Mitigation of augmented extrasynaptic NMDAR signaling and apoptosis in cortico-striatal co-cultures from Huntington's disease mice

Milnerwood, Austen J.; Kaufman, Alexandra M.; Sepers, Marja D.; Gladding, Clare M.; Zhang, Lily; Wang, Liang; Fan, Jing; Coquinco, Ainsley; Qiao, Joy Yi; Lee, Hwan; Wang, Yu Tian; Cynader, Max S.; Raymond, Lynn A.

doi:10.1016/j.nbd.2012.05.013

Cited by 64 publications

(65 citation statements)

References 59 publications

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“…Only heterozygous KI mice that faithfully reproduce the human condition were included and are herein referred to as KI. Cultures were prepared as previously described (Milnerwood et al, 2012); briefly, cortical neurons were isolated from pups at E16.5, brains were removed and placed on ice in Hank's Balanced Salt Solution (HBSS, GIBCO). 24-well plates were seeded at 115 k cells/well in 1 ml plating medium (PM, 2% B27+1/100 penicillin/streptomycin, Invitrogen; 0.5 mM α-glutamine; neurobasal medium, GIBCO).…”

Section: Experimental Methodsmentioning

confidence: 99%

“…Whole-cell patch-clamp recordings were performed on cortical cells at DIV21–26 in voltage clamp at Vh −70 mV and the membrane test function was used to determine intrinsic membrane properties ~1 min after obtaining whole-cell configuration, as described previously (Tapia et al, 2011; Kaufman et al, 2012; Milnerwood et al, 2012; Brigidi et al, 2014). Briefly, neurons were perfused at room temperature with extracellular solution (ECS) containing (in mM unless stated): 167 NaCl, 2.4 KCl, 1 MgCl 2 , 10 glucose, 10 HEPES, 2 CaCl 2 , pH 7.4, 300 mOsm.…”

Section: Experimental Methodsmentioning

confidence: 99%

“…Tolerance for series resistance (Rs) was <25 MΩ and uncompensated; ΔRs tolerance cut-off was <10%. mEPSCs and mIPSCs were analyzed with experimenter blind to genotype using Clampfit10 (threshold 5pA mEPSC, 10pA mIPSC); all events were checked by eye and monophasic events were used for amplitude and decay kinetics, while others were suppressed but included in frequency counts, as in Tapia et al (2011), Kaufman et al (2012), Milnerwood et al (2012), Brigidi et al (2014). Data are presented as mean ± s.e.m.…”

Section: Experimental Methodsmentioning

confidence: 99%

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Synaptic function is modulated by LRRK2 and glutamate release is increased in cortical neurons of G2019S LRRK2 knock-in mice

Beccano-Kelly

Kuhlmann

Tatarnikov

et al. 2014

Front. Cell. Neurosci.

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101

129

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Mutations in Leucine-Rich Repeat Kinase-2 (LRRK2) result in familial Parkinson's disease and the G2019S mutation alone accounts for up to 30% in some ethnicities. Despite this, the function of LRRK2 is largely undetermined although evidence suggests roles in phosphorylation, protein interactions, autophagy and endocytosis. Emerging reports link loss of LRRK2 to altered synaptic transmission, but the effects of the G2019S mutation upon synaptic release in mammalian neurons are unknown. To assess wild type and mutant LRRK2 in established neuronal networks, we conducted immunocytochemical, electrophysiological and biochemical characterization of >3 week old cortical cultures of LRRK2 knock-out, wild-type overexpressing and G2019S knock-in mice. Synaptic release and synapse numbers were grossly normal in LRRK2 knock-out cells, but discretely reduced glutamatergic activity and reduced synaptic protein levels were observed. Conversely, synapse density was modestly but significantly increased in wild-type LRRK2 overexpressing cultures although event frequency was not. In knock-in cultures, glutamate release was markedly elevated, in the absence of any change to synapse density, indicating that physiological levels of G2019S LRRK2 elevate probability of release. Several pre-synaptic regulatory proteins shown by others to interact with LRRK2 were expressed at normal levels in knock-in cultures; however, synapsin 1 phosphorylation was significantly reduced. Thus, perturbations to the pre-synaptic release machinery and elevated synaptic transmission are early neuronal effects of LRRK2 G2019S. Furthermore, the comparison of knock-in and overexpressing cultures suggests that one copy of the G2019S mutation has a more pronounced effect than an ~3-fold increase in LRRK2 protein. Mutant-induced increases in transmission may convey additional stressors to neuronal physiology that may eventually contribute to the pathogenesis of Parkinson's disease.

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Section: Experimental Methodsmentioning

confidence: 99%

Section: Experimental Methodsmentioning

confidence: 99%

Section: Experimental Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Synaptic function is modulated by LRRK2 and glutamate release is increased in cortical neurons of G2019S LRRK2 knock-in mice

Beccano-Kelly

Kuhlmann

Tatarnikov

et al. 2014

Front. Cell. Neurosci.

Self Cite

101

129

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“…Corticostriatal Co-cultures and Transfections-Neuronal culture preparation and maintenance were similar to those described previously (21,22). Briefly, striatal and cortical tissues were dissected from Day 17-18 WT FVB/N or YAC18 embryos in ice-cold Hanks' balanced salt solution (Invitrogen), digested in trypsin, and further dissociated in trypsin inhibitor solution.…”

Section: Methodsmentioning

confidence: 99%

Bidirectional Control of Postsynaptic Density-95 (PSD-95) Clustering by Huntingtin

Parsons

Kang

Buren

et al. 2014

Journal of Biological Chemistry

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Background: Wild-type huntingtin (HTT) interacts with synaptic proteins, yet its role in synaptic function is unclear. Results: HTT bidirectionally influences PSD-95 clustering in striatal spiny projection neurons. Conclusion: HTT plays a role in synaptic protein organization. Significance: HTT-reducing strategies are being tested as a treatment for Huntington disease. It is therefore critical to understand the role of HTT in cellular and synaptic function.

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“…112 Although many cell lines have been used, they might reveal different aspects in comparison with primary cells. Therefore, primary neurons prepared from HD transgenic mice are frequently used: neocortical or striatal cultures from HdhQ111 mice that have 111 CAG repeats in exon 1 of the mHtt gene [115][116][117][118][119] ; neostriatal cultures of the YAC46 (668 line) and YAC72 (2511 line) mice, which express the full-length mutant huntingtin containing 46 or 72 glutamine repeats (46Q or 72Q) 120 ; YAC128 (line 55) mice expressing full-length human mHtt containing 128 CAG repeats 121 ; transgenic BACHD mice that express a fulllength mHtt with 97 glutamine repeats. 122 Besides the neuronal cells, mHtt may also be transfected to non-neuronal cells, 95 such as HeLa cells, [123][124][125][126] human embryonic kidney cell-line 293T (HEK293T), 93,123,126,127 and monkey kidney cell lines (COS-7).…”

Section: -89mentioning

confidence: 99%

Studying neurodegenerative diseases in culture models

Schlachetzki

Saliba

Oliveira

2013

Rev. Bras. Psiquiatr.

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Neurodegenerative diseases are pathological conditions that have an insidious onset and chronic progression. Different models have been established to study these diseases in order to understand their underlying mechanisms and to investigate new therapeutic strategies. Although various in vivo models are currently in use, in vitro models might provide important insights about the pathogenesis of these disorders and represent an interesting approach for the screening of potential pharmacological agents. In the present review, we discuss various in vitro and ex vivo models of neurodegenerative disorders in mammalian cells and tissues.

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Mitigation of augmented extrasynaptic NMDAR signaling and apoptosis in cortico-striatal co-cultures from Huntington's disease mice

Cited by 64 publications

References 59 publications

Synaptic function is modulated by LRRK2 and glutamate release is increased in cortical neurons of G2019S LRRK2 knock-in mice

Synaptic function is modulated by LRRK2 and glutamate release is increased in cortical neurons of G2019S LRRK2 knock-in mice

Bidirectional Control of Postsynaptic Density-95 (PSD-95) Clustering by Huntingtin

Studying neurodegenerative diseases in culture models

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