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Main Title Incidence of Chromosome Aberrations in Mammalian Sperm Stained with Hoechst 33342 and UV-Laser Irradiated during Flow Sorting.
Author Libbus, B. L. ; Perreault, S. D. ; Johnson, L. A. ; Pinkel, D. ;
CORP Author Health Effects Research Lab., Research Triangle Park, NC. ;Agricultural Research Service, Beltsville, MD. ;Lawrence Livermore National Lab., CA. Biomedical Sciences Div.
Year Published 1987
Report Number EPA/600/J-87/268;
Stock Number PB88-171251
Additional Subjects Radiation effects ; Chromosomes ; Mammals ; Sperm ; Reprints ; Ultraviolet laser radiation ; Flow sorting ; Hoechst 33342
Library Call Number Additional Info Location Last
NTIS  PB88-171251 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 12p
The separation of two sperm populations is possible using the technique of flow sorting, provided that a significant difference exists in the DNA content of X- and Y-bearing sperm. In order to ascertain whether or not chromosome damage was induced in sorted sperm, chromosome preparations were made from isolated sperm that had been microinjected into hamster eggs. While egg chromosomes exhibited a low frequency of chromosome aberrations, ranging from 4 to 7%, a large proportion of sperm cells exhibited chromosome damage. Between 29% of unstained and unsorted sperm and 38% of stained and unsorted sperm exhibited some type of chromosomal abnormality and this proportion increased to 50% in sorted sperm. If only damaged sperm nuclei are considered, the two unsorted sperm groups had a mean of 0.6 breaks, 0.8 triradial exchanges, and 0.2 quadriradial exchanges per nucleus. However, sorted sperm, which were stained with a fluorochrome and exposed to UV-laser irradiation, exhibited a mean of 2.9 breaks, 2.6 triradial, and 1.9 quadriradial exchanges per nucleus in which damage occurred. These observations indicate that the treatments and manipulations to which sperm nuclei are subjected during flow sorting cause chromosomal aberrations, and that exposure of the cells to UV-laser irradiation contributes substantially to the chromosome damage observed.