Protocols in Current Issue
    Cranioplastic Surgery and Acclimation Training for Awake Mouse fMRI
    Authors:  Tomokazu Tsurugizawa, Kota Tamada, Clement Debacker, Andrew Zalesky and Toru Takumi, date: 04/05/2021, view: 181, Q&A: 0

    MRI is a promising tool for translational research to link brain function and structure in animal models of disease to patients with neuropsychiatric disorders. However, given that mouse functional MRI (fMRI) typically relies on anesthetics to suppress head motion and physiological noise, it has been difficult to directly compare brain fMRI in

    Transcardiac Perfusion of the Mouse for Brain Tissue Dissection and Fixation
    Authors:  Jinyun Wu, Yuqi Cai, Xiaoyang Wu, Yue Ying, Yilin Tai and Miao He, date: 03/05/2021, view: 773, Q&A: 0
    [Abstract] Transcardiac perfusion with saline followed by 4% paraformaldehyde (PFA) is widely used to clear blood and preserve brain for immunostaining or in situ hybridization. PFA breaks into formaldehyde in solution, which cross-link protein and DNA molecules to preserve tissue and cell structure. Here we provide a step by step guide for ...
    Tracing Nitrogen Metabolism in Mouse Tissues with Gas Chromatography-Mass Spectrometry
    Authors:  Rong Xu, Yekai Wang and Jianhai Du, date: 02/20/2021, view: 905, Q&A: 0

    Nitrogen-containing metabolites including ammonia, amino acids, and nucleotides, are essential for cell metabolism, growth, and neural transmission. Nitrogen metabolism is tightly coordinated with carbon metabolism in the breakdown and biosynthesis of amino acids and nucleotides. Both nuclear magnetic resonance spectroscopy and mass spectrometry

    Polyamine Transport Assay Using Reconstituted Yeast Membranes
    Authors:  Sarah van Veen, Shaun Martin, Marleen Schuermans and Peter Vangheluwe, date: 01/20/2021, view: 1049, Q&A: 0

    ATP13A2/PARK9 is a late endo-/lysosomal P5B transport ATPase that is associated with several neurodegenerative disorders. We recently characterized ATP13A2 as a lysosomal polyamine exporter, which sheds light on the molecular identity of the unknown mammalian polyamine transport system. Here, we describe step by step a protocol to measure

    Low- and High-resolution Dynamic Analyses for Magnetic Resonance Spectroscopy Data
    Author:  Reuben Rideaux, date: 01/20/2021, view: 516, Q&A: 0

    Magnetic resonance spectroscopy (MRS) can be used to measure in vivo concentrations of neurometabolites. This information can be used to identify neurotransmitter involvement in healthy (e.g., perceptual and cognitive processes) and unhealthy brain function (e.g., neurological and psychiatric illnesses). The standard approach for analyzing MRS

    Preparation of Synaptoneurosomes to Study the Synapse in the Murine Cerebral Cortex
    Authors:  Ariel Diaz, Enrique Torre and Manuel Yepes, date: 01/20/2021, view: 822, Q&A: 1

    The synapse is a complex structure where the transmission of information takes place. Synaptic dysfunction is one of the earliest pathophysiological events in several diseases, such as traumatic brain injury, cerebral ischemia, and neurodegenerative diseases. Thus, a methodology to study synaptic structure and function is crucial for the

    Using the Parafilm-assisted Microdissection (PAM) Method to Sample Rodent Nucleus Accumbens
    [Abstract] Microdissection techniques are very important for anatomical and functional studies focused on neuroscience, where it is often necessary microdissect specific brain areas to perform molecular or anatomical analyses. The parafilm®-assisted microdissection (PAM) was previously described and involves the microdissection of tissue ...
    Preparing Viable Hippocampal Slices from Adult Mice for the Study of Sharp Wave-ripples
    Authors:  Linhua Liu, Xiaojing Zhou and Jian-young Wu, date: 10/05/2020, view: 1016, Q&A: 0
    [Abstract] We describe a protocol for preparing acute brain slices which can produce robust hippocampal sharp wave-ripples (SWRs) in vitro. The protocol is optimized for its simplicity and reliability for the preparation of solutions, slicing, and recovery incubation. Most slices in almost every mouse prepared though the protocol expressed vigorous ...
    Method for Prolonged Incubation of Brain Slices
    Authors:  Orsolya Kekesi and Yossi Buskila, date: 07/20/2020, view: 1271, Q&A: 0
    [Abstract] Slices of neuronal tissue maintain a high degree of topographical and functional properties of neurons and glia and therefore are extensively used for measurements of neuronal activity at the molecular, cellular and network levels. However, the lifespan of slice preparations is narrow, averaging of 6-8 hours. Moreover, the average viability of ...
    Optic Nerve Crush in Mice to Study Retinal Ganglion Cell Survival and Regeneration
    [Abstract] In diseases such as glaucoma, the failure of retinal ganglion cell (RGC) neurons to survive or regenerate their optic nerve axons underlies partial and, in some cases, complete vision loss. Optic nerve crush (ONC) serves as a useful model not only of traumatic optic neuropathy but also of glaucomatous injury, as it similarly induces RGC cell death ...

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