TY  - THES
A1  - Dilling, Christina
T1  - Auswirkungen des Protocadherin-gamma-C3-Knockouts auf die Barriereeigenschaften der Blut-Hirn-Schranke
T1  - Impact of Protocadherin gamma C3 knock-out on the barrier properties of the blood-brain-barrier
N2  - Protocadherine spielen eine wichtige Rolle bei der Entwicklung des Nervensystems und sind an Prozessen der Zellmigration und -differenzierung, sowie der Hemmung von Zellwachstum beteiligt. 
Um die Funktion und Regulation von Protocadherin gamma C3 (PcdhγC3) an mikrovaskulären Endothelzellen des Großhirns (cEND) und des Kleinhirns (cerebEND) zu untersuchen, wurden die PcdhγC3-Knock-out (KO) Zelllinien mit der CRISPR/Cas9 Methode etabliert.
Der KO führt zu verminderten Barriereeigenschaften der Blut-Hirn-Schranke (BHS), was sich in einer erhöhten Permeabilität für Fluoreszein und einem verringerten transendothelialen elektrischen Widerstand (TEER) widerspiegelt. 
Es konnte eine Veränderung der Wachstumsrate und dem Adhäsionsverhalten der KO-Zellen nachgewiesen werden. Auch die Expression der Tight-Junction-Proteine, sowie einiger Komponenten des Wnt und mTOR Signalwegs wurden durch den KO von PcdhgC3 beeinflusst.
N2  - Protocadherins (Pcdhs) play an important role in neuronal development, cell migration and differentiation, as well as inhibition of cell growth. 
To investigate the function and regulation of Protocadherin gamma C3 (PcdhgC3) on microvascular endothelial cells of the cerebrum (cEND) and cerebellum (cerebEND), PcdhgC3 knock-out (KO) cell lines were generated, using the CRIPS/Cas9 method.
The Knock-out lowers the barrier function oft he blood-brain-barrier (BBB), which can be seen in lower transendothelial electrical resistance (TEER) and higher permeabilitiy for fluorescein.
We detectet a change in the growth rate and the cellular adhesion of the KO-cells. Also the expression of tight-junction proteins, as well as parts oft the Wnt and mTOR signalling pathway are altered by the KO of PcdhgC3.
KW  - Blut-Hirn-Schranke
KW  - Knockout
KW  - Tight junction
KW  - Claudine
KW  - Occludin
KW  - Protocadherin gamma C3
KW  - Barriereeigenschaften
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-260137
ER  - 
TY  - JOUR
A1  - Gabbert, Lydia
A1  - Dilling, Christina
A1  - Meybohm, Patrick
A1  - Burek, Malgorzata
T1  - Deletion of Protocadherin Gamma C3 Induces Phenotypic and Functional Changes in Brain Microvascular Endothelial Cells In Vitro
JF  - Frontiers in Pharmacology
N2  - Inflammation of the central nervous system (CNS) is associated with diseases such as multiple sclerosis, stroke and neurodegenerative diseases. Compromised integrity of the blood-brain barrier (BBB) and increased migration of immune cells into the CNS are the main characteristics of brain inflammation. Clustered protocadherins (Pcdhs) belong to a large family of cadherin-related molecules. Pcdhs are highly expressed in the CNS in neurons, astrocytes, pericytes and epithelial cells of the choroid plexus and, as we have recently demonstrated, in brain microvascular endothelial cells (BMECs). Knockout of a member of the Pcdh subfamily, PcdhgC3, resulted in significant changes in the barrier integrity of BMECs. Here we characterized the endothelial PcdhgC3 knockout (KO) cells using paracellular permeability measurements, proliferation assay, wound healing assay, inhibition of signaling pathways, oxygen/glucose deprivation (OGD) and a pro-inflammatory cytokine tumor necrosis factor alpha (TNFα) treatment. PcdhgC3 KO showed an increased paracellular permeability, a faster proliferation rate, an altered expression of efflux pumps, transporters, cellular receptors, signaling and inflammatory molecules. Serum starvation led to significantly higher phosphorylation of extracellular signal-regulated kinases (Erk) in KO cells, while no changes in phosphorylated Akt kinase levels were found. PcdhgC3 KO cells migrated faster in the wound healing assay and this migration was significantly inhibited by respective inhibitors of the MAPK-, β-catenin/Wnt-, mTOR- signaling pathways (SL327, XAV939, or Torin 2). PcdhgC3 KO cells responded stronger to OGD and TNFα by significantly higher induction of interleukin 6 mRNA than wild type cells. These results suggest that PcdhgC3 is involved in the regulation of major signaling pathways and the inflammatory response of BMECs.
KW  - blood-brain barrier
KW  - protocadherin gamma C3
KW  - inflammation
KW  - oxygen/glucose deprivation
KW  - stroke
KW  - tumor necrosis factor-α
KW  - proliferation
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-219828
SN  - 1663-9812
VL  - 11
ER  -