Mammalia

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    Protocols in Current Issue
    Translating Ribosome Affinity Purification (TRAP) of Cell Type-specific mRNA from Mouse Brain Lysates
    Authors:  Catherine L. Salussolia, Kellen D. Winden and Mustafa Sahin, date: 05/05/2022, view: 459, Q&A: 0
    [Abstract]

    Mammalian tissues are highly heterogenous and complex, posing a challenge in understanding the molecular mechanisms regulating protein expression within various tissues. Recent studies have shown that translation at the level of the ribosome is highly regulated, and can vary independently of gene expression observed at a transcriptome level, as

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    Immunoprecipation Assay to Quantify the Amount of tRNAs associated with Their Interacting Proteins in Tissue and Cell Culture
    Authors:  Sarada Das, Amila Zuko, Robin Thompson, Erik Storkebaum and Zoya Ignatova, date: 02/20/2022, view: 1116, Q&A: 1
    [Abstract]

    Transfer RNAs (tRNAs) are highly abundant species and, along their biosynthetic and functional path, they establish interactions with a plethora of proteins. The high number of nucleobase modifications in tRNAs renders conventional RNA quantification approaches unsuitable to study protein-tRNA interactions and their associated functional roles in

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    An in vitro Blood-brain Barrier Model to Study the Penetration of Nanoparticles
    Authors:  Sezin Aday, Wen Li, Jeffrey M. Karp and Nitin Joshi, date: 02/20/2022, view: 2578, Q&A: 0
    [Abstract]

    The blood-brain barrier (BBB), a crucial protection mechanism in the central nervous system (CNS), is a selective barrier comprised of endothelial cells. It hampers the development of therapeutic and diagnostic tools for neurological diseases due to the poor penetration of most of these agents. Rationally engineered nanoparticles (NP) can

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    Subcellular RNA-seq for the Analysis of the Dendritic and Somatic Transcriptomes of Single Neurons
    Authors:  Julio D. Perez and Erin M. Schuman, date: 01/05/2022, view: 1383, Q&A: 0
    [Abstract]

    In neurons, local translation in dendritic and axonal compartments allows for the fast and on-demand modification of the local proteome. As the last few years have witnessed dramatic advancements in our appreciation of the brain’s neuronal diversity, it is increasingly relevant to understand how local translation is regulated according to

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    Delivery of AAV for Expression of Fluorescent Biosensors in Juvenile Mouse Hippocampus
    [Abstract]

    Genetically encoded fluorescent biosensors are versatile tools for studying brain metabolism and function in live tissue. The genetic information for these biosensors can be delivered into the brain by stereotaxic injection of engineered adeno-associated viruses (AAVs), which can selectively target different cell types depending on the capsid

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    Conditional Gene Editing in Presynaptic Extinction-ensemble Cells via the CRISPR-SaCas9 System
    Authors:  Haojie Sun, Ming Yi and You Wan, date: 12/05/2021, view: 921, Q&A: 0
    [Abstract]

    The CRISPR-Cas9 enables efficient gene editing in various cell types, including post-mitotic neurons. However, neuronal ensembles in the same brain region can still be functionally or anatomically different, and such heterogeneity requires gene editing in specific neuronal populations. We recently developed a CRISPR-SaCas9 system-based technique.

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    A Retro-orbital Sinus Injection Mouse Model to Study Early Events and Reorganization of the Astrocytic Network during Pneumococcal Meningitis
    Authors:  Chakir Bello, Martine Cohen-Salmon and Guy Tran Van Nhieu, date: 12/05/2021, view: 783, Q&A: 0
    [Abstract]

    Pneumococcal (PN) meningitis is a life-threatening disease with high mortality rates that leads to permanent neurological sequelae. Studies of the process of bacterial crossing of the blood brain barrier (BBB) are hampered by the lack of relevant in vitro and in vivo models of meningitis that recapitulate the human disease. PN meningitis involves

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    Cortical Laminar Recording of Multi-unit Response to Distal Forelimb Electrical Stimulation in Rats
    Authors:  Charles-Francois V. Latchoumane, Rameen Forghani and Lohitash Karumbaiah, date: 11/20/2021, view: 1499, Q&A: 0
    [Abstract]

    Severe traumatic brain injury (sTBI) survivors experience permanent functional disabilities due to significant volume loss and the brain’s poor capacity to regenerate. Chondroitin sulfate glycosaminoglycans (CS-GAGs) are key regulators of growth factor signaling and neural stem cell homeostasis in the brain. In this protocol, we describe how

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    Knockoff: Druggable Cleavage of Membrane Proteins
    Authors:  John H. Rinald, Jason D. Vevea and Edwin R. Chapman, date: 11/20/2021, view: 1165, Q&A: 0
    [Abstract]

    Comparative cell biology relies on methods that disrupt protein function. Traditional approaches target the gene that encodes the protein of interest via conventional knockout (KO) methodology, conditional Cre-lox system, or recently, flexible protocols based on CRISPR/Cas9. However, these technologies lack precise temporal control (hours),

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    Macroscopic Structural and Connectome Mapping of the Mouse Brain Using Diffusion Magnetic Resonance Imaging
    Authors:  Tanzil Mahmud Arefin, Choong Heon Lee, Jordon D. White, Jiangyang Zhang and Arie Kaffman, date: 11/20/2021, view: 1715, Q&A: 0
    [Abstract]

    Translational work in rodents elucidates basic mechanisms that drive complex behaviors relevant to psychiatric and neurological conditions. Nonetheless, numerous promising studies in rodents later fail in clinical trials, highlighting the need for improving the translational utility of preclinical studies in rodents. Imaging of small rodents

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