Recapitulation of zebrafish sncga expression pattern and labeling the habenular complex in transgenic zebrafish using green fluorescent protein reporter gene.
Human synuclein family consists of alpha-, beta-, and gamma-synucleins. Here, we cloned three genes, sncb, sncga and sncgb from zebrafish. They encode beta-, gamma1-, and gamma2-synucleins, respectively. The zSyn-beta, zSyn-gamma1, and zSyn-gamma2 proteins display 69%, 47%, and 50% identity to human beta-synuclein and gamma-synuclein, respectively. By reverse transcriptase-polymerase chain reaction, we demonstrated that sncb and sncga mRNA were abundant in brain and eye, while sncgb expression was moderate in brain, kidney, ovary and testis. The 1.8-kb 5'-upstream/promoter region of the sncga gene was sufficient to direct green fluorescent protein (GFP) expression in the central nervous system and cranial ganglions. A transgenic line, Tg(sncga:GFP), was generated and its GFP expression is similar to that of endogenous sncga mRNA. Moreover, this line also labels the habenular complex and the domain of GFP expression is larger in the left than in the right habenula. Thus, this line can be used to study sncga gene regulation and for left-right asymmetry study in zebrafish brain.
Chen
Yi-Chung
YC
Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan.
Cheng
Chia-Hsiung
CH
Chen
Gen-Der
GD
Hung
Chin-Chun
CC
Yang
Chung-Hsiang
CH
Hwang
Sheng-Ping L
SP
Kawakami
Koichi
K
Wu
Bo-Kai
BK
Huang
Chang-Jen
CJ
D000375:Aging / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004625:Embryo, Nonmammalian / Q000196:embryology / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D019262:Habenula / Q000196:embryology / Q000254:growth & development / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D016415:Sequence Alignment; D051843:Synucleins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish / Q000196:embryology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
C540271:Sncga protein, zebrafish; D051843:Synucleins; D029961:Zebrafish Proteins; D049452:Green Fluorescent Proteins