- Transcriptomics supports local sensory regulation in the antenna of the kissing- bug Rhodnius prolixus. - New nuclear receptor and takeout gene sequences were identified for this species, as well as those of enzymes involved in the biosynthesis and processing of neuropeptides and biogenic amines.. - Conclusions: We report a broad repertoire of neuromodulatory and neuroendocrine-related genes expressed in the antennae of R. - Diverse neuropeptide precursor genes showed consistent expression in the antennae of all stages studied. - In fact, variations in gene expression depending on nutritional status or de- velopment have been described for olfactory co- receptors in the antennae of R. - In this sense, the main objective of this study is to characterize modulatory components potentially involved in the local regulation of antennal sensory function using R. - Furthermore, we looked at gene expression in the antennae of 5 th instar larvae and, male and female adults to characterize changes in this regulatory gene repertoire potentially associated with imaginal moult and sex.. - Besides, IDLSRF-like peptide, glycoprotein hor- mones alpha-2 (GPA2) and beta-5 (GPB5), and bursicon- beta (also known as the partner of bursicon- Burs) genes were identified in the R. - prolixus adipokinetic hormone (AKH) precur- sor gene, originated through alternative splicing, was iden- tified in the antennal assemblies. - Both AKH isoforms share the signal peptide and the active conserved peptide, but differ in the C-terminal region. - some fragments were located in the opposite strand or were ab- sent from the genome assembly.. - In the case of Family A neuropeptide receptor genes, a total of 15 gene models based on Ons et al. - [30] were included in the GFF file of the R. - Besides, 11 gene models of this receptor family were edited in the existing GFF file of the genome. - Most of the Family B neuropeptide receptor gene models were fixed and included in the modified version of the R. - All enzymes known to mediate biogenic amine biosyn- thesis in other insects were annotated in the last version of the R. - 2) DOPA decarboxylase (Ddc), involved in the synthesis of dopamine from L-DOPA. - The neuropeptide processing enzymes were not previ- ously annotated in the R. - 4) the carboxypeptidase M - CPM (two new isoforms were identified in the antennal assemblies with differences in the 3′ region) and D (known as silver, which trims C-terminal Arg and Lys after Furins/PC2 cleavage reaction). - To date, the ecdysone receptor (EcR) gene was the only nuclear receptor annotated in the R. - melanoga- ster (Additional file 7: Figure S2), we have annotated 20 additional nuclear receptor genes in the R. - The orthologs of knirps (kni) and hormone receptor-like 83 (HR83) genes were not identified either in the R. - humanus genomes, while the gene named eagle (ea) is ab- sent in the genomes of R. - Interestingly, two paralogs of the hepatocyte nuclear factor 4 ( HNF4) gene (A and B) were found in the R. - Three takeout (to) genes had been previously identified in the R. - two conserved cysteine resi- dues in the N-terminal region and two conserved charac- teristic motifs [34]. - Data from our RNA-Seq ex- periment were used to previously report changes in the expression of genes with sensory function, e.g., ORs [17].. - Out of the 44 neuropeptide precursor genes annotated in the R. - Allatostatin-CC (AstCC), allatostatin-CCC (AstCCC), ITG-like, IDLSRF-like peptide and OK were the most highly expressed neuropeptide genes in the an- tennae of R. - 0.05) was also observed in the antennae of both adult stages when compared to larvae. - Interestingly, the expression of K receptor 2 (K-R2) increased significantly in the antennae of adults (FDR <. - In the case of Family B neuropeptide receptor genes, only CT/DH-R2 showed FPKM values lower than 1 (Additional file 11: Data file S1). - In fact, its expression showed a significant increase in the antennae of males (FDR <. - Regarding opsin expression, transcripts of UV opsin and long-wave sensitive opsin 1 (LWS1) were detected in the three librar- ies (Fig. - Tyrosine kinase and guanylyl cyclase type receptors The neuropeptide-like precursor 1 putative receptor or NPLP1-R (tyrosine kinase-type) and the potential neuro- parsin receptor or NP-R (guanylyl cyclase receptor) were found to be expressed in the antennae of R. - All enzymes involved in neuropeptide processing, except prohormone convertase 1, were expressed in the antennae of R. - Expression of at least 16 out of 20 biogenic amine recep- tor genes was detected in the antennae of R. - All genes encoding for enzymes involved in the bio- synthetic pathway of biogenic amines were detected in the antennae of R. - 10 in the three libraries (Fig. - 1 in the three libraries) in the R. - 1 Heat map comparing the expression levels of (a) neuropeptide precursor genes, (b) G protein-coupled receptor genes, and (c) neuropeptide processing enzymes in the antennae of R. - In the first place, Rproto11 gene showed a signifi- cant decrease after imaginal moult (FDR <. - Our study has characterized the expression profile of a diverse set of genes encoding different modulatory elements (neuropeptides, GPCRs, nuclear receptors and to genes) in the antenna of R. - Our results have proven that neuropeptide gene tran- scripts are produced in the antennae of kissing-bugs (a total of 31 neuropeptide genes were found to be expressed). - The expression of neuropeptide gene tran- scripts has already been reported in the antennae of a few insect species . - prolixus nuclear receptor genes in the antennae of larvae (L), female (F) and male (M) adults. - prolixus to genes in the antennae of larvae (L), female (F) and male (M) adults.. - The significantly augmented expression of K-R2 observed in the antennae of adults (Fig. - A similar in- creased adult expression profile was previously observed in the antennae of R. - Therefore, it would be interesting to study whether these are functionally connected in the adult phase. - The sig- nificant decrease observed in the expression of the AKH- R gene in female antennae may suggest a relation to the modulation of pheromone perception and production as it was observed for D. - It would be interest- ing to analyse its functional role in the antennae of. - Actually, in situ hybridization allowed detecting octopamine and tyramine receptor gene transcripts in the vicinity of sensory recep- tor neurons of different insects [22, 47]. - Consistent with these findings, a diverse set of transcripts of biogenic amine receptors was identified in the antennal transcriptome of R. - As observed for neuropep- tides, most of the genes coding for enzymes involved in the biosynthesis of biogenic amines seem to be expressed in R. - Interestingly, this receptor modulates sex pheromone sensi- tivity in the antennal lobe of male moths [50]. - Here we show that most insect nuclear receptors are expressed in the antennae of an insect (Fig. - melanogaster and control the expression of clock genes, playing an im- portant role in the maintenance of locomotor rhythms [52, 53]. - [56] reported 19 nuclear hor- mone receptor genes in the genome of R. - [57], we have also found two knrl genes of the NR0 sub- family in the R. - Furthermore, we con- firm here that HR83 is not present in the genomes of R.. - Fifteen to genes were identified in the R. - [59] identified 25 to transcripts in the transcriptome of the kissing-bug Triatoma brasiliensis.. - This putative function could explain the high expression ob- served in the three antennal libraries (Fig. - 4), however, functional studies need to be performed to be able to confirm these roles in the antennae of kissing-bugs. - Heat maps showing gene expression (expressed as Log10 FPKM value + 1) of the different protein families in the conditions tested were prepared using the gplot package in R.. - In the case of neuropeptide precursor and GPCR genes that were manually corrected/extended, new GFF files were created and included in the RproC3.3 version of the R. - The protein sequences of all genes analysed and the edited GFF files are included in the Add- itional file 13: Data file S2 and Additional file 14: Data file S3, respectively).. - were used in tBLASTn searches in the R. - prolixus [15] were used as queries to search for new sequences in the genome. - Fasta sequences from different in- sects used in the phylogenetic analyses of CT/DH – CRF/DH and nuclear receptor genes are included in Additional file 15: Data file S4.. - number in the RproC3 genome assembly, prefix is RPRC. - AAs – number of encoded amino acids in the protein. - Comments – comments on the OGS gene model and repairs performed in the genome assembly based on BLAST against de novo antennal assemblies. - VectorBase code – the official gene number in the RproC3 genome assembly, prefix is RPRC. - Com- ments – comments on the OGS gene model and repairs performed in the genome assembly (available on VectorBase) based on BLAST against de novo antennal assemblies. - Details of enzymes involved in the biogenic amine synthesis. - VectorBase code – the official gene number in the RproC3 genome assembly, prefix is RPRC;. - FPKM values of target genes in the three libraries.. - Fasta sequences from different insects used in the phylogenetic analyses of CT/DH – CRF/DH and nuclear receptor genes.. - Gabriel da Rocha Fernandes and Fausto Gonçalves dos Santos for their help and support in the bioinformatic analyses.. - All these funds provided financial support in the design of the study and col- lection, analysis, and interpretation of data and in writing the manuscript.. - Circadian organization of locomotion activity in the. - Serotonin: a coordinator of feeding-related physiological events in the blood-gorging bug, Rhodnius prolixus. - Behavioural and physiological state dependency of host seeking in the blood-sucking insect Rhodnius prolixus. - Orcokinin neuropeptides regulate ecdysis in the hemimetabolous insect Rhodnius prolixus. - RNA-seq analyses of blood-induced changes in gene expression in the mosquito vector species, Aedes aegypti. - Neuromodulation of olfactory sensitivity in the peripheral olfactory organs of the American cockroach, Periplaneta americana. - Cloning and expression of long neuropeptide F and the role of FMRFamide-like peptides in regulating egg production in the Chagas vector, Rhodnius prolixus. - Ultrastructural analysis of neurosecretory cells in the antennae of the mosquito, Culex salinarius (Diptera: Culicidae). - Neuroendocrine modulation of olfactory sensory neuron signal reception via axo-dendritic synapses in the antennae of the mosquito, Aedes aegypti. - Adipokinetic hormone signalling system in the Chagas disease vector, Rhodnius prolixus. - Adipokinetic hormone receptor gene identification and its role in triacylglycerol metabolism in the blood-sucking insect Rhodnius prolixus. - Age-and behaviour-related changes in the expression of biogenic amine receptor genes in the antennae of honey bees (Apis mellifera). - Immunocytochemical localization of serotonin in the central and peripheral chemosensory system of mosquitoes.. - Neuropeptide precursor gene discovery in the Chagas disease vector Rhodnius prolixus.. - Neuropeptides in the regulation of Rhodnius prolixus physiology. - Peptidomics and processing of regulatory peptides in the fruit fly Drosophila. - annotation and expression in the adult brain. - Sexual dimorphic expression of putative antennal carrier protein genes in the malaria vector Anopheles gambiae
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