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Daily rhythms in antennal protein and olfactory sensitivity in the malaria mosquito Anopheles gambiae.

Rund SS, Bonar NA, Champion MM, Ghazi JP, Houk CM, Leming MT, Syed Z, Duffield GE - Sci Rep (2013)

Bottom Line: Further, electrophysiological investigations demonstrate time-of-day specific differences in olfactory sensitivity of antennae to major host-derived odorants.The pre-dusk/dusk peaks in OBPs and takeout gene expression correspond with peak protein abundance at night, and in turn coincide with the time of increased olfactory sensitivity to odorants requiring OBPs and times of increased blood-feeding behavior.This suggests an important role for OBPs in modulating temporal changes in odorant sensitivity, enabling the olfactory system to coordinate with the circadian niche of An. gambiae.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biological Sciences and Eck Institute for Global Health, Galvin Life Science Center, University of Notre Dame, Notre Dame, IN 46556 [2].

ABSTRACT
We recently characterized 24-hr daily rhythmic patterns of gene expression in Anopheles gambiae mosquitoes. These include numerous odorant binding proteins (OBPs), soluble odorant carrying proteins enriched in olfactory organs. Here we demonstrate that multiple rhythmically expressed genes including OBPs and takeout proteins, involved in regulating blood feeding behavior, have corresponding rhythmic protein levels as measured by quantitative proteomics. This includes AgamOBP1, previously shown as important to An. gambiae odorant sensing. Further, electrophysiological investigations demonstrate time-of-day specific differences in olfactory sensitivity of antennae to major host-derived odorants. The pre-dusk/dusk peaks in OBPs and takeout gene expression correspond with peak protein abundance at night, and in turn coincide with the time of increased olfactory sensitivity to odorants requiring OBPs and times of increased blood-feeding behavior. This suggests an important role for OBPs in modulating temporal changes in odorant sensitivity, enabling the olfactory system to coordinate with the circadian niche of An. gambiae.

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Antennal protein rhythms correspond with RNA rhythms.(a) Quantitative proteomics reveals rhythms in antennal protein abundance that correspond highly with RNA expression profiles from whole-heads. Proteins are grouped into OBPs, non-OBP chemosensory proteins (AGAP007286, SAPP1, A10, TO1, and TO2/3) and non-olfactory proteins (CYP6P3, RFeSP, PPO6, and VATI). All gene symbols as listed in VectorBase except for the following genes: RFeSP (homologue to Drosophila Rieske iron-sulfur protein, AGAP008955), A10 (homologue to Drosophilla antennal protein 10, AGAP008055), VATI (predicted V-type proton ATPase catalytic subunit I, AGAP001587) and AGAP007286 (Ae. aegypti OBP43 homologue). Protein abundance normalized with tubulin. See Fig. S3 for protein rhythms in the second biological replicate antennae run and THAs. (b) Histogram of peak RNA expression phases in total heads and the corresponding peak antennal protein levels as determined by cosinor analysis from genes/proteins in panel A. (c) Cosinor analysis of antennae OBP protein levels from panel A (p < 0.001; acrophase ZT17.7); note OBP10 excluded as it has antiphasic expression compared to the other OBPs.
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f2: Antennal protein rhythms correspond with RNA rhythms.(a) Quantitative proteomics reveals rhythms in antennal protein abundance that correspond highly with RNA expression profiles from whole-heads. Proteins are grouped into OBPs, non-OBP chemosensory proteins (AGAP007286, SAPP1, A10, TO1, and TO2/3) and non-olfactory proteins (CYP6P3, RFeSP, PPO6, and VATI). All gene symbols as listed in VectorBase except for the following genes: RFeSP (homologue to Drosophila Rieske iron-sulfur protein, AGAP008955), A10 (homologue to Drosophilla antennal protein 10, AGAP008055), VATI (predicted V-type proton ATPase catalytic subunit I, AGAP001587) and AGAP007286 (Ae. aegypti OBP43 homologue). Protein abundance normalized with tubulin. See Fig. S3 for protein rhythms in the second biological replicate antennae run and THAs. (b) Histogram of peak RNA expression phases in total heads and the corresponding peak antennal protein levels as determined by cosinor analysis from genes/proteins in panel A. (c) Cosinor analysis of antennae OBP protein levels from panel A (p < 0.001; acrophase ZT17.7); note OBP10 excluded as it has antiphasic expression compared to the other OBPs.

Mentions: In order to determine if proteins identified by qualitative proteomics had rhythms specific to sensory tissues, we utilized targeted quantitative MRM proteomics to both identify and determine concentration in antennae and THAs252627282930. Biological material was collected every 4 hr for 24 hr from adult females maintained under strict LD cycle conditions (12 hr light: 12 hr dark including 1 hr dawn/dusk transitions). In antennae and THAs, we were able to defensibly quantify 21 of 25 targeted proteins, respectively, including 11 OBPs and 5 other putative chemosensory proteins (Fig. 2, Fig. S3). The profiles of protein abundance for these proteins were found to be rhythmic. Detected OBPs include OBP1 (AGAP003309), known to bind indole and DEET1115, and OBP20 (AGAP005208), known to bind DEET, 6-methyl-5-heptone-2-one and indole14. Detected and rhythmic in our analysis, OBP1, 2 (AGAP003306), 3 (AGAP001409), 20, 22 (AGAP010409), 25 (AGAP012320), 26 (AGAP012321) and 47 (AGAP007287) are reported to have gene expression enriched in female heads/olfactory organs relative to male tissues and/or female bodies101213. A recent RNA-seq experiment also revealed that many of these OBP genes are down-regulated in antennae following blood-feeding and as the mosquito transitions to oviposition behavior16. These studies provide further evidence that these OBPs in particular are potentially involved in olfactory host-seeking behavior in females. Two other chemosensory proteins, SAPP1 and A10, were also found to be rhythmic. Additionally, the takeout proteins TO1 (AGAP004263) and TO2/3 (AGAP012703/AGAP004262), which have been implicated in chemoreception and in blood-feeding behavior119, were detected and rhythmic in antennae and THAs. Many protein rhythms were robust, with 3.2, 8.4, 4.5 and 5.3 peak-to-trough fold changes observed for OBP1, OBP26, TO1 and SAPP1 antennae protein abundance, respectively.


Daily rhythms in antennal protein and olfactory sensitivity in the malaria mosquito Anopheles gambiae.

Rund SS, Bonar NA, Champion MM, Ghazi JP, Houk CM, Leming MT, Syed Z, Duffield GE - Sci Rep (2013)

Antennal protein rhythms correspond with RNA rhythms.(a) Quantitative proteomics reveals rhythms in antennal protein abundance that correspond highly with RNA expression profiles from whole-heads. Proteins are grouped into OBPs, non-OBP chemosensory proteins (AGAP007286, SAPP1, A10, TO1, and TO2/3) and non-olfactory proteins (CYP6P3, RFeSP, PPO6, and VATI). All gene symbols as listed in VectorBase except for the following genes: RFeSP (homologue to Drosophila Rieske iron-sulfur protein, AGAP008955), A10 (homologue to Drosophilla antennal protein 10, AGAP008055), VATI (predicted V-type proton ATPase catalytic subunit I, AGAP001587) and AGAP007286 (Ae. aegypti OBP43 homologue). Protein abundance normalized with tubulin. See Fig. S3 for protein rhythms in the second biological replicate antennae run and THAs. (b) Histogram of peak RNA expression phases in total heads and the corresponding peak antennal protein levels as determined by cosinor analysis from genes/proteins in panel A. (c) Cosinor analysis of antennae OBP protein levels from panel A (p < 0.001; acrophase ZT17.7); note OBP10 excluded as it has antiphasic expression compared to the other OBPs.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3756343&req=5

f2: Antennal protein rhythms correspond with RNA rhythms.(a) Quantitative proteomics reveals rhythms in antennal protein abundance that correspond highly with RNA expression profiles from whole-heads. Proteins are grouped into OBPs, non-OBP chemosensory proteins (AGAP007286, SAPP1, A10, TO1, and TO2/3) and non-olfactory proteins (CYP6P3, RFeSP, PPO6, and VATI). All gene symbols as listed in VectorBase except for the following genes: RFeSP (homologue to Drosophila Rieske iron-sulfur protein, AGAP008955), A10 (homologue to Drosophilla antennal protein 10, AGAP008055), VATI (predicted V-type proton ATPase catalytic subunit I, AGAP001587) and AGAP007286 (Ae. aegypti OBP43 homologue). Protein abundance normalized with tubulin. See Fig. S3 for protein rhythms in the second biological replicate antennae run and THAs. (b) Histogram of peak RNA expression phases in total heads and the corresponding peak antennal protein levels as determined by cosinor analysis from genes/proteins in panel A. (c) Cosinor analysis of antennae OBP protein levels from panel A (p < 0.001; acrophase ZT17.7); note OBP10 excluded as it has antiphasic expression compared to the other OBPs.
Mentions: In order to determine if proteins identified by qualitative proteomics had rhythms specific to sensory tissues, we utilized targeted quantitative MRM proteomics to both identify and determine concentration in antennae and THAs252627282930. Biological material was collected every 4 hr for 24 hr from adult females maintained under strict LD cycle conditions (12 hr light: 12 hr dark including 1 hr dawn/dusk transitions). In antennae and THAs, we were able to defensibly quantify 21 of 25 targeted proteins, respectively, including 11 OBPs and 5 other putative chemosensory proteins (Fig. 2, Fig. S3). The profiles of protein abundance for these proteins were found to be rhythmic. Detected OBPs include OBP1 (AGAP003309), known to bind indole and DEET1115, and OBP20 (AGAP005208), known to bind DEET, 6-methyl-5-heptone-2-one and indole14. Detected and rhythmic in our analysis, OBP1, 2 (AGAP003306), 3 (AGAP001409), 20, 22 (AGAP010409), 25 (AGAP012320), 26 (AGAP012321) and 47 (AGAP007287) are reported to have gene expression enriched in female heads/olfactory organs relative to male tissues and/or female bodies101213. A recent RNA-seq experiment also revealed that many of these OBP genes are down-regulated in antennae following blood-feeding and as the mosquito transitions to oviposition behavior16. These studies provide further evidence that these OBPs in particular are potentially involved in olfactory host-seeking behavior in females. Two other chemosensory proteins, SAPP1 and A10, were also found to be rhythmic. Additionally, the takeout proteins TO1 (AGAP004263) and TO2/3 (AGAP012703/AGAP004262), which have been implicated in chemoreception and in blood-feeding behavior119, were detected and rhythmic in antennae and THAs. Many protein rhythms were robust, with 3.2, 8.4, 4.5 and 5.3 peak-to-trough fold changes observed for OBP1, OBP26, TO1 and SAPP1 antennae protein abundance, respectively.

Bottom Line: Further, electrophysiological investigations demonstrate time-of-day specific differences in olfactory sensitivity of antennae to major host-derived odorants.The pre-dusk/dusk peaks in OBPs and takeout gene expression correspond with peak protein abundance at night, and in turn coincide with the time of increased olfactory sensitivity to odorants requiring OBPs and times of increased blood-feeding behavior.This suggests an important role for OBPs in modulating temporal changes in odorant sensitivity, enabling the olfactory system to coordinate with the circadian niche of An. gambiae.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biological Sciences and Eck Institute for Global Health, Galvin Life Science Center, University of Notre Dame, Notre Dame, IN 46556 [2].

ABSTRACT
We recently characterized 24-hr daily rhythmic patterns of gene expression in Anopheles gambiae mosquitoes. These include numerous odorant binding proteins (OBPs), soluble odorant carrying proteins enriched in olfactory organs. Here we demonstrate that multiple rhythmically expressed genes including OBPs and takeout proteins, involved in regulating blood feeding behavior, have corresponding rhythmic protein levels as measured by quantitative proteomics. This includes AgamOBP1, previously shown as important to An. gambiae odorant sensing. Further, electrophysiological investigations demonstrate time-of-day specific differences in olfactory sensitivity of antennae to major host-derived odorants. The pre-dusk/dusk peaks in OBPs and takeout gene expression correspond with peak protein abundance at night, and in turn coincide with the time of increased olfactory sensitivity to odorants requiring OBPs and times of increased blood-feeding behavior. This suggests an important role for OBPs in modulating temporal changes in odorant sensitivity, enabling the olfactory system to coordinate with the circadian niche of An. gambiae.

Show MeSH
Related in: MedlinePlus