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Role of the protocadherin-gamma protein family at the blood-brain barrier
Germany (bvmd) - Julius-Maximilians Universitaet, Wuerzburg
University Hospital W
Prof. Dr. Dr. h.c. Norbert Roewer
PD Dr. rer. nat. Malgorzata Burek
Type of Research Project
- Basic science
What is the background of the project?
Protocadherins are known to regulate neuronal survival, development and cell to cell communication. In addition to neurons, the expression of protocadherins has been characterized in astrocytes and in the choroid plexus. The protocadherin expression was documented in the brain vasculature, but this subject has not been further explored. Recently, we determined the expression of multiple members of the protocadherin family in human and mouse endothelial cells isolated from different vascular beds. Especially in the brain microvascular endothelial cells, which are a major component of the blood-brain barrier, the high levels of protocadherins were detected. In order to study the cellular functions of protocadherins at the blood-brain barrier in detail, we generated brain endothelial knockout cell lines deficient for one member of protocadherin protein family, PcdhgC3, using an innovative CRISPR/Cas9 system (Dilling et al., JCBFM in press). Knockout of this molecule led to the changes in expression of junctional proteins leading to the conclusion that PcdhgC3 might play a decisive role in endothelial cell signaling either at cerebral vessels or at the vasculature of other organs.
What is the aim of the project?
The aim of the project is to elucidate the function of PcdhgC3 on endothelial characteristics by analyzing the wild type brain endothelial cells and endothelial cells lacking this molecule (PcdhgC3 KO). The aspects of the physical, enzymatic and transport barrier will be analyzed in both cell types.
What techniques and methods are used?
Cell culture, cellular assays (viability, apoptosis, cell growth) Western Blot, quantitative real time polymerase chain reactions (qPCR), immunocytochemistry
What is the role of the student?
What are the tasks expected to be accomplished by the student?
The project will apply a row of biochemical, cell and molecular biology techniques. Following tasks will be performed on his/her own: culturing of immortalized and primary brain endothelial cells, isolation of proteins, isolation of RNA, performing of Western blot, performing of real-time polymerase chain reaction (PCR), staining of the cells and imaging using an immunofluorescence, analyzing of the results, preparing the Power point presentation of the results and literature.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The student will obtain theoretical manuals and tutorials considering the laboratory techniques to be applied as well as the project-related literature. The student will attend lab »Work in progeress« and »Journal Club« seminars as well as will present their own results and one »Journal Club« seminar. Other seminars and lectures offered in the department and at the university are optional.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a poster - The student will prepare a presentation - The student will prepare a scientific report - The student will prepare an abstract - The student’s name will be mentioned in a future publication - The student will have the opportunity to present the results together with the supervisor at a conference
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
A basic knowledge of biochemisty and cell biolgogy is required. Highly motivated students enjoying the team work are welcomed.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Pre-Medical students from the American-British system - Students in biomedical fields - Dental medicine students (IADS members)
- Dilling C; Roewer N; Förster CY; Burek M (2017) Multiple protocadherins are expressed in brain microvascular endothelial cells and might play a role in tight junction protein regulation. JCBFM in press
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