Development of a dose-response bioassay for stanniocalcin in fish

Stanniocalcin (STC) is a polypeptide hormone that was first discovered in fish and recently identified in mammals. In fish, STC is released into the bloodstream in classical endocrine fashion and has well established regulatory effects on calcium and phosphate homeostasis. However, there are no suitable dose-response bioassays for STC and consequently no methods for assigning units of potency to preparations of the hormone. All the available in vitro bioassays are too complex from a technical standpoint to readily accommodate the large number of samples required in dose-response bioassays. Most in vivo bioassays are hampered by the fact that fish have natural rhythms governing plasma STC levels which tend to make them variably sensitive to the injected hormone. In this report we have developed a new in vivo bioassay for STC using rainbow trout. The key feature of the bioassay involves suppressing plasma STC levels to the extent that fish are always receptive to injected hormone. This has been accomplished by phosphate-loading the animals, which lowers their plasma calcium levels, removes the stimulus for STC secretion and brings about a reduction in resting plasma hormone levels. The net effect is an animal that is always responsive to injected STC. With this bioassay we have been able to obtain sensitive and reproducible, dose-related effects of salmon STC on gill calcium transport.