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Biomedical subjects

E E Generoso

Publications and source records attributed to E E Generoso.

10 recordsLinked to original sources
PubMed1996

Detailed comparative map of human chromosome 19q and related regions of the mouse genome.

One of the larger contiguous blocks of mouse-human genomic homology includes the proximal portion of mouse chromosome 7 and the long arm of human chromosome 19. Previous studies have demonstrated the close relationship between the two regions, but have also indicated significant rearrangements in the relative orders of homologous mouse and human genes. Here we present the genetic locations of the homologs of 42 human chromosome 19q markers in the mouse, with an emphasis on genes also included in the human chromosome 19 physical map. Our results demonstrate that despite an overall inversion of sequences relative to the centromere, apparent "transpositions" of three gene-rich segments, and a local inversion of markers mapping near the 19q telomere, gene content, order, and spacing are remarkably well conserved throughout the lengths of these related mouse and human regions. Although most human 19q markers have remained genetically linked in mouse, one small human segment forms a separate region of homology between human chromosome 19q and mouse chromosome 17. Three of the four rearrangements of mouse versus human 19q sequences involve segments that are located directly adjacent to each other in 19q13.3-q13.4, suggesting either the coincident occurrence of these events or their common association with unstable DNA sequences. These data permit an unusually in-depth examination of this large region of mouse-human genomic homology and provide an important new tool to aid in the mapping of genes and associated phenotypes in both species.

Animals↗
PubMed1991

Ethylene oxide inhalation at different exposure-rates affects binding levels in mouse germ cells and hemoglobin. Possible explanation for the effect.

Male mice were exposed to [3H]EtO by inhalation at different exposure rates (300 parts per million (ppm) of EtO for 1 h: 150 ppm for 2 h: 75 ppm for 4 h). The total exposure was fixed at 300 ppm-h. The amount of EtO binding to developing spermatogenic stages, to sperm DNA, to testis DNA and to hemoglobin was then measured as a function of the EtO exposure rate. Generally, as the exposure rate increased there was an increase in the amount of EtO binding to the targets. For example, alkylation of sperm from the caudal epididymides 6 d posttreatment, of DNA from the vas sperm (averaged over 4 time points), of testis DNA (90 min posttreatment), and of hemoglobin (averaged over 4 time points), was 2.0 +/- 0.2 (SD), 1.8 +/- 0.4, 2.9 +/- 0.3, and 1.5 +/- 0.1 times greater, respectively, after an exposure to 300 ppm for 1 h than after an exposure to 75 ppm for 4 h. The testicular DNA from animals exposed to 300 ppm of [3H]EtO for 1 h was also analyzed for the presence of N7-hydroxyethylguanine (N7HEG) and O6-hydroxyethylguanine (O6HEG). The half-life (T1 2) of the N7HEG in the testis DNA was calculated to be 2.8 d. This lesion was removed relatively rapidly from the testis DNA and was probably excised by enzymatic repair. No formation of O6HEG was detected in any of the testis DNA samples analyzed. Additional experiments showed that the exposure rate effect was the result of less total EtO being taken in by the mice over long exposure times compared to that taken in during shorter exposure times at higher concentrations. This result argues against the idea that the exposure rate effect is the result of physiological/enzymological changes affecting transport or metabolism of the chemical within the animals under different exposure rate conditions.

Administration, Inhalation↗
PubMed1990

Acrylamide exposure induces a delayed unscheduled DNA synthesis in germ cells of male mice that is correlated with the temporal pattern of adduct formation in testis DNA.

A study of meiotic and postmeiotic germ-cell-stage sensitivity of male mice to induction of unscheduled DNA synthesis (UDS) by acrylamide showed that DNA repair could be detected in early spermatocytes (after the last scheduled DNA synthesis) through about mid-spermatid stages. No DNA repair could be detected in later stages. The maximum UDS response was observed 6 hr after i.p. exposure and was about 5 times greater than the response measured immediately after treatment. This is the longest delay between chemical treatment and maximum UDS response yet observed in mouse germ cells. There was a linear relationship between the UDS response and acrylamide exposure from 7.8 to 125 mg/kg. By using 14C-labeled acrylamide it was determined that the temporal pattern of adduct formation in testes DNA paralleled that of the UDS response, with maximum binding occurring 4 to 6 hr after exposure. In contrast, the temporal pattern of adduct formation in liver DNA showed maximum binding within 1 to 2 hr after exposure and was an order of magnitude greater than that found for the testis DNA.

Acrylamide↗
PubMed1990

Measurement of DNA breakage in specific germ-cell stages of male mice exposed to acrylamide, using an alkaline-elution procedure.

DNA breakage in spermiogenic stages of the mouse was studied after exposure to acrylamide (AA), using an alkaline-elution technique. At daily intervals over a 3-week period following i.p. injection of 100 mg AA/kg, mature spermatozoa were recovered from treated ([3H]dThd-labeled) and control ([14C]dThd-labeled) animals, and were lysed together on polycarbonate filters; the DNA was eluted with a high-pH (12.0) buffer. Elution of germ-cell DNA from AA-exposed animals increased (more DNA-strand breaks) in stages sensitive to the dominant-lethal effects of AA (late spermatids to early spermatozoa) (Shelby et al., 1986). The stage-related pattern of AA-induced DNA breakage also paralleled the pattern of sperm alkylation and protamine alkylation found to be produced by AA (Sega et al., 1989). While dominant-lethal damage from AA exposure is greatest in the spermatids and early spermatozoa, no such damage was observed in pachytene spermatocytes and early spermatids (Shelby et al., 1986). Therefore, AA-induced DNA breakage was also studied directly in pachytene spermatocytes and in early spermatids at short intervals (up to 4 days) after exposure. DNA breakage was clearly detected in these cell stages, with maximum breakage occurring at 1 day after treatment. At later times, the breakage gradually decreased, presumably as a result of DNA repair. By the time these cell stages gave rise to functional spermatozoa, DNA breaks that could have produced dominant-lethal events had apparently been reduced to a level where no genetic effect could be observed.

Acrylamides↗
PubMed1988

Measurement of DNA breakage in spermiogenic germ-cell stages of mice exposed to ethylene oxide, using an alkaline elution procedure.

DNA breakage in spermiogenic stages of the mouse was studied after exposure to ethylene oxide (EtO), using an alkaline elution technique. At daily intervals over a 23-day period following i.p. injection of 100 mg EtO/kg, mature spermatozoa were recovered from treated ([3H]dThd-labeled) and control ([14C]dThd-labeled) animals, lysed together on polycarbonate filters, and the DNA was eluted with a high pH (12.2) buffer. Elution of germ-cell DNA from EtO-exposed animals increased (more DNA strand breaks) in stages sensitive to the genetic effects of EtO (late spermatids to early spermatozoa). The stage-related pattern of EtO-induced DNA breakage paralleled the pattern of sperm alkylation and protamine alkylation found to be produced by EtO in an earlier study (Sega and Owens, 1987). At 9 days posttreatment (sperm sampled were in late-spermatid stages at the time of EtO exposure) the amount of sperm DNA eluted did not change significantly over a pH range of 11.6-12.8, indicating that, at the time of assay, DNA breaks were already present in the sperm.

Animals↗
PubMed1980

Time of recombination in the Drosophila melanogaster oocyte. II. Electron microscopic and genetic studies of a temperature-sensitive recombination mutant.

A test has been carried out to determine if the restrictive temperature (31 degrees) acts to reduce recombination in the temperature-sensitive recombination-deficient genotype rec-l26/rec-l16 by reducing or eliminating the synaptonemal complex. Measurements of the length of synaptonemal complexes in heat-treated and untreated stage 1 oocytes, following termination of the temperature-sensitive period, reveal less than a 5% difference, with the greater length present in the treated oocytes. Alterations are not observed in synaptonemal complex distribution within the nucleus or in its fine structure. Parallel genetic studies confirm earlier observations that the restrictive temperature, whose action is confined to a 36-h sensitive period virtually coextensive with premeiotic-S, drastically reduces recombination to approximately 10% of normal. The results are most simply interpreted to mean that the restrictive temperatures acts directly on the recombination process.

Animals↗
PubMed1980

Synaptonemal complexes at premeiotic interphase in the mouse spermatocyte.

Male mice were injected intraperitoneally with 125 microCi (1 Ci = 3.7 X 10(10) becquerels) of [3H]thymidine at 1-hr intervals and killed 1 hr after the second injection. Testes were prepared for bright-field and electron microscopic autoradiography. Primary spermatocytes, identified by light microscopy to be at the premeiotic interphase stage, were found to be heavily labeled. Electron microscopic examination disclosed the coincidental occurrence of synaptonemal complexes and label within the nuclei of premeiotic interphase spermatocytes, indicating synapses of homologues had begun during the S phase. The significance of this finding for the traditional view of meiosis is discussed.

Animals↗
PubMedDate not supplied

The use of alkaline elution procedures to measure DNA damage in spermiogenic stages of mice exposed to methyl methanesulfonate.

The ability of methyl methanesulfonate (MMS) to induce DNA breakage in spermiogenic stages of the mouse was studied using an alkaline elution technique. At daily intervals over a 3-week period following i.p. injection of 50 mg MMS/kg, mature spermatozoa were recovered from treated (3H-labeled) and control (14C-labeled) animals, lysed together on polycarbonate filters, and eluted with a high pH (12.2) buffer. Elution of germ-cell DNA from MMS-treated animals was found to increase in stages in which genetic damage from MMS is greatest. In general, the pattern of DNA elution from treated, spermiogenic stages paralleled the pattern of sensitivity to dominant lethals, specific-locus mutations and heritable translocations found by other investigators. It also paralleled the pattern of sperm-head methylation and protamine methylation measured in an earlier study (Sega and Owens, 1983). At 9 days post treatment (sperm sampled were in mid-to late-spermatid stages at the time of MMS exposure) the elution of sperm DNA did not change significantly over a pH range of 11.6-12.8, suggesting that, at the time of assay, DNA breaks were already present in the sperm. Because of the parallelism found between increased sperm DNA elution and increased genetic damage after mutagen treatment, alkaline elution may prove useful in monitoring potential genetic damage in human sperm.

Animals↗
PubMedDate not supplied

Inhalation exposure-rate of ethylene oxide affects the level of DNA breakage and unscheduled DNA synthesis in spermiogenic stages of the mouse.

The effect of ethylene oxide (EtO) inhalation-exposure rate on the induction of DNA breakage in late spermatids and on unscheduled DNA synthesis (UDS) in early spermatids was studied. The exposures were 450 parts per million (ppm) for 4 h, 900 ppm for 2 h, and 1800 ppm for 1 h. Thus, the total exposure was always 1800 ppm-h. Both DNA breakage and UDS were found to increase by a factor of approximately 3 in going from the low to high EtO concentration, suggesting that the molecular dose of EtO to the testis had increased by a similar factor. Our results are consistent with the EtO exposure-rate effect found by Generoso et al. (1986) for induction of dominant-lethal mutations in late spermatids and early spermatozoa.

Administration, Inhalation↗