Induced pluripotent stem cell - derived neurons for the study of spinocerebellar ataxia type 3.
The neurodegenerative disease spinocerebellar ataxia type 3 (SCA3) is caused by a CAG-repeat expansion in the ATXN3 gene. In this study, induced pluripotent stem cell (iPSC) lines were established from two SCA3 patients. Dermal fibroblasts were reprogrammed using an integration-free method and the resulting SCA3 iPSCs were differentiated into neurons. These neuronal lines harbored the disease causing mutation, expressed comparable levels of several neuronal markers and responded to the neurotransmitters, glutamate/glycine, GABA and acetylcholine. Additionally, all neuronal cultures formed networks displaying synchronized spontaneous calcium oscillations within 28days of maturation, and expressed the mature neuronal markers NeuN and Synapsin 1 implying a relatively advanced state of maturity, although not comparable to that of the adult human brain. Interestingly, we were not able to recapitulate the glutamate-induced ataxin-3 aggregation shown in a previously published iPSC-derived SCA3 model. In conclusion, we have generated a panel of SCA3 patient iPSCs and a robust protocol to derive neurons of relatively advanced maturity, which could potentially be valuable for the study of SCA3 disease mechanisms.
Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Groennegårdsvej 7, 1870 Frb C, Denmark; H. Lundbeck A/S, Ottiliavej 9, Valby 2500, Denmark. Electronic address: sus.khansen@gmail.com.
Susanne K
SK
Hansen
H. Lundbeck A/S, Ottiliavej 9, Valby 2500, Denmark.
Tina C
TC
Stummann
H. Lundbeck A/S, Ottiliavej 9, Valby 2500, Denmark.
Helena
H
Borland
Institute of Cellular and Molecular Medicine, University of Copenhagen, Blegdamsvej 3B, 2200 N, Denmark.
Lis F
LF
Hasholt
Applied Human Molecular Genetics, Kennedy Center, Department of Clinical Genetics, Copenhagen University Hospital, Rigshospitalet, Gl. Landevej 7, Glostrup 2600, Denmark.
Zeynep
Z
Tümer
Institute of Cellular and Molecular Medicine, University of Copenhagen, Blegdamsvej 3B, 2200 N, Denmark; Neurogenetics Clinic & Research Laboratory, Danish Dementia Research Centre, Rigshospitalet, University of Copenhagen, Blegdamsvej 9, 2100 Copenhagen, Denmark.
Jørgen E
JE
Nielsen
Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Groennegårdsvej 7, 1870 Frb C, Denmark.
Mikkel A
MA
Rasmussen
Neurogenetics Clinic & Research Laboratory, Danish Dementia Research Centre, Rigshospitalet, University of Copenhagen, Blegdamsvej 9, 2100 Copenhagen, Denmark.
Troels T
TT
Nielsen
H. Lundbeck A/S, Ottiliavej 9, Valby 2500, Denmark.
Justus C A
JC
Daechsel
H. Lundbeck A/S, Ottiliavej 9, Valby 2500, Denmark.
Karina
K
Fog
Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Groennegårdsvej 7, 1870 Frb C, Denmark.
Poul
P
Hyttel
D034961:Antigens, Nuclear / Q000235:genetics / Q000378:metabolism; D000067699:Ataxin-3 / Q000235:genetics; D001921:Brain / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D065150:Cellular Reprogramming; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D015759:Ionomycin / Q000494:pharmacology; D059785:Karyotype; D017827:Machado-Joseph Disease / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008856:Microscopy, Fluorescence; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D066329:Protein Aggregates / Q000187:drug effects; D012097:Repressor Proteins / Q000235:genetics; D016704:Synapsins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D034961:Antigens, Nuclear; D009419:Nerve Tissue Proteins; D066329:Protein Aggregates; D012097:Repressor Proteins; D016704:Synapsins; D014157:Transcription Factors; C506808:neuronal nuclear antigen NeuN, human; D015759:Ionomycin; C092341:ATXN3 protein, human; D000067699:Ataxin-3; D002118:Calcium