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Potential mosquito repellent compounds of Ocimum species against 3N7H and 3Q8I of Anopheles gambiae.

ABSTRACT: Mosquitoes are exceptionally efficient in detecting their hosts for blood meal using odorant binding proteins, viz. 3N7H and 3Q8I and spread several dreadful diseases. DEET is a synthetic mosquito repellent widely used all over world for protection against mosquito bite. Reports reveal that, synthetic mosquito repellents may pose health problems in considerably large population. In view of the above fact, we made an attempt to discover efficient and novel natural mosquito repellent compounds with least impact on human health. Methanolic leaf extracts of Ocimum basilicum Linn. var. pilosum (willd.)-Benth and Ocimum tenuiflorum var. CIM-AYU were subjected to GC-MS analysis and obtained 35 phytochemical constituents. Repellent potentiality of the Ocimum compounds was assessed against 3Q8I and 3N7H of Anopheles gambiae. PDB structures of mosquito odorant binding proteins were downloaded, processed and docking studies were performed along with reference ligand DEET using Schrodinger MAESTRO 9.2 software. Molecular docking results reveal that phenol, 2-methoxy-3-(2-propenyl)-, licopersin, gamma sitosterol and benzene, 1,2-dimethoxy-4-(2-propenyl)- from O. tenuiflorum var. CIM-AYU are strongly bound with 3N7H. Whereas, 4h-1-benzopyran-4-one, 5-hydroxy-6,7-dimethoxy-2-(4-methoxyphenyl)-, catechol and monoacetin from O. basilicum Linn. var. pilosum (willd.)-Benth. show high binding affinity with odorant binding protein 3Q8I. All natural compounds tested in the present study display better docking scores than DEET. The results further substantiate that the 12 out of 35 compounds of the two Ocimum species found to be ideal candidates for design and development of potential mosquito repellents. ADME properties of the tested compounds further confirm that bioactive compounds of Ocimum species were found to be in acceptable range. Synchronized application of at least two different natural compounds (with best docking scores) which target 3N7H and 3Q8I (Odorant Binding Proteins of mosquito) proteins may provide enhanced protection against mosquitoes bite. Based on the ADME properties, natural compounds of Ocimum species can be considered for design and development of safe mosquito repellents.

SUBMITTER: Gaddaguti V

PROVIDER: S-EPMC4711284 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature

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Potential mosquito repellent compounds of Ocimum species against 3N7H and 3Q8I of Anopheles gambiae.

Gaddaguti Venugopal V Venkateswara Rao Talluri T Prasada Rao Allu A

3 Biotech 20160111 1

Mosquitoes are exceptionally efficient in detecting their hosts for blood meal using odorant binding proteins, viz. 3N7H and 3Q8I and spread several dreadful diseases. DEET is a synthetic mosquito repellent widely used all over world for protection against mosquito bite. Reports reveal that, synthetic mosquito repellents may pose health problems in considerably large population. In view of the above fact, we made an attempt to discover efficient and novel natural mosquito repellent compounds wit ...[more]

PMID: 28330095

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Project description:The goal of this study is to analyze the gene expression profile of Anopheles gambiae Ag55 cells and compare it with gene expression profile of blood fed female Anopheles gambiae adult to establish Ag55 cells as a potential model to study mosquito-pathogen interactions Methods: The transcript profiles of Ag55 cells were generated by RNA-sequencing, in triplicate, using Illumina HiSeq 2000 v3 platform. The 7 samples (paired-end reads, including replicates ( 3 samples from Ag55 cells and 4 samples from blood-fed female adults downloaded from NCBI GEO (Series GSE55453, SRA id : SRP039058))) were independently mapped on to A. gambiae genome (Anopheles-gambiae-PEST_CHROMOSOMES_AgamP4.fa.gz) downloaded from vectorbase by using TopHat followed by Cufflinks to estimate the expression values of the transcripts in FPKM (Fragments Per Kilobase per Million mapped reads) with the cuffdiff2 default geometric normalization. Differentially expressed genes (FDR<0.05 after Benjamini-Hochberg correction for multiple-testing) were identified for Ag55 cells relative to Blood fed female adults Results: We provide detailed gene expression and proteome profiles of Ag55 cells. We further compare the gene expression profiles of Ag55 cells and blood fed female adults. Gene Ontology enrichment analysis of our transcriptomic data suggested that Ag55 cells have phagocytic properties, a hypothesis which we confirmed using confocal imaging. Transcriptomic data further backed by proteomic data suggest that Ag55 cells express hemocyte like properties, and are immune competent. Conclusion: As Ag55 cells are immune competent and express hemocyte like properties they can be used as a model to study vector-pathogen immune response. Furthermore, the availability of transcriptomic and proteomic data of Ag55 cells will help researchers use and engineer the Ag55 cell line in an efficient way, for example by developing strategies to make it more suitable for studies of interactions with Plasmodium and other microbes.

Project description:BackgroundGenome sequencing of Anopheles gambiae was completed more than ten years ago and has accelerated research on malaria transmission. However, annotation needs to be refined and verified experimentally, as most predicted transcripts have been identified by comparative analysis with genomes from other species. The mosquito midgut-the first organ to interact with Plasmodium parasites-mounts effective antiplasmodial responses that limit parasite survival and disease transmission. High-throughput Illumina sequencing of the midgut transcriptome was used to identify new genes and transcripts, contributing to the refinement of An. gambiae genome annotation.ResultsWe sequenced ~223 million reads from An. gambiae midgut cDNA libraries generated from susceptible (G3) and refractory (L35) mosquito strains. Mosquitoes were infected with either Plasmodium berghei or Plasmodium falciparum, and midguts were collected after the first or second Plasmodium infection. In total, 22,889 unique midgut transcript models were generated from both An. gambiae strain sequences combined, and 76% are potentially novel. Of these novel transcripts, 49.5% aligned with annotated genes and appear to be isoforms or pre-mRNAs of reference transcripts, while 50.5% mapped to regions between annotated genes and represent novel intergenic transcripts (NITs). Predicted models were validated for midgut expression using qRT-PCR and microarray analysis, and novel isoforms were confirmed by sequencing predicted intron-exon boundaries. Coding potential analysis revealed that 43% of total midgut transcripts appear to be long non-coding RNA (lncRNA), and functional annotation of NITs showed that 68% had no homology to current databases from other species. Reads were also analyzed using de novo assembly and predicted transcripts compared with genome mapping-based models. Finally, variant analysis of G3 and L35 midgut transcripts detected 160,742 variants with respect to the An. gambiae PEST genome, and 74% were new variants. Intergenic transcripts had a higher frequency of variation compared with non-intergenic transcripts.ConclusionThis in-depth Illumina sequencing and assembly of the An. gambiae midgut transcriptome doubled the number of known transcripts and tripled the number of variants known in this mosquito species. It also revealed existence of a large number of lncRNA and opens new possibilities for investigating the biological function of many newly discovered transcripts.

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Potential mosquito repellent compounds of Ocimum species against 3N7H and 3Q8I of Anopheles gambiae.

Publications

Potential mosquito repellent compounds of Ocimum species against 3N7H and 3Q8I of Anopheles gambiae.

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