TY  - JOUR
A1  - Krajka, Victor
A1  - Naujock, Maximilian
A1  - Pauly, Martje G.
A1  - Stengel, Felix
A1  - Meier, Britta
A1  - Stanslowsky, Nancy
A1  - Klein, Christine
A1  - Seibler, Philip
A1  - Wegner, Florian
A1  - Capetian, Philipp
T1  - Ventral Telencephalic Patterning Protocols for Induced Pluripotent Stem Cells
JF  - Frontiers in Cell and Developmental Biology
N2  - The differentiation of human induced pluripotent stem cells (hiPSCs) into specific cell types for disease modeling and restorative therapies is a key research agenda and offers the possibility to obtain patient-specific cells of interest for a wide range of diseases. Basal forebrain cholinergic neurons (BFCNs) play a particular role in the pathophysiology of Alzheimer’s dementia and isolated dystonias. In this work, various directed differentiation protocols based on monolayer neural induction were tested for their effectiveness in promoting a ventral telencephalic phenotype and generating BFCN. Ventralizing factors [i.e., purmorphamine and Sonic hedgehog (SHH)] were applied at different time points, time intervals, and concentrations. In addition, caudal identity was prevented by the use of a small molecule XAV-939 that inhibits the Wnt-pathway. After patterning, gene expression profiles were analyzed by quantitative PCR (qPCR). Rostro-ventral patterning is most effective when initiated simultaneously with neural induction. The most promising combination of patterning factors was 0.5 μM of purmorphamine and 1 μM of XAV-939, which induces the highest expression of transcription factors specific for the medial ganglionic eminence, the source of GABAergic inter- and cholinergic neurons in the telencephalon. Upon maturation of cells, the immune phenotype, as well as electrophysiological properties were investigated showing the presence of marker proteins specific for BFCN (choline acetyltransferase, ISL1, p75, and NKX2.1) and GABAergic neurons. Moreover, a considerable fraction of measured cells displayed mature electrophysiological properties. Synaptic boutons containing the vesicular acetylcholine transporter (VACHT) could be observed in the vicinity of the cells. This work will help to generate basal forebrain interneurons from hiPSCs, providing a promising platform for modeling neurological diseases, such as Alzheimer’s disease or Dystonia.
KW  - induced pluripotent stem cells
KW  - medial ganglionic eminence
KW  - Sonic hedgehog
KW  - XAV-939
KW  - purmorphamine
KW  - basal forebrain cholinergic neurons
KW  - GABAergic neurons
KW  - electrophysiology
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-244607
SN  - 2296-634X
VL  - 9
ER  - 
TY  - JOUR
A1  - Song, Ning-Ning
A1  - Xiu, Jian-Bo
A1  - Huang, Ying
A1  - Chen, Jia-Yin
A1  - Zhang, Lei
A1  - Gutknecht, Lise
A1  - Lesch, Klaus Peter
A1  - Li, He
A1  - Ding, Yu-Qiang
T1  - Adult Raphe-Specific Deletion of Lmx1b Leads to Central Serotonin Deficiency
JF  - PLoS ONE
N2  - The transcription factor Lmx1b is essential for the differentiation and survival of central serotonergic (5-HTergic) neurons during embryonic development. However, the role of Lmx1b in adult 5-HTergic neurons is unknown. We used an inducible Cre-LoxP system to selectively inactivate Lmx1b expression in the raphe nuclei of adult mice. Pet1-CreER(T2) mice were generated and crossed with Lmx1b(flox/flox) mice to obtain Pet1-CreER(T2); Lmx1b(flox/flox) mice (which termed as Lmx1b iCKO). After administration of tamoxifen, the level of 5-HT in the brain of Lmx1b iCKO mice was reduced to 60% of that in control mice, and the expression of tryptophan hydroxylase 2 (Tph2), serotonin transporter (Sert) and vesicular monoamine transporter 2 (Vmat2) was greatly down-regulated. On the other hand, the expression of dopamine and norepinephrine as well as aromatic L-amino acid decarboxylase (Aadc) and Pet1 was unchanged. Our results reveal that Lmx1b is required for the biosynthesis of 5-HT in adult mouse brain, and it may be involved in maintaining normal functions of central 5-HTergic neurons by regulating the expression of Tph2, Sert and Vmat2.
KW  - Molecular-genetics
KW  - Sonic hedgehog
KW  - Neurons
KW  - Mice
KW  - Brain
KW  - Expression
KW  - System
KW  - PET-1
KW  - Transporter
KW  - Disorders
Y1  - 2011
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-133581
VL  - 6
IS  - 1
ER  -