Adriamycin induces H2AX phosphorylation in human spermatozoa.
نویسندگان
چکیده
AIM To investigate whether adriamycin induces DNA damage and the formation of gammaH2AX (the phosphorylated form of histone H2AX) foci in mature spermatozoa. METHODS Human spermatozoa were treated with adriamycin at different concentrations. gammaH2AX was analyzed by immunofluorescent staining and flow cytometry and double-strand breaks (DSB) were detected by the comet assay. RESULTS The neutral comet assay revealed that the treatment with adriamycin at 2 microg/mL for different times (0.5, 2, 8 and 24 h), or for 8 h at different concentrations (0.4, 2 and 10 microg/mL), induced significant DSB in spermatozoa. Immunofluorent staining and flow cytometry showed that the expression of gH2AX was increased in a dose-dependent and time-dependant manner after the treatment of adriamycin. Adriamycin also induced the concurrent appearance of DNA maintenance/repair proteins RAD50 and 53BP1 with gammaH2AX in spermatozoa. Wortmannin, an inhibitor of the phosphatidylinositol 3-kinase (PI3K) family, abolished the co-appearance of these two proteins with gammaH2AX. CONCLUSION Human mature spermatozoa have the same response to DSB-induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/repair proteins as somatic cells.
منابع مشابه
O-7: Insulin Exerts a Prosurvival Effect on Human Spermatozoa via Mechanisms That Involve the Stimulation of Akt Phosphorylation
Background: The purpose of this study was to examine the impact of Insulin (INS) on human sperm function, in light of a recent proteomic analysis indicating that these cells express the INS receptor (INSR). Materials and Methods: We used Western blot and Immunocytochemical analyses. Results: Immunocytochemical analyses confirmed the presence of INSR in human spermatozoa and localized this recep...
متن کاملIn vivo Analysis of H2AX Phosphorylation Induced by γ-Radiation
ABSTRACT Background and Objectives: Exposure to ionizing radiation in modern societies is inevitable and can cause a variety of adverse health effects such as cancer and birth defects. Therefore, a reliable, repeatable and sensitive method is required for evaluation of radiation exposure. The aim of this study was to determine the amount of hist...
متن کاملSerum starvation induces H2AX phosphorylation to regulate apoptosis via p38 MAPK pathway.
Phosphorylation of H2AX is believed to be associated with the repair of damaged DNA. Recent findings suggest a novel function of H2AX in cellular apoptosis. Specifically, it was shown that ultraviolet A-activated JNK phosphorylates H2AX to regulate apoptosis. Here we show that serum starvation induces H2AX phosphorylation and apoptosis independent of the JNK pathway. Serum starvation induced p3...
متن کاملMST1 promotes apoptosis through phosphorylation of histone H2AX.
MST1 (mammalian STE20-like kinase 1) is a serine/threonine kinase that is cleaved and activated by caspases during apoptosis. Overexpression of MST1 induces apoptotic morphological changes such as chromatin condensation, but the mechanism is not clear. Here we show that MST1 induces apoptotic chromatin condensation through its phosphorylation of histone H2AX at Ser-139. During etoposide-induced...
متن کاملPerturbed gap-filling synthesis in nucleotide excision repair causes histone H2AX phosphorylation in human quiescent cells.
Human histone H2AX is rapidly phosphorylated on serine 139 in response to DNA double-strand breaks and plays a crucial role in tethering the factors involved in DNA repair and damage signaling. Replication stress caused by hydroxyurea or UV also initiates H2AX phosphorylation in S-phase cells, although UV-induced H2AX phosphorylation in non-cycling cells has recently been observed. Here we stud...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Asian journal of andrology
دوره 10 5 شماره
صفحات -
تاریخ انتشار 2008