Exogenous cMHM regulates the expression of DMRT1 and ER alpha in avian testes.
The male hypemethylated (MHM) region, associated with sex-specific DNA methylation and histone acetylation, encodes non-coding RNAs that are expressed exclusively on Z chromosome in female chickens. The function of the MHM non-coding RNAs is unclear yet. In order to analyze the regulation effect of MHM on mRNA expression of sex-dependent genes, exogenous plasmid containing chicken MHM (cMHM) was constructed and injected into left testes of 13-week-old roosters. Comb color of the roosters in the pEGFP-N1-cMHM group became much paler, an indication of change in sex hormones, as compared with that of the negative control groups at the 10 days after injection. Real-time PCR was used to determine mRNA expression of sex-dependent genes in testes tissue. Transcription of ER alpha gene was significantly up-regulated in the pEGFP-N1-cMHM group compared with that of the pEGFP-N1 and normal saline groups (P < 0.01), while expression of DMRT1 mRNA declined severely in the pEGFP-N1-cMHM group compared with that of normal saline groups (P < 0.05). Nevertheless, expression of other candidate genes, such as AMH, FOXL2, SOX9, and P450arom did not change after injection of exogenous cMHM. In conclusion, down-regulation of DMRT1 and up-regulation of ER alpha in rooster testes treated with exogenous pEGFP-N1-cMHM plasmid might indicate a novel cMHM non-coding RNA-mediated mechanism in avian sex differentiation.
Yang
Xiurong
X
State Key Laboratory for Agrobiotechnology and Department of Animal Genetics and Breeding, College of Animal Science and Technology, China Agricultural University, 100193, Beijing, China.
Zheng
Jiangxia
J
Xu
Guiyun
G
Qu
Lujiang
L
Chen
Sirui
S
Li
Junying
J
Yang
Ning
N
D000818:Animals; D002645:Chickens / Q000235:genetics*; D019175:DNA Methylation / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics; D047628:Estrogen Receptor alpha / Q000235:genetics* / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008297:Male; D010641:Phenotype; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012727:Sex Characteristics; D013737:Testis / Q000201:enzymology*; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
C110547:DMRT1 protein; D004274:DNA, Recombinant; D047628:Estrogen Receptor alpha; D012333:RNA, Messenger; D014157:Transcription Factors; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins