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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

Heat map displaying the relative increases in gene expression for selected genes for the initial 72h post-eclosion for adult female Cx. quinquefasciatus, strain HAmCqG8, and for 72h post blood meal. Females offered a blood meal were 6 days old at the time of the blood feeding.
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Figure 4: Heat map displaying the relative increases in gene expression for selected genes for the initial 72h post-eclosion for adult female Cx. quinquefasciatus, strain HAmCqG8, and for 72h post blood meal. Females offered a blood meal were 6 days old at the time of the blood feeding.

Mentions: In order to estimate the accuracy of our RNA Seq results, and to determine the expression of selected genes throughout the pre blood-feeding and into the post blood-feeding time periods of Cx. quinquefasciatus, we selected a total of 36 genes from the RNA Seq data that showed the different levels of expression during 72 h post-eclosion time periods. The genes selected included genes that were involved in: host finding or feeding behavior (eg/ odorant-binding proteins) 23, 24, the maturation of the pharate female to the adult (cuticular proteins), the taking of the blood meal (salivary proteins) 28, the digestion of the blood meal (trypsins, collagenase, lipases, uricase) 29, 30, the provisioning of nutrients to the egg (vitellogenins, adipophilin/perilipin) 31, embryogenesis (wnt inhibitor, oskar) 32 as well as proteins with other functions involved in metabolism and development, including cytochrome P450s, calbindins, and oxidoreductases. The expression of these 36 genes was investigated from 2 to 72 h post-eclosion to be consistent with/validate our RNA Seq data. In addition, we further examined the expression of these 36 genes from 2 to 72 h post blood-feeding to evaluate the relationship of their expression changes following the blood feeding. Our qRT-PCR results showed that the expression of these genes was mostly consistent with the RNA Seq data - with similar traces of up- down-, or no change patterns during the different time points of 72 h after eclosion (Fig. 4). Our study showed that, according to the temporal expression, the genes can be grouped into early, middle, and late expression after eclosion and blood feeding. Our results also showed that all three general odorant-binding proteins (CPIJ012716, CPIJ012719, CPIJ012721) had their highest expression at 2 h post-eclosion except CPIJ012716 which also showed high expression at 12 h, indicating that females initially expressed the odorant-binding proteins to aid for food searching (sugar or blood). Other genes of interest were the salivary genes (CPIJ002046 and CPIJ019052) which reached maximal gene expression values at 24 and 48 h, respectively, and then remained at low expression levels afterwards (Fig. 4). Trypsin CPIJ007079 was up-regulated immediately following blood feeding, while trypsin CPIJ004660 and chymotrypsin CPIJ003915 were up-regulated at 48 h and 60 h post blood-feeding, respectively. These proteases are likely involved in the digestion of the blood meal, with the first gene representing early trypsin, and the two later two representing late trypsins. Another gene involved in the processing of the blood meal, CPIJ003456 (uricase) 30 showed the highest level of gene expression at 20 h post blood-feeding, which decreased by 60 h post blood-feeding. Among the genes that were predicted to be up-regulated throughout the post-eclosion and pre-blooding time period, the vitellogenin genes CPIJ001357/CPIJ001358 reached maximal gene expression at 60 h post blood-feeding, while the vitellogenin genes CPIJ010190/CPIJ010191 and CPIJ005473 reached maximal expression at 36 h post blood-feeding (Fig. 5). Following the blood meal, two trypsins (CPIJ007079, CPIJ004660) and one chymotrypsin (CPIJ003915) were up-regulated (Fig. 4).


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)

Heat map displaying the relative increases in gene expression for selected genes for the initial 72h post-eclosion for adult female Cx. quinquefasciatus, strain HAmCqG8, and for 72h post blood meal. Females offered a blood meal were 6 days old at the time of the blood feeding.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 4: Heat map displaying the relative increases in gene expression for selected genes for the initial 72h post-eclosion for adult female Cx. quinquefasciatus, strain HAmCqG8, and for 72h post blood meal. Females offered a blood meal were 6 days old at the time of the blood feeding.
Mentions: In order to estimate the accuracy of our RNA Seq results, and to determine the expression of selected genes throughout the pre blood-feeding and into the post blood-feeding time periods of Cx. quinquefasciatus, we selected a total of 36 genes from the RNA Seq data that showed the different levels of expression during 72 h post-eclosion time periods. The genes selected included genes that were involved in: host finding or feeding behavior (eg/ odorant-binding proteins) 23, 24, the maturation of the pharate female to the adult (cuticular proteins), the taking of the blood meal (salivary proteins) 28, the digestion of the blood meal (trypsins, collagenase, lipases, uricase) 29, 30, the provisioning of nutrients to the egg (vitellogenins, adipophilin/perilipin) 31, embryogenesis (wnt inhibitor, oskar) 32 as well as proteins with other functions involved in metabolism and development, including cytochrome P450s, calbindins, and oxidoreductases. The expression of these 36 genes was investigated from 2 to 72 h post-eclosion to be consistent with/validate our RNA Seq data. In addition, we further examined the expression of these 36 genes from 2 to 72 h post blood-feeding to evaluate the relationship of their expression changes following the blood feeding. Our qRT-PCR results showed that the expression of these genes was mostly consistent with the RNA Seq data - with similar traces of up- down-, or no change patterns during the different time points of 72 h after eclosion (Fig. 4). Our study showed that, according to the temporal expression, the genes can be grouped into early, middle, and late expression after eclosion and blood feeding. Our results also showed that all three general odorant-binding proteins (CPIJ012716, CPIJ012719, CPIJ012721) had their highest expression at 2 h post-eclosion except CPIJ012716 which also showed high expression at 12 h, indicating that females initially expressed the odorant-binding proteins to aid for food searching (sugar or blood). Other genes of interest were the salivary genes (CPIJ002046 and CPIJ019052) which reached maximal gene expression values at 24 and 48 h, respectively, and then remained at low expression levels afterwards (Fig. 4). Trypsin CPIJ007079 was up-regulated immediately following blood feeding, while trypsin CPIJ004660 and chymotrypsin CPIJ003915 were up-regulated at 48 h and 60 h post blood-feeding, respectively. These proteases are likely involved in the digestion of the blood meal, with the first gene representing early trypsin, and the two later two representing late trypsins. Another gene involved in the processing of the blood meal, CPIJ003456 (uricase) 30 showed the highest level of gene expression at 20 h post blood-feeding, which decreased by 60 h post blood-feeding. Among the genes that were predicted to be up-regulated throughout the post-eclosion and pre-blooding time period, the vitellogenin genes CPIJ001357/CPIJ001358 reached maximal gene expression at 60 h post blood-feeding, while the vitellogenin genes CPIJ010190/CPIJ010191 and CPIJ005473 reached maximal expression at 36 h post blood-feeding (Fig. 5). Following the blood meal, two trypsins (CPIJ007079, CPIJ004660) and one chymotrypsin (CPIJ003915) were up-regulated (Fig. 4).

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