PBAN regulation

Another technique that is utilized to help ensure that label is incorporated into the pheromone is to apply the precursor at the same time as pheromone biosynthesis activating neuropeptide (PBAN). PBAN is a peptide hormone that regulates pheromone biosynthesis in most, but not all, moths. So, first it must be demonstrated that PBAN regulates pheromone production. In the case of the cabbage looper,... 

PBAN regulation of pheromone biosynthesis in female moths... 

Fang N., Teal R E. A. and Tumlinson J. H. (1995) PBAN regulation of pheromone biosynthesis in female tobacco hornworm moths, Manduca sexta (L.). Arch. Insect Biochem. Physiol. 29, 35 -4. 

Raina A. K., Jaffe H., Kempe T. G., Keim P., Blacher R. W., Fales H. M., Riley C. T., Klun J. A., Ridgway R. L. and Hayes D. K. 1989, Science 244, 796-798. Rafaeli A, Jurenka J. PBAN regulation of pheromone biosynthesis in female moths. In Blomquist G, Vogt R, editors. Insect pheromone biochemistry and molecular biology. NY Academic Press 2003. p. 107—36. 

Rafaeli A, Jurenka J. PBAN regulation of pheromone biosynthesis in female moths. In Blomquist G, Vogt R, editors. Insect pheromone biochemistry and molecular biology. NY Academic Press 2003. p. 107-36. 

Keywords Pheromone Biosynthetic pathways Hormonal regulation PBAN ... 

The role of the nervous system in pheromone biosynthesis in moths is not clearly understood. Christensen and co-workers [208-211] proposed that the neurotransmitter octopamine may be involved as an intermediate messenger during the stimulation of sex pheromone production in H. virescens. These workers suggested that octopamine was involved in the regulation of pheromone production and that PBAN s role lies in the stimulation of octopamine release at nerve endings. However, contradicting results concerning octopa-mine-stimulated pheromone production were reported in the same species as well as other moth species [163,172,212-214]. 

In moths, it was discovered in Helicoverpa zea that a peptide produced in the subesophageal ganglion portion of the brain complex regulates pheromone production in female moths (19). This factor has been purified and characterized in three species, Helicoverpa zea (20), Bombyx mori (21, 22), and Lymantria dispar (23). They are all a 33- or 34-amino acid peptide (named pheromone biosynthesis activating neuropeptide, PBAN) and have in common an amidated C-terminal 5-amino acid sequence (FXPRL-amide), which is the minimum peptide fragment required for pheromon-tropic activity. In the redbanded leafroller moth, it was shown that PBAN from the brain stimulates the release of a different peptide from the bursae copulatrix that is used to stimulate pheromone production in the pheromone gland found at the posterior tip of the abdomen (24). 

The three hormones that regulate pheromone production in insects are shown in Figure 1.1. PBAN alters enzyme activity through second messengers at one or more steps during or subsequent to fatty acid synthesis during pheromone production (Rafaeli and Jurenka, Chapter 5). In contrast, 20-hydroxyecdysone and JH induce or repress the synthesis of specific enzymes at the transcription level (Tittiger, Chapter 7 Blomquist, Chapter 9). 

Figure 1.1 The three major types of hormones that regulate pheromone production in insects. A Juvenile Hormone III (C16 JH), B 20-Hydroxyecdysone and C PBANs from the corn earworm, Helicoverpa zea (Raina et al., 1989), the silkworm moth Bombyx mori (Kitamura et al., 1989) and the gypsy moth, Lymantira dispar (Master et al., 1994). The minimum sequence (pentapeptide) required for activity is indicated.

In no model pheromone biosynthetic system is the molecular mechanism of hormonal regulation completely understood. The mechanism of action of JH and the nature of its receptor remain one of the mysteries of insect science, and the clear-cut action of JH by itself in inducing specific genes in pheromone production in bark beetles offers an excellent model for study. A better understanding of the PBAN receptor and the second messenger system it triggers as well as the steps regulated in pheromone biosynthesis is also needed. The next several years should see some of the key questions answered in model insects. 

In this chapter we will review the current state of knowledge about how pheromone production is regulated in female moths. Discussion of PBAN identification and localization within the nervous system will be followed by how PBAN acts to stimulate pheromone biosynthesis. The final major topic will be a discussion of mediators and inhibitors of PBAN action. A considerable amount of information has accumulated with regard to regulation of pheromone biosynthesis in moths since Pheromone Biochemistry (Prestwich and Blomquist, 1987) was first published, and this chapter is not all inclusive. Further information can also be obtained in several reviews (Raina, 1993 Jurenka, 1996 Teal et al., 1996 Rafaeli et al., 1997b Raina, 1997 Rafaeli, 2002). 

Once it was established that pheromone biosynthesis was regulated by a peptide produced in the SEG, the next goal was to identify the peptide. In the purification of any biologically active factor, each purification step requires a sensitive bioassay to measure the active material. In the purification of PBAN, the bioassay consisted of head ligated females that were injected with bioactive fractions. After a 1-3 h period of incubation, the pheromone gland was excised and titers of pheromone determined by gas chromatography (GC). The first PBAN was purified and identified from H. zea (Raina el ah, 1989). Dissection of about 5000 brain-SEG complexes followed by several steps of HPLC purification resulted in a pure peptide that could be sequenced. It was found to be a 33 amino acid peptide with a C-terminal amide (Table 5.1). The peptide was synthesized and was shown to be active in the bioassay in a dose as low as 2 pmol (Raina et al., 1989). In the same year, a PBAN from B. mori was purified and sequenced (Kitamura et al.,... 

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