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Chemosensillum immunolocalization and ligand specificity of chemosensory proteins in the alfalfa plant bug Adelphocoris lineolatus (Goeze).

Sun L, Zhou JJ, Gu SH, Xiao HJ, Guo YY, Liu ZW, Zhang YJ - Sci Rep (2015)

Bottom Line: Further cellular investigation of their immunolocalization revealed their dynamic protein expression profiles among different types of antennal sensilla.In ad`dition, a cooperative interaction was observed between two co-expressed CSPs resulting in an increase of the binding affinities by a mixture of AlinCSP5 and AlinCSP6 to terpenoids which do not bind to individual CSPs.These findings in combination with our previous data for AlinCSP1-3 indicate a possible functional differentiation of CSPs in the A. lineolatus olfactory system.

View Article: PubMed Central - PubMed

Affiliation: 1] State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China [2] Key Laboratory of Tea Plants Biology and Resources Utilization of Agriculture Ministry, Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310008, China [3] Key Laboratory of Integrated Management of Crop Diseases and Pests (Ministry of Education), College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China.

ABSTRACT
Insect chemosensory proteins (CSPs) are a family of small soluble proteins. To date, their physiological functions in insect olfaction remain largely controversial in comparison to odorant binding proteins (OBPs). In present study, we reported the antenna specific expression of three CSPs (AlinCSP4-6) from Adelphocoris lineolatus, their distinct chemosensillum distribution as well as ligand binding capability thus providing the evidence for the possible roles that they could play in semiochemical detection of the plant bug A. lineolatus. The results of qRT-PCR and western blot assay clearly showed that all of these three CSPs are highly expressed in the adult antennae, the olfactory organ of insects. Further cellular investigation of their immunolocalization revealed their dynamic protein expression profiles among different types of antennal sensilla. In a fluorescence competitive binding assay, the selective ligand binding was observed for AlinCSP4-6. In ad`dition, a cooperative interaction was observed between two co-expressed CSPs resulting in an increase of the binding affinities by a mixture of AlinCSP5 and AlinCSP6 to terpenoids which do not bind to individual CSPs. These findings in combination with our previous data for AlinCSP1-3 indicate a possible functional differentiation of CSPs in the A. lineolatus olfactory system.

No MeSH data available.


Immunocytochemistry analysis demonstrates the co-localization of AlinCSP4-6 in the different antennal sensilla of A. lineolatus.A, E, and I show the serial sections that were incubated with anti-AlinCSP4, anti-AlinCSP5, and anti-AlinCSP6 antiserum, respectively. The images shown in B, C, D; F, G, H; and J, K, L are higher-magnification images of the ssba, lssch, and mlsba shown in A, E, and I, respectively. The results clearly demonstrate that AlinCSP5 and AlinCSP6 can be co-expressed in the outer sensilla lymph of ssba. The few grains found over the cuticles and the dendrites represent the non-specific background. The dilution of each primary antibody was 1:2000, and the secondary antibody was anti-rabbit IgG conjugated with 10-nm colloidal gold granules at a dilution of 1:20.
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f7: Immunocytochemistry analysis demonstrates the co-localization of AlinCSP4-6 in the different antennal sensilla of A. lineolatus.A, E, and I show the serial sections that were incubated with anti-AlinCSP4, anti-AlinCSP5, and anti-AlinCSP6 antiserum, respectively. The images shown in B, C, D; F, G, H; and J, K, L are higher-magnification images of the ssba, lssch, and mlsba shown in A, E, and I, respectively. The results clearly demonstrate that AlinCSP5 and AlinCSP6 can be co-expressed in the outer sensilla lymph of ssba. The few grains found over the cuticles and the dendrites represent the non-specific background. The dilution of each primary antibody was 1:2000, and the secondary antibody was anti-rabbit IgG conjugated with 10-nm colloidal gold granules at a dilution of 1:20.

Mentions: Using continuous sections, we also investigated the possible co-expression of AlinCSP4-6 in antennal sensilla of A. lineolatus. The results shown in Figure 7 indicate again that AlinCSP4 protein specifically expressed in the lymph of the mlsba sensillum (Figure 7B and 7D). The co-expression of AlinCSP5 and AlinCSP6 was found in the outer lymph of the ssba sensillum (Figure 7F and 7J). The labeling of anti-AlinCSP4 in the outside ring of the ssba sensillium was most likely unspecific labelling (Figure 4G and Figure 7B) as demonstrated in a repeated experiment (data not shown).


Chemosensillum immunolocalization and ligand specificity of chemosensory proteins in the alfalfa plant bug Adelphocoris lineolatus (Goeze).

Sun L, Zhou JJ, Gu SH, Xiao HJ, Guo YY, Liu ZW, Zhang YJ - Sci Rep (2015)

Immunocytochemistry analysis demonstrates the co-localization of AlinCSP4-6 in the different antennal sensilla of A. lineolatus.A, E, and I show the serial sections that were incubated with anti-AlinCSP4, anti-AlinCSP5, and anti-AlinCSP6 antiserum, respectively. The images shown in B, C, D; F, G, H; and J, K, L are higher-magnification images of the ssba, lssch, and mlsba shown in A, E, and I, respectively. The results clearly demonstrate that AlinCSP5 and AlinCSP6 can be co-expressed in the outer sensilla lymph of ssba. The few grains found over the cuticles and the dendrites represent the non-specific background. The dilution of each primary antibody was 1:2000, and the secondary antibody was anti-rabbit IgG conjugated with 10-nm colloidal gold granules at a dilution of 1:20.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4308698&req=5

f7: Immunocytochemistry analysis demonstrates the co-localization of AlinCSP4-6 in the different antennal sensilla of A. lineolatus.A, E, and I show the serial sections that were incubated with anti-AlinCSP4, anti-AlinCSP5, and anti-AlinCSP6 antiserum, respectively. The images shown in B, C, D; F, G, H; and J, K, L are higher-magnification images of the ssba, lssch, and mlsba shown in A, E, and I, respectively. The results clearly demonstrate that AlinCSP5 and AlinCSP6 can be co-expressed in the outer sensilla lymph of ssba. The few grains found over the cuticles and the dendrites represent the non-specific background. The dilution of each primary antibody was 1:2000, and the secondary antibody was anti-rabbit IgG conjugated with 10-nm colloidal gold granules at a dilution of 1:20.
Mentions: Using continuous sections, we also investigated the possible co-expression of AlinCSP4-6 in antennal sensilla of A. lineolatus. The results shown in Figure 7 indicate again that AlinCSP4 protein specifically expressed in the lymph of the mlsba sensillum (Figure 7B and 7D). The co-expression of AlinCSP5 and AlinCSP6 was found in the outer lymph of the ssba sensillum (Figure 7F and 7J). The labeling of anti-AlinCSP4 in the outside ring of the ssba sensillium was most likely unspecific labelling (Figure 4G and Figure 7B) as demonstrated in a repeated experiment (data not shown).

Bottom Line: Further cellular investigation of their immunolocalization revealed their dynamic protein expression profiles among different types of antennal sensilla.In ad`dition, a cooperative interaction was observed between two co-expressed CSPs resulting in an increase of the binding affinities by a mixture of AlinCSP5 and AlinCSP6 to terpenoids which do not bind to individual CSPs.These findings in combination with our previous data for AlinCSP1-3 indicate a possible functional differentiation of CSPs in the A. lineolatus olfactory system.

View Article: PubMed Central - PubMed

Affiliation: 1] State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China [2] Key Laboratory of Tea Plants Biology and Resources Utilization of Agriculture Ministry, Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310008, China [3] Key Laboratory of Integrated Management of Crop Diseases and Pests (Ministry of Education), College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China.

ABSTRACT
Insect chemosensory proteins (CSPs) are a family of small soluble proteins. To date, their physiological functions in insect olfaction remain largely controversial in comparison to odorant binding proteins (OBPs). In present study, we reported the antenna specific expression of three CSPs (AlinCSP4-6) from Adelphocoris lineolatus, their distinct chemosensillum distribution as well as ligand binding capability thus providing the evidence for the possible roles that they could play in semiochemical detection of the plant bug A. lineolatus. The results of qRT-PCR and western blot assay clearly showed that all of these three CSPs are highly expressed in the adult antennae, the olfactory organ of insects. Further cellular investigation of their immunolocalization revealed their dynamic protein expression profiles among different types of antennal sensilla. In a fluorescence competitive binding assay, the selective ligand binding was observed for AlinCSP4-6. In ad`dition, a cooperative interaction was observed between two co-expressed CSPs resulting in an increase of the binding affinities by a mixture of AlinCSP5 and AlinCSP6 to terpenoids which do not bind to individual CSPs. These findings in combination with our previous data for AlinCSP1-3 indicate a possible functional differentiation of CSPs in the A. lineolatus olfactory system.

No MeSH data available.