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A novel peptide in the AKH/RPCH family isolated from the corpora cardiaca of the Emperor dragonfly, Anax imperator.

Using a heterologous (in locusts and cockroaches) and a homologous bioassay, we have isolated the neuropeptide pGlu-Val-Asn-Phe-Ser-Pro-Ser-Trp-NH2 from extracts of corpora cardiaca of the Emperor dragonfly, Anax imperator. The sequence elucidation was achieved by Edman degradation of the deblocked peptide and by electrospray mass spectrometry. Low concentrations of the synthetic peptide injected into the Emperor dragonfly increased the hemolymph lipid concentration, suggesting a possible role of the peptide in lipid homeostasis during flight. Therefore, it is named Ani-AKH, Anax imperator adipokinetic hormone.

Amino Acid Sequence↗

Insect NMDA receptors mediate juvenile hormone biosynthesis.

In vertebrates, the N-methyl-D-aspartate subtype of glutamate receptors (NMDAR) appears to play a role in neuronal development, synaptic plasticity, memory formation, and pituitary activity. However, functional NMDAR have not yet been characterized in insects. We have now demonstrated immunohistochemically glutamatergic nerve terminals in the corpora allata of an adult female cockroach, Diploptera punctata. Cockroach corpus allatum (CA) cells, exposed to NMDA in vitro, exhibited elevated cytosolic [Ca(2+)], but not in culture medium nominally free of calcium or containing NMDAR-specific channel blockers: MK-801 and Mg(2+). Sensitivity of cockroach corpora allata to NMDA changed cyclically during the ovarian cycle. Highly active glands of 4-day-old mated females, exposed to 3 microM NMDA, produced 70% more juvenile hormone (JH) in vitro, but the relatively inactive glands of 8-day-old mated females showed little response to the agonist. The stimulatory effect of NMDA was eliminated by augmenting the culture medium with MK-801, conantokin, or high Mg(2+). Having obtained substantive evidence of functioning NMDAR in insect corpora allata, we used reverse transcription PCR to demonstrate two mRNA transcripts, DNMDAR1 and DNMDAR2, in the ring gland and brain of last-instar Drosophila melanogaster. Immunohistochemical labeling, using mouse monoclonal antibody against rat NMDAR1, showed that only one of the three types of endocrine cells in the ring gland, CA cells, expressed rat NMDAR1-like immunoreactive protein. This antibody also labeled two brain neurons in the lateral protocerebrum, one neuron per brain hemisphere. Finally, we used the same primers for DNMDAR1 to demonstrate a fragment of putative NMDA receptor in the corpora allata of Diploptera punctata. Our results suggest that the NMDAR has a role in regulating JH synthesis and that ionotropic-subtype glutamate receptors became specialized early in animal evolution.

Animals↗

Effects of an allatostatin and a myosuppressin on midgut carbohydrate enzyme activity in the cockroach Diploptera punctata.

Neuropeptides of the cockroach allatostatin (AST) family are known for their ability to inhibit the production of juvenile hormone by the corpora allata of cockroaches. Since their discovery, they have also been shown to modulate myotropic activity in a range of insect species as well as to act as neurotransmitters in Crustaceans and possibly in insects. The midgut of cockroaches contains numerous endocrine cells, some of which produce AST whereas others produce the FMRFamide-related peptide, leucomyosuppressin (LMS). We have determined if ASTs and LMS are also able to influence carbohydrate-metabolizing enzyme activity in the midgut of the cockroach, Diploptera punctata. Dippu-AST 7 stimulates activity of both invertase and alpha-amylase in a dose-dependent fashion in the lumen contents of ligatured midguts in vitro, but not in midgut tissue, whereas the AST analog AST(b)phi2, a cyclopropyl-ala, hydrocinnamic acid analog of Dippu-AST 6, has no effect. Leucomyosuppressin also stimulates enzyme activity in lumen contents only, although the EC50 is considerably greater than for Dippu-AST. Dippu-AST is also able to inhibit proctolin-induced contractions of midgut muscle, and this action had already been described for LMS [18]. Thus, in this organ, AST and LMS have at least two distinct physiological effects.

Animals↗

Suppressive effects of Calendula micrantha essential oil and gibberelic acid (PGR) on repro ductive potential of the Mediterranean fruit fly Ceratitis capitata Wied. (Diptera: Tephritidae).

The volatile oil of Calendula micrtantha plant was extracted and the components were identified by Gc/Ms. Adulticidal efficiency of the volatile oil and gibberelic acid "plant growth promoting hormone" as well as their mixture was assessed against the Mediterranean fruit fly Ceratitis capitata. The result showed that the two compounds capable have characteristic resembling to insect juvenile hormones and have suppressive effect on reproductive potential. They induced the significant disturbances in the ovarian protein fraction and the amino acids patterns.

Animals↗

Structure-activity relationships for the lipid-mobilising action of locust adipokinetic hormone. Synthesis and activity of a series of hormone analogues.

A series of compounds structurally related to adipokinetic hormone, the decapeptide neurohormone less than Glu-Leu-Asn-Phe-Thr-Pro-Asn-Trp-Gly-Thr-NH2, have been prepared by synthesis and by enzymic cleavages of synthetic hormone. Their relative agonist activities in mobilising lipids over a fixed time interval (1 h) in locusts were assessed. The similar time courses for lipid release shown by two of the peptide analogues and adipokinetic hormone suggest that the analogues and the hormone are transported to the receptors on the fat body cells, and are also degraded, at similar rates. Consequently, the analogue activities can be correlated with the structural requirements of the locust fat body hormone receptors. The requirements for activity demonstrated in this study are as follows. Residues 1--8 from the N-terminus are necessary to elicit some activity (20%). Residues 5 and 7 in the octapeptide can be changed without affecting activity but L-pyroglutamic acid as the N-terminal residue is necessary formaximum activity both in the octapeptide and the decapeptide. Full activity is achieved only by adding the dipeptide glycyl threonine amide to the active octapeptide 'core'. In the decapeptide, residues cannot be interchanged to the same extent as in the octapeptide without reducing activity. The peptide probably has to be uncharged. Inactive analogues of seven residues or less do not interfere in the hormone-receptor interaction.

Animals↗

Two hexameric cyanoprotein subunits from an insect, Riptortus clavatus. Sequence, phylogeny and developmental and juvenile hormone regulation.

Hemolymph of a hemipteran insect, Riptortus clavatus, contains four distinct hexameric proteins (cyanoproteins) composed of two distinct subunits, CP alpha and CP beta, which show profiles of abundance depending on developmental stage, diapause status controlled by juvenile hormone, and sex. We have isolated cDNA clones encoding the two subunits and determined the complete sequences. The nucleotide sequences predict polypeptides of 693 and 691 residues for CP alpha and CP beta, respectively, including identical 16-residue signal peptides. The deduced amino acid sequences of both CP alpha and CP beta have significant similarity to other hemocyanin-related proteins, indicating that the cyanoproteins represent hexamerins. Phylogenetic analyses show that the cyanoprotein subunits can be grouped together with other hexamerins from exopterygote insects. Developmental Northern-blot analyses suggest that the expression of the cyanoproteins is regulated at the level of transcript abundance. In addition, the expression of the CP alpha subunit in female adults has been shown to be enhanced by juvenile hormone (JH) while the expression of the CP beta subunit is suppressed by the same hormone. To our knowledge, the CP alpha subunit of R. clavatus is the first case of a JH-enhanceable hexamerin whose sequence has been determined.

Amino Acid Sequence↗

The neuroendocrine regulation of Drosophila aging.

The rate of aging is regulated by hormones in insects and, most likely, in mammals. Mutations of the insulin-signaling pathway extend lifespan in the fruit fly and influence the level of other hormones, specifically juvenile hormone and the sterol ecdysone, each of which may directly influence senescence. With new genetic and genomic tools in Drosophila biology we are now exploring how the neuroendocrine system responds to environmental conditions to modify insulin action, how these signals control secondary hormones, and how these messages together modulate animal aging.

Aging↗