Establishment of immortalized gba1 mouse cortical neurons - What can we learn from this model?

Friday, November 08, 2013 — Poster Session III
10:00 a.m. – 12:00 p.m.	FAES Academic Center (Upper-Level Terrace)	NHGRI	GEN-31

Authors

M. Siebert
R. Tamargo
R. Burnett
B. Berhe
N. Tayebi
W. Westbroek
E. Sidransky

Abstract

Glucocerebrosidase (GCase), a lysosomal hydrolase encoded by GBA1, is involved in the breakdown of glucosylceramide. GCase deficiency is caused by mutations in GBA1, resulting in Gaucher disease (GD), a recessive lysosomal storage disorder. Mutations in GBA1 are also a common genetic risk factor for Parkinson disease (PD). While primary rodent neurons and neuroblastoma cell lines are used to study the GBA1 and PD link, these models have limited utility because of challenges in culturing cells and/or manipulating levels of GCase expression. To overcome these difficulties, we immortalized GCase deficient neurons from gba knock-out mice for further studies. Initially, we immortalized cortical neurons from the null allele gba-/- mouse by infecting differentiated primary cortical neurons with EF1α-SV40T lentivirus. After extensive selection with puromycin, the immortalized neurons were characterized. Gba-/- neurons showed no GCase enzyme activity or expression of GCase protein compared to WT neurons. The immortalized neurons were positive for neuronal markers including TUJ-1 and MAP-2, but negative for GFAP. The SV40T immortalized lines did not show tumorigenicity in nude mice. Immortalized and characterized cortical neurons from gba mouse models can be utilized for studies of the pathogenesis of neuronopathic Gaucher disease, the glucocerebrosidase-PD association and the evaluation of therapeutics.