Chapter title |
The CamKKβ Inhibitor STO609 Causes Artefacts in Calcium Imaging and Selectively Inhibits BKCa in Mouse Carotid Body Type I Cells
|
---|---|
Chapter number | 3 |
Book title |
Arterial Chemoreceptors in Physiology and Pathophysiology
|
Published in |
Advances in experimental medicine and biology, January 2015
|
DOI | 10.1007/978-3-319-18440-1_3 |
Pubmed ID | |
Book ISBNs |
978-3-31-918439-5, 978-3-31-918440-1
|
Authors |
Jennifer G. Jurcsisn, Richard L. Pye, Jon Ali, Barbara L. Barr, Christopher N. Wyatt |
Abstract |
It has previously been reported that AMP-activated protein kinase (AMPK) may be critical for hypoxic chemotransduction in carotid body type I cells. This study sought to determine the importance of the regulatory upstream kinase of AMPK, CamKKβ, in the acute response to hypoxia in isolated mouse type I cells.Initial data indicated several previously unreported artefacts associated with using the CamKKβ inhibitor STO609 and Ca(2+) imaging techniques. Most importantly Fura-2 and X-Rhod1 imaging revealed that STO609 quenched emission fluorescence even in the absence of intracellular Ca(2+) ([Ca(2+)]i). Furthermore, STO609 (100 μM) rapidly inhibited outward macroscopic currents and this inhibition was abolished in the presence of the selective BKCa inhibitor paxilline.Taken together these data suggest that ST0609 should be used with caution during Ca(2+) imaging studies as it can directly interact with Ca(2+) binding dyes. The rapid inhibitory effect of STO609 on BKCa was unexpected as the majority of studies using this compound required an incubation of approximately 10 min to inhibit the kinase. Furthermore, as AMPK activation inhibits BKCa, inhibiting AMPK's upstream kinases would, if anything, be predicted to have the opposite effect on BKCa. Future work will determine if the inhibition of BKCa is via CamKKβ or via an off target action of STO609 on the channel itself. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 5 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Bachelor | 2 | 40% |
Professor > Associate Professor | 1 | 20% |
Professor | 1 | 20% |
Unknown | 1 | 20% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 1 | 20% |
Psychology | 1 | 20% |
Medicine and Dentistry | 1 | 20% |
Unknown | 2 | 40% |