Molecular cloning and characterization of the germ cell-related nuclear orphan receptor in chickens.
Nuclear receptor subfamily 6, group A, member 1 (NR6A1), also known as germ cell nuclear factor/retinoid receptor-related testis-associated receptor and neuronal cell nuclear factor, is a member of the nuclear orphan receptor superfamily. NR6A1 has been cloned in various species including humans and mice, but it has been scarcely investigated in avian species. In the present study, we cloned the chicken NR6A1 (cNR6A1) from a testis cDNA library. The cloned cNR6A1 sequence was mapped to chromosome 17 and contained an open reading frame of 1.4 kb encoding 445 amino acids. Multiple alignment analysis of the cNR6A1 protein-coding sequence with NR6A1s from humans, mice, boars, rats, zebrafish, and Xenopus showed high degrees of homology, 89%, 90%, 89%, 88%, 83%, and 87%, respectively. Using RNA interference, changes in the expression of pluripotency-, germ cell-, and differentiation-related key genes by silencing of cNR6A1 were validated in chicken blastoderm-derived embryonic stem cells. Among those genes, the relative expression levels of POU5F1, CRIPTO, DAZL, DDX4, BMP15, GSC, and SOX7 changed significantly compared to the control group. We also confirmed that the activity of alkaline phosphatase, known as a pluripotency marker, was maintained by cNR6A1 gene silencing in chicken blastodermal cells. Collectively, our data suggest that cNR6A1 may play an important role during chicken embryonic development and differentiation.
Lee
Sang In
SI
WCU Biomodulation Major, Department of Agricultural Biotechnology and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul, Korea.
Kim
Jin Kyoo
JK
Park
Hyun Jeong
HJ
Jang
Hyun Jun
HJ
Lee
Hyung Chul
HC
Min
Taesun
T
Song
Gwonhwa
G
Han
Jae Yong
JY
D000469:Alkaline Phosphatase / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001756:Blastoderm / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D002645:Chickens / Q000235:genetics*; D002874:Chromosome Mapping / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods; D004285:Dogs; D053595:Embryonic Stem Cells / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D008969:Molecular Sequence Data; D057136:Nuclear Receptor Subfamily 6, Group A, Member 1 / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D039904:Pluripotent Stem Cells / Q000378:metabolism; D016133:Polymerase Chain Reaction; D034622:RNA Interference / Q000502:physiology*; D051381:Rats; D015203:Reproducibility of Results; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D057136:Nuclear Receptor Subfamily 6, Group A, Member 1; D000469:Alkaline Phosphatase