OX2 orexin/hypocretin receptor signal transduction in recombinant Chinese hamster ovary cells.

There are two subtypes of orexin receptors, OX1 and OX2. Signalling pathways have been mapped in much higher detail for OX1 receptors than OX2 receptors. Almost all the detailed studies have been performed in Chinese hamster ovary cells, and we thus chose the same cell background for the studies on human OX2 receptors to allow comparison to human OX1 receptors. Adenylyl cyclase, phospholipase A2, C and D and diacylglycerol lipase activities were assessed by precursor radiolabelling and chromatographic separation (ion exchange, affinity or thin layer), calcium by a fluorescent method, and receptor binding with [(125)I]-orexin-A. Upon activation with orexin-A, OX2 receptors stimulated phospholipase A2, C and D, diacylglycerol lipase and calcium elevation, and both inhibited and stimulated adenylyl cyclase; i.e., the responses to OX2 activation by orexin-A were principally like those of OX1, in contrast to some previous suggestions. The responses occurred mostly in the same concentration range as those for OX1 activation and via the same signal cascades. However, some responses were weaker, suggesting a partially differential coupling to some cascades. In summary, OX2 receptor signalling is principally similar to OX1 receptor signalling suggesting also a physiologically similar coupling, though this needs to be verified in physiological contexts. Some (relatively weak) differences between the receptors may be investigated in further studies.