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Real‐time electrochemical detection of endogenous substance release in freshly isolated organs
Author(s) -
Palygin Oleg,
Levchenko Vladislav,
Ilatovskaya Daria V,
Pavlov Tengis S,
Ryan Robert P,
Cowley Allen W,
Staruschenko Alexander
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.910.16
Subject(s) - chemistry , kidney , in vivo , endogeny , ex vivo , microdialysis , medulla , perfusion , losartan , medicine , renal cortex , endocrinology , amperometry , antagonist , biosensor , receptor , biochemistry , angiotensin ii , biology , in vitro , extracellular , electrode , electrochemistry , microbiology and biotechnology
We established a novel approach to measure acute changes in endogenous substances in the freshly isolated kidney. This method is based on enzymatic microelectrode biosensors and utilizes amperometry technique. In addition to quantitation of basal levels this technique allows real‐time detection of variety of substances, reflecting changes and fluctuations in their concentrations in freshly isolated organs during vehicle or drug perfusion. Basal levels of H 2 O 2 in the kidney cortex and medulla of Sprague Dawley (SD) rats were measured by biosensors and conventional interstitial fluid collection method. Real‐time changes in ATP and H 2 O 2 concentrations in response to angiotensin II (Ang II) were determined. Freshly isolated kidneys of SD and Dahl salt‐sensitive (SS) rats were perfused with 1 μM Ang II under constant laminar flow. This produced a rapid simultaneous release of both ATP and H 2 O 2 in the kidney cortex area with a strong enhancement in SS rats, especially when animals were fed a high‐salt diet. Biosensors specificity revealed by kidney perfusion with AT 1 receptor antagonist losartan and decomposition of H 2 O 2 with catalase enzyme. Established method represents a unique powerful approach for the real‐time monitoring of the endogenous substance level in organs or tissue and could be used for ex vivo and in vivo measurements during normal or pathological conditions. This research was supported by NHLBI.