Aneuploidy in late-step spermatids of mice detected by two-chromosome fluorescencein situ hybridization
| Autor: | D. Pinkel, Jack B. Bishop, Xiu Lowe, Andrew J. Wyrobek |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
Male
endocrine system X Chromosome Centromere Aneuploidy Bone Marrow Cells Haploidy Biology Chromosomes Mice Species Specificity Genetics medicine Animals Humans Lymphocytes Metaphase In Situ Hybridization Fluorescence Meiotic metaphase II Mice Inbred C3H medicine.diagnostic_test urogenital system Hybridization probe Chromosome Cell Biology medicine.disease Spermatids Molecular biology Sperm Mice Inbred C57BL Meiosis Phenotype Organ Specificity DNA Probes Molecular probe Developmental Biology Fluorescence in situ hybridization |
| Zdroj: | Molecular Reproduction and Development. 40:259-266 |
| ISSN: | 1098-2795 1040-452X |
| DOI: | 10.1002/mrd.1080400216 |
| Popis: | A multicolor procedure employing fluorescence in situ hybridization is described for detecting chromosomal domains and germinal aneuploidy in late-step spermatids in mice using DNA probes specific for repetitive sequences near the centromeres of chromosomes 8 and X. These probes were nick-translated with biotin- or digoxigenin-labeled nucleotides, and were detected with FITC or rhodamine. Probe and hybridization specificities were confirmed using metaphase chromosomes from spleen and bone marrow cells as well as from primary and secondary spermatocytes. Late-step spermatids, identified in testicular preparations by their hooked shape, yielded compact fluorescence domains in ∼ 50% and > 99% of cells when hybridized with probes for chromosomes X and 8, respectively. In a survey of > 80,000 late-step spermatids from 8 healthy young adult C57BL/6 or B6C3F1 mice, ∼ 3/10,000 spermatids had fluorescence phenotypes indicative of X-X or 8–8 hyperhaploidy. These frequencies are consistent with published frequencies of aneuploidy in meiotic metaphase II and first cleavage metaphases of the mouse, providing preliminary validation of sperm hybridization for the detection of aneuploidy. No significant animal or strain differences were observed. In addition, the hyperhaploidy frequencies for murine spermatids were indistinguishable for those for sperm from healthy men obtained by a similar hybridization procedure. These procedures for detecting aneuploid male gametes are examples of “bridging biomarkers” between human and animal studies. They have promising applications for investigations of the genetic, reproductive, and toxicological factors leading to abnormal reproductive outcomes of paternal origin. © 1995 Wiley-Liss, Inc. |
| Databáze: | OpenAIRE |
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