Subcellular Fractionation for the Isolation of Synaptic Components from the Murine Brain DOI Open Access
Sofia Massaro Tieze, Sreeganga S. Chandra, D. J. Vidyadhara

et al.

Journal of Visualized Experiments, Journal Year: 2022, Volume and Issue: 187

Published: Sept. 14, 2022

Synaptic terminals are the primary sites of neuronal communication. dysfunction is a hallmark many neuropsychiatric and neurological disorders. The characterization synaptic sub-compartments by biochemical isolation is, therefore, powerful method to elucidate molecular bases processes, both in health disease. This protocol describes from mouse brains subcellular fractionation. First, sealed terminal structures, known as synaptosomes, isolated following brain tissue homogenization. Synaptosomes pre- post-synaptic compartments with pinched-off membranes. These structures retain metabolically active state valuable for studying structure function. synaptosomes then subjected hypotonic lysis ultracentrifugation obtain enriched vesicles, cytosol, plasma membrane. Fraction purity confirmed electron microscopy enrichment analysis proteins specific sub-synaptic compartments. presented straightforward tool structural functional characteristics synapse etiology various

Language: Английский

Cocaine-regulated trafficking of dopamine transporters in cultured neurons revealed by a pH sensitive reporter DOI Creative Commons
Jacqueline Saenz,

Oscar Yao,

Elnaz Khezerlou

et al.

iScience, Journal Year: 2022, Volume and Issue: 26(1), P. 105782 - 105782

Published: Dec. 9, 2022

Cocaine acts by inhibiting plasma membrane dopamine transporter (DAT) function and altering its surface expression. The precise manner mechanism which cocaine regulates DAT trafficking, especially at neuronal processes, are poorly understood. In this study, we engineered validated the use of DAT-pHluorin for studying localization dynamic trafficking processes cultured mouse midbrain neurons. We demonstrate that unlike soma dendrites, contain a majority DATs in weakly acidic intracellular compartments, axonal both shafts boutons primarily (75%) localized to membrane, whereas large varicosities abundant within structures. also exposure leads Synaptojanin1-sensitive internalization process followed reinsertion lasts days. Thus, our study reveals previously unknown dynamics molecular regulation cocaine-regulated processes.

Language: Английский

Citations

7
Sofalcone attenuates neurodegeneration in MPTP-induced mouse model of Parkinson’s disease by inhibiting oxidative stress and neuroinflammation DOI
Mulan Chen, Xin He,

Yepeng Fan

et al.

Molecular Biology Reports, Journal Year: 2024, Volume and Issue: 51(1)

Published: Aug. 14, 2024

Language: Английский

Citations

1
Decoding transcriptomic signatures of Cysteine String Protein alpha-mediated synapse maintenance DOI Creative Commons
Na Wang, Biqing Zhu, Mary Alice Allnutt

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Oct. 3, 2023

Synapse maintenance is essential for generating functional circuitry and decrement in this process a hallmark of neurodegenerative disease. While we are beginning to understand the basis synapse formation, much less known about

Language: Английский

Citations

2
Impaired pre-synaptic plasticity and visual responses in auxilin-knockout mice DOI Creative Commons
Xi Cheng, Yu Tang, D. J. Vidyadhara

et al.

iScience, Journal Year: 2023, Volume and Issue: 26(10), P. 107842 - 107842

Published: Sept. 6, 2023

Highlights•Pre-synaptic short-term plasticity is impaired in Aux-KO mice•Orientation selectivity and visual mismatch responses are reduced mice•Visual familiarity-induced oscillations mice•Optokinetic miceSummaryAuxilin (DNAJC6/PARK19), an endocytic co-chaperone, essential for maintaining homeostasis the readily releasable pool (RRP) by aiding clathrin-mediated uncoating of synaptic vesicles. Its loss-of-function mutations, observed familial Parkinson's disease (PD), lead to basal ganglia motor deficits cortical dysfunction. We discovered that auxilin-knockout (Aux-KO) mice exhibited pre-synaptic layer 4 2/3 pyramidal cell synapses primary cortex (V1), including facilitation depression. Computational modeling revealed increased RRP refilling during short repetitive stimulation, which diminished prolonged stimulation. Silicon probe recordings V1 demonstrated disrupted circuit responses, orientation selectivity, compromised negativity, shorter familiarity-evoked theta oscillations. Pupillometry analysis optokinetic response. Auxilin-dependent endocytosis dysfunction was associated with plasticity, functions, eye movement prodromally or at early stage symptoms.Graphical abstract

Language: Английский

Citations

2
Subcellular Fractionation for the Isolation of Synaptic Components from the Murine Brain DOI Open Access
Sofia Massaro Tieze, Sreeganga S. Chandra, D. J. Vidyadhara

et al.

Journal of Visualized Experiments, Journal Year: 2022, Volume and Issue: 187

Published: Sept. 14, 2022

Synaptic terminals are the primary sites of neuronal communication. dysfunction is a hallmark many neuropsychiatric and neurological disorders. The characterization synaptic sub-compartments by biochemical isolation is, therefore, powerful method to elucidate molecular bases processes, both in health disease. This protocol describes from mouse brains subcellular fractionation. First, sealed terminal structures, known as synaptosomes, isolated following brain tissue homogenization. Synaptosomes pre- post-synaptic compartments with pinched-off membranes. These structures retain metabolically active state valuable for studying structure function. synaptosomes then subjected hypotonic lysis ultracentrifugation obtain enriched vesicles, cytosol, plasma membrane. Fraction purity confirmed electron microscopy enrichment analysis proteins specific sub-synaptic compartments. presented straightforward tool structural functional characteristics synapse etiology various

Language: Английский

Citations

3