title: Human peripheral blood lymphocytes and fibroblasts as Notch3 expression models.
abstract: Notch3 is a single pass transmembrane protein belonging to the Notch receptor family. Notch proteins are involved in a very conserved signaling system (Notch signaling) with a broad spectrum of functions, from cell proliferation and differentiation to apoptosis. Mutations in Notch3 gene are linked to cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), a disorder characterized by stroke and dementia in young adults. Studies evaluating Notch3 expression in human differentiated cells and adult tissues have shown high Notch3 levels only in vascular smooth muscle cells (VSMC). However, it has been hypothesized that Notch3 is ubiquitously expressed in adult human tissues. Our aim was to evaluate Notch3 expression in human peripheral blood lymphocytes (PBLs) and fibroblasts from normal healthy subjects. In both cell types, we examined the expression of Notch3 by reverse transcriptase-polymerase chain reaction (RT-PCR) and quantitative real-time polymerase chain reaction (qRT-PCR). Moreover, we assessed Notch3 protein expression by Western blot analysis. RT-PCR and qRT-PCR analysis showed the presence of Notch3 mRNA in both cell types. Western blot analysis confirmed Notch3 protein expression in PBLs and fibroblasts though showing different profiles. Our data support the expression of Notch3 in adult human cell types, and suggests that PBLs and fibroblasts could provide readily available cells for the study of the role of Notch3 expression in the pathogenetic mechanisms leading to different human disease.
authors.affiliation: Department of Neurological, Neurosurgical and Behavioural Sciences, University of Siena, Policlinico Le Scotte, Siena, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Di Maio
authors.forename: Patrizia
authors.initials: P
authors.lastname: Formichi
authors.forename: Elena
authors.initials: E
authors.lastname: Radi
authors.forename: Silvia
authors.initials: S
authors.lastname: Bianchi
authors.forename: Anna
authors.initials: A
authors.lastname: Onnis
authors.forename: Ermelinda
authors.initials: E
authors.lastname: Tarquini
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Leoncini
authors.forename: Antonio
authors.initials: A
authors.lastname: Federico
mesh_terms: D000328:Adult; D001483:Base Sequence; D015153:Blotting, Western; D005260:Female; D005347:Fibroblasts / Q000378:metabolism*; D020869:Gene Expression Profiling; D006801:Humans; D066298:In Vitro Techniques; D019169:Jurkat Cells; D008214:Lymphocytes / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D008954:Models, Biological; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D000071656:Receptor, Notch3; D051880:Receptors, Notch / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: C499374:NOTCH3 protein, human; D012333:RNA, Messenger; D000071656:Receptor, Notch3; D051880:Receptors, Notch