Abstract
The lens plays an important role in the development of the optic cup[1,2]. Using the zebrafish as a model organism, questions regarding lens development can be addressed. The zebrafish is useful for genetic studies due to several advantageous characteristics, including small size, high fecundity, short lifecycle, and ease of care. Lens development occurs rapidly in zebrafish. By 72 hpf, the zebrafish lens is functionally mature [3]. Abundant genetic and molecular resources are available to support research in zebrafish. In addition, the similarity of the zebrafish eye to those of other vertebrates provides basis for its use as an excellent animal model of human defects[4-7]. Several zebrafish mutants exhibit lens abnormalities, including high levels of cell death, which in some cases leads to a complete degeneration of lens tissues [8]. To determine whether lens abnormalities are due to intrinsic causes or to defective interactions with the surrounding tissues, transplantation of a mutant lens into a wild-type eye is performed. Using fire-polished metal needles, mutant or wild-type lenses are carefully dissected from the donor animal, and transferred into the host. To distinguish wild-type and mutant tissues, a transgenic line is used as the donor. This line expresses membrane-bound GFP in all tissues, including the lens. This transplantation technique is an essential tool in the studies of zebrafish lens mutants.
Highlights
The lens plays an important role in the development of the optic cup[1,2]
The zebrafish is useful for genetic studies due to several advantageous characteristics, including small size, high fecundity, short lifecycle, and ease of care
Several zebrafish mutants exhibit lens abnormalities, including high levels of cell death, which in some cases leads to a complete degeneration of lens tissues [8]
Summary
Insert a glass capillary into the Pasteur pipette tip so that about half of its length is inside it. This will make it easier to immobilize the tungsten wire by melting it into the glass in the step. 3. insert a thin tungsten wire into the open end of the capillary. Melt the glass over the wire with a Bunsen burner. Using forceps, hold steady the glass end with the metal wire while twisting the other end of the needle with your hand. To create a sharpened needle, hold the tungsten wire over a Bunsen burner for 1-1.5 minutes, burning off the metal so a very fine tip is created. Check the quality of tip by microscropy.It is a good idea to sterilize both needles in the flame for a few seconds immediately before transplantation
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