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Updated: Nov 4, 2025
High Resolution Whole Mount In Situ Hybridization within Zebrafish Embryos to Study Gene Expression and Function
Published on: October 19, 2013
Mvda is required for zebrafish early development
Wenghong Wong1, Yike Huang1,2, Zhuanbin Wu3
1Department of Dermatology, Huashan Hospital, Shanghai Medical College of Fudan University, Shanghai, China.
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.
Insights
The MVD gene
Area of Science:
- Developmental Biology
- Genetics
- Cell Biology
Background:
- Mutations in the MVD gene are linked to porokeratosis, a skin disorder.
- The precise biological function of the MVD gene is not well understood.
- The mvda gene in zebrafish is orthologous to the human MVD gene.
Purpose of the Study:
- To investigate the function of the mvda gene in zebrafish development.
- To understand the role of mvda in epidermal morphology and angiogenesis.
Main Methods:
- Morpholino antisense oligonucleotide technique for mvda knockdown.
- RT-PCR and Sanger sequencing for confirmation of knockdown.
- Electron microscopy for epidermal morphology analysis.
- Tg(fli1a:EGFP) transgenic zebrafish for angiogenesis studies.
- Acridine orange staining for apoptosis detection.
Main Results:
- mvda knockdown resulted in abnormal epidermal morphology in zebrafish.
- Ectopic sprouts and impaired caudal vein plexus formation were observed in mvda morphants.
- Increased apoptosis was detected in the tail, heart, and eyes of mvda-deficient zebrafish.
Conclusions:
- The mvda gene plays a crucial role in zebrafish early development.
- This study provides the first in vivo knockdown data for zebrafish mvda.
- Findings may offer insights into the function of the human MVD gene in porokeratosis.
