वयस्क Zebrafish में बायोकेमिकल विश्लेषण के लिए रक्त संग्रह
Summary
इस कागज Zebrafish की पृष्ठीय महाधमनी से रक्त का संग्रह के लिए एक तकनीक प्रस्तुत करता है. यह भी जैव रासायनिक विश्लेषण में कोलेस्ट्रॉल और ट्राइग्लिसराइड का स्तर निर्धारित करने के लिए परीक्षण के रूप में उपयोग के लिए सीरम प्राप्त करने के लिए निर्देश प्रदान करता है.
Abstract
The zebrafish has been used as an animal model for studies of several human diseases. It can serve as a powerful preclinical platform for studies of molecular events and therapeutic strategies as well as for evaluating the physiological mechanisms of some pathologies1.
There are relatively few publications related to adult zebrafish physiology of organs and systems2, which may lead researchers to infer that the basic techniques needed to allow the exploration of zebrafish systems are lacking3. Hematologic biochemical values of zebrafish were first reported in 2003 by Murtha and colleagues4 who employed a blood collection technique first described by Jagadeeswaran and colleagues in 1999. Briefly, blood was collected via a micropipette tip through a lateral incision, approximately 0.3 cm in length, in the region of the dorsal aorta5. Because of the minute dimensions involved, this is a high-precision technique requiring a highly skilled practitioner. The same technique was used by the same group in another publication in that same year6. In 2010, Eames and colleagues assessed whole blood glucose levels in zebrafish7. They gained access to the blood by performing decapitations with scissors and then inserting a heparinized microcapillary collection tube into the pectoral articulation. They mention difficulties with hemolysis that were solved with an appropriate storage temperature based on the work Kilpatrick et al.8. When attempting to use Jagadeeswaran’s technique in our laboratory, we found that it was difficult to make the incision in precisely the right place as not to allow a significant amount of blood to be lost before collection could be started.
Recently, Gupta et al.9 described how to dissect adult zebrafish organs, Kinkle et al.10 described how to perform intraperitoneal injections, and Pugach et al.11 described how to perform retro-orbital injections. However, more work is needed to more fully explore basic techniques for research in zebrafish.
The small size of zebrafish presents challenges for researchers using it as an experimental model. Furthermore, given this smallness of scale, it is important that simple techniques are developed to enable researchers to explore the advantages of the zebrafish model.
Protocol
Discussion
यह पत्र एक सरल तकनीक है कि आगे और zebrafish प्रयोगों में सीरम विश्लेषण रक्त की अनुमति देता है प्रस्तुत करता है. इस तकनीक को भविष्य zebrafish hematologic रक्त पैरामीटर डेटा की आवश्यकता के अध्ययन के लिए योगदान करने की क्षमत?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
/ FIPE HCPA है Fundo डे Incentivo एक Pesquisa ई Eventos
Capes – Coordenação डे Aperfeiçoamento डे Pessoal डे Nível सुपीरियर
Materials
| Name of the reagent | Company | Catalogue number | Comments |
| Low retention tips | Applied Biosystems | 022493020 | |
| Eppendorf Centrifuge 5415D | Eppendorf | Discontinued |
References
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- Kinkel, M. D., Eames, S. C., Philipson, L. H., Prince, V. E. Intraperitoneal Injection into Adult Zebrafish. J. Vis. Exp. (42), e2126 (2010).
- Pugach, E. K., Li, P., White, R., Zon, L. Retro-orbital Injection in Adult Zebrafish. J. Vis. Exp. (34), e1645 (2009).
