In this report is described a method for the measurement of the endogenous poly(ADP-ribose) polymerase (pADPRP) activity in permeabilized lymphocytes isolated from peripheral blood samples. The potential use of the data obtained by this technique is proposed for biomonitoring human exposure to genotoxic agents. This multifunctional nuclear catalytic protein is involved in basic cell functions, but a major role is associated with DNA synthesis and DNA repair activity. In fact, this enzyme is promptly stimulated by DNA strand breaks or by free ends induced by endogenous or exogenous agents. This rapid increase of the basal activity level can be used to reveal biological modifications early after exposure to physical or chemical agents. During isolation and permeabilization of blood lymphocytes, DNA structure remained unaltered. Therefore, no artificial activation of the endogenous pADPRP activity was observed. This technique combines the radiometric assay of pADPRP with the isopycnic isolation of blood lymphocytes; results are reliable, reproducible, and easily applicable to small blood specimens from treated animals or humans. The data presented in this report add confidence to the correlation between endogenous pADPRP level and DNA damage extent in hepatocytes and peripheral blood lymphocytes after in vivo exposure. Preliminary data on lymphocytes from humans are also reported.
- DNA damage
- Poly(ADP-ribose) polymerase
ASJC Scopus subject areas
- Health, Toxicology and Mutagenesis