Determination of locust AKH-I by radioimmunoassay and the identification of an AKH-like factor in the locust brain

Abstract

The N-terminal glutamic acid analog of the decapeptide locust adipokinetic hormone-I, [Glu¹]AKH-I, was prepared by solid-phase synthesis and derivatized to [4-hydroxyphenyl propionyl-Glu¹]AKH-I. This derivative was radioiodinated on the 4-hydroxyphenyl function ([¹²⁵I]AKH analog). [Glu¹]AKH-I was coupled to bovine serum albumin to produce rabbit antiserum. This, together with the [¹²⁵I]AKH analog was utilized to develop a highly selective radioimmunoassay procedure (RIA) for detecting AKH-I in locusts. Methanol extracts of locust brain contain two factors (BI and BII) that inhibit protein synthesis in excised fat body tissue of vitellogenic female locusts. BI also elicits lipid mobilization when injected into adult male locusts. The HPLC retention time of BI on an RP-18 column is identical to that of locust adipokinetic hormone (AKH-I), both synthetic and derived from adult corpora cardiaca (CC). The second brain factor (BII) does not exhibit lipid mobilizing activity. One tissue-equivalent of BI contains 33 ng of AKH-I as determined by RIA, compared to 450 ng native HPLC-separated AKH-I in locust CC.