ABSTRACT
In present work we studied DNA damage in human and bovine lymphocytes and spermatozoa by means of single cell gel electrophoresis followed by silver staining. The spontaneous frequency of DNA damage estimated manually in spermatozoa from healthy donors did not exceed 9% (on average -- 4.8 +/- 1.2%). The frequency of DNA damages in bull sperm after short (less than a year) and long period (more than 20 years) of cryopreservation was assessed as 3.1 +/- 0.9 and 4.3 +/- 0.5%, correspondingly. The comparative estimation of DNA damages in lymphocytes followed by silver staining is a valuable tool to estimate DNA damage in populations of somatic and reproductive cells.
Subject(s)
DNA Damage , Lymphocytes/chemistry , Spermatozoa/chemistry , Animals , Cattle , Comet Assay , Cryopreservation , DNA Breaks , Humans , Male , Silver Staining , Time FactorsSubject(s)
Blood Pressure , Cardiovascular Diseases/radiotherapy , Infrared Rays/therapeutic use , Low-Level Light Therapy , Nitric Oxide Synthase/genetics , Peptidyl-Dipeptidase A/genetics , Blood Pressure/genetics , Blood Pressure/radiation effects , Cardiovascular Diseases/enzymology , Cardiovascular Diseases/genetics , Erythrocytes/enzymology , Erythrocytes/metabolism , Gene Expression/genetics , Genes/genetics , Humans , Hypertension/enzymology , Hypertension/genetics , Hypertension/radiotherapy , Lymphocytes/enzymology , Lymphocytes/metabolism , Myocardial Ischemia/enzymology , Myocardial Ischemia/genetics , Myocardial Ischemia/radiotherapy , Nitric Oxide Synthase Type III , Polymorphism, GeneticABSTRACT
It has been recently shown that okadaic acid (OA), an inhibitor of dephosphorilation, is capable of inducing changes in the nucleolar organizer regions (AgNORs) of some mammalian cells. Our work was focused on studying the structural changes in AgNORs of tumour cells of rat rhabdomyosarcoma RA-23 by their exposure to 100 nM OA. A standard silver staining procedure of interphase AgNORs in tumour cells was used. We measured no less than 100 tumour cells in each clone. In the examined tumour cell populations, the index of interphase AgNORs varied from 1.54 to 4.35. A clear structure and form of AgNORs was not observed in 30% okadaic acid-treated tumour cells, as opposed to 10% of the control ones. AgNORs in these cells looked like a mixture of thin threads encompassing some dark dots lying, mostly, separately. Additional procedures of tumour cell staining with Giemsa and fluorescent dye Hoechst 33,258, respectively, revealed that such structures were not chromosomes. Meanwhile, the frequency of cells at the stage of prophase exceeded 3%, as opposed to the control, where the frequency of cells at this stage was less than 0.5%. Thus, we can conclude that we have detected specific changes in AgNORs and chromatin structure of okadaic acid-treated tumour cells.
