.45%）	42 626 253（79.31%）
GF A3	57 431 858	56 874 014	48 683 278（85.60%）	3 484 909（6.13%）	45 198 369（79.47%）

样品名 Sample name	总序列数 Raw reads	干净序列数 Clean reads	总对比到的序列数 Total mapped reads	多重对比的序列数 Multiple mapped reads	唯一对比的序列数 Uniquely mapped reads
Normal A1	60 328 616	59 694 488	50 892 462（85.25%）	4 145 489（6.94%）	46 746 973（78.31%）
Normal A2	53 878 956	53 400 340	45 855 826（85.87%）	4 209 183（7.88%）	41 646 643（77.99%）
Normal A3	65 393 020	64 761 370	55 361 124（85.48%）	5 082 167（7.85%）	50 278 957（77.64%）
GF A1	48 721 914	48 260 760	41 315 919（85.61%）	2 945 880（6.10%）	38 370 039（79.51%）
GF A2	54 262 278	53 749 144	46 093 576（85.76%）	3 467 323（6.45%）	42 626 253（79.31%）
GF A3	57 431 858	56 874 014	48 683 278（85.60%）	3 484 909（6.13%）	45 198 369（79.47%）

基因名称Gene Name	log2FC	变化Change	功能Function
TNS4	4.756	Up	Function unknown
TMP	4.587	Up	Function unknown
NRIP3	4.305	Up	Proteolysis,aspartic-type endopeptidase activity
MAPKAPK	4.088	Up	ATP binding,protein kinase activity
PRG4	4.076	Up	Receptor-mediated endocytosis
IRF5	4.051	Up	Function unknown
IRF6	4.007	Up	Keratinocyte proliferation
ITIH3	3.668	Up	Hyaluronan metabolic process
3,5,3’,5’- tetraiodothyronine	3.566	Up	Hormone biosynthetic process
Phenylalanine 4-monooxygenase	3.55	Up	Amino acid transport and metabolism,oxidoreductase activity
C4BPA	-7.537	Down	Positive regulation of protein catabolic
SQRDL	-6.269	Down	Oxidoreductase activity
PIK3C2G	-4.847	Down	Phosphatidylinositol 3-kinase（PI3K）complex
KCNIP2	-4.293	Down	Cell cycle control,myosin light chain
LIPG	-4.149	Down	Lipid metabolic,high-density lipoprotein particle remodeling
MAP6	-3.894	Down	Lysosome localization
CES1	-3.625	Down	Lipid transport and metabolism,Carboxylesterase
CYP1A1	-3.588	Down	Steroid metabolic,Steroid hormone biosynthesis
SucCβ	-3.498	Down	Fatty acid biosynthetic process,acetyl-CoA biosynthetic process
LRFN2	-3.452	Down	Function unknown

基因名称Gene Name	log2FC	变化Change	功能Function
TNS4	4.756	Up	Function unknown
TMP	4.587	Up	Function unknown
NRIP3	4.305	Up	Proteolysis,aspartic-type endopeptidase activity
MAPKAPK	4.088	Up	ATP binding,protein kinase activity
PRG4	4.076	Up	Receptor-mediated endocytosis
IRF5	4.051	Up	Function unknown
IRF6	4.007	Up	Keratinocyte proliferation
ITIH3	3.668	Up	Hyaluronan metabolic process
3,5,3’,5’- tetraiodothyronine	3.566	Up	Hormone biosynthetic process
Phenylalanine 4-monooxygenase	3.55	Up	Amino acid transport and metabolism,oxidoreductase activity
C4BPA	-7.537	Down	Positive regulation of protein catabolic
SQRDL	-6.269	Down	Oxidoreductase activity
PIK3C2G	-4.847	Down	Phosphatidylinositol 3-kinase（PI3K）complex
KCNIP2	-4.293	Down	Cell cycle control,myosin light chain
LIPG	-4.149	Down	Lipid metabolic,high-density lipoprotein particle remodeling
MAP6	-3.894	Down	Lysosome localization
CES1	-3.625	Down	Lipid transport and metabolism,Carboxylesterase
CYP1A1	-3.588	Down	Steroid metabolic,Steroid hormone biosynthesis
SucCβ	-3.498	Down	Fatty acid biosynthetic process,acetyl-CoA biosynthetic process
LRFN2	-3.452	Down	Function unknown