Wenghong Wong1, Yike Huang1,2, Zhuanbin Wu3, Yu Kong4, Jing Luan1, Qiaoan Zhang1, Jiewen Pan1, Kexiang Yan5, Zhenghua Zhang6. 1. Department of Dermatology, Huashan Hospital, Shanghai Medical College of Fudan University, Shanghai, China. 2. Department of Dermatology, Xiamen Chang Gung Hospital, Xiamen, China. 3. Shanghai Model Organisms Center Inc, Shanghai, China. 4. Institute of Neuroscience, Chinese Academy of Science, Shanghai, China. 5. Department of Dermatology, Huashan Hospital, Shanghai Medical College of Fudan University, Shanghai, China. ykx2292002@aliyun.com. 6. Department of Dermatology, Huashan Hospital, Shanghai Medical College of Fudan University, Shanghai, China. verzhang@foxmail.com.
Abstract
BACKGROUND: The MVD gene mutations are identified in porokeratosis, which is considered a skin-specific autoinflammatory keratinization disease. However, the biological function of MVD gene remains largely unknown. Therefore, we analyzed the function of mvda gene, orthologous to the human MVD gene, in developing zebrafish. METHODS: Morpholino antisense oligonucleotide technique was used to generate mvda loss-of-function phenotypes. Knockdown of mvda was confirmed by RT-PCR and Sanger sequencing. Scanning and transmission electron microscopy were performed to analyze the morphology of the epidermis. Angiogenesis study was presented using the Tg(fli1a:EGFP)y1 transgenic strain. In addition, acridine orange staining was used to examine the apoptotic cells in vivo. RESULTS: As expected, the mvda morphants showed abnormal morphology of the epidermis. Moreover, we observed ectopic sprouts in trunk angiogenesis and impaired formation of the caudal vein plexus in the mvda-deficient zebrafish. Besides, increased apoptosis was found throughout the tail, heart, and eyes in mvda zebrafish morphants. CONCLUSIONS: These findings indicated the essential role of mvda in the early development of zebrafish. This was the first in vivo knockdown study of the zebrafish mvda gene, which might offer insight into the biological function of the human MVD gene.
BACKGROUND: The MVD gene mutations are identified in porokeratosis, which is considered a skin-specific autoinflammatory keratinization disease. However, the biological function of MVD gene remains largely unknown. Therefore, we analyzed the function of mvda gene, orthologous to the humanMVD gene, in developing zebrafish. METHODS:Morpholino antisense oligonucleotide technique was used to generate mvda loss-of-function phenotypes. Knockdown of mvda was confirmed by RT-PCR and Sanger sequencing. Scanning and transmission electron microscopy were performed to analyze the morphology of the epidermis. Angiogenesis study was presented using the Tg(fli1a:EGFP)y1 transgenic strain. In addition, acridine orange staining was used to examine the apoptotic cells in vivo. RESULTS: As expected, the mvda morphants showed abnormal morphology of the epidermis. Moreover, we observed ectopic sprouts in trunk angiogenesis and impaired formation of the caudal vein plexus in the mvda-deficient zebrafish. Besides, increased apoptosis was found throughout the tail, heart, and eyes in mvdazebrafish morphants. CONCLUSIONS: These findings indicated the essential role of mvda in the early development of zebrafish. This was the first in vivo knockdown study of the zebrafishmvda gene, which might offer insight into the biological function of the humanMVD gene.