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Blood, 15 April 2005, Vol. 105, No. 8, pp. 3278-3285.
Prepublished online as a Blood First Edition Paper on December 23, 2004; DOI 10.1182/blood-2004-08-3073.
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NEOPLASIA
Suppression of apoptosis by bcl-2 overexpression in lymphoid cells of transgenic zebrafish
David M. Langenau,
Cicely Jette,
Stephane Berghmans,
Teresa Palomero,
John P. Kanki,
Jeffery L. Kutok, and
A. Thomas Look
From the Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School; and the Department of Pathology, Brigham & Women's Hospital, Boston, MA.
The zebrafish is an attractive vertebrate model for genetic studies of development, apoptosis, and cancer. Here we describe a transgenic zebrafish line in which T- and B-lymphoid cells express a fusion transgene that encodes the zebrafish bcl-2 protein fused to the enhanced green fluorescence protein (EGFP). Targeting EGFP-bcl-2 to the developing thymocytes of transgenic fish resulted in a 2.5-fold increase in thymocyte numbers and a 1.8-fold increase in GFP-labeled B cells in the kidney marrow. Fluorescent microscopic analysis of living rag2-EGFP-bcl-2 transgenic fish showed that their thymocytes were resistant to irradiation- and dexamethasone-induced apoptosis, when compared with control rag2-GFP transgenic zebrafish. To test the ability of bcl-2 to block irradiation-induced apoptosis in malignant cells, we compared the responsiveness of Myc-induced leukemias with and without EGFP-bcl-2 expression in living transgenic zebrafish. T-cell leukemias induced by the rag2-EGFP-Myc transgene were ablated by irradiation, whereas leukemias in double transgenic fish expressing both Myc and EGFP-bcl-2 were resistant to irradiation-induced apoptotic cell death. The forward genetic capacity of the zebrafish model system and the ability to monitor GFP-positive thymocytes in vivo make this an ideal transgenic line for modifier screens designed to identify genetic mutations or small molecules that modify bcl-2-mediated antiapoptotic pathways. (Blood. 2005;105:3278-3285)
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