TY - JOUR
T1 - Serotonin-specific neurons differentiated from human iPSCs form distinct subtypes with synaptic protein assembly
AU - Jansch, C.
AU - Ziegler, G.C.
AU - Forero, A.
AU - Gredy, S.
AU - Waldchen, S.
AU - Vitale, M.R.
AU - Svirin, E.
AU - Zoller, J.E.M.
AU - Waider, J.
AU - Gunther, K.
AU - Edenhofer, F.
AU - Sauer, M.
AU - Wischmeyer, E.
AU - Lesch, K.P.
N1 - Funding Information:
This work was supported by ERA-Net NEURON/RESPOND, No. 01EW1602B, ERA-Net NEURON/DECODE, No. FKZ01EW1902, the European Union’s Seventh Framework Programme under Grant No. 602805 (Aggressotype), the Horizon 2020 Research and Innovation Programme under Grant No. 728018 (Eat2beNICE), the 5-100 Russian Academic Excellence Project, the German Research Foundation (DFG: CRU 125, CRC TRR 58 A1/A5, No. 44541416, and Project No. 413657723 Clinician Scientist-Programme UNION CVD) and the University of Wuerzburg in the funding programme Open Access Publishing. We also would like to thank Julia Merk for her excellent technical support.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/2/1
Y1 - 2021/2/1
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.
AB - 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 - cadherin-13 (cdh13)
KW - human induced pluripotent stem cell (hipsc)
KW - median and dorsal raphe
KW - neuropsychiatric disorders
KW - serotonin-specific neurons
KW - synapse formation
KW - Synapse formation
KW - Neuropsychiatric disorders
KW - Serotonin-specific neurons
KW - Cadherin-13 (CDH13)
KW - Human induced pluripotent stem cell (hiPSC)
KW - Median and dorsal raphe
U2 - 10.1007/s00702-021-02303-5
DO - 10.1007/s00702-021-02303-5
M3 - Article
C2 - 33560471
SN - 0300-9564
VL - 128
SP - 225
EP - 241
JO - Journal of Neural Transmission
JF - Journal of Neural Transmission
IS - 2
ER -