Figure 1: Orexin A-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/OX1 cells. The cells were loaded with Calcium-4 prior to being stimulated with OX1 receptor agonist, orexin A. The intracellular calcium change was measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses (5-fold dilution) of orexin A (Mean ± SD, n = 2). The EC50 of orexin A on CHO-K1/OX1 cells was 7.4 nM.
Note:
1. EC50 value is calculated with four parameter logistic equation:
Y=Bottom + (Top-Bottom)/ (1+10^ ((LogEC50-X)*HillSlope))
X is the logarithm of concentration.
Y is the response and starts at Bottom and goes to Top with a sigmoid shape.
CHO-K1/OX1 Stable Cell Line
The orexin/hypocretin peptides orexin A and orexin B (also known as hcrt-1 and hcrt-2) are 33- and 28-amino acid peptides, respectively. They are preferentially expressed in hypothalamus. The orexins have a range of physiological functions including feeding control, energy metabolism, neuroendocrine function modulation, and sleep-wake cycle regulation. The two orexin receptor subtypes OX1 and OX2 both mediate the action of orexin-A and orexin-B and couple efficiently through Gq/11 to activate phospholipase C and lead to elevation of intracellular calcium.
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