Radiochemical nitric oxide synthase activity determination in rat brain with fast and accurate HPLC analysis.
In the central nervous system, NOS activity is involved in several physiological events, such as refinement of afferent connections in development, or linking cerebral blood flow to neural activity in adulthood, and also in many pathological events, such as cell death in brain ischemia and regulation of vasospasm in hemorrhage. Therefore, we studied NOS activity in the CNS. We describe a fast and accurate method in which we use HPLC analysis to identify and quantify citrulline eluted by ion-exchange chromatography, thus implementing the current method to evaluate NOS activity. This technique could be readily applied for NOS activity determination not only in brain, but also in all other tissues.
Repici
M
M
Department of Anatomy, Pharmacology and Forensic Medicine, University of Torino Medical School, Via Michelangelo 27, 10126, Turin, Italy. mariaelena.repici@unito.it
Cabella
C
C
Colombatto
S
S
Vercelli
A
A
D000818:Animals; D001921:Brain / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards*; D002852:Chromatography, Ion Exchange / Q000379:methods* / Q000592:standards*; D002956:Citrulline; D019001:Nitric Oxide Synthase / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results
D002956:Citrulline; D019001:Nitric Oxide Synthase