NIDDK, NIH

3 ARTICLES PUBLISHED IN JoVE

JoVE Journal

Measuring Nitrite and Nitrate, Metabolites in the Nitric Oxide Pathway, in Biological Materials using the Chemiluminescence Method

Barbora Piknova¹, Ji Won Park¹, Katelyn S. Cassel¹, Cameron N. Gilliard¹, Alan N. Schechter¹

¹Molecular Medicine Branch, NIDDK, NIH

Nitric oxide (NO) is an important signaling molecule in vascular homeostasis. NO production in vivo is too low for direct measurement. Chemiluminescence provides useful insight into NO cycle via measuring its precursors and oxidation products, nitrite and nitrate. Nitrite / nitrate determination in body tissues and fluids is explained.

Medicine

Platelet-based Detection of Nitric Oxide in Blood by Measuring VASP Phosphorylation

Sirada Srihirun^1,2, Alan N. Schechter², Barbora Piknova²

¹Department of Pharmacology, Faculty of Dentistry, Mahidol University, ²Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health

Here, we present a protocol to address the potential use of platelets as a highly sensitive nitric oxide sensor in blood. It describes initial platelet preparation and the use of nitrite and deoxygenated red blood cells as nitric oxide generators.

Biology

Preparation of Rat Skeletal Muscle Homogenates for Nitrate and Nitrite Measurements

Ji Won Park^*1, Samantha M. Thomas^*1, Lee J. Wylie², Andrew M. Jones², Anni Vanhatalo², Alan N. Schechter¹, Barbora Piknova¹

¹Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, ²Sport and Health Sciences, College of Life and Environmental Sciences, St Luke’s Campus, University of Exeter

We present protocols for three different methods for the homogenization of four different muscle groups of rat skeletal muscle tissue to measure and compare the levels of nitrate and nitrite. Furthermore, we compare different sample weights to investigate whether tissue sample size affects the results of homogenization.