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Temporal Gene Expression Profiles of Pre Blood-Fed Adult Females Immediately Following Eclosion in the Southern House Mosquito Culex Quinquefasciatus.

Reid WR, Zhang L, Liu N - Int. J. Biol. Sci. (2015)

Bottom Line: We found that adult females required a minimum of 48 h post-eclosion before they freely took their first blood meal.In addition, Trypsin was found to be up-regulated immediately following blood feeding, while trypsin and chymotrypsin were up-regulated at 48 h and 60 h post blood-feeding, respectively, suggesting that these proteases are likely involved in the digestion of the blood meal.Overall, this study reviewed multiple genes that might be involved in the adult female competency for blood meal acquisition in mosquitoes.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Entomology and Plant Pathology, Auburn University, Auburn, AL 36849, USA ; 2. Current address: UDSA-ARS Center for Medical Veterinary and Agricultural Entomology, Mosquito and Fly Research Unit, Gainesville, FL 32608, USA.

ABSTRACT
Prior to acquisition of the first host blood meal, the anautogenous mosquito Culex quinquefasciatus requires a period of time in order to prepare for the blood feeding and, later, vitellogenesis. In the current study, we conducted whole transcriptome analyses of adult female Culex mosquitoes to identify genes that may be necessary for both taking of the blood meal, and processing of the blood meal in adult female mosquitoes Cx. quinquefasciatus. We examined temporal expression of genes for the periods of post eclosion and prior to the female freely taking a blood meal. We further evaluated the temporal expression of certain genes for the periods after the taking of a blood meal to identify genes that may be necessary for both the taking of the blood meal, and the processing of the blood meal. We found that adult females required a minimum of 48 h post-eclosion before they freely took their first blood meal. We hypothesized that gene expression signatures were altered in the mosquitoes before blood feeding in preparation for the acquisition of the blood meal through changes in multiple gene expression. To identify the genes involved in the acquisition of blood feeding, we quantified the gene expression levels of adult female Cx. quinquefasciatus using RNA Seq throughout a pre-blooding period from 2 to 72 h post eclosion at 12 h intervals. A total of 325 genes were determined to be differentially-expressed throughout the pre-blooding period, with the majority of differentially-expressed genes occurring between the 2 h and 12 h post-eclosion time points. Among the up-regulated genes were salivary proteins, cytochrome P450s, odorant-binding proteins, and proteases, while the majority of the down-regulated genes were hypothetical or cuticular genes. In addition, Trypsin was found to be up-regulated immediately following blood feeding, while trypsin and chymotrypsin were up-regulated at 48 h and 60 h post blood-feeding, respectively, suggesting that these proteases are likely involved in the digestion of the blood meal. Overall, this study reviewed multiple genes that might be involved in the adult female competency for blood meal acquisition in mosquitoes.

No MeSH data available.


Related in: MedlinePlus

Total proportions of cumulative gene expression levels within the Structural Classification of Proteins (SCOP) general function categories for adult sugar-fed female Culex quinquefasciatus, strain HAmCqG8, for the initial 72 h post-eclosion. Gene expression values expressed are summed within each SCOP category to provide an overall profile of the complete distribution of all gene expression within the mosquitoes.
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Figure 2: Total proportions of cumulative gene expression levels within the Structural Classification of Proteins (SCOP) general function categories for adult sugar-fed female Culex quinquefasciatus, strain HAmCqG8, for the initial 72 h post-eclosion. Gene expression values expressed are summed within each SCOP category to provide an overall profile of the complete distribution of all gene expression within the mosquitoes.

Mentions: According to the pre-determination of the first blood meal time period, we conducted RNA Seq to characterize the genes that were involved in the taken of blood meal from mosquitoes. Seven time points of mosquitoes post eclosion, i.e., 2, 12, 24, 36 48, 60, and 72 h, were selected for the RNA Seq analysis, covering the time period from eclosion to the first sign of the blood feeding (i.e., by 48 h post-eclosion) and to reaching to the maximum mating period of Culex mosquitoes, i.e., Cx. quinquefasciatus has been shown to begin mating in 24 h after eclosion and reach a maximum by 72 h 2. A total of 200 females collected from each of the time points were pooled for the RNA extraction. Except for the 2 h time point, in which mosquito pupae were allowed to eclose over the 2 h period only, mosquito pupae at all-time points were allowed to eclose over the 12 h period and the females were collected at 12, 24, 36, 48, 60, and 72 h time points after eclosion. Overall, the depths of sequencing for the sample time points ranged from 26 to 51 million paired-end reads (Table 1) and after mapping the reads to the Cx. quinquefasciatus genome, the genes that were identified as expressed (i.e., those genes had an FPKM >1 14), were divided among the Structural Classification Of Proteins (SCOP) general function categories of metabolism, regulation, extra-cellular processes, intra-cellular processes, information, general, other, and no annotation 16-19. When the genes were sorted into their respective SCOP general function categories and their FPKM gene expression values were summed to estimate the total proportion of gene expression within each of the SCOP general function categories for each of the time points, the pattern revealed that there was an overall decrease in total gene expression among the “No annotation” and the “General” SCOP general function categories occurring from 2 to 12 h post-eclosion time points, with a respective increase in total gene expression for the categories of “regulation” and “other” for the 12 h time point (Fig. 2). Beyond the 12 h post-eclosion time point, the total cumulative gene expression profiles within each of the SCOP general function categories were similar up to the 72 h post-eclosion time point that had been tested (Fig. 2). These results suggested that the major global changes in gene expression of adult female Cx. quinquefasciatus during the initial 12 h post-eclosion may be involved in the post eclosion process as well as possibly in the preparation of the female for the taking of a blood meal.


Temporal Gene Expression Profiles of Pre Blood-Fed Adult Females Immediately Following Eclosion in the Southern House Mosquito Culex Quinquefasciatus.

Reid WR, Zhang L, Liu N - Int. J. Biol. Sci. (2015)

Total proportions of cumulative gene expression levels within the Structural Classification of Proteins (SCOP) general function categories for adult sugar-fed female Culex quinquefasciatus, strain HAmCqG8, for the initial 72 h post-eclosion. Gene expression values expressed are summed within each SCOP category to provide an overall profile of the complete distribution of all gene expression within the mosquitoes.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4582154&req=5

Figure 2: Total proportions of cumulative gene expression levels within the Structural Classification of Proteins (SCOP) general function categories for adult sugar-fed female Culex quinquefasciatus, strain HAmCqG8, for the initial 72 h post-eclosion. Gene expression values expressed are summed within each SCOP category to provide an overall profile of the complete distribution of all gene expression within the mosquitoes.
Mentions: According to the pre-determination of the first blood meal time period, we conducted RNA Seq to characterize the genes that were involved in the taken of blood meal from mosquitoes. Seven time points of mosquitoes post eclosion, i.e., 2, 12, 24, 36 48, 60, and 72 h, were selected for the RNA Seq analysis, covering the time period from eclosion to the first sign of the blood feeding (i.e., by 48 h post-eclosion) and to reaching to the maximum mating period of Culex mosquitoes, i.e., Cx. quinquefasciatus has been shown to begin mating in 24 h after eclosion and reach a maximum by 72 h 2. A total of 200 females collected from each of the time points were pooled for the RNA extraction. Except for the 2 h time point, in which mosquito pupae were allowed to eclose over the 2 h period only, mosquito pupae at all-time points were allowed to eclose over the 12 h period and the females were collected at 12, 24, 36, 48, 60, and 72 h time points after eclosion. Overall, the depths of sequencing for the sample time points ranged from 26 to 51 million paired-end reads (Table 1) and after mapping the reads to the Cx. quinquefasciatus genome, the genes that were identified as expressed (i.e., those genes had an FPKM >1 14), were divided among the Structural Classification Of Proteins (SCOP) general function categories of metabolism, regulation, extra-cellular processes, intra-cellular processes, information, general, other, and no annotation 16-19. When the genes were sorted into their respective SCOP general function categories and their FPKM gene expression values were summed to estimate the total proportion of gene expression within each of the SCOP general function categories for each of the time points, the pattern revealed that there was an overall decrease in total gene expression among the “No annotation” and the “General” SCOP general function categories occurring from 2 to 12 h post-eclosion time points, with a respective increase in total gene expression for the categories of “regulation” and “other” for the 12 h time point (Fig. 2). Beyond the 12 h post-eclosion time point, the total cumulative gene expression profiles within each of the SCOP general function categories were similar up to the 72 h post-eclosion time point that had been tested (Fig. 2). These results suggested that the major global changes in gene expression of adult female Cx. quinquefasciatus during the initial 12 h post-eclosion may be involved in the post eclosion process as well as possibly in the preparation of the female for the taking of a blood meal.

Bottom Line: We found that adult females required a minimum of 48 h post-eclosion before they freely took their first blood meal.In addition, Trypsin was found to be up-regulated immediately following blood feeding, while trypsin and chymotrypsin were up-regulated at 48 h and 60 h post blood-feeding, respectively, suggesting that these proteases are likely involved in the digestion of the blood meal.Overall, this study reviewed multiple genes that might be involved in the adult female competency for blood meal acquisition in mosquitoes.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Entomology and Plant Pathology, Auburn University, Auburn, AL 36849, USA ; 2. Current address: UDSA-ARS Center for Medical Veterinary and Agricultural Entomology, Mosquito and Fly Research Unit, Gainesville, FL 32608, USA.

ABSTRACT
Prior to acquisition of the first host blood meal, the anautogenous mosquito Culex quinquefasciatus requires a period of time in order to prepare for the blood feeding and, later, vitellogenesis. In the current study, we conducted whole transcriptome analyses of adult female Culex mosquitoes to identify genes that may be necessary for both taking of the blood meal, and processing of the blood meal in adult female mosquitoes Cx. quinquefasciatus. We examined temporal expression of genes for the periods of post eclosion and prior to the female freely taking a blood meal. We further evaluated the temporal expression of certain genes for the periods after the taking of a blood meal to identify genes that may be necessary for both the taking of the blood meal, and the processing of the blood meal. We found that adult females required a minimum of 48 h post-eclosion before they freely took their first blood meal. We hypothesized that gene expression signatures were altered in the mosquitoes before blood feeding in preparation for the acquisition of the blood meal through changes in multiple gene expression. To identify the genes involved in the acquisition of blood feeding, we quantified the gene expression levels of adult female Cx. quinquefasciatus using RNA Seq throughout a pre-blooding period from 2 to 72 h post eclosion at 12 h intervals. A total of 325 genes were determined to be differentially-expressed throughout the pre-blooding period, with the majority of differentially-expressed genes occurring between the 2 h and 12 h post-eclosion time points. Among the up-regulated genes were salivary proteins, cytochrome P450s, odorant-binding proteins, and proteases, while the majority of the down-regulated genes were hypothetical or cuticular genes. In addition, Trypsin was found to be up-regulated immediately following blood feeding, while trypsin and chymotrypsin were up-regulated at 48 h and 60 h post blood-feeding, respectively, suggesting that these proteases are likely involved in the digestion of the blood meal. Overall, this study reviewed multiple genes that might be involved in the adult female competency for blood meal acquisition in mosquitoes.

No MeSH data available.


Related in: MedlinePlus