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Ixodes ricinus and Its Endosymbiont Midichloria mitochondrii: A Comparative Proteomic Analysis of Salivary Glands and Ovaries.

Di Venere M, Fumagalli M, Cafiso A, De Marco L, Epis S, Plantard O, Bardoni A, Salvini R, Viglio S, Bazzocchi C, Iadarola P, Sassera D - PLoS ONE (2015)

Bottom Line: We detected 21 spots showing significant differences in the relative abundance between the OT and SG, ten of which showed 4- to 18-fold increase/decrease in density.Additionally, we were able to use an immunoproteomic approach to detect a protein from the symbiont.Finally, the method here developed will pave the way for future studies on the proteomics of I. ricinus, with the goals of better understanding the biology of this vector and of its symbiont M. mitochondrii.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Medicine, University of Pavia, Pavia, Italy.

ABSTRACT
Hard ticks are hematophagous arthropods that act as vectors of numerous pathogenic microorganisms of high relevance in human and veterinary medicine. Ixodes ricinus is one of the most important tick species in Europe, due to its role of vector of pathogenic bacteria such as Borrelia burgdorferi and Anaplasma phagocytophilum, of viruses such as tick borne encephalitis virus and of protozoans as Babesia spp. In addition to these pathogens, I. ricinus harbors a symbiotic bacterium, Midichloria mitochondrii. This is the dominant bacteria associated to I. ricinus, but its biological role is not yet understood. Most M. mitochondrii symbionts are localized in the tick ovaries, and they are transmitted to the progeny. M. mitochondrii bacteria have however also been detected in the salivary glands and saliva of I. ricinus, as well as in the blood of vertebrate hosts of the tick, prompting the hypothesis of an infectious role of this bacterium. To investigate, from a proteomic point of view, the tick I. ricinus and its symbiont, we generated the protein profile of the ovary tissue (OT) and of salivary glands (SG) of adult females of this tick species. To compare the OT and SG profiles, 2-DE profiling followed by LC-MS/MS protein identification were performed. We detected 21 spots showing significant differences in the relative abundance between the OT and SG, ten of which showed 4- to 18-fold increase/decrease in density. This work allowed to establish a method to characterize the proteome of I. ricinus, and to detect multiple proteins that exhibit a differential expression profile in OT and SG. Additionally, we were able to use an immunoproteomic approach to detect a protein from the symbiont. Finally, the method here developed will pave the way for future studies on the proteomics of I. ricinus, with the goals of better understanding the biology of this vector and of its symbiont M. mitochondrii.

No MeSH data available.


Related in: MedlinePlus

(A) 2-DE maps of three different pools of salivary gland (SG, left) and ovarian tissue (OT, right) of I. ricinus, obtained by performing IEF on 7 cm IPG strips with 3–10 NL pH range and SDS-PAGE in the second dimension on 8x6 cm slabs, 12.5% T gels. (B) SG and OT master gels obtained merging the three gels for each sample type. (C) 2-DE High Master Gel created comparing the SG and OT gels.
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pone.0138842.g001: (A) 2-DE maps of three different pools of salivary gland (SG, left) and ovarian tissue (OT, right) of I. ricinus, obtained by performing IEF on 7 cm IPG strips with 3–10 NL pH range and SDS-PAGE in the second dimension on 8x6 cm slabs, 12.5% T gels. (B) SG and OT master gels obtained merging the three gels for each sample type. (C) 2-DE High Master Gel created comparing the SG and OT gels.

Mentions: The master gels from both SG and OT pools showed similar patterns of proteins such that they could be matched to each other. This facilitated the correlation of gels and the creation of a virtual image, indicated as high master gel (HMG), comprehensive of all matched spots derived from master gels. The procedure described is summarized in Fig 1.


Ixodes ricinus and Its Endosymbiont Midichloria mitochondrii: A Comparative Proteomic Analysis of Salivary Glands and Ovaries.

Di Venere M, Fumagalli M, Cafiso A, De Marco L, Epis S, Plantard O, Bardoni A, Salvini R, Viglio S, Bazzocchi C, Iadarola P, Sassera D - PLoS ONE (2015)

(A) 2-DE maps of three different pools of salivary gland (SG, left) and ovarian tissue (OT, right) of I. ricinus, obtained by performing IEF on 7 cm IPG strips with 3–10 NL pH range and SDS-PAGE in the second dimension on 8x6 cm slabs, 12.5% T gels. (B) SG and OT master gels obtained merging the three gels for each sample type. (C) 2-DE High Master Gel created comparing the SG and OT gels.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4580635&req=5

pone.0138842.g001: (A) 2-DE maps of three different pools of salivary gland (SG, left) and ovarian tissue (OT, right) of I. ricinus, obtained by performing IEF on 7 cm IPG strips with 3–10 NL pH range and SDS-PAGE in the second dimension on 8x6 cm slabs, 12.5% T gels. (B) SG and OT master gels obtained merging the three gels for each sample type. (C) 2-DE High Master Gel created comparing the SG and OT gels.
Mentions: The master gels from both SG and OT pools showed similar patterns of proteins such that they could be matched to each other. This facilitated the correlation of gels and the creation of a virtual image, indicated as high master gel (HMG), comprehensive of all matched spots derived from master gels. The procedure described is summarized in Fig 1.

Bottom Line: We detected 21 spots showing significant differences in the relative abundance between the OT and SG, ten of which showed 4- to 18-fold increase/decrease in density.Additionally, we were able to use an immunoproteomic approach to detect a protein from the symbiont.Finally, the method here developed will pave the way for future studies on the proteomics of I. ricinus, with the goals of better understanding the biology of this vector and of its symbiont M. mitochondrii.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Medicine, University of Pavia, Pavia, Italy.

ABSTRACT
Hard ticks are hematophagous arthropods that act as vectors of numerous pathogenic microorganisms of high relevance in human and veterinary medicine. Ixodes ricinus is one of the most important tick species in Europe, due to its role of vector of pathogenic bacteria such as Borrelia burgdorferi and Anaplasma phagocytophilum, of viruses such as tick borne encephalitis virus and of protozoans as Babesia spp. In addition to these pathogens, I. ricinus harbors a symbiotic bacterium, Midichloria mitochondrii. This is the dominant bacteria associated to I. ricinus, but its biological role is not yet understood. Most M. mitochondrii symbionts are localized in the tick ovaries, and they are transmitted to the progeny. M. mitochondrii bacteria have however also been detected in the salivary glands and saliva of I. ricinus, as well as in the blood of vertebrate hosts of the tick, prompting the hypothesis of an infectious role of this bacterium. To investigate, from a proteomic point of view, the tick I. ricinus and its symbiont, we generated the protein profile of the ovary tissue (OT) and of salivary glands (SG) of adult females of this tick species. To compare the OT and SG profiles, 2-DE profiling followed by LC-MS/MS protein identification were performed. We detected 21 spots showing significant differences in the relative abundance between the OT and SG, ten of which showed 4- to 18-fold increase/decrease in density. This work allowed to establish a method to characterize the proteome of I. ricinus, and to detect multiple proteins that exhibit a differential expression profile in OT and SG. Additionally, we were able to use an immunoproteomic approach to detect a protein from the symbiont. Finally, the method here developed will pave the way for future studies on the proteomics of I. ricinus, with the goals of better understanding the biology of this vector and of its symbiont M. mitochondrii.

No MeSH data available.


Related in: MedlinePlus