Biosynthesis of ecdysteroid hormones by crustacean Y-organs: conversion of cholesterol to 7-dehydrocholesterol is suppressed by a steroid 5α-reductase inhibitor

Abstract

A pair of glands (Y-organs) in crustaceans synthesize and secrete ecdysteroid hormones; the obligate precursor for synthesis is circulating cholesterol. Ecdysteroid output by the Y-organs is regulated negatively by an eyestalk neurosecretory peptide, molt-inhibiting hormone (MIH). The question was addressed, does MIH suppress ecdysteroid synthesis by decreasing cholesterol supply (uptake) or its utilization or both? Experiments were conducted with Y-organs in vitro from the crab, Menippe mercenaria, in the presence of labeled cholesterol, with or without the steroid 5α-reductase inhibitor, L-645390 (Merck). Other experiments superimposed the presence or absence of eyestalk extract containing MIH activity. L-645390 greatly depressed incorporation of cholesterol into an early intermediate, 7-dehydrocholesterol and into secreted ecdysteroids. At the same time, cholesterol accumulated in the Y-organs, to levels significantly higher than in untreated controls. MIH alone depressed both cholesterol uptake and incorporation. MIH together with L-645390 produced the greatest depression of cholesterol incorporation while also preventing the cholesterol accumulation seen with L-645390 alone. These results indicate that cholesterol uptake and its metabolic utilization in Y-organs are separable events representing separate sites of regulation by MIH.