Lens Transplantation in Zebrafish and its Application in the Analysis of Eye Mutants

Yan Zhang&#160;; Kyle McCulloch; Jarema Malicki

doi:10.3791/1258

Lens Transplantation in Zebrafish and its Application in the Analysis of Eye Mutants

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Lens Transplantation in Zebrafish and its Application in the Analysis of Eye Mutants

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10:39 min

June 1, 2009

DOI: 10.3791/1258-v

Yan Zhang ^1,2, Kyle McCulloch², Jarema Malicki²

¹The Second Teaching Hospital of Jilin University, ²Department of Ophthalmology,Harvard Medical School

Lens development involves interactions with other tissues. Several zebrafish eye mutants are characterized by an abnormally small lens size. Here we demonstrate a lens transplantation experiment to determine whether this phenotype is due to intrinsic causes or defective interactions with tissues that surround the lens.

To begin lens transplantation in zebrafish GFP expressing transgenic donors and wild tap recipient hosts are first incubated for 30 minutes in separate pet dishes containing calcium free zebrafish ringer solution. Fish larvae are then embedded in two parallel lines in low melting point agros. One line A is for donors and the other B.For hosts, the entire donor eye is removed while only the lens is removed from the host.

The tissues surrounding the donor lens are removed before placing the lens in the host eye. After transplantation, fis are stored in labeled 24 well plates. The transplanted lens should dis display GFP fluorescence in the host.

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