title: Detection of DNA damage caused by cryopreservation using a modified SCGE in large yellow croaker, Pseudosciaena crocea.
abstract: We used single-cell gel electrophoresis (SCGE) to detect the integrity of sperm DNA of the teleost large yellow croaker, Pseudosciaena crocea, cryopreserved with Cortland solution and a range of 5% to 30% DMSO concentrations in order to test how sperm cryopreservation affected the DNA stability of nuclei. Electrophoresis was conducted for 60 min at 130 mA and 15 V. The comet images were analyzed with software CometScore 1.5, and parameters such as comet length, tail length and percentage DNA in the tail were obtained. Then the comet rate and damage coefficient were calculated. Results demonstrated that there were no significant differences in motility, comet rate and damage coefficient between fresh sperm and cryopreserved sperm stored in 5%, 10%, 15% and 20% DMSO, while the sperm cryopreserved with 25% and 30% DMSO had a lower motility, higher comet length and damage coefficients than those of fresh sperm. There was a positive correlation between comet rate of cryopreserved sperm and the concentration of DMSO. Our results demonstrate that toxicity of the cryoprotectant is the main cause of DNA damage in cryopreserved sperm nuclei.
authors.affiliation: Zhejiang University School of Medicine Department of Reproductive Endocrinology, Key Laboratory of Reproductive Genetics, Women's Hospital 1 Xueshi Road Hangzhou 310006 China.
authors.forename: Xiang-Rong
authors.initials: XR
authors.lastname: Xu
authors.affiliation: Zhejiang University The First Affiliated Hospital, College of Medicine Hangzhou 310003 China.
authors.forename: Fu-Qing
authors.initials: FQ
authors.lastname: Tan
authors.affiliation: Ningbo University Key Laboratory of Applied Marine Biotechnology by the Ministry of Education Ningbo 315211 China.
authors.forename: Jun-Quan
authors.initials: JQ
authors.lastname: Zhu
authors.affiliation: Ningbo University Key Laboratory of Applied Marine Biotechnology by the Ministry of Education Ningbo 315211 China.
authors.forename: Ting
authors.initials: T
authors.lastname: Ye
authors.affiliation: Ningbo University Key Laboratory of Applied Marine Biotechnology by the Ministry of Education Ningbo 315211 China.
authors.forename: Chun-Lin
authors.initials: CL
authors.lastname: Wang
authors.affiliation: Ningbo University Key Laboratory of Applied Marine Biotechnology by the Ministry of Education Ningbo 315211 China.
authors.forename: Yi-Feng
authors.initials: YF
authors.lastname: Zhu
authors.affiliation: KMU - Kaohsiung Medical University Dept. of Biomedical Science and Environmental Biology No. 100, Shin-Chuan 1st Road Kaohsiung 80708 Taiwan R.O.C.
authors.forename: Hans-Uwe
authors.initials: HU
authors.lastname: Dahms
authors.affiliation: Zhejiang University School of Medicine Department of Reproductive Endocrinology, Key Laboratory of Reproductive Genetics, Women's Hospital 1 Xueshi Road Hangzhou 310006 China.
authors.forename: Fan
authors.initials: F
authors.lastname: Jin
authors.affiliation: Zhejiang University The Sperm Laboratory, College of Life Science Hangzhou 310012 China.
authors.forename: Wan-Xi
authors.initials: WX
authors.lastname: Yang
mesh_terms: D000818:Animals; D020552:Comet Assay; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology* / Q000633:toxicity; D004249:DNA Damage; D004121:Dimethyl Sulfoxide / Q000494:pharmacology* / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D008297:Male; D010473:Perciformes / Q000235:genetics* / Q000378:metabolism; D012662:Semen Preservation / Q000009:adverse effects / Q000379:methods*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology
chemical_list: D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide
keywords: Pseudosciaena crocea; cryopreservation; genetic damage; single-cell gel electrophoresis; sperm