title: Localization of the bacterial symbiont Candidatus Midichloria mitochondrii within the hard tick Ixodes ricinus by whole-mount FISH staining.
abstract: Here, we present an investigation on the spatial distribution of the bacterial symbiont Candidatus Midichloria mitochondrii within Ixodes ricinus, by whole mount fluorescence in situ hybridization (FISH). M. mitochondrii is a peculiar, recently discovered bacterium that resides in the mitochondria of female ticks. We applied a rapid and specific FISH protocol with oligonucleotide probes targeted on the 16S rRNA of M. mitochondrii, 12S rRNA of tick mitochondria, and a probe revealing active mitochondria. In this report that represents the first application of whole mount FISH on ticks, we observed strong, specific fluorescence signals in all the examined life stages, as the optimized protocol allowed us to overcome the autofluorescence interference of the cuticle. Cellular localization and quantification of the symbionts were also assessed with electron microscopy and specific real-time PCR, respectively.
authors.affiliation: Dipartimento di Scienze Veterinarie e Sanità Pubblica, Sezione di Parassitologia, Università degli Studi di Milano, Via Celoria 10, Milan, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Epis
authors.forename: Mauro
authors.initials: M
authors.lastname: Mandrioli
authors.forename: Marco
authors.initials: M
authors.lastname: Genchi
authors.forename: Matteo
authors.initials: M
authors.lastname: Montagna
authors.forename: Luciano
authors.initials: L
authors.lastname: Sacchi
authors.forename: Dario
authors.initials: D
authors.lastname: Pistone
authors.forename: Davide
authors.initials: D
authors.lastname: Sassera
mesh_terms: D020561:Alphaproteobacteria / Q000145:classification* / Q000302:isolation & purification*; D000818:Animals; D005260:Female; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D018884:Ixodes / Q000382:microbiology* / Q000648:ultrastructure; D012313:RNA / Q000235:genetics; D000077278:RNA, Mitochondrial; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D025604:Rhodopsins, Microbial; D013559:Symbiosis
chemical_list: D000077278:RNA, Mitochondrial; D012336:RNA, Ribosomal, 16S; D025604:Rhodopsins, Microbial; D012313:RNA