title: Germ cell responses to doxorubicin exposure in vitro.
abstract: Anthracyclines such as doxorubicin (Dox), widely used to treat various types of tumours, may result in induced testicular toxicity and oxidative stress. The present investigation was designed to determine whether exposure of isolated and purified mouse germ cells to Dox induces DNA damage in the form of strand breaks (presumably) resulting in apoptosis and to investigate the relative sensitivity of specific cell types. DNA damage was assessed using the Comet assay and the presence of apoptosis was determined by TUNEL assay. Isolated mouse germ cells were treated with different concentrations (0.05, 0.5 and 1mM, respectively) of Dox, and fixed 1h after treatment. The incidences of both DNA damage shown by single cell gel-electrophoresis and of apoptosis increased significantly in each specific cell type in a concentration-dependent manner. The DNA damage and apoptosis incidences gradually increased with concentration from 0.05 to 1mM with Dox. Our results indicate that apoptosis plays a vital role in the induction of germ cell phase-specific toxicity caused by Dox with pre-meiotically and meiotically dividing spermatogonia and spermatocytes respectively as highly susceptible target cells.
authors.affiliation: School of Medical Sciences, Faculty of Life Sciences, University of Bradford, Bradford, Richmond Road, West Yorkshire, BD7 1DP, UK.
authors.forename: Khaled
authors.initials: K
authors.lastname: Habas
authors.affiliation: School of Medical Sciences, Faculty of Life Sciences, University of Bradford, Bradford, Richmond Road, West Yorkshire, BD7 1DP, UK.
authors.forename: Diana
authors.initials: D
authors.lastname: Anderson
authors.affiliation: School of Medical Sciences, Faculty of Life Sciences, University of Bradford, Bradford, Richmond Road, West Yorkshire, BD7 1DP, UK. Electronic address: m.h.brinkworth@bradford.ac.uk.
authors.forename: Martin H
authors.initials: MH
authors.lastname: Brinkworth
mesh_terms: D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D020552:Comet Assay; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000633:toxicity*; D020287:In Situ Nick-End Labeling; D008297:Male; D008815:Mice, Inbred Strains; D018384:Oxidative Stress / Q000187:drug effects; D013087:Spermatids / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D013090:Spermatocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D013093:Spermatogonia / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D013094:Spermatozoa / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology
chemical_list: D000903:Antibiotics, Antineoplastic; D004317:Doxorubicin
keywords: Apoptosis; Comet assay; DNA damage; Doxorubicin; Male germ cells; TUNEL assay