Project description:As a model hemimetabolous insect species and an invasive urban pest that is globally distributed, the American cockroach, Periplaneta americana, is of great interest in both basic and applied research. Previous studies on P. americana neuropeptide identification have been based on biochemical isolation and molecular cloning. In the present study, an integrated approach of genomics- and peptidomics-based discovery was performed for neuropeptide identification in this insect species. Using large-scale peptidomic analysis of peptide extracts from 4 different tissues (the central nervous system, corpora cardiac and corpora allata complex, midgut, and male accessory gland), 35 conserved (predicted) and a potential (novel) neuropeptides were then identified. Subsequent experiments revealed the tissue distribution, sex difference, and developmental patterns of 2 conserved neuropeptides (allatostatin B and short neuropeptide F) and a novel neuropeptide PaOGS36577. Our study shows a comprehensive neuropeptidome and detailed spatiotemporal distribution patterns, providing a solid basis for future functional studies of neuropeptides in the American cockroach.
Project description:Background: Germ-free or axenic organisms are valuable tools for studying immunity, digestion, and development in different hosts. Although most of these studies have been conducted on mice, recently, germ-free invertebrate models (e.g. Drosophila and Apis) are used due to their easy husbandry, low cost for production, maintenance and the high number of individuals per generation they produce. However, a limitation of using these insects is the simple bacterial community present in their guts. The gut of the American cockroach Periplaneta americana displays a complex gut bacterial community composed of hundreds of species. Using P. americana, we developed a germ-free omnivorous invertebrate model to investigate how gut bacteria stimulate and shape normal gut development and metabolism. To determine if the insect host is directly affected by the presence of specific members of their bacterial community, gnotobiotic cockroaches were generated by inoculating a set of various P. americana gut-endemic Gram-negative (Bacteroidetes; n=11) and Gram-positive (Firmicutes; n=2) bacterial strains into germ-free insects. Additionally, we were able to recover the ‘normal’ bacterial-induced gut phenotype by co-housing germ-free cockroaches with wildtype P. americana to produce gut-bacteria conventionalized insects. Changes in gene expression profiles from two distinct regions (midgut and hindgut) of P. americana guts were quantified by RNA-Seq analysis of the germfree, gnotobiotic and conventionalized insects. Basic transcriptomics description: High-resolution transcriptome profiling of germ-free, gnotobiotic, and conventionalized treated P. americana midgut and hindguts. Ca. 43 million reads were obtained for each treatment. A de-novo assembly of all sequence reads was performed by Trinity assembler. Transcriptome assembly yielded 369,082 gene models and 554,155 isoforms. After running Trinotate pipeline, 65,047 (12 %) these transcripts matched an annotated product in at least one of the reference databases used (Uniprot, pfam, KEGG, COG). Additionally, 1,008 putative bacterial genes were annotated in the P. americana genome and ultimately excluded from these analyses. After bacteria decontamination, 553,147 assembled isoforms were used for transcript quantification and differential expression analysis using the DESeq2 pipeline. DESeq2 analysis detected 6,730 and 3,958 differentially expressed transcripts among the germ-free, gnotobiotic and conventionalized treatments in P. americana hindgut and midgut, respectively.
Project description:Various organic and natural pesticides play a vital role in insect control, leading to variation in their protein profile (Abubakar et al., 2019; James & Xu, 2012). The sex, size, weight, and growth conditions could be the possible factors associated with fluctuations in the concentration of protein production in the cockroach (Bouchebtiet al., 2022) The current study of Perplanata americana under pesticide activity considered to observe the proteomics of Brain and protein expression under stressed conditions. 15 cockroaches were exposed to 5g of Laxman Rekha (11:1 ratio) to assess the impact of pesticide on protein synthesis. National and international research conducted on aqueous and alcoholic extraction of gut, exoskeleton and brain protein of Periplaneta spp. and Blatta spp have revealed 10 potential antibacterial proteins between 18 kDa to 72kDa range. These proteins were purified using chromatographic techniques like single dimension reducing PAGE, FPLC-MS, LC-MS, MALDI TOF-MS etc. The current study is aimed at isolation, identification and characterisation of proteins from cockroach brain tissue lysate using Liquid chromatography and Orbitrap Mass spectrometer technique. The data analysis was done using Thermo Proteome Discoverer software version 2.2. 0.388. The database used was Uniprot-Periplaneta americana.
2024-06-27 | PXD052356 | Pride
Project description:Raw Genome Sequences of American cockroach (Periplaneta americana)