Calcium Toolkit genes expression in the zebrafish brain

Zebrafish are well suited for in vivo calcium imaging because the transparency of their larvae and the ability to express calcium probes in various cell subtypes. This model organism has been used extensively to study brain development, neuronal function, and network activity. However, only a few studies have investigated calcium homeostasis and signaling in zebrafish neurons, and little is known about the proteins that are involved in these processes. Using bioinformatics analysis and available databases, the present study identified 491 genes of the zebrafish Calcium Toolkit (CaTK). Using RNA-sequencing, we then evaluated the expression of these genes in the adult zebrafish brain and found 380 hits that belonged to the CaTK. Based on quantitative real-time polymerase chain reaction arrays, we estimated the relative mRNA levels in the brain of CaTK genes at two developmental stages. In both 5 dpf larvae and adult zebrafish, the highest relative expression was observed for tmbim4, which encodes a Golgi membrane protein. The present data on CaTK genes will contribute to future applications of zebrafish as a model for in vivo and in vitro studies of Ca2+ signaling. qPCR gene expression profiling. 3 different samples of adult brain RNA and 3 different samples of larvae head RNA were used in the analysis. cDNA (25 ng) was used for each reaction. Actin beta 1 (actb1), glyceraldehyde-3-phosphate dehydrogenase (gapdh), tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide (yhwaz), and ribosomal protein S18 (rps18) were used as reference genes. No-templete-control samples were included. All the PCR reactions were run in the duplicates.