Isolation of sockeye salmon growth hormone utilizing serum triiodothyronine enhancement in rainbow trout to monitor biological activity.
Growth hormone (GH) was isolated from sockeye salmon (Oncorhynchus nerka) pituitary glands using established techniques of affinity and gel filtration chromatography, and preparative polyacrylamide gel electrophoresis. The GH activity was followed throughout the fractionation procedure with a bioassay based on the increase of serum triiodothyronine (T3) in rainbow trout (Oncorhynchus mykiss), measured by radioimmunoassay (RIA). Amino-terminal amino acid sequence analysis and subsequent comparison with established GH sequences from other Oncorhynchus sp. were used to confirm the isolation of sockeye salmon GH (ssGH). The bioassay was sensitive to a dose of 55 ng of purified ssGH/g fish. Monomeric GHs, located in the carbohydrate-poor protein fraction, were the only pituitary components that elevated serum T3. Twenty-four hours after GH injection was an appropriate and practical time to blood sample, allowing completion of the bioassay, including RIA, in 3 days. The generic homology, between the source of pituitaries and the bioassay animals used in this study, should permit the bioassay to be useful during GH isolation from pituitaries of all Oncorhynchus sp.
Nagler
J J
JJ
Marine Sciences Research Laboratory, Memorial University of Newfoundland, St. John's, Canada.
Hwang
S J
SJ
Idler
D R
DR
D000704:Analysis of Variance; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D013006:Growth Hormone / Q000302:isolation & purification* / Q000494:pharmacology; D010902:Pituitary Gland / Q000378:metabolism; D011863:Radioimmunoassay; D012474:Salmon; D012680:Sensitivity and Specificity; D014284:Triiodothyronine / Q000096:biosynthesis*; D014337:Trout
D014284:Triiodothyronine; D013006:Growth Hormone