title: Proteomic analysis for testis of mice exposed to carbon ion radiation.
abstract: This paper investigates the mechanism of action of heavy ion radiation (HIR) on mouse testes. The testes of male mice subjected to whole body irradiation with carbon ion beam (0.5 and 4Gy) were analyzed at 7days after irradiation. A two-dimensional gel electrophoresis approach was employed to investigate the alteration of protein expression in the testes. Spot detection and matching were performed using the PDQuest 8.0 software. A difference of more than threefold in protein quantity (normalized spot volume) is the standard for detecting differentially expressed protein spots. A total of 11 differentially expressed proteins were found. Protein identification was performed using matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF-TOF). Nine specific proteins were identified by searching the protein sequence database of the National Center for Biotechnology Information. These proteins were found involved in molecular chaperones, metabolic enzymes, oxidative stress, sperm function, and spermatogenic cell proliferation. HIR decreased glutathione activity and increased malondialdehyde content in the testes. Given that Pin1 is related to the cell cycle and that proliferation is affected by spermatogenesis, we analyzed testicular histological changes and Pin1 protein expression through immunoblotting and immunofluorescence. Alterations of multiple pathways may be associated with HIR toxicity to the testes. Our findings are essential for studies on the development, biology, and pathology of mouse testes after HIR in space or radiotherapy.
authors.affiliation: Department of Heavy Ion Radiation Medicine, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000, China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
authors.forename: Yi
authors.initials: Y
authors.lastname: Xie
authors.forename: Yuxuan
authors.initials: Y
authors.lastname: He
authors.forename: Guoying
authors.initials: G
authors.lastname: Miao
authors.forename: Lina
authors.initials: L
authors.lastname: Yang
authors.forename: Cuixia
authors.initials: C
authors.lastname: Di
authors.forename: Yang
authors.initials: Y
authors.lastname: He
mesh_terms: D000818:Animals; D002244:Carbon / Q000633:toxicity*; D018797:Cell Cycle Proteins / Q000096:biosynthesis / Q000235:genetics; D002454:Cell Differentiation / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling / Q000379:methods*; D005978:Glutathione / Q000032:analysis; D020450:Heavy Ions / Q000009:adverse effects*; D015227:Lipid Peroxidation / Q000528:radiation effects; D008297:Male; D008315:Malondialdehyde / Q000032:analysis; D051379:Mice; D008856:Microscopy, Fluorescence; D018832:Molecular Chaperones / Q000096:biosynthesis / Q000235:genetics; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D018384:Oxidative Stress / Q000235:genetics / Q000528:radiation effects; D019696:Peptidylprolyl Isomerase / Q000096:biosynthesis / Q000235:genetics; D014176:Protein Biosynthesis / Q000528:radiation effects*; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013091:Spermatogenesis / Q000235:genetics; D013382:Subtraction Technique; D013737:Testis / Q000378:metabolism / Q000528:radiation effects* / Q000648:ultrastructure; D014916:Whole-Body Irradiation
chemical_list: D018797:Cell Cycle Proteins; D018832:Molecular Chaperones; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D008315:Malondialdehyde; D002244:Carbon; D019696:Peptidylprolyl Isomerase; C000606850:Pin1 protein, mouse; D005978:Glutathione
keywords: 2-DE; CCB; CCT; Carbon ion radiation; DAB; GSH; GSTM7; HIR; HIRFL; HRP; HSPa4L; Heavy Ion Research Facility in Lanzhou; MALDI-TOF-TOF; MDA; PDC; PDH-E1; PDIA3; PEBPS; PGP; Pin1; Proteomics; TBS; Testis; chaperonin containing TCP-1; colloidal coomassie blue G 250; diaminobenzidine; glutathione; glutathione S-transferase Mu 7; heat shock 70kDa protein 4L; heavy ions radiation; horseradish peroxidase; malondialdehyde; matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry; peptidyl-prolylcis-trans isomerase1; phosphatidylethanolamine binding proteins; phosphoglycolate phosphatase; protein disulfide-isomerase A3 precursor; pyruvate dehydrogenase E1; pyruvate dehydrogenase complex; tris buffer saline; two-dimensional gel electrophoresis