lncRNA TCONS_00815878对猪骨骼肌卫星细胞分化的影响

doi:10.16288/j.yczz.19-146

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Hereditas(Beijing) ›› 2019, Vol. 41 ›› Issue (12): 1119-1128.doi: 10.16288/j.yczz.19-146

• Research Article • Previous Articles Next Articles

The effect of lncRNA TCONS_00815878 on differentiation of porcine skeletal muscle satellite cells

Ziying Huang¹, Long Li¹, Qianqian Li¹, Xiangdong Liu^1,², Changchun Li^1,²()

^1. Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China
^2. Guangxi Yangxiang Co., Ltd. Production Center, Guigang 537131, China

Received:2019-05-21 Revised:2019-11-19 Online:2019-12-20 Published:2019-12-05
Contact: Li Changchun E-mail:lichangchun@mail.hzau.edu.cn
Supported by:
Supported by the National Natural Science Foundation of China No(31872322);Central University Innovation Research Fund No(2662017PY030)

Abstract

Abstract:

Porcine skeletal muscle development is a complex biological process, and differentiation of skeletal muscle satellite cells is an important part of skeletal muscle development. In recent years, it has been found that lncRNA plays an important role in the differentiation of skeletal muscle satellite cells. Here we investigate the effect of lncRNA TCONS_00815878 on the differentiation of porcine skeletal muscle satellite cells. We first used qRT-PCR to detect the expression levels of TCONS_00815878 in six tissues (heart, spleen, lung, kidney, back muscles and leg muscles) of Yorkshire piglets within seven days of birth. At the same time, the expression levels of TCONS_00815878 at five different time points from the embryonic stage to the postnatal stage (35 d, 45 d, 55 d of embryos, and 7 d, 200 d of postpartum leg muscles) were examined. The expression of the differentiation marker genes MyoD, MyoG and MyHC was examined by knocking down TCONS_00815878 in porcine skeletal muscle satellite cells using antisense oligonucleotides (ASO). The target gene of TCONS_00815878 was predicted by bioinformatics analysis, and the function and pathway of its target gene were predicted online using DAVID software. The results showed that TCONS_00815878 had the highest expression level in pig myocardium and leg muscles. Within seven days after birth, TCONS_00815878 increased in the muscle tissue of pigs, and reached the peak of expression level on the 7th day. During the process of proliferation and differentiation of porcine skeletal muscle satellite cells, the expression level of TCONS_00815878 increased during the differentiation stage and peaked at 30 h of differentiation. After knocking down TCONS_00815878, the expression levels of MyoD, MyoG and MyHC were decreased, but the expression level of MyoD was significantly decreased (P<0.05). In addition, functional predictions revealed that the target gene of TCONS_00815878 is enriched in multiple biological processes, such as glycolysis and pyruvate metabolism, related to skeletal muscle satellite cell differentiation. This study speculates that lncRNA TCONS_00815878 may promote the differentiation of porcine skeletal muscle satellite cells.

Key words: lncRNA TCONS_00815878, porcine skeletal muscle satellite cells, differentiation, knockdown, target gene

Ziying Huang, Long Li, Qianqian Li, Xiangdong Liu, Changchun Li. The effect of lncRNA TCONS_00815878 on differentiation of porcine skeletal muscle satellite cells[J]. Hereditas(Beijing), 2019, 41(12): 1119-1128.

Figures/Tables 7

Table 1

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Supplementary Table 1

List of target genes"

基因名称	基因名称	基因名称	基因名称	基因名称	基因名称
ARFGAP3	ADGRL4	POLR3K	SLC25A26	LAMC2	ARHGEF10
IFT27	PGM1	KCTD5	MITF	INTS7	PLXNA3
PRPF40B	KIF2C	ADRA2B	GXYLT2	LPGAT1	REEP1
IGFBP6	SMOC2	MAP4K4	PPARG	HHAT	TECTA
NCKAP1L	CNKSR3	TBC1D8	GK5	MGAT5	SLC16A3
STAT2	NT5DC1	CHST10	P2RY14	ADGRA2	ADAMTS2
BAZ2A	SNAP91	ANKRD23	SENP5	HERC2	ELOVL5
RASSF8	PSTPIP2	SMYD1	PARP14	SCN3A	CES3
KCNJ8	CTIF	TMSB10	CD47	CALCRL	KDM4B
P3H3	ZNF106	DOK1	OTC	TFPI	BBS9
GAPDH	TYRO3	DGUOK	ATP6AP2	TMEFF2	MPI
TNFRSF1A	RPAP1	PREB	SLC9A7	PLCL1	PPIP5K2
CRACR2A	ATP10A	NBAS	WDR45	CARF	GREB1
ITFG2	ZNF516	CD38	SYNJ2	PARD3B	PTPN6
BID	SERPINB2	KLHL2	PGK1	BARD1	PXDC1
SLC22A23	DPP8	GUCY1B1	ZMAT1	CYP27A1	KDSR
SERPINB1	ARF6	SLC4A4	ELF4	PRKAG3	ST6GAL1
JARID2	KANK1	ARHGAP10	ENOX2	GPC1	NOA1
PHF1	NPR2	ANXA5	FMR1	HDLBP	IGFBP5
VEGFA	TMEM8B	STPG2	TMEM185A	SNED1	SRPK2
ZXDC	ZCCHC7	MAPK10	RENBP	NEB	SLC25A40
TMEM266	FKTN	PAQR3	NADSYN1	CAV2	SCAP
LINGO1	OLFML2A	FGF9	BRMS1	DBNL	ACSM5
EDC3	DNM1	EBPL	EHBP1L1	ZNF282	PADI2
AKAP5	EXD3	PHF11	MAP4K2	CDK3	DCUN1D2
GALNT16	FBXL6	MPHOSPH8	MARK2	GALK1	CARHSP1
IFT43	MYC	DNAH10	DTX4	TANC2	B4GALT2
VASH1	NCOA2	TMEM120B	KBTBD4	FKBP10	CTC1
IRF2BPL	SCYL3	PPTC7	ABTB2	TBKBP1	LMAN2L
SETD3	F5	ACAD10	LMO2	TSPOAP1	CASP10
WDR20	PBX1	HSPB8	MPPED2	ABR	STK36
MBTPS1	CD84	NOS1	TEAD1	ANKFY1	BDKRB2
THAP11	SLAMF8	MIF	SCAMP4	SMTNL2	MED31
SLC9A5	SYT11	LZTR1	ANKRD24	ENO3	CENPH
YIF1B	CERS2	DGCR2	CHAF1A	BCL6B	PANK4
ARHGEF1	FMO5	SIPA1L2	PNPLA6	MYH8	ESYT3
IRF2BP1	ZNF697	ARID5B	ZNF608	SLC5A10	OBSL1
PLEKHA4	TTF2	SPOCK2	PDLIM4	ARHGAP23	CASP8
BCAT2	CTTNBP2NL	CEP55	IL5	PFKM	FLYWCH1
BAX	ARHGAP29	TBC1D12	TRPC7	OXTR	VPS37C
CACNG6	GLMN	SORBS1	FZD4	KCNA5	PARVA
PUSL1	TGFBR3	ZDHHC6	ARHGAP42	CEP19	PHACTR4
MMEL1	GBP2	EIF3A	HINFP	DUS2	RASIP1
CLCN6	HSPA12B	PPP1R12B	SORL1	DIS3L2	OLFM1
NPPA	TTI1	RPP25L	NTM	HS6ST1	CCDC112
VPS13D	KCNB1	CACNB2	PRELP	STK38L	HS1BP3
DHRS3	GUSB	CREM	SRI	WDR73	AHR
FHAD1	CTF1	TRAK1	CALCR	IL20RB	TBC1D23
HMGN2	SETD1A	PTH1R	CROT	CALM3	LYSMD4
EPB41	REXO5	TEX264	PHTF2	WDR4	PDIK1L
CLSPN	ZNF174	NISCH	LAMB1	NFIA
MOCOS	RHBDF1	GLT8D1	KIAA1614	TIA1

Supplementary Table 1

Supplementary Fig. 1

References 30

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The effect of lncRNA TCONS_00815878 on differentiation of porcine skeletal muscle satellite cells

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目的基因	引物序列( 5′→3′)	复性温度(℃)	产物大小(bp)
TCONS_00815878	F: ACACCCTTTCCCAAAATCAA	56	93
TCONS_00815878	R: GCAGACTGTCCAAATCTACCCT	56	93
PFKM	F: GAGAGCGTTTCCATGATGCTTC	60	221
PFKM	R: AATCAAAGAGGGTGCCATCCAT	60	221
ELOVL5	F: ATATGAAGATCATCCGCGTGCT	60	59
ELOVL5	R: GTGATCTGGTGGTTGTTCTTGC	60	59
MBTPS1	F: CCTCAACAGTGGTGGAATACGA	60	179
MBTPS1	R: TTGGGATGATCTTCAAGCGTCA	60	179
MyoD	F: GGCTGCCCAAGGTGGAAATC	60	170
MyoD	R: TGCGTCTGAGTCACCGCTGTAG	60	170
MyoG	F: ATGAGACATCCCCCTACTTCTACCA	60	160
MyoG	R: GTCCCCAGCCCCTTATCTTCC	60	160
Pax7	F: GGAGTACAAGAGGGAGAACCC	60	122
Pax7	R: TTCTGAGCACGCGGCTAATC	60	122
Myf5	F: GAATGCCATCCGCTACAT	60	125
Myf5	R: AACTGCTGCTCTTTCTGG	60	125
MyHC	F: GTTCAGAGAAAGGCATCCCAAA	60	135
MyHC	R: GAGAGTGACCGACACCACAAGTG	60	135
FKBP10	F: ATGTGTCCTGGAGAGAGAAGGA	60	209
FKBP10	R: AATCAAAGAGGGTGCCATCCAT	60	209
18S rRNA	F: TCCCGACGTGACTGCTC	60	133
18S rRNA	R: GGTGACAGCGGGGTGG	60	133

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