Articles by Huijia Wang in JoVE
Microinjeção de embriões Medaka para uso como um organismo modelo genético Sean R. Porazinski1, Huijia Wang1, Makoto Furutani-Seiki1 1Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath Medaka e peixe-zebra são complementares para análise genética de funções genoma dos vertebrados. Este protocolo destaca os pontos chave para a microinjeção de sucesso em embriões medaka, uma técnica importante para a análise embriológica e genética utilizando medaka zebrafish e em um laboratório.
Dechorionation de Embriões Medaka e Transplante de Células de Geração de Quimeras Sean R. Porazinski1, Huijia Wang1, Makoto Furutani-Seiki1 1Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath Devido ao córion duro e macio embriões, manipulação de embriões medaka está mais envolvido do que em zebrafish. Este vídeo mostra passo-a-passo os procedimentos de como manipular embriões medaka, incluindo dechorionation, montagem em agarose para a imagem latente e transplante de células para a produção de quimeras. Estes procedimentos são essenciais para a utilização medaka zebrafish e em um laboratório para tirar o máximo proveito de seus recursos complementares para a análise genética de funções genoma dos vertebrados.
Other articles by Huijia Wang on PubMed
Insufficiency of BUBR1, a Mitotic Spindle Checkpoint Regulator, Causes Impaired Ciliogenesis in Vertebrates Human Molecular Genetics. May, 2011 | Pubmed ID: 21389084 Budding uninhibited by benzimidazole-related 1 (BUBR1) is a central molecule of the spindle assembly checkpoint. Germline mutations in the budding uninhibited by benzimidazoles 1 homolog beta gene encoding BUBR1 cause premature chromatid separation (mosaic variegated aneuploidy) [PCS (MVA)] syndrome, which is characterized by constitutional aneuploidy and a high risk of childhood cancer. Patients with the syndrome often develop Dandy-Walker complex and polycystic kidneys; implying a critical role of BUBR1 in morphogenesis. However, little is known about the function of BUBR1 other than mitotic control. Here, we report that BUBR1 is essential for the primary cilium formation, and that the PCS (MVA) syndrome is thus a novel ciliopathy. Morpholino knockdown of bubr1 in medaka fish also caused ciliary dysfunction characterized by defects in cerebellar development and perturbed left-right asymmetry of the embryo. Biochemical analyses demonstrated that BUBR1 is required for ubiquitin-mediated proteasomal degradation of cell division cycle protein 20 in the G0 phase and maintains anaphase-promoting complex/cyclosome-CDC20 homolog 1 activity that regulates the optimal level of dishevelled for ciliogenesis.
Essential Techniques for Introducing Medaka to a Zebrafish Laboratory--towards the Combined Use of Medaka and Zebrafish for Further Genetic Dissection of the Function of the Vertebrate Genome Methods in Molecular Biology (Clifton, N.J.). 2011 | Pubmed ID: 21805266 The medaka, Oryzias latipes, a small egg-laying freshwater fish, is one of the three vertebrate model organisms in which genome-wide phenotype-driven mutant screens have been carried out. Despite a number of large-scale screens in zebrafish, a substantial number of mutants with new distinct phenotypes were identified in similar large-scale screens in the medaka. This observed difference in phenotype is due to the two species having a unique combination of genetic, biological and evolutional properties. The two genetic models share a whole-genome duplication event over that of tetrapods; however, each has independently specialized or lost the function of one of the two paralogues. The two fish species complement each other as genetic systems as straightforward comparison of phenotypes, ease of side-by-side analysis using the same techniques and simple and inexpensive husbandry of mutants make these small teleosts quite powerful in combination. Furthermore, both have draft genome sequences and bioinformatic tools available that facilitate further genetic dissection including whole-genome approaches. Together with the gene-driven approach to generate gene knockout mutants of the fish models, the two fish models complement the mouse in genetically dissecting vertebrate genome functions. The external embryogenesis and transparent embryos of the fish allow systematic isolation of embryonic lethal mutations, the most difficult targets in mammalian mutant screens. This chapter will describe how to work with both medaka and zebrafish almost as one species in a lab, focusing on medaka and highlighting the differences between the medaka and zebrafish systems.