TY  - JOUR
A1  - Jansch, Charline
A1  - Ziegler, Georg C.
A1  - Forero, Andrea
A1  - Gredy, Sina
A1  - Wäldchen, Sina
A1  - Vitale, Maria Rosaria
A1  - Svirin, Evgeniy
A1  - Zöller, Johanna E. M.
A1  - Waider, Jonas
A1  - Günther, Katharina
A1  - Edenhofer, Frank
A1  - Sauer, Markus
A1  - Wischmeyer, Erhard
A1  - Lesch, Klaus-Peter
T1  - Serotonin-specific neurons differentiated from human iPSCs form distinct subtypes with synaptic protein assembly
JF  - Journal of Neural Transmission
N2  - Human induced pluripotent stem cells (hiPSCs) have revolutionized the generation of experimental disease models, but the development of protocols for the differentiation of functionally active neuronal subtypes with defined specification is still in its infancy. While dysfunction of the brain serotonin (5-HT) system has been implicated in the etiology of various neuropsychiatric disorders, investigation of functional human 5-HT specific neurons in vitro has been restricted by technical limitations. We describe an efficient generation of functionally active neurons from hiPSCs displaying 5-HT specification by modification of a previously reported protocol. Furthermore, 5-HT specific neurons were characterized using high-end fluorescence imaging including super-resolution microscopy in combination with electrophysiological techniques. Differentiated hiPSCs synthesize 5-HT, express specific markers, such as tryptophan hydroxylase 2 and 5-HT transporter, and exhibit an electrophysiological signature characteristic of serotonergic neurons, with spontaneous rhythmic activities, broad action potentials and large afterhyperpolarization potentials. 5-HT specific neurons form synapses reflected by the expression of pre- and postsynaptic proteins, such as Bassoon and Homer. The distribution pattern of Bassoon, a marker of the active zone along the soma and extensions of neurons, indicates functionality via volume transmission. Among the high percentage of 5-HT specific neurons (~ 42%), a subpopulation of CDH13 + cells presumably designates dorsal raphe neurons. hiPSC-derived 5-HT specific neuronal cell cultures reflect the heterogeneous nature of dorsal and median raphe nuclei and may facilitate examining the association of serotonergic neuron subpopulations with neuropsychiatric disorders.
KW  - neuropsychiatric disorders
KW  - human induced pluripotent stem cell (hiPSC)
KW  - serotonin-specific neurons
KW  - median and dorsal raphe
KW  - synapse formation
KW  - Cadherin-13 (CDH13)
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-268519
SN  - 1435-1463
VL  - 128
IS  - 2
ER  - 
TY  - JOUR
A1  - Vitale, Maria Rosaria
A1  - Zöller, Johanna Eva Maria
A1  - Jansch, Charline
A1  - Janz, Anna
A1  - Edenhofer, Frank
A1  - Klopocki, Eva
A1  - van den Hove, Daniel
A1  - Vanmierlo, Tim
A1  - Rivero, Olga
A1  - Kasri, Nael Nadif
A1  - Ziegler, Georg Christoph
A1  - Lesch, Klaus-Peter
T1  - Generation of induced pluripotent stem cell (iPSC) lines carrying a heterozygous (UKWMPi002-A-1) and null mutant knockout (UKWMPi002-A-2) of Cadherin 13 associated with neurodevelopmental disorders using CRISPR/Cas9
JF  - Stem Cell Research
N2  - Fibroblasts isolated from a skin biopsy of a healthy 46-year-old female were infected with Sendai virus containing the Yamanaka factors to produce transgene-free human induced pluripotent stem cells (iPSCs). CRISPR/Cas9 was used to generate isogenic cell lines with a gene dose-dependent deficiency of CDH13, a risk gene associated with neurodevelopmental and psychiatric disorders. Thereby, a heterozygous CDH13 knockout (CDH13\(^{+/-}\)) and a CDH13 null mutant (CDH13\(^{-/-}\)) iPSC line was obtained. All three lines showed expression of pluripotency-associated markers, the ability to differentiate into cells of the three germ layers in vitro, and a normal female karyotype.
KW  - CRISPR-Cas Systems
KW  - cadherins
KW  - female
KW  - heterozygote
KW  - humans
KW  - Induced Pluripotent Stem Cells
KW  - middle aged
KW  - neurodevelopmental disorders / genetics
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-260331
VL  - 51
ER  -