Líffræðifélag Íslands
Líffræðiráðstefnan 2015
Erindi/veggspjald / Talk/poster V39
Srdanović, S., 1,2 Þorsteinsson, H.,1,2 Ágústsdóttir A.B., 1,2 & Karlsson, K.Æ.,2,3
1 Department of Biotechnology, University of Rijeka, Rijeka Croatia 2 3Z Pharmaceuticals, Reykjavik Iceland 3 Department of Biomedical Engineering, School of Science and Engineering, Reykjavik University, Reykjavik Iceland
Kynnir / Presenter: Srdanović, S.
Tengiliður / Corresponding author: Karlsson, K.Æ., (karlsson@ru.is)
Protein kinase inhibitors are a large class of enzyme inhibitors that block the action of kinases, thus blocking the addition of a phosphate group to proteins or other organic molecules, and turning proteins off. Currently we describe the effects of 190 item kinase inhibitor library (SYNLibrary 190) on sleep and wake in larval zebrafish. Fertilized embryos were incubated at 28.5° C for 120 hrs and then placed in 96 well-plates. Next, twenty-four 96 well-plates were placed in light and temperature controlled recording apparatus which was fitted with 24 separate cameras and a Perkin-Elmer microfluidics dispenser. After a 24 hr acclimation period, drugs were dispensed to the wells, and the larvae´s behavior was recorded for 24 hrs. In a typical run each group comprised 16 larvae. Each drug was applied in three dose sizes (1, 10 and 20 µM), in addition to 3 control groups. First, behavior was dichotomized into moving and not-moving (movement cut-off 1.0 cm/s), next sleep was defined as immobility period exceeding 6 s, and the remainder was classified as wake. Finally, the behavior was analyzed for sleep percentage, sleep fragmentation, average sleep and wake bout length, and sleep latency, using custom written software. We show that sleep active and sedative drugs are easily identified using the assay. 24 drugs were identified that decrease sleep and increase sleep fragmentation, while only a single novel compound was found to increase sleep and decrease sleep fragmentation. We postulate that the sleep increase is mediated via decreased Akt signaling, previously implicated in shortened circadian rhythm cycles. We identify a novel sleep-promoting action of a protein kinase inhibitor.