title: Gonadal Mosaicism Induced by Chemical Treatment of Sperm in Drosophila melanogaster.
abstract: Accurate interpretation of forward genetic screens of chromosomes exposed in mature spermatozoa to a mutagenic chemical requires understanding-incomplete to date-of how exposed chromosomes and their replicas proceed through early development stages from the fertilized ovum to establishment of the germline of the treated male's offspring. We describe a model for early embryonic development and establishment of the germline of Drosophila melanogaster and a model-validating experiment. Our model proposes that, barring repair, DNA strands modified by treatment with alkylating agents are stable and mutagenic. Each replication of an alkylated strand can result in misreplication and a mutant-bearing daughter nucleus. Daughter nuclei thenceforth replicate faithfully and their descendants comprise the embryonic syncytium. Of the 256 nuclei present after the eighth division, several migrate into the polar plasm at the posterior end of the embryo to found the germline. Based upon distribution of descendants of the alkylated strands, the misreplication rate, and the number of nuclei selected as germline progenitors, the frequency of gonadal mosaicism is predictable. Experimentally, we tracked chromosomes 2 and 3 from EMS-treated sperm through a number of generations, to characterize autosomal recessive lethal mutations and infer gonadal genetic content of the sons of treated males. Over 50% of 106 sons bore germlines that were singly, doubly, or triply mosaic for chromosome 2 or chromosome 3. These findings were consistent with our model, assuming a rate of misreplication between 0.65 and 0.80 at each replication of an alkylated strand. Crossing treated males to mismatch-repair-deficient females had no apparent effect on mutation rate.
authors.affiliation: Department of Cell and Developmental Biology, University of California, San Diego, California 92093.
authors.forename: Dan L
authors.initials: DL
authors.lastname: Lindsley
authors.affiliation: Department of Cell and Developmental Biology, University of California, San Diego, California 92093.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Hardy
authors.affiliation: Universidad Autónoma, de Madrid, 98049, Madrid, Spain.
authors.forename: Pedro
authors.initials: P
authors.lastname: Ripoll
authors.affiliation: Cisco Systems, San Jose, California 95134-1718 dlindsley@ucsd.edu.
authors.forename: Dart
authors.initials: D
authors.lastname: Lindsley
mesh_terms: D000477:Alkylating Agents / Q000494:pharmacology*; D000818:Animals; D003433:Crosses, Genetic; D053843:DNA Mismatch Repair; D004331:Drosophila melanogaster; D005020:Ethyl Methanesulfonate / Q000494:pharmacology*; D005260:Female; D005804:Genes, Lethal; D018095:Germ-Line Mutation; D007178:Inbreeding; D008297:Male; D008957:Models, Genetic; D009030:Mosaicism / Q000187:drug effects*; D016296:Mutagenesis; D009153:Mutagens / Q000494:pharmacology*; D013094:Spermatozoa / Q000187:drug effects*; D013234:Stem Cells
chemical_list: D000477:Alkylating Agents; D009153:Mutagens; D005020:Ethyl Methanesulfonate
keywords: male sterility; meiotic mutants; paternal effect mutants; spermatogenesis