The paracrine effect of exogenous growth hormone alleviates dysmorphogenesis caused by tbx5 deficiency in zebrafish (Danio rerio) embryos

Journal of Biomedical Science

Jen-Her Lu⁴  Hsin-Yu Lee²  Ren-Bing Tang⁴  Shu-Yu Yeh¹  Sie-Lin Choo¹  Jen-Kann Lu¹  Tzu-Chun Tsai³ 

[1] Laboratory of Molecular Biology, Institute of Aquaculture, National Taiwan Ocean University, Keelung, Taiwan, Republic of China;Institute of Zoology, National Taiwan University, Taipei, Taiwan, Republic of China;School of Medicine, National Yang Ming University, Taipei, Taiwan, Republic of China;Department of Pediatrics, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China

关键词: Zebrafish;    Embryogenesis;    Apoptosis;    Growth hormone;    tbx5;   

Others  :  824818
DOI  :  10.1186/1423-0127-19-63

 received in 2012-02-07, accepted in 2012-07-09,  发布年份 2012

【 摘 要 】

Background

Dysmorphogenesis and multiple organ defects are well known in zebrafish (Danio rerio) embryos with T-box transcription factor 5 (tbx5) deficiencies, mimicking human Holt-Oram syndrome.

Methods

Using an oligonucleotide-based microarray analysis to study the expression of special genes in tbx5 morphants, we demonstrated that GH and some GH-related genes were markedly downregulated. Zebrafish embryos microinjected with tbx5-morpholino (MO) antisense RNA and mismatched antisense RNA in the 1-cell stage served as controls, while zebrafish embryos co-injected with exogenous growth hormone (GH) concomitant with tbx5-MO comprised the treatment group.

Results

The attenuating effects of GH in tbx5-MO knockdown embryos were quantified and observed at 24, 30, 48, 72, and 96 h post-fertilization. Though the understanding of mechanisms involving GH in the tbx5 functioning complex is limited, exogenous GH supplied to tbx5 knockdown zebrafish embryos is able to enhance the expression of downstream mediators in the GH and insulin-like growth factor (IGF)-1 pathway, including igf1, ghra, and ghrb, and signal transductors (erk1, akt2), and eventually to correct dysmorphogenesis in various organs including the heart and pectoral fins. Supplementary GH also reduced apoptosis as determined by a TUNEL assay and decreased the expression of apoptosis-related genes and proteins (bcl2 and bad) according to semiquantitative reverse-transcription polymerase chain reaction and immunohistochemical analysis, respectively, as well as improving cell cycle-related genes (p27 and cdk2) and cardiomyogenetic genes (amhc, vmhc, and cmlc2).

Conclusions

Based on our results, tbx5 knockdown causes a pseudo GH deficiency in zebrafish during early embryonic stages, and supplementation of exogenous GH can partially restore dysmorphogenesis, apoptosis, cell growth inhibition, and abnormal cardiomyogenesis in tbx5 knockdown zebrafish in a paracrine manner.

【 授权许可】

   
2012 Tsai et al.; licensee BioMed Central Ltd.

【 图 表 】

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