Effect of the urease-derived peptide Jaburetox on the central nervous system of Triatoma infestans (Insecta: Heteroptera).

BACKGROUND: Triatoma infestans is the main vector of Chagas'disease in Southern Cone countries. In triatomines, symptoms suggesting neurotoxicity were observed after treatment with Jaburetox (Jbtx), the entomotoxic peptide obtained from jackbean urease. Here, we study its effect in the central nervous system (CNS) of this species. METHODS: Immunohistochemistry, Western blots, immunoprecipitation, two-dimensional electrophoresis, tandem mass spectrometry and enzymatic assays were performed. RESULTS: Anti-Jbtx antibody labeled somata of the antennal lobe only in Jbtx-treated insects. Western blot assays of nervous tissue using the same antibody reacted with a 61kDa protein band only in peptide-injected insects. Combination of immunoprecipitation, two-dimensional electrophoresis and tandem mass spectrometry identified UDP-N-acetylglucosamine pyrophosphorylase (UDP-GlcNAcP) as a molecular target for Jbtx. The activity of UDP-GlcNAcP increased significantly in the CNS of Jbtx-treated insects. The effect of Jbtx on the activity of nitric oxide synthase (NOS) and NO production was investigated as NO is a recognized messenger molecule in the CNS of T. infestans. NOS activity and NO levels decreased significantly in CNS homogenates of Jbtx-treated insects. CONCLUSIONS: UDP-GlcNAcP is a molecular target of Jbtx. Jbtx impaired the activity of T. infestans nitrergic system, which may be related with early behavioral effects. GENERAL SIGNIFICANCE: We report that the CNS of Triatoma infestans is a target for the entomotoxic peptide and propose that a specific area of the brain is involved. Besides potentially providing tools for control strategies of Chagas' disease vectors our data may be relevant in various fields of research as insect physiology, neurobiology and protein function.

Distribution and morphometric studies of flagellar sensilla in Emphorini bees (Hymenoptera, Apoidea).

The tribe Emphorini is a group of pollen-collecting solitary bees with a geographical distribution restricted to the western hemisphere. Most of the Emphorini bees collect Page 10 linepollen from a few specific plant families and display specialized behaviors for constructing their nests. Insect sensilla are the basic structural and functional units of cuticle receptors, serving mainly mechano- and chemo-receptor functions. The external morphology of the antennal sensilla has been well characterized in species of different families of Apoidea, however there is scarce information about this issue in solitary bees of the family Apidae. For a better understanding of the association between the external sensory system and several types of behaviors which emerged along the evolutionary history of bees, it is important to characterize the antennal receptors in several representative species of this tribe. The distribution of the antennal sensilla on the dorsal flagella of 18 taxa was studied in insects of both sexes, using light and scanning electron microscopy. There were six types of sensilla and setae on the antennae, which were identified as sensilla placodea, trichodea, basiconica, coeloconica, coelocapitular and ampullacea. The sensilla trichodea were classified into subtypes, A, B, C-D. Sensilla subtype A were the most abundant sensilla and were distributed over the entire antennae, while sensilla placodea and sensilla trichodea type B, showed a restricted distribution on specific areas of the flagella. We have recognized four patterns of spatial distribution of setae on dorsal flagella. Species having setae on the distal part of the flagellomeres tended to contain a low density of sensilla trichodea type A. Females showed a higher number of sensilla subtypes B and C-D than males; instead sensilla trichodea A were more abundant in males. No significant difference was found in the number of sensilla placodea, ampullacea, coeloconica and coelocapitular. Sensilla basiconica were found only in females. Our results showed that gustative and tactile sensilla were more abundant in female bees, as well as, olfactory receptors predominate in the antennal system of males. The possible coevolution of flagellar sensilla in males and females of solitary bees is discussed in light of previous reports. Patterns of distribution of setae determine the relative abundance of the types of sensilla in the flagellum.

Localization of cholecystokinin-like immunoreactivity in the central nervous system of Triatoma infestans (Insecta: Heteroptera).

The distribution of cholecystokinin-like immunoreactivity was studied in the central nervous system of the heteropteran insect Triatoma infestans using high-sensitivity immunocytochemistry. In the protocerebrum, CCK-IR somata were observed in the anteromedial, anterolateral and posterior cell-body layers. The neuropils displayed different densities of immunoreactive neurites. Few immunoreactive somata were found in the optic lobe in both the medial and lateral soma rinds, as well as in the proximal optic lobe. Immunoreactive fibers were present in the medulla and lobula neuropils. The sensory deutocerebrum contained a higher number of immunopositive perikarya than the antennal mechanosensory and motor center. The antennal lobe glomeruli displayed a moderate density of immunoreactive fibers. With regard to the subesophageal ganglion, numerous CCK-IR somata were found close to the root of the mandibular nerve; others were present in the soma rind of the remaining neuromeres. CCK-IR perikarya were present in both thoracic ganglia, with the abdominal neuromeres containing the highest number of positive somata. The neuropils of both ganglia showed moderate densities of immunopositive processes. The distribution of CCK-LI in somata and neuropils of central nervous system of T. infestans is widespread suggesting that a CCK-like peptide may act mainly as a neuromodulator in the integration of information from distinct sensory receptors.