Molecular cloning and expression of a smooth muscle-specific gene SM22alpha in zebrafish.
SM22alpha is a kind of 22-kDa protein which is exclusively expressed in smooth muscle containing tissues of the vertebrates. Here we report molecular cloning of a novel zebrafish SM22alpha gene. The full length of zebrafish SM22alpha cDNA is 1296bp and it encodes a polypeptide of 201 amino acids which shares 69.2%, 69.7%, 69.2%, 67.2%, and 61.2% overall identity with human, mouse, rat, chicken, and bovine SM22alpha, respectively. Characterization of zebrafish SM22alpha genomic sequence reveals that it spans 7.7kb and contains five exons and four introns. The expression pattern of SM22alpha in zebrafish embryonic development is studied by whole-mount in situ hybridization. Strong expression is observed in vascular, gut, swim bladder, branchial arches, and fin epidermis. Furthermore, we carry out gene knock-down by antisense morpholino oligonucleotide, which results in disappearance of yolk extension, caudal fin aberrance, and deficiency of circulation system in zebrafish embryo. Cross-section of SM22alpha-deficient embryo suggests that SM22alpha may play roles in smooth muscle cell morphology transform.
Yang
Xue-yan
XY
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, 200433, PR China.
Yao
Ji-hua
JH
Cheng
Lu
L
Wei
Dong-wang
DW
Xue
Jing-lun
JL
Lu
Da-ru
DR
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002645:Chickens; D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D006801:Humans; D017403:In Situ Hybridization; D051379:Mice; D008840:Microfilament Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D009130:Muscle, Smooth / Q000196:embryology / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000235:genetics; D009928:Organ Specificity; D051381:Rats; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D015027:Zebrafish / Q000196:embryology* / Q000378:metabolism*
D008840:Microfilament Proteins; D009124:Muscle Proteins; D016376:Oligonucleotides, Antisense; C058992:Tagln protein, mouse; C051615:transgelin