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BMC Physiology (1)
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Ibarra, Cristian (2)
Uhlén, Per (2)
Andäng, Michael (1)
Ernfors, Patrik (1)
Fritz, Nicolas (1)
Kitambi, Satish Srinivas (1)
Nilsson, Erik S (1)
Sekyrova, Petra (1)
Tekeoh, Gilbert Nyah (1)
Zhang, Songbai (1)
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Small molecule screening platform for assessment of cardiovascular toxicity on adult zebrafish heart
Kitambi, Satish Srinivas
Nilsson, Erik S
Tekeoh, Gilbert Nyah
Cardiovascular toxicity is a major limiting factor in drug development and requires multiple cost-effective models to perform toxicological evaluation. Zebrafish is an excellent model for many developmental, toxicological and regenerative studies. Using approaches like morpholino knockdown and electrocardiogram, researchers have demonstrated physiological and functional similarities between zebrafish heart and human heart. The close resemblance of the genetic cascade governing heart development in zebrafish to that of humans has propelled the zebrafish system as a cost-effective model to conduct various genetic and pharmacological screens on developing embryos and larvae. The current report describes a methodology for rapid isolation of adult zebrafish heart, maintenance ex vivo, and a setup to perform quick small molecule throughput screening, including an in-house implemented analysis script.
Adult zebrafish were anesthetized and after rapid decapitation the hearts were isolated. The short time required for isolation of hearts allows dissection of multiple fishes, thereby obtaining a large sample size. The simple protocol for ex vivo culture allowed maintaining the beating heart for several days. The in-house developed script and spectral analyses allowed the readouts to be presented either in time domain or in frequency domain. Taken together, the current report offers an efficient platform for performing cardiac drug testing and pharmacological screens.
The new methodology presents a fast, cost-effective, sensitive and reliable method for performing small molecule screening. The variety of readouts that can be obtained along with the in-house developed analyses script offers a powerful setup for performing cardiac toxicity evaluation by researchers from both academics and industry.
Heart; Screening; Zebrafish; Small molecule; Ex vivo; Ca2+ signaling
Inositol 1,4,5-Trisphosphate Receptor Subtype-Specific Regulation of Calcium Oscillations
Oscillatory fluctuations in the cytosolic concentration of free calcium ions (Ca2+) are considered a ubiquitous mechanism for controlling multiple cellular processes. Inositol 1,4,5-trisphosphate (IP3) receptors (IP3R) are intracellular Ca2+ release channels that mediate Ca2+ release from endoplasmic reticulum (ER) Ca2+ stores. The three IP3R subtypes described so far exhibit differential structural, biophysical, and biochemical properties. Subtype specific regulation of IP3R by the endogenous modulators IP3, Ca2+, protein kinases and associated proteins have been thoroughly examined. In this article we will review the contribution of each IP3R subtype in shaping cytosolic Ca2+ oscillations.
Inositol 1,4,5-trisphosphate receptor; Inositol 1,4,5-trisphosphate receptor-associated protein; Calcium signaling; Calcium oscillations
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