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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 map of OT obtained by performing IEF on a 4–7 linear pH range and SDS-PAGE on a constant 12,5% T in the second dimension, to separate proteins clustered in the single spot shown in Fig 4. (B) Immunoblotting of the gel slab indicated in Panel A.Arrow points to spot originated from separation and identified as FliD.
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pone.0138842.g005: (A) 2-DE map of OT obtained by performing IEF on a 4–7 linear pH range and SDS-PAGE on a constant 12,5% T in the second dimension, to separate proteins clustered in the single spot shown in Fig 4. (B) Immunoblotting of the gel slab indicated in Panel A.Arrow points to spot originated from separation and identified as FliD.

Mentions: In an effort to overcome the limitations indicated above and to definitively establish (or exclude) the presence of FliD under the spot(s) examined in the OT pool, we worked on the optimization of the electrophoretic conditions. After performing a extensive set of trials with various electrophoretic conditions, the best option was found to be the application of a narrow range pH gradient (linear pH 4–7). This provided a better resolution of proteins, minimizing potential spot overlaps (Fig 5A). The immunoreactive protein spot (indicated by an arrow in PVDF membrane, B) was evidenced in OT profile obtained under the experimental conditions mentioned above.


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 map of OT obtained by performing IEF on a 4–7 linear pH range and SDS-PAGE on a constant 12,5% T in the second dimension, to separate proteins clustered in the single spot shown in Fig 4. (B) Immunoblotting of the gel slab indicated in Panel A.Arrow points to spot originated from separation and identified as FliD.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0138842.g005: (A) 2-DE map of OT obtained by performing IEF on a 4–7 linear pH range and SDS-PAGE on a constant 12,5% T in the second dimension, to separate proteins clustered in the single spot shown in Fig 4. (B) Immunoblotting of the gel slab indicated in Panel A.Arrow points to spot originated from separation and identified as FliD.
Mentions: In an effort to overcome the limitations indicated above and to definitively establish (or exclude) the presence of FliD under the spot(s) examined in the OT pool, we worked on the optimization of the electrophoretic conditions. After performing a extensive set of trials with various electrophoretic conditions, the best option was found to be the application of a narrow range pH gradient (linear pH 4–7). This provided a better resolution of proteins, minimizing potential spot overlaps (Fig 5A). The immunoreactive protein spot (indicated by an arrow in PVDF membrane, B) was evidenced in OT profile obtained under the experimental conditions mentioned above.

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