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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.


Immunocytochemical localization of AlinCSP6 in different antennal sensilla of male adults.The outer sensillum lymph of ssba and lcsch and the sub-cuticular spaces (ss) were heavily labelled, whereas other sensilla, such as str, mlsba, and lssch, were not labelled (the longitudinal sections are shown in A, D, G, and J, and the cross sections are shown in B, C, E, F, H, I, K, and L). The distribution of AlinCSP6 in the different antennal sensilla of the females was similar in that found in the males. The few grains found over the cuticle and the dendrites represent the non-specific background. The dilution of the 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. The abbreviations are similar to those used in Figure 4.
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f6: Immunocytochemical localization of AlinCSP6 in different antennal sensilla of male adults.The outer sensillum lymph of ssba and lcsch and the sub-cuticular spaces (ss) were heavily labelled, whereas other sensilla, such as str, mlsba, and lssch, were not labelled (the longitudinal sections are shown in A, D, G, and J, and the cross sections are shown in B, C, E, F, H, I, K, and L). The distribution of AlinCSP6 in the different antennal sensilla of the females was similar in that found in the males. The few grains found over the cuticle and the dendrites represent the non-specific background. The dilution of the 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. The abbreviations are similar to those used in Figure 4.

Mentions: The immunolocalization of AlinCSP4-6 proteins in different antennal sensilla of both sexes was examined with ultrathin sections of the antennae subjected to colloidal gold post-embedding and immunocytochemical labelling. In general, each of AlinCSP4-6 proteins was shown to be expressed in distinct antennal sensilla of A. lineolatus, and similar expression patterns were observed in both sexes. AlinCSP4 was found strongly expressed in sensilla trichodea (str) and middle long sensilla basiconic (mlsba) in the sensillum lymph, the hair shaft and the cavity below the sensilla hair base (Figure 4). No positive signals were detected in short sensilla basiconica (ssba) and in two types of sensilla chaetica: long curved sensilla chaetica (lcsch) and long straight sensilla chaetica (lssch). Interestingly, not all of the str sensilla were labelled by the anti-AlinCSP4 antiserum, but nearly all of the mlsba sensilla were labelled. Anti-AlinCSP5 and anti-AlinCSP6 antiserum exhibited different labelling distributions compared with anti-AlinCSP4 antiserum. The short sensilla basiconica (ssba) were intensely stained by both anti-AlinCSP5 and anti-AlinCSP6 antisera with strong expression of both proteins in the outer sensillum lymph (Figure 5H, 5I and 6H, 6I) but not in the inner sensillum lymph where the neuronal dendrites are located16. Anti-AlinCSP6 also showed intensive labelling in the lcsch sensilla (Figure 6K). However, no labelling signals were observed with the neuronal dendrites for all three CSPs (Figure 4,5,6).


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)

Immunocytochemical localization of AlinCSP6 in different antennal sensilla of male adults.The outer sensillum lymph of ssba and lcsch and the sub-cuticular spaces (ss) were heavily labelled, whereas other sensilla, such as str, mlsba, and lssch, were not labelled (the longitudinal sections are shown in A, D, G, and J, and the cross sections are shown in B, C, E, F, H, I, K, and L). The distribution of AlinCSP6 in the different antennal sensilla of the females was similar in that found in the males. The few grains found over the cuticle and the dendrites represent the non-specific background. The dilution of the 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. The abbreviations are similar to those used in Figure 4.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f6: Immunocytochemical localization of AlinCSP6 in different antennal sensilla of male adults.The outer sensillum lymph of ssba and lcsch and the sub-cuticular spaces (ss) were heavily labelled, whereas other sensilla, such as str, mlsba, and lssch, were not labelled (the longitudinal sections are shown in A, D, G, and J, and the cross sections are shown in B, C, E, F, H, I, K, and L). The distribution of AlinCSP6 in the different antennal sensilla of the females was similar in that found in the males. The few grains found over the cuticle and the dendrites represent the non-specific background. The dilution of the 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. The abbreviations are similar to those used in Figure 4.
Mentions: The immunolocalization of AlinCSP4-6 proteins in different antennal sensilla of both sexes was examined with ultrathin sections of the antennae subjected to colloidal gold post-embedding and immunocytochemical labelling. In general, each of AlinCSP4-6 proteins was shown to be expressed in distinct antennal sensilla of A. lineolatus, and similar expression patterns were observed in both sexes. AlinCSP4 was found strongly expressed in sensilla trichodea (str) and middle long sensilla basiconic (mlsba) in the sensillum lymph, the hair shaft and the cavity below the sensilla hair base (Figure 4). No positive signals were detected in short sensilla basiconica (ssba) and in two types of sensilla chaetica: long curved sensilla chaetica (lcsch) and long straight sensilla chaetica (lssch). Interestingly, not all of the str sensilla were labelled by the anti-AlinCSP4 antiserum, but nearly all of the mlsba sensilla were labelled. Anti-AlinCSP5 and anti-AlinCSP6 antiserum exhibited different labelling distributions compared with anti-AlinCSP4 antiserum. The short sensilla basiconica (ssba) were intensely stained by both anti-AlinCSP5 and anti-AlinCSP6 antisera with strong expression of both proteins in the outer sensillum lymph (Figure 5H, 5I and 6H, 6I) but not in the inner sensillum lymph where the neuronal dendrites are located16. Anti-AlinCSP6 also showed intensive labelling in the lcsch sensilla (Figure 6K). However, no labelling signals were observed with the neuronal dendrites for all three CSPs (Figure 4,5,6).

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.