Insect Biochemistry and Molecular Biology Insect Biochemistry and Molecular Biology 34 (2004) 1215–1224 Selectivity and neuroendocrine regulation of the precursor uptake by pheromone glands from hemolymph in geometrid female moths, which secrete epoxyalkenyl sex pheromones Wei Wei a , Masanobu Yamamoto a , Tetsuhiro Asato a , Takeshi Fujii a , Guan-Qin Pu b , Tetsu Ando a,Ã a Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan b Biotechnology College, Su Zhou University, Su Zhou 215151, P. R. China Received 18 June 2004; received in revised form 12 August 2004; accepted 17 August 2004 Abstract Macrolepidopteran female moths in families such as Geometridae produce epoxyalkenyl sex pheromones, which are biosynthesized via epoxidation of polyunsaturated hydrocarbons in their pheromone glands. The precursors, however, are expected to be produced outside of the pheromone glands, probably in oenocytes or in the fat body, and transported to the glands via hemolymph. Based on these facts, the selectivity of the epoxidation substrates and of the precursor uptake by pheromone glands was examined with two geometrid species, Hemerophila artilineata and Ascotis selenaria cretacea, using binary mixtures of deuterated precursors and their analogs, which were topically applied to the pheromone glands or injected into the abdomen. GC–MS measurements of pheromone extracts showed equal epoxidation of two polyenes, indicating a low selectivity for both processes, while the epoxidation proceeded at only one double bond specific to each species. This result makes it possible to conclude that the formation of species-specific epoxyalkenyl pheromones results from the rigid formation of polyunsaturated precursors and their epoxidation at a fixed position. Next, the neuroendocrine regulation of these processes was studied with in vivo and in vitro experiments using decapitated females. The epoxy pheromones disappeared completely within 36h of decapitation, and epoxidation of the injected precursors was not detected in the decapitated females, which restarted the reaction by treatment with a pheromone biosynthesis-activating neuropeptide (PBAN). The precursors topically applied to glands of the decapitated females, however, were converted into epoxy pheromones without PBAN, indicating that this neuropeptide hormone accelerated the precursor uptake by pheromone glands but not the epoxidation already underway in the glands. r 2004 Elsevier Ltd. All rights reserved. Keywords: Sex pheromone biosynthesis; Epoxidation of (Z,Z,Z)-3,6,9-triene; Uptake by pheromone gland; Pheromone biosynthesis-activating neuropeptide (PBAN); Mulberry looper (H. artilineata); Japanese giant looper (A. s. cretacea); Geometridae; Lepidoptera; GC–MS analysis 1. Introduction For sexual communication to promote species-specific mating, females of many lepidopteran insects secrete special blends composed of C 10 –C 18 unsaturated fatty alcohols and/or their derivatives (Type 1 compounds). The biosynthesis of these sex pheromones has been investigated in detail (Jurenka, 2003, 2004; Tillman et al., 1999) and understood at the level of molecular biology (Knipple and Roelofs, 2003). Recently, cDNA clones encoding enzymes for desaturation introducing a double bond (Knipple et al., 1998; Roelofs and Rooney, 2003) and for the reduction of an acyl intermediate into alcohol (Moto et al., 2003) were isolated and character- ized. In addition to the Type 1 compounds, polyunsa- turated hydrocarbons and their epoxy derivatives with a C 17 –C 23 straight chain (Type 2 compounds) have been ARTICLE IN PRESS www.elsevier.com/locate/ibmb 0965-1748/$-see front matter r 2004 Elsevier Ltd. All rights reserved. doi:10.1016/j.ibmb.2004.08.004 Ã Corresponding author. Tel./fax: +81-423-887-278. E-mail address: antetsu@cc.tuat.ac.jp (T. Ando).