Characterization of F-spondin in Japanese flounder (Paralichthys olivaceus) and its role in the nervous system development of teleosts

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• 作者：Hongshuang Hu ; Nian Xin ; Jinxiang Liu ; Mengmeng Liu ; Zhenwei Wang ; Wenji Wang ; Quanqi Zhang ; Jie Qi ; ^{qijie@ouc.edu.cn" class="auth_mail" title="E-mail the corresponding author}
• 关键词：ECM ; extracellular matrix ; TSRs ; thrombospondin repeats ; APP ; amyloid precursor protein ; AD ; Alzheimer disease ; ISH ; in situ hybridization ; DIG ; digoxigenin ; SEM ; standard error of the mean ; ORF ; open reading frame ; TSR1 ; thrombospondin type 1 repeat
• 刊名：Gene
• 出版年：2016
• 出版时间：10 January 2016
• 年：2016
• 卷：575
• 期：2
• 页码：623-631
• 全文大小：1567 K

文摘

F-spondin was originally isolated from the developing embryonic floor plate of vertebrates, secreting numerous kinds of neuron-related molecules. The protein performs a positive function in nervous system development, which is attributed to the high conservation of F-spondin protein, an extracellular matrix (ECM) protein in several species. However, its precise function remains unknown, especially in marine fish. In this study, the F-spondin of Japanese flounder (Paralichthys olivaceus).

was cloned, and its expression pattern and structural characteristics were analyzed. The 2421 bp-long cDNA ORF of PoF-spondin was obtained and divided into 14 exons spread over 61,496 bp of the genomic sequence. Phylogenetic analysis showed that PoF-spondin was actually the ortholog of the human spon1 gene and shared high identities with other teleost spon1a genes. Quantitative RT-PCR analysis showed that PoF-spondin was maternally expressed, and transcripts were present from one-cell stage to hatching stage, peaking at tailbud stage. Tissue distribution analysis indicated that PoF-spondin was detectable mainly in the gonads (especially in the ovary) and the brain. Whole mount in situ hybridization analysis revealed that the PoF-spondin transcription distributed throughout the cleavage of the ball in the early stage and expressed at a high level in the floor plate of the trunk at tailbud and pre-hatching stages. Furthermore, the expression of genes related to nervous system development (spon1b, foxo3b, and foxj1a) was significantly increased after the injection of PoF-spondin into the embryos of wild-type zebrafish. Furthermore, PoF-spondin significantly suppressed the expression of the chordamesoderm marker gene ntl, increased the expression of otx2/krox20, ectoderm mark genes, and left the expression of dorsal mesodermal marker gene gsc unaffected at 50% epiboly stage in zebrafish. In short, our results suggest that PoF-spondin functions in the development of the teleost nervous system.