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1.
Pest Manag Sci ; 80(9): 4495-4504, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38676657

RESUMEN

BACKGROUND: Cockroaches are widely acknowledged as significant vectors of pathogenic microorganisms. The Periplaneta fuliginosa densovirus (PfDNV) infects the smoky-brown cockroach P. fuliginosa and causes host mortality, which identifies the PfDNV as a species-specific and environmentally friendly biopesticide. However, although the biochemical characterization of PfDNV has been extensively studied, the immune response against PfDNV remains largely unclear. RESULTS: Here, we investigated the replication of PfDNV and its associated pathological phenotype in the foregut and hindgut. Consequently, we dissected and performed transcriptome sequencing on the foregut, midgut, and hindgut separately. We revealed the up-regulation of immune response signaling pathway c-Jun N-terminal kinase (JNK) and apoptosis in response to viral infection. Furthermore, knockdown of the JNK upstream gene Ben resulted in a decrease in virus titer and delayed host mortality. CONCLUSION: Taken together, our findings provide evidence that the Ben-JNK signaling plays a crucial role in PfDNV infection, leading to excessive apoptosis in intestinal tissues and ultimately resulting in the death of the host. Our results indicated that the host response to PfDNV fosters viral infection, thereby increasing host lethality. This underscores the potential of PfDNV as a viable, environmentally friendly biopesticide. © 2024 Society of Chemical Industry.


Asunto(s)
Densovirus , Periplaneta , Animales , Periplaneta/virología , Periplaneta/fisiología , Periplaneta/genética , Densovirus/fisiología , Densovirus/genética , Proteínas de Insectos/metabolismo , Proteínas de Insectos/genética , Sistema de Señalización de MAP Quinasas , Apoptosis , Transducción de Señal
2.
Sci China Life Sci ; 67(7): 1455-1467, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38523236

RESUMEN

Volatile sex pheromones are vital for sexual communication between males and females. Females of the American cockroach, Periplaneta americana, produce and emit two sex pheromone components, periplanone-A (PA) and periplanone-B (PB). Although PB is the major sex attractant and can attract males, how it interacts with PA in regulating sexual behaviors is still unknown. In this study, we found that in male cockroaches, PA counteracted PB attraction. We identified two odorant receptors (ORs), OR53 and OR100, as PB/PA and PA receptors, respectively. OR53 and OR100 were predominantly expressed in the antennae of sexually mature males, and their expression levels were regulated by the sex differentiation pathway and nutrition-responsive signals. Cellular localization of OR53 and OR100 in male antennae further revealed that two types of sensilla coordinate a complex two-pheromone-two-receptor pathway in regulating cockroach sexual behaviors. These findings indicate distinct functions of the two sex pheromone components, identify their receptors and possible regulatory mechanisms underlying the male-specific and age-dependent sexual behaviors, and can guide novel strategies for pest management.


Asunto(s)
Periplaneta , Receptores Odorantes , Atractivos Sexuales , Conducta Sexual Animal , Animales , Masculino , Atractivos Sexuales/metabolismo , Femenino , Receptores Odorantes/metabolismo , Receptores Odorantes/genética , Periplaneta/metabolismo , Periplaneta/fisiología , Periplaneta/genética , Conducta Sexual Animal/fisiología , Antenas de Artrópodos/metabolismo , Antenas de Artrópodos/fisiología , Comunicación Animal , Proteínas de Insectos/metabolismo , Proteínas de Insectos/genética , Receptores de Feromonas/metabolismo , Receptores de Feromonas/genética
3.
Bull Entomol Res ; 113(6): 787-793, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-38037350

RESUMEN

Periplaneta americana, one of the most widely distributed insects all over the world, can survive and reproduce in harsh environment which may be closely related to the critical roles of intestinal microorganisms in its multiple physiological functions. However, the composition and structure of gut microbiota throughout different life stages and its effects on the strong resilient and environmental adaptability of P. americana remain unclear. In this study, the gut microbiota across life stages including ootheca (embryos), nymph and adult of P. americana were investigated by 16S rRNA high-throughput sequencing. Multivariate statistical analysis showed the richness and diversity of bacterial communities were significantly different among ootheca, nymph and adult stage of P. americana. Taxonomic analysis showed Blattabacterium was the dominant genus in bacterial community of ootheca while the nutrient absorption-related genera including Christensenellaceae and Ruminococcaceae showed high relative abundance in nymph samples. Moreover, functional prediction analysis showed the metabolic categories in ootheca might have more influence on the basic life activities of the host than improved production and viability, while it was more associated to the society activities, reproduction and development of host in nymph and adult. It was suggested that the gut microbiota in each life stage might meet the requirements for environmental adaptability and survival of P. americana via transforming the composition and structure with specific metabolic capabilities. Overall, these results provided a novel sight to better understand the strong vitality and adaptability throughout life stages of P. americana.


Asunto(s)
Microbioma Gastrointestinal , Periplaneta , Animales , Periplaneta/genética , Periplaneta/microbiología , ARN Ribosómico 16S/genética , Bacterias
4.
Insect Mol Biol ; 32(1): 46-55, 2023 02.
Artículo en Inglés | MEDLINE | ID: mdl-36214335

RESUMEN

Insects are the most widely distributed and successful animals on the planet. A large number of insects are capable of flight with functional wings. Wing expansion is an important process for insects to achieve functional wings after eclosion and healthy genital morphology is crucial for adult reproduction. Myofilaments are functional units that constitute sarcomeres and trigger muscle contraction. Here, we identified four myofilament proteins, including Myosin, Paramyosin, Tropomyosin and Troponin T, from the wing pads of nymphs in the American cockroach, Periplaneta americana. RNAi-mediated knockdown of Myosin, Paramyosin, Tropomyosin and Troponin T in the early stage of final instar nymphs caused a severely curly wing phenotype in the imaginal moult, especially in the Paramyosin and Troponin T knockdown groups, indicating that these myofilament proteins are involved in controlling wing expansion behaviours during the nymph-adult transition. In addition, the knockdown resulted in abnormal external genitalia, caused ovulation failure, and affected male accessory gland development. Interestingly, the expression of myofilament genes was induced by methoprene, a juvenile hormone (JH) analogue, and decreased by the depletion of the JH receptor gene Met. Altogether, we have determined that myofilament genes play an important role in promoting wing expansion and maintaining adult genitalia morphology, and their expression is induced by JH signalling. Our data reveal a novel mechanism by which wing expansion is regulated by myofilaments and the functions of myofilaments are involved in maintaining genitalia morphology.


Asunto(s)
Periplaneta , Femenino , Masculino , Animales , Periplaneta/genética , Periplaneta/metabolismo , Miofibrillas , Tropomiosina/genética , Tropomiosina/metabolismo , Troponina T/metabolismo , Metamorfosis Biológica , Insectos , Hormonas Juveniles/metabolismo , Ninfa
5.
Int J Mol Sci ; 23(3)2022 Jan 31.
Artículo en Inglés | MEDLINE | ID: mdl-35163598

RESUMEN

Biogenic amines constitute an important group of neuroactive substances that control and modulate various neural circuits. These small organic compounds engage members of the guanine nucleotide-binding protein coupled receptor (GPCR) superfamily to evoke specific cellular responses. In addition to dopamine- and 5-hydroxytryptamine (serotonin) receptors, arthropods express receptors that are activated exclusively by tyramine and octopamine. These phenolamines functionally substitute the noradrenergic system of vertebrates Octopamine receptors that are the focus of this study are classified as either α- or ß-adrenergic-like. Knowledge on these receptors is scarce for the American cockroach (Periplaneta americana). So far, only an α-adrenergic-like octopamine receptor that primarily causes Ca2+ release from intracellular stores has been studied from the cockroach (PaOctα1R). Here we succeeded in cloning a gene from cockroach brain tissue that encodes a ß-adrenergic-like receptor and leads to cAMP production upon activation. Notably, the receptor is 100-fold more selective for octopamine than for tyramine. A series of synthetic antagonists selectively block receptor activity with epinastine being the most potent. Bioinformatics allowed us to identify a total of 19 receptor sequences that build the framework of the biogenic amine receptor clade in the American cockroach. Phylogenetic analyses using these sequences and receptor sequences from model organisms showed that the newly cloned gene is an ß2-adrenergic-like octopamine receptor. The functional characterization of PaOctß2R and the bioinformatics data uncovered that the monoaminergic receptor family in the hemimetabolic P. americana is similarly complex as in holometabolic model insects like Drosophila melanogaster and the honeybee, Apis mellifera. Thus, investigating these receptors in detail may contribute to a better understanding of monoaminergic signaling in insect behavior and physiology.


Asunto(s)
Adenilil Ciclasas , Señalización del Calcio , Proteínas de Insectos , Periplaneta , Receptores de Amina Biogénica , Adenilil Ciclasas/genética , Adenilil Ciclasas/metabolismo , Animales , AMP Cíclico/genética , AMP Cíclico/metabolismo , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Octopamina/metabolismo , Periplaneta/genética , Periplaneta/metabolismo , Receptores de Amina Biogénica/genética , Receptores de Amina Biogénica/metabolismo
6.
Insect Biochem Mol Biol ; 131: 103551, 2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33556555

RESUMEN

Matrix metalloproteinases (MMPs) are the major proteinases that process or degrade numerous extracellular matrix (ECM) components and are evolutionarily conserved from nematodes to humans. During molting in insects, the old cuticle is removed and replaced by a new counterpart. Although the regulatory mechanisms of hormones and nutrients in molting have been well studied, very little is known about the roles of ECM-modifying enzymes in this process. Here, we found that MMPs are necessary for imaginal molting of the American cockroach, Periplaneta americana. Inhibition of Mmp activity via inhibitor treatment led to the failure of eclosion and wing expansion. Five Mmps genes were identified from the P. americana genome, and PaMmp2 played the dominant roles during molting. Further microscopic investigations showed that newly formed adult cuticles were attenuated and that then chitin content was reduced upon Mmp inhibition. Transcriptomic analysis of the integument demonstrated that multiple signaling and metabolic pathways were changed. Microscopic investigation of the wings showed that epithelial cells were restrained together because they were incapable of degrading the ECM upon Mmp inhibition. Transcriptomic analysis of the wing identified dozens of possible genes functioned in wing expansion. This is the first study to show the essential roles of Mmps in the nymph-adult transition of hemimetabolous insects.


Asunto(s)
Metaloproteinasas de la Matriz , Periplaneta , Alas de Animales , Animales , Quitina/metabolismo , Perfilación de la Expresión Génica , Genes de Insecto , Larva/metabolismo , Metaloproteinasas de la Matriz/genética , Metaloproteinasas de la Matriz/metabolismo , Metamorfosis Biológica , Muda , Ninfa/metabolismo , Periplaneta/embriología , Periplaneta/genética , Periplaneta/metabolismo , Periplaneta/fisiología , Alas de Animales/embriología , Alas de Animales/metabolismo
7.
J Vis Exp ; (178)2021 12 17.
Artículo en Inglés | MEDLINE | ID: mdl-34978287

RESUMEN

Cockroaches, a sanitary pest, are essential species in insect developmental and metamorphic studies due to their easy feeding and hemimetabolous characteristics. Altogether with well-annotated genome sequences, these advantages have made American cockroach, Periplaneta americana, an important hemimetabolous insect model. Limited by the shortage of knockout strategy, effective RNA interference (RNAi)-based gene knockdown becomes an indispensable technique in functional gene research of P. americana. The present protocol describes the RNAi operation techniques in P. americana. The protocol includes (1) selection of the P. americana at proper developmental stages, (2) preparation for the injection setting, (3) dsRNA injection, and (4) gene knockdown efficiency detection. RNAi is a powerful reverse genetic tool in P. americana. The majority of P. americana tissues are sensitive to extracellular dsRNA. Its simplicity allows researchers to quickly obtain dysfunctional phenotypes under one or multiple targeting dsRNA injections, enabling researchers to better use the P. americana for developmental and metamorphic studies.


Asunto(s)
Cucarachas , Periplaneta , Animales , Cucarachas/genética , Insectos/genética , Periplaneta/genética , Interferencia de ARN , ARN Bicatenario/genética
8.
J Proteome Res ; 20(2): 1217-1228, 2021 02 05.
Artículo en Inglés | MEDLINE | ID: mdl-33166158

RESUMEN

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. First, 67 conserved neuropeptide or neurohormone precursor genes were predicted via an in silico analysis of the P. americana genome and transcriptome. Using a large-scale peptidomic analysis of peptide extracts from four different tissues (the central nervous system, corpora cardiac and corpora allata complex, midgut, and male accessory gland), 35 conserved (predicted) neuropeptides and a potential (novel) neuropeptide were then identified. Subsequent experiments revealed the tissue distribution, sex difference, and developmental patterns of two 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 (data are available via ProteomeXchange with identifier PXD021660).


Asunto(s)
Neuropéptidos , Periplaneta , Secuencia de Aminoácidos , Animales , Femenino , Genómica , Masculino , Neuropéptidos/genética , Péptidos/genética , Periplaneta/genética
9.
Peptides ; 136: 170466, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33253775

RESUMEN

The SMYamide genes are paralogs of the SIFamide genes and code for neuropeptides that are structurally similar to SIFamide. In the American cockroach, Periplanea americana, the SMYamide gene is specifically expressed in the SN2 neurons that innervate the salivary glands and are known to produce action potentials during feeding. The SN2 axon terminals surround rather than directly innervate the salivary gland acini. Therefore one may expect that on activation of these neurons significant amounts of SMYamide will be released into the hemolymph, thus suggesting that SMYamide may also have a hormonal function. In the Periplaneta genome there are two putative SIFamide receptors and these are both expressed not only in the central nervous system and the salivary gland, but also in the gonads and other peripheral tissues. This reinforces the hypothesis that SMYamide also has an endocrine function in this species.


Asunto(s)
Neuronas/metabolismo , Neuropéptidos/genética , Receptores de Neuropéptido/genética , Glándulas Salivales/metabolismo , Animales , Ganglios de Invertebrados/metabolismo , Regulación de la Expresión Génica/genética , Neuropéptidos/metabolismo , Periplaneta/genética , Periplaneta/metabolismo , Glándulas Salivales/inervación
10.
DNA Cell Biol ; 38(10): 1078-1087, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31524500

RESUMEN

The globally distributed American cockroach (Periplaneta americana) is considered a pest, but it has been widely used in traditional Chinese medicine. In the past, the American cockroach's genome and transcriptomes were sequenced, but the differential expression transcripts between developmental stages were unavailable. We performed the de novo assembly and analysis of American cockroach transcriptomes from four developmental stages. Approximately 200 million high-quality paired-end reads were generated by using Illumina Hiseq 2000 sequencer. The assembly produced 291,250 transcripts with an average length of 714 bp. In addition, 38,052 microsatellites and 11,060,020 transposable elements were identified. Based on sequence homology, 53,262 transcripts were annotated. After calculating the expression levels of all the transcripts, we found that 13 transcripts were highly expressed in all the samples and at least two, p10 and actin-related protein 1, played important roles during development. A total of 7954 differentially expressed transcripts (DETs) were identified. The adult had the largest number of DETs when compared to other samples (4818), while the 3rd and 8th larva had the least number of DETs (1332). We performed gene enrichment analysis with the DETs, and some interesting results were detected in the different groups. For example, chitin is the major component of the insect exoskeleton, and the chitin-related genes in larvae and new molted samples had higher expression levels than in adults. In addition, the enrichment analysis detected many chitin-related pathways. Our study performed the first large-scale comparative transcriptomics between the developmental stages of American cockroach, which could provide useful gene expression data for future studies.


Asunto(s)
Genoma de los Insectos , Estadios del Ciclo de Vida/genética , Redes y Vías Metabólicas/genética , Periplaneta/genética , Transcriptoma , Animales , Elementos Transponibles de ADN , Femenino , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Ontología de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Masculino , Repeticiones de Microsatélite , Anotación de Secuencia Molecular , Periplaneta/clasificación , Periplaneta/crecimiento & desarrollo , Periplaneta/metabolismo , Filogenia
11.
Biochem J ; 476(8): 1267-1284, 2019 04 26.
Artículo en Inglés | MEDLINE | ID: mdl-30962256

RESUMEN

Communications between various organelle-organelles play an essential role in cell survival. The cross-talk between mitochondria and vacuoles comes up with the vital roles of the intercompartmental process. In this study, we found a couple of cell death features, membrane damage, and apoptosis using antimicrobial peptide from American Cockroach. Periplanetasin-4 (LRHKVYGYCVLGP-NH2) is a 13-mer peptide derived from Periplaneta americana and exhibits phosphatidylserine exposure and caspase activation without DNA fragmentation. Apoptotic features without DNA damage provide evidence that this peptide did not interact with DNA directly and exhibited dysfunction of mitochondria and vacuoles. Superoxide radicals were generated from mitochondria and converted to hydrogen peroxide. Despite the enhancement of catalase and total glutathione contents, oxidative damage disrupted intracellular contents. Periplanetasin-4 induced cell death associated with the production of superoxide radicals, calcium uptake in mitochondria and disorder of vacuoles, such as increased permeability and alkalization. While calcium movement from vacuoles to the mitochondria occurred, the cross-talk with these organelles proceeded and the inherent functionality was impaired. To sum up, periplanetasin-4 stimulates superoxide signal along with undermining the mitochondrial functions and interfering in communication with vacuoles.


Asunto(s)
Péptidos Catiónicos Antimicrobianos/metabolismo , Proteínas de Insectos/metabolismo , Mitocondrias/metabolismo , Periplaneta/metabolismo , Vacuolas/metabolismo , Animales , Péptidos Catiónicos Antimicrobianos/genética , Apoptosis/genética , Señalización del Calcio/genética , Fragmentación del ADN , Proteínas de Insectos/genética , Mitocondrias/genética , Periplaneta/genética , Vacuolas/genética
12.
Artículo en Inglés | MEDLINE | ID: mdl-31022468

RESUMEN

Juvenile hormones (JH) regulate wide-ranging physiological and developmental processes in insects. However, molecular mechanisms underlying JH signaling remain to be determined. Vitellogenin (Vg) is primarily an egg-yolk protein, but recently proposed to serve many functions in insects. In the female American cockroach (Periplaneta americana), vitellogenin (Vg) genes are activated by JH III and suppressed by 20-hydroxyecdysone (20E) via cis-regulatory elements in a dose-dependent manner. In the present study, the upstream promoter region (935 bp) of Vg1 was cloned to elucidate the action of these hormones. A luciferase reporter assay identified an 81 bp region in the promoter region of Vg1 (-120 to -39 bp) that we found to be critical for JH III activation and 20E suppression. This 81 bp region contains a direct repeat separated by a 2-nucleotide spacer-designated Vg1HRE- that is similar to the Drosophila ecdysone response element direct repeat 4. Moreover, nuclear proteins isolated from nymphs, males, females, and Sf9 cells successfully bound to Vg1HRE, while binding was outcompeted by a 100-fold excess of cold probe or dephosphorylated nuclear protein extracts. In addition, binding was outcompeted by other ecdysone and JH response elements with similar half-site sequences (direct repeats) but to varying extents. Ultimately, we postulate that JH III indirectly activates Vg expression by interfering with or inhibiting the phosphorylation of nuclear proteins bound to Vg1HRE. Involvement of JH III in both induction of Vg1 and control of nuclear proteins binding to Vg1HRE suggest the latter to play an important role in JH signaling.


Asunto(s)
Regulación de la Expresión Génica , Proteínas de Insectos , Periplaneta , Secuencias Repetitivas de Ácidos Nucleicos , Elementos de Respuesta , Vitelogeninas , Animales , Femenino , Proteínas de Insectos/biosíntesis , Proteínas de Insectos/genética , Hormonas Juveniles/genética , Hormonas Juveniles/metabolismo , Masculino , Ninfa , Periplaneta/genética , Periplaneta/metabolismo , Transducción de Señal/genética , Vitelogeninas/biosíntesis , Vitelogeninas/genética
13.
Ann Clin Lab Sci ; 48(3): 323-327, 2018 May.
Artículo en Inglés | MEDLINE | ID: mdl-29970435

RESUMEN

OBJECTIVE: To clone, express, purify the Per a 4 gene encoding an allergen of Periplaneta Americana and prepare monoclonal antibodies against the recombinant allergen. METHODS: The total RNA was extracted from P. Americana, and the target gene was amplified by RT-PCR and cloned into pMD18-T vector. After being confirmed by nucleotide sequencing, the gene was then inserted into pGEX-3X to construct the express vector pGEX-3X-Per a 4. Further, the pGEX-3X-Per a 4 was transformed into E. coli BL21 (DE3), and induced for expression by IPTG. By affinity chromatography, the recombinant allergen was purified and identified by SDS-PAGE and Western blotting. The BALB/c mouse was immunized with the recombinant allergen to prepare the specific monoclonal antibodies, which was then identified by co-immunoprecipitatin and western blotting. RESULTS: The full-length cDNA encoding Per a 4 of P. Americana was obtained with 552 bp in length, which had 99.4% similarity with the reference sequence (GenBank AY792948). The constructed vector pGEX-3X-Per a 4 was transformed in E. coli BL21 (DE3), expressed with the induction of IPTG. By SDS-PAGE, a band of about 49 KD was present. Further, the western-blotting showed that the prepared monoclonal antibodies can bind the serum antibodies in patients allergic to P. Americana. CONCLUSIONS: Both the recombinant allergen Per a 4 and its monoclonal antibodies were obtained.


Asunto(s)
Alérgenos/inmunología , Anticuerpos Monoclonales/inmunología , Proteínas de Insectos/inmunología , Periplaneta/inmunología , Proteínas Recombinantes/inmunología , Alérgenos/genética , Alérgenos/aislamiento & purificación , Animales , Clonación Molecular , Proteínas de Insectos/genética , Proteínas de Insectos/aislamiento & purificación , Ratones , Ratones Endogámicos BALB C , Periplaneta/genética , Periplaneta/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación
14.
Insect Sci ; 25(5): 751-764, 2018 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-28548451

RESUMEN

The small Rab GTPases are key regulators of membrane vesicle trafficking. Ovaries of Periplaneta americana (Linnaeus) (Blattodea: Blattidae) have small molecular weight GTP/ATP-binding proteins during early and late vitellogenic periods of oogenesis. However, the identification and characterization of the detected proteins have not been yet reported. Herein, we cloned a cDNA encoding Rab5 from the American cockroach, P. americana, ovaries (PamRab5). It comprises 796 bp, encoding a protein of 213 amino acid residues with a predicted molecular weight of 23.5 kDa. PamRab5 exists as a single-copy gene in the P. americana genome, as revealed by Southern blot analysis. An approximate 2.6 kb ovarian mRNA was transcribed especially at high levels in the previtellogenic ovaries, detected by Northern blot analysis. The muscle and head tissues also showed high levels of PamRab5 transcript. PamRab5 protein was localized, via immunofluorescence labeling, to germline-derived cells of the oocytes, very early during oocyte differentiation. Immunoblotting detected a ∼25 kDa signal as a membrane-associated form revealed after application of detergent in the extraction buffer, and 23 kDa as a cytosolic form consistent with the predicted molecular weight from amino acid sequence in different tissues including ovary, muscles and head. The PamRab5 during late vitellogenic periods is required to regulate the endocytotic machinery during oogenesis in this cockroach. This is the first report on Rab5 from a hemimetabolan, and presents an inaugural step in probing the molecular premises of insect oocyte endocytotic trafficking important for oogenesis and embryonic development.


Asunto(s)
Variaciones en el Número de Copia de ADN , Proteínas de Insectos/genética , Oogénesis/genética , Periplaneta/fisiología , Transcripción Genética , Proteínas de Unión al GTP rab5/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Femenino , Perfilación de la Expresión Génica , Proteínas de Insectos/química , Proteínas de Insectos/metabolismo , Ovario/metabolismo , Periplaneta/genética , Filogenia , ARN Mensajero/genética , Alineación de Secuencia , Proteínas de Unión al GTP rab5/química , Proteínas de Unión al GTP rab5/metabolismo
15.
Insect Sci ; 25(6): 978-990, 2018 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28556457

RESUMEN

The physiological functions of insect foregut, especially in xenobiotic detoxification, are scarcely reported because of unimportance in appearance and insufficient molecular information. The cockroach Periplaneta americana, an entomological model organism, provides perfect material to study physiological functions of foregut tissue due to its architectural feature. Through Illumina sequencing of foregut tissue from P. americana individuals (control) or insects treated with cycloxaprid, as a novel neonicotinoid insecticide, 54 193 166 clean reads were obtained and further assembled into 53 853 unigenes with an average length of 366 bp. Furthermore, the number of unigenes involved in xenobiotic detoxification was analyzed, mainly including 70 cytochrome P450s, 12 glutathione S-transferases (GSTs), seven carboxylesterases (CarEs) and seven adenosine triphosphate-binding cassette (ABC) transporters. Compared to control, the expression of 22 xenobiotic detoxification unigenes was up-regulated after cycloxaprid application, mainly containing 18 P450s, one GST, two CarEs and one ABC adenosine triphosphate transporter, indicating that the oxidation-reduction was the major reactive process to cycloxaprid application. Through quantitative real-time polymerase chain reaction analysis, the expression of selected unigenes (six P450s, one GST and one CarE) was up-regulated at least two-fold following cycloxaprid treatment, and was generally in agreement with transcriptome data. Compared to the previous midgut transcriptome of P. americana, it looks like the expressive abundance of the xenobiotic detoxification unigenes might be important factors to the detoxifying functional differences between foregut and midgut. In conclusion, insect foregut would also play important roles in the physiological processes related to xenobiotic detoxification.


Asunto(s)
Tracto Gastrointestinal/metabolismo , Perfilación de la Expresión Génica , Inactivación Metabólica , Periplaneta/genética , Periplaneta/metabolismo , Xenobióticos/metabolismo , Animales , Periplaneta/anatomía & histología
16.
Sci Rep ; 7(1): 12483, 2017 10 02.
Artículo en Inglés | MEDLINE | ID: mdl-28970561

RESUMEN

Evolution of resistance among insects to action of pesticides has led to the discovery of several insecticides (neonicotinoids and organophosphates) with new targets in insect nervous system. Present study evaluates the mode of inhibition of acetylchlonesterase (AChE), biochemical efficacy, and molecular docking of 2,3-dimethylmaleic anhydride, against Periplaneta americana and Sitophilus oryzae. The knockdown activity of 2,3-dimethylmaleic anhydride was associated with in vivo inhibition of AChE. At KD99 dosage, the 2,3-dimethylmaleic anhydride showed more than 90% inhibition of AChE activity in test insects. A significant impairment in antioxidant system was observed, characterized by alteration in superoxide dismutase and catalase activities along with increase in reduced glutathione levels. Computational docking programs provided insights in to the possible interaction between 2,3-dimethylmaleic anhydride and AChE of P. americana. Our study reveals that 2,3-dimethylmaeic anhydride elicits toxicity in S. oryzae and P. americana primarily by AChE inhibition along with oxidative stress.


Asunto(s)
Acetilcolinesterasa/química , Inhibidores de la Colinesterasa/farmacología , Proteínas de Insectos/antagonistas & inhibidores , Insecticidas/farmacología , Anhídridos Maleicos/farmacología , Periplaneta/efectos de los fármacos , Gorgojos/efectos de los fármacos , Acetilcolinesterasa/genética , Acetilcolinesterasa/metabolismo , Secuencia de Aminoácidos , Animales , Catalasa/antagonistas & inhibidores , Catalasa/metabolismo , Dominio Catalítico , Inhibidores de la Colinesterasa/química , Relación Dosis-Respuesta a Droga , Expresión Génica , Glutatión/antagonistas & inhibidores , Glutatión/metabolismo , Proteínas de Insectos/química , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Insecticidas/química , Cinética , Anhídridos Maleicos/química , Simulación del Acoplamiento Molecular , Sistema Nervioso/efectos de los fármacos , Sistema Nervioso/enzimología , Oryza/parasitología , Estrés Oxidativo/efectos de los fármacos , Periplaneta/enzimología , Periplaneta/genética , Periplaneta/crecimiento & desarrollo , Unión Proteica , Conformación Proteica en Hélice alfa , Conformación Proteica en Lámina beta , Dominios y Motivos de Interacción de Proteínas , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Superóxido Dismutasa/antagonistas & inhibidores , Superóxido Dismutasa/metabolismo , Termodinámica , Gorgojos/enzimología , Gorgojos/genética , Gorgojos/crecimiento & desarrollo
17.
Int J Mol Sci ; 18(11)2017 Oct 30.
Artículo en Inglés | MEDLINE | ID: mdl-29084141

RESUMEN

The catecholamines norepinephrine and epinephrine regulate important physiological functions in vertebrates. In insects; these neuroactive substances are functionally replaced by the phenolamines octopamine and tyramine. Phenolamines activate specific guanine nucleotide-binding (G) protein-coupled receptors (GPCRs). Type 1 tyramine receptors are better activated by tyramine than by octopamine. In contrast; type 2 tyramine receptors are almost exclusively activated by tyramine. Functionally; activation of type 1 tyramine receptors leads to a decrease in the intracellular concentration of cAMP ([cAMP]i) whereas type 2 tyramine receptors can mediate Ca2+ signals or both Ca2+ signals and effects on [cAMP]i. Here; we report that the American cockroach (Periplaneta americana) expresses a second type 1 tyramine receptor (PeaTAR1B) in addition to PeaTAR1A (previously called PeaTYR1). When heterologously expressed in flpTM cells; activation of PeaTAR1B by tyramine leads to a concentration-dependent decrease in [cAMP]i. Its activity can be blocked by a series of established antagonists. The functional characterization of two type 1 tyramine receptors from P. americana; PeaTAR1A and PeaTAR1B; which respond to tyramine by changing cAMP levels; is a major step towards understanding the actions of tyramine in cockroach physiology and behavior; particularly in comparison to the effects of octopamine.


Asunto(s)
Periplaneta/genética , Periplaneta/metabolismo , Receptores de Amina Biogénica/genética , Receptores de Amina Biogénica/metabolismo , Secuencia de Aminoácidos , Animales , Clonación Molecular , ADN Complementario/química , ADN Complementario/genética , Relación Dosis-Respuesta a Droga , Expresión Génica , Periplaneta/clasificación , Filogenia , Receptores de Amina Biogénica/agonistas , Receptores de Amina Biogénica/química , Tiramina/farmacología
18.
J Agric Food Chem ; 65(36): 7865-7873, 2017 Sep 13.
Artículo en Inglés | MEDLINE | ID: mdl-28820587

RESUMEN

A novel chemotype insecticide flupyrimin (FLP) [N-[(E)-1-(6-chloro-3-pyridinylmethyl)pyridin-2(1H)-ylidene]-2,2,2-trifluoroacetamide], discovered by Meiji Seika Pharma, has unique biological properties, including outstanding potency to imidacloprid (IMI)-resistant rice pests together with superior safety toward pollinators. Intriguingly, FLP acts as a nicotinic antagonist in American cockroach neurons, and [3H]FLP binds to the multiple high-affinity binding components in house fly nicotinic acetylcholine (ACh) receptor (nAChR) preparation. One of the [3H]FLP receptors is identical to the IMI receptor, and the alternative is IMI-insensitive subtype. Furthermore, FLP is favorably safe to rats as predicted by the very low affinity to the rat α4ß2 nAChR. Structure-activity relationships of FLP analogues in terms of receptor potency, featuring the pyridinylidene and trifluoroacetyl pharmacophores, were examined, thereby establishing the FLP molecular recognition at the Aplysia californica ACh-binding protein, a suitable structural surrogate of the insect nAChR. These FLP pharmacophores account for the excellent receptor affinity, accordingly revealing differences in its binding mechanism from IMI.


Asunto(s)
Insecticidas/química , Insecticidas/farmacología , Antagonistas Nicotínicos/química , Antagonistas Nicotínicos/farmacología , Receptores Nicotínicos/química , Animales , Aplysia/efectos de los fármacos , Aplysia/metabolismo , Sitios de Unión , Proteínas de Insectos/química , Proteínas de Insectos/metabolismo , Cinética , Periplaneta/efectos de los fármacos , Periplaneta/genética , Periplaneta/metabolismo , Ratas , Receptores Nicotínicos/metabolismo , Relación Estructura-Actividad
19.
Insect Mol Biol ; 26(6): 687-701, 2017 12.
Artículo en Inglés | MEDLINE | ID: mdl-28719023

RESUMEN

The American cockroach (Periplaneta americana) is an urban pest with a precise chemosensory system that helps it achieve complex physiological behaviours, including locating food and mating. However, its chemosensory mechanisms have not been well studied. Here, we identified 71 putative odorant carrier protein genes in P. americana, including 57 new odorant-binding proteins (OBPs) and 11 chemosensory proteins (CSPs). To identify their physiological functions, we investigated their tissue expression patterns in antennae, mouthparts, legs, and the remainder of the body of both sexes, and determined that most of these genes were expressed in chemosensory organs. A phylogenetic tree showed that the putative pheromone-binding proteins of P. americana were in different clades from those of moths. Two genes, PameOBP24 and PameCSP7, were expressed equally in antennae of both sexes and highly expressed amongst the OBPs and CSPs. These genes were expressed in Escherichia coli and the resultant proteins were purified. The binding affinities of 74 common odorant compounds were tested with recombinant PameOBP24 and PameCSP7. Both proteins bound a variety of ligands. Our findings provide a foundation for future research into the chemosensory mechanisms of P. americana and help in identifying potential target genes for managing this pest.


Asunto(s)
Proteínas de Insectos/genética , Periplaneta/genética , Receptores Odorantes/genética , Olfato/fisiología , Animales , Antenas de Artrópodos/metabolismo , Femenino , Expresión Génica , Proteínas de Insectos/aislamiento & purificación , Proteínas de Insectos/metabolismo , Masculino , Percepción Olfatoria , Periplaneta/metabolismo , Filogenia , Receptores Odorantes/aislamiento & purificación , Receptores Odorantes/metabolismo
20.
Int J Mol Med ; 38(6): 1795-1805, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-27840974

RESUMEN

Per a 9 is a major allergen of the American cockroach (CR), which has been recognized as an important cause of imunoglobulin E-mediated type I hypersensitivity worldwide. However, it is not neasy to obtain a substantial quantity of this allergen for use in functional studies. In the present study, the Per a 9 gene was cloned and expressed in Escherichia coli (E. coli) systems. It was found that 13/16 (81.3%) of the sera from patients with allergies caused by the American CR reacted to Per a 9, as assessed by enzyme-linked immunosorbent assay, confirming that Per a 9 is a major allergen of CR. The induction of the expression of CD63 and CCR3 in passively sensitized basophils (from sera of patients with allergies caused by the American CR) by approximately 4.2-fold indicated that recombinant Per a 9 was functionally active. Three immunoinformatics tools, including the DNAStar Protean system, Bioinformatics Predicted Antigenic Peptides (BPAP) system and the BepiPred 1.0 server were used to predict the potential B cell epitopes, while Net-MHCIIpan-2.0 and NetMHCII-2.2 were used to predict the T cell epitopes of Per a 9. As a result, we predicted 11 peptides (23-28, 39-46, 58-64, 91-118, 131-136, 145-154, 159-165, 176-183, 290-299, 309-320 and 338-344) as potential B cell linear epitopes. In T cell prediction, the Per a 9 allergen was predicted to have 5 potential T cell epitope sequences, 119-127, 194-202, 210-218, 239-250 and 279-290. The findings of our study may prove to be useful in the development of peptide-based vaccines to combat CR-induced allergies.


Asunto(s)
Alérgenos/genética , Alérgenos/inmunología , Arginina Quinasa/genética , Arginina Quinasa/inmunología , Clonación Molecular , Epítopos de Linfocito B/inmunología , Expresión Génica , Inmunoglobulina E/inmunología , Periplaneta/genética , Periplaneta/inmunología , Alérgenos/química , Secuencia de Aminoácidos , Animales , Arginina Quinasa/química , Secuencia de Bases , Basófilos/inmunología , Sitios de Unión , Epítopos de Linfocito B/química , Humanos , Inmunoglobulina E/sangre , Modelos Moleculares , Filogenia , Conformación Proteica , Proteínas Recombinantes , Rinitis Alérgica/sangre , Rinitis Alérgica/inmunología
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