Quantification of PARP Activity in Human Tissues: Ex Vivo Assays in Blood Cells and Immunohistochemistry in Human Biopsies

Abstract

Poly (ADP-ribosyl)ation of proteins is a posttranslational modification mediated by poly (ADP-ribose) polymerases (PARPs) that uses NAD+ as substrate to form the negatively charged polymer of poly (ADP-ribose) (PAR). After DNA damage, PARP-1 is responsible for approximately 90% of the total cellular PARylation activity. Numerous studies showed activation of PARP-1 in various conditions associated with oxidative and nitrosative stress, such as ischemia–reperfusion injury, diabetes mellitus, and inflammation, and also proved the beneficial effects of PARP inhibitors. Pharmacological inhibitors of PARP move toward clinical testing for a variety of indications, including cardioprotection and malignant tumors. Some of the compounds are already in clinical trials. These advances necessitate the detection of PARP activation in human tissues. In the present chapter, we review specific methods used to detect PARP activation in human circulating leukocytes and human tissue sections.