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Role of astroglia in Down's syndrome revealed by patient-derived human-induced pluripotent stem cells.

Chen C, Jiang P, Xue H, Peterson SE, Tran HT, McCann AE, Parast MM, Li S, Pleasure DE, Laurent LC, Loring JF, Liu Y, Deng W - Nat Commun (2014)

Bottom Line: DS astroglia exhibit higher levels of reactive oxygen species and lower levels of synaptogenic molecules.Transplantation studies show that DS astroglia do not promote neurogenesis of endogenous neural stem cells in vivo.Finally, we show that the FDA-approved antibiotic drug, minocycline, partially corrects the pathological phenotypes of DS astroglia by specifically modulating the expression of S100B, GFAP, inducible nitric oxide synthase, and thrombospondins 1 and 2 in DS astroglia.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, California 95817, USA [2] Institute for Pediatric Regenerative Medicine, Shriners Hospitals for Children, Sacramento, California 95817, USA [3] Department of Neurology, Institute of Neurology, Tianjin General Hospital, Tianjin Medical University, Tianjin 300070, China [4].

ABSTRACT
Down's syndrome (DS), caused by trisomy of human chromosome 21, is the most common genetic cause of intellectual disability. Here we use induced pluripotent stem cells (iPSCs) derived from DS patients to identify a role for astrocytes in DS pathogenesis. DS astroglia exhibit higher levels of reactive oxygen species and lower levels of synaptogenic molecules. Astrocyte-conditioned medium collected from DS astroglia causes toxicity to neurons, and fails to promote neuronal ion channel maturation and synapse formation. Transplantation studies show that DS astroglia do not promote neurogenesis of endogenous neural stem cells in vivo. We also observed abnormal gene expression profiles from DS astroglia. Finally, we show that the FDA-approved antibiotic drug, minocycline, partially corrects the pathological phenotypes of DS astroglia by specifically modulating the expression of S100B, GFAP, inducible nitric oxide synthase, and thrombospondins 1 and 2 in DS astroglia. Our studies shed light on the pathogenesis and possible treatment of DS by targeting astrocytes with a clinically available drug.

No MeSH data available.


Related in: MedlinePlus

Electrophysiological properties of DS neurons.(a–c) Quantification of membrane capacitance, inputresistance (Rin) and resting membrane potential (RMP)recorded from control (Cont) neurons (Neu), DS neurons, Cont neurons fedwith Cont ACM or DS ACM, and DS neurons fed with DS ACM, Cont ACM orDS-Mino ACM. One-wayanalysis of variance test, *P<0.05. n=10. (d)Summarized data showing that more neurons display synaptic activity when fedwith Cont ACM or DS-MinoACM than those without being fed with ACM or fed with DS ACM. (e)Representative tracing showing that synaptic activities were recorded fromCont neurons fed with Cont ACM, DS neurons fed with Cont ACM orDS-Mino ACM, but notfrom Cont neuron alone or Cont neurons fed with DS ACM, and DS neuron aloneor DS neuron fed with DS ACM. (f) Representative tracing showing atRMP, DS neurons fed with Cont ACM or DS-Mino ACM fire action potentials more robustly than DSneurons alone.
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f4: Electrophysiological properties of DS neurons.(a–c) Quantification of membrane capacitance, inputresistance (Rin) and resting membrane potential (RMP)recorded from control (Cont) neurons (Neu), DS neurons, Cont neurons fedwith Cont ACM or DS ACM, and DS neurons fed with DS ACM, Cont ACM orDS-Mino ACM. One-wayanalysis of variance test, *P<0.05. n=10. (d)Summarized data showing that more neurons display synaptic activity when fedwith Cont ACM or DS-MinoACM than those without being fed with ACM or fed with DS ACM. (e)Representative tracing showing that synaptic activities were recorded fromCont neurons fed with Cont ACM, DS neurons fed with Cont ACM orDS-Mino ACM, but notfrom Cont neuron alone or Cont neurons fed with DS ACM, and DS neuron aloneor DS neuron fed with DS ACM. (f) Representative tracing showing atRMP, DS neurons fed with Cont ACM or DS-Mino ACM fire action potentials more robustly than DSneurons alone.

Mentions: A previous study37 showed that hESC-derived neurons could matureover time in culture and exhibited passive and active electrophysiologicalproperties. During brain development, astroglia have crucial roles in promotingneuronal maturation38, and astrocyte-secreted factors powerfullyinduce the formation of functional synapses between neurons1617. To investigate whether DS astroglia regulate the maturation of neurons invitro, under directed neuronal differentiation condition, we added ACM(control ACM, DS ACM, DS S100BsiRNA ACM and DS-Mino ACM) to the 4-week-old control orDS neurons, and examined the electrophysiological properties of neurons at 6- to7-week time point. We first examined the electrophysiological properties of theneurons cultured in the presence and absence of different ACM by measuring cellmembrane capacitance (Cm), input resistance(Rin), resting membrane potential (RMP) and actionpotential evoked by depolarizing current pulses. As shown in Fig.4a–c, in the absence of ACM, DS neurons had similarCm, Rin and RMP to control neurons(Cm=20.8±2.4 and 18.1±1.5 pF;Rin=3.6±0.5 and3.0±0.5 GΩ; and RMP= −30.8±3.2and −32.1±2.3 mV for control neuron only and DSneuron only, respectively; P>0.05; n=10 for each group).Adding DS ACM did not significantly change these properties compared withcontrol neurons alone or DS neurons alone(Cm=19.1±1.7 pF,Rin=3.4±0.7 GΩ and RMP=−33.4±1.9 for control neurons fed with DS ACM;Cm=16.5±1.8 pF,Rin=3.8±0.6 GΩ and RMP=−130.7±2.7 mV for DS neurons fed with DS ACM;P>0.05; n=10 for each group). Interestingly, fed withcontrol ACM or DS-Mino ACM,the neurons had larger Cm, higher Rin andmore negative RMP compared with control neurons alone, DS neurons alone and DSneurons fed with DS ACM (Cm=40.3±6.5 pF,Rin=1.1±0.5 GΩ andRMP=−46.4±5.7 mV for DS neurons fed with controlACM; Cm=41.6±5.3 pF,Rin=1.0±0.5 GΩ andRMP=−44.6±6.2 mV for control neurons fed withcontrol ACM; Cm=33.4±3.8 pF,Rin=1.1±0.2 GΩ andRMP=−45.3±2.1 mV for DS neurons fed withDS-Mino ACM,respectively; P<0.05; n=10 for each group). However, we didnot observe any significant changes in these properties of neurons when addingDS S100BsiRNA ACM, indicating that overexpression of S100B in DS astroglia may not affectthe maturation of electrophysiological properties of DS neurons. Next, werecorded the spontaneous synaptic activity of neurons cultured in the presenceof different ACM. As shown in Fig. 4d,e, no synapticactivities were recorded from 6-week-old control neurons, consistent with aprevious study on the synaptic activity of hESC-derived neurons37. Similarly, DS neurons, DS neurons fed with DS ACM and control neurons fedwith DS ACM did not show any synaptic activities. Interestingly,~81.8% (9 of 11) of control neurons fed with control ACM, 87.5% (7 of8) of DS neurons fed with control ACM, and 54.5% (6 of 11) of DS neurons fedwith DS-Mino ACM showedsynaptic activities. Moreover, DS neurons fed with control ACM andDS-Mino ACM showed morerobust action potential firings compared with control neurons alone, controlneurons fed with DS ACM, DS neurons alone and DS neurons fed with DS ACM (Fig. 4f and Supplementary Fig. 6).


Role of astroglia in Down's syndrome revealed by patient-derived human-induced pluripotent stem cells.

Chen C, Jiang P, Xue H, Peterson SE, Tran HT, McCann AE, Parast MM, Li S, Pleasure DE, Laurent LC, Loring JF, Liu Y, Deng W - Nat Commun (2014)

Electrophysiological properties of DS neurons.(a–c) Quantification of membrane capacitance, inputresistance (Rin) and resting membrane potential (RMP)recorded from control (Cont) neurons (Neu), DS neurons, Cont neurons fedwith Cont ACM or DS ACM, and DS neurons fed with DS ACM, Cont ACM orDS-Mino ACM. One-wayanalysis of variance test, *P<0.05. n=10. (d)Summarized data showing that more neurons display synaptic activity when fedwith Cont ACM or DS-MinoACM than those without being fed with ACM or fed with DS ACM. (e)Representative tracing showing that synaptic activities were recorded fromCont neurons fed with Cont ACM, DS neurons fed with Cont ACM orDS-Mino ACM, but notfrom Cont neuron alone or Cont neurons fed with DS ACM, and DS neuron aloneor DS neuron fed with DS ACM. (f) Representative tracing showing atRMP, DS neurons fed with Cont ACM or DS-Mino ACM fire action potentials more robustly than DSneurons alone.
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Related In: Results  -  Collection

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f4: Electrophysiological properties of DS neurons.(a–c) Quantification of membrane capacitance, inputresistance (Rin) and resting membrane potential (RMP)recorded from control (Cont) neurons (Neu), DS neurons, Cont neurons fedwith Cont ACM or DS ACM, and DS neurons fed with DS ACM, Cont ACM orDS-Mino ACM. One-wayanalysis of variance test, *P<0.05. n=10. (d)Summarized data showing that more neurons display synaptic activity when fedwith Cont ACM or DS-MinoACM than those without being fed with ACM or fed with DS ACM. (e)Representative tracing showing that synaptic activities were recorded fromCont neurons fed with Cont ACM, DS neurons fed with Cont ACM orDS-Mino ACM, but notfrom Cont neuron alone or Cont neurons fed with DS ACM, and DS neuron aloneor DS neuron fed with DS ACM. (f) Representative tracing showing atRMP, DS neurons fed with Cont ACM or DS-Mino ACM fire action potentials more robustly than DSneurons alone.
Mentions: A previous study37 showed that hESC-derived neurons could matureover time in culture and exhibited passive and active electrophysiologicalproperties. During brain development, astroglia have crucial roles in promotingneuronal maturation38, and astrocyte-secreted factors powerfullyinduce the formation of functional synapses between neurons1617. To investigate whether DS astroglia regulate the maturation of neurons invitro, under directed neuronal differentiation condition, we added ACM(control ACM, DS ACM, DS S100BsiRNA ACM and DS-Mino ACM) to the 4-week-old control orDS neurons, and examined the electrophysiological properties of neurons at 6- to7-week time point. We first examined the electrophysiological properties of theneurons cultured in the presence and absence of different ACM by measuring cellmembrane capacitance (Cm), input resistance(Rin), resting membrane potential (RMP) and actionpotential evoked by depolarizing current pulses. As shown in Fig.4a–c, in the absence of ACM, DS neurons had similarCm, Rin and RMP to control neurons(Cm=20.8±2.4 and 18.1±1.5 pF;Rin=3.6±0.5 and3.0±0.5 GΩ; and RMP= −30.8±3.2and −32.1±2.3 mV for control neuron only and DSneuron only, respectively; P>0.05; n=10 for each group).Adding DS ACM did not significantly change these properties compared withcontrol neurons alone or DS neurons alone(Cm=19.1±1.7 pF,Rin=3.4±0.7 GΩ and RMP=−33.4±1.9 for control neurons fed with DS ACM;Cm=16.5±1.8 pF,Rin=3.8±0.6 GΩ and RMP=−130.7±2.7 mV for DS neurons fed with DS ACM;P>0.05; n=10 for each group). Interestingly, fed withcontrol ACM or DS-Mino ACM,the neurons had larger Cm, higher Rin andmore negative RMP compared with control neurons alone, DS neurons alone and DSneurons fed with DS ACM (Cm=40.3±6.5 pF,Rin=1.1±0.5 GΩ andRMP=−46.4±5.7 mV for DS neurons fed with controlACM; Cm=41.6±5.3 pF,Rin=1.0±0.5 GΩ andRMP=−44.6±6.2 mV for control neurons fed withcontrol ACM; Cm=33.4±3.8 pF,Rin=1.1±0.2 GΩ andRMP=−45.3±2.1 mV for DS neurons fed withDS-Mino ACM,respectively; P<0.05; n=10 for each group). However, we didnot observe any significant changes in these properties of neurons when addingDS S100BsiRNA ACM, indicating that overexpression of S100B in DS astroglia may not affectthe maturation of electrophysiological properties of DS neurons. Next, werecorded the spontaneous synaptic activity of neurons cultured in the presenceof different ACM. As shown in Fig. 4d,e, no synapticactivities were recorded from 6-week-old control neurons, consistent with aprevious study on the synaptic activity of hESC-derived neurons37. Similarly, DS neurons, DS neurons fed with DS ACM and control neurons fedwith DS ACM did not show any synaptic activities. Interestingly,~81.8% (9 of 11) of control neurons fed with control ACM, 87.5% (7 of8) of DS neurons fed with control ACM, and 54.5% (6 of 11) of DS neurons fedwith DS-Mino ACM showedsynaptic activities. Moreover, DS neurons fed with control ACM andDS-Mino ACM showed morerobust action potential firings compared with control neurons alone, controlneurons fed with DS ACM, DS neurons alone and DS neurons fed with DS ACM (Fig. 4f and Supplementary Fig. 6).

Bottom Line: DS astroglia exhibit higher levels of reactive oxygen species and lower levels of synaptogenic molecules.Transplantation studies show that DS astroglia do not promote neurogenesis of endogenous neural stem cells in vivo.Finally, we show that the FDA-approved antibiotic drug, minocycline, partially corrects the pathological phenotypes of DS astroglia by specifically modulating the expression of S100B, GFAP, inducible nitric oxide synthase, and thrombospondins 1 and 2 in DS astroglia.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, California 95817, USA [2] Institute for Pediatric Regenerative Medicine, Shriners Hospitals for Children, Sacramento, California 95817, USA [3] Department of Neurology, Institute of Neurology, Tianjin General Hospital, Tianjin Medical University, Tianjin 300070, China [4].

ABSTRACT
Down's syndrome (DS), caused by trisomy of human chromosome 21, is the most common genetic cause of intellectual disability. Here we use induced pluripotent stem cells (iPSCs) derived from DS patients to identify a role for astrocytes in DS pathogenesis. DS astroglia exhibit higher levels of reactive oxygen species and lower levels of synaptogenic molecules. Astrocyte-conditioned medium collected from DS astroglia causes toxicity to neurons, and fails to promote neuronal ion channel maturation and synapse formation. Transplantation studies show that DS astroglia do not promote neurogenesis of endogenous neural stem cells in vivo. We also observed abnormal gene expression profiles from DS astroglia. Finally, we show that the FDA-approved antibiotic drug, minocycline, partially corrects the pathological phenotypes of DS astroglia by specifically modulating the expression of S100B, GFAP, inducible nitric oxide synthase, and thrombospondins 1 and 2 in DS astroglia. Our studies shed light on the pathogenesis and possible treatment of DS by targeting astrocytes with a clinically available drug.

No MeSH data available.


Related in: MedlinePlus