title: Application of highly sensitive saturation labeling to the analysis of differential protein expression in infected ticks from limited samples.
abstract: BACKGROUND
  Ticks are vectors of pathogens that affect human and animal health worldwide. Proteomics and genomics studies of infected ticks are required to understand tick-pathogen interactions and identify potential vaccine antigens to control pathogen transmission. One of the limitations for proteomics research in ticks is the amount of protein that can be obtained from these organisms. In the work reported here, individual naturally-infected and uninfected Rhipicephalus spp. ticks were processed using a method that permits simultaneous extraction of DNA, RNA and proteins. This approach allowed using DNA to determine pathogen infection, protein for proteomics studies and RNA to characterize mRNA levels for some of the differentially expressed proteins. Differential protein expression in response to natural infection with different pathogens was characterized by two-dimensional (2-D) differential in gel electrophoresis (DIGE) saturation labeling in combination with mass spectrometry analysis. To our knowledge, this is the first report of the application of DIGE saturation labeling to study tick proteins.
  
  
  RESULTS
  Questing and feeding Rhipicephalus spp. adult ticks were collected in 27 farms located in different Sicilian regions. From 300 collected ticks, only 16 were found to be infected: R. sanguineus with Rickettsia conorii and Ehrlichia canis; R. bursa with Theileria annulata; and R. turanicus with Anaplasma ovis. The proteomic analysis conducted from a limited amount of proteins allowed the identification of host, pathogen and tick proteins differentially expressed as a consequence of infection.
  
  
  CONCLUSIONS
  These results showed that DIGE saturation labeling is a powerful technology for proteomics studies in small number of ticks and provided new information about the effect of pathogen infection in ticks.
authors: Instituto de Investigación en Recursos Cinegéticos IREC (CSIC-UCLM-JCCM), Ronda de Toledo s/n, 13005 Ciudad Real, Spain. margaritam.villar@uclm.es.
authors: Margarita
authors: M
authors: Villar
authors: Alessandra
authors: A
authors: Torina
authors: Yolanda
authors: Y
authors: Nuñez
authors: Zorica
authors: Z
authors: Zivkovic
authors: Anabel
authors: A
authors: Marina
authors: Angela
authors: A
authors: Alongi
authors: Salvatore
authors: S
authors: Scimeca
authors: Giuseppa
authors: G
authors: La Barbera
authors: Santo
authors: S
authors: Caracappa
authors: Jesús
authors: J
authors: Vázquez
authors: José de la
authors: Jde L
authors: Fuente