Our major goal is to understand how genes control the structure, function, and development of the nervous system in a vertebrate. Initially, we plan to give the highest priority to the isolation and study of mutations which affect the structures of photoreceptor cells in the retina of the eye of the zebra fish (Brachydanio rerio.) A corollary goal of our work is to develop rapid and efficient procedures that will permit a variety of genetic analyses in a vertebrate at the level of the whole organism. Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. We have perfected methods for making homozygotes in zebra fish: Sperm whose genetic contribution has been eliminated by ultraviolet irradiation are used to activate eggs. The maternal haploid set of chromosomes is allowed to replicate once, and partitioning of the duplicated chromosomes into two cells is prevented by hydrostatic pressure or heat shock. A cell with two identical sets of chromosomes is thereby produced. We propose to mutagenize mother fish during their early development with ethylnitrosourea (ENU). Among the progeny, clones of mutants will be recognized by use of a vision-dependent feeding test. Mutants will be characterized through optomotor and optokinetic tests, flicker fusion frequency, electroretinogram shape, and retinal anatomy. In addition to the direct screening of mutagenized progeny, gamma-ray-induced mutations (possibly long deletions) with specific retinal lesions will be used to select ENU-induced point mutations. Through the study of these mutants we hope to learn what kinds of alterations are specific to retinal photoreceptor cells, what the developmental, structural, functional, and behavioral consequences of the mutations are, how mutation affects other cell types, and what the molecular nature of the mutations are.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM022731-10
Application #
3271274
Study Section
Genetics Study Section (GEN)
Project Start
1979-03-01
Project End
1987-02-28
Budget Start
1985-03-01
Budget End
1987-02-28
Support Year
10
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Oregon
Department
Type
Graduate Schools
DUNS #
948117312
City
Eugene
State
OR
Country
United States
Zip Code
97403
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Grunwald, D J; Streisinger, G (1992) Induction of recessive lethal and specific locus mutations in the zebrafish with ethyl nitrosourea. Genet Res 59:103-16
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Grunwald, D J; Kimmel, C B; Westerfield, M et al. (1988) A neural degeneration mutation that spares primary neurons in the zebrafish. Dev Biol 126:115-28
Kljavin, I J (1987) Early development of photoreceptors in the ventral retina of the zebrafish embryo. J Comp Neurol 260:461-71

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