Characterization and expression of serotonin transporter genes in zebrafish.
To understand the development of serotonergic neurons in vertebrates, we used zebrafish as a model system. In this study we cloned two cDNAs (complementary DNAs) coding for serotonin transporter (SERT) from the zebrafish, named serta and sertb. The serta cDNA encodes a protein of 693 amino acids and showed high level of sequence identity with rat and human SERTs. In situ hybridization showed serta to be expressed in raphe nuclei, ventral posterior tuberculum and pineal organ. The expression of serta in raphe and ventral posterior tuberculum overlapped with the location of serotonin and expression of tryptophan hydroxylase, which is a key enzyme for serotonin synthesis. In the pineal organ serta is expressed in the cells in the vicinity of tryptophan hydroxylase-positive cells. We also cloned another zebrafish serotonin transporter, sertb, and found to be expressed in the medulla oblongata and in the inner nuclear layer of retina. The existence of two sert genes in the zebrafish genome indicates the gene was duplicated in the process of evolution as can be seen in other genes in the teleosts including zebrafish. The expression of the serta cDNA in cultured cells conferred a serotonin transport activity, thus indicating the validity of the cloned cDNA. We have established the expression system of zebrafish serotonin transporter in the cell culture in the present study, which is useful for the pharmacological analysis to determine the important residues for the interaction with serotonin and inhibitors. The expression system in the cell culture can be used to determine the effective concentration of inhibitors and addictive drugs. These information might be useful to evaluate the effect of those chemicals on serotonin neuron development and behavior of the animal.
Department of Anatomy, Biology and Medicine, Oita University School of Medicine, Japan.
Yuhong
Y
Wang
Rie
R
Takai
Hidekatsu
H
Yoshioka
Komei
K
Shirabe
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D017124:Conserved Sequence; D004625:Embryo, Nonmammalian; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D010802:Phylogeny; D010870:Pineal Gland / Q000166:cytology / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D011903:Raphe Nuclei / Q000166:cytology / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D012160:Retina / Q000166:cytology / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012249:Rhombencephalon / Q000166:cytology / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012701:Serotonin / Q000032:analysis / Q000378:metabolism; D050486:Serotonin Plasma Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014365:Tryptophan Hydroxylase / Q000378:metabolism; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D029961:Zebrafish Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D050486:Serotonin Plasma Membrane Transport Proteins; D029961:Zebrafish Proteins; D049452:Green Fluorescent Proteins; D012701:Serotonin; D014365:Tryptophan Hydroxylase