ASGSB 1998 Annual Meeting Abstracts

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METHOD FOR THE ISOLATION AND PURIFICATION OF THE JELLYFISH THYROXINE-LIKE HORMONE (Jf-T4).    D.B. Spangenberg*, M.C.Castle and F.A. Lattanzio. Depts. of Pathology & Anatomy* and Physiological Sciences, Eastern Virginia Medical School, Norfolk, Virginia.

Results from the Jellyfish-in-Space Experiment from the SLS-1 mission indicated that the hormone (Jf-T4) utilized by the jellyfish for metamorphosis induction may be synthesized or utilized differently in micro-g than on Earth. Therefore, research was begun to develop methods for the measurement of the hormone and its receptors in jellyfish flown in micro-g and their ground controls. An experiment for this purpose was proposed and accepted for a flight experiment utilizing ARF hardware.

The hormone measurement techniques (although constrained by a maximum sample of 30 polyps per group) were developed accordingly. Polyps are homogenized and the resulting homogenate is extracted with ethanolic- NaOH. The ethanolic-NaOH mixture is homogenized at 4^oC and the solution centrifuged at 10,000g for 10 minutes. The supernatant is removed and the pellet is re-extracted two more times. The pooled supernatant is then diluted with aqueous HCl. To extract hormone from artificial sea water (ASW), the ASW is adjusted to an identical ethanol- HCl concentration. The ethanol-HCl mixture is run through columns of Sephadex LH-20 resin. The columns are washed consecutively with HCL, H20, ethanol: NH₄OH, and NH₄OH in ethanol The first portion of the NH₄OH in ethanol contains the iodinated compounds of interest. These fractions are combined and dried under nitrogen and then separated by HPLC. The Jf-T4 was eluted from HPLC as a single peak as detected by both radiochemical and UV detectors. This peak did not coincide identically with T4 (thyroxine) nor did isolated Jf-T4 cross react with several different sources of T4 antibodies, so it is likely that the compound is different from T4. Of particular significance is the fact that the parent compound and metabolites can be measured by this method using mass spectrometry. This method can be used to obtain the information proposed in the ARF-2 experiment through another future Jellyfish-in-Space experiment. Sponsored by NASA grant #NAG10-0178 and an EVMS Institutional grant.

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