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Evaluation of Muscle Performance in Mice by Treadmill Exhaustion Test and Whole-limb Grip Strength Assay

Featured protocol,  Authors: Beatriz Castro
Beatriz CastroAffiliation: Department of Animal Sciences, Purdue University, West Lafayette, USA
For correspondence: beacb@purdue.edu
Bio-protocol author page: a4386
 and Shihuan Kuang
Shihuan KuangAffiliation 1: Department of Animal Sciences, Purdue University, West Lafayette, USA
Affiliation 2: Center for Cancer Research, Purdue University, West Lafayette, USA
For correspondence: skuang@purdue.edu
Bio-protocol author page: a2572
date: 4/20/2017, 93 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2237.

Brief version appeared in Elife, Sep 2016
In vivo muscle function testing has become of great interest as primary phenotypic analysis of muscle performance. This protocol provides detailed procedures to perform the treadmill exhaustion test and the whole-limb grip strength assay, two methods commonly used in the neuromuscular research field.

The Object Context-place-location Paradigm for Testing Spatial Memory in Mice

Featured protocol,  Authors: Edith Lesburguères
Edith LesburguèresAffiliation: Center for Neural Science, New York University, New York, USA
Bio-protocol author page: a4246
Panayiotis Tsokas
Panayiotis TsokasAffiliation 1: Department of Physiology and Pharmacology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Affiliation 2: Department of Anesthesiology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Bio-protocol author page: a4243
Todd Charlton Sacktor
Todd Charlton SacktorAffiliation 1: Department of Physiology and Pharmacology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Affiliation 2: Department of Neurology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Bio-protocol author page: a4244
 and André Antonio Fenton
André Antonio FentonAffiliation 1: Center for Neural Science, New York University, New York, USA
Affiliation 2: Department of Physiology and Pharmacology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
For correspondence: afenton@nyu.edu
Bio-protocol author page: a4245
date: 4/20/2017, 92 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2231.

Brief version appeared in Elife, May 2016
This protocol was originally designed to examine long-term spatial memory in PKMζ knockout (i.e., PKMζ-null) mice (Tsokas et al., 2016). Our main goal was to test whether the ability of these animals to maintain previously acquired spatial information was sensitive to the type and complexity of the spatial information that needs to be remembered. Accordingly, we modified and combined into a single protocol, three novelty-preference tests, specifically the object-in-context, object-in-place and object-in-location tests, adapted from previous studies in rodents (Mumby et al., 2002; Langston and Wood, 2010; Barker and Warburton, 2011). During the training (learning) phase of the procedure, mice are repeatedly exposed to three different environments in which they learn the spatial arrangement of an environment-specific set of non-identical objects. After this learning phase is completed, each mouse receives three different memory tests configured as environment mismatches, in which the previously learned objects-in-space configurations have been modified from the original training situation. The mismatch tests differ in their cognitive demands due to the type of spatial association that is manipulated, specifically evaluating memory for object-context and object-place associations. During each memory test, the time differential spent exploring the novel (misplaced) and familiar objects is computed as an index of novelty discrimination. This index is the behavioral measure of memory recall of the previously acquired spatial associations.

Preparation of Primary Astrocyte Culture Derived from Human Glioblastoma Multiforme Specimen

Featured protocol,  Authors: Mansoureh Hashemi
Mansoureh HashemiAffiliation: Functional Neurosurgery Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
For correspondence: mansoureh.hashemi@sbmu.ac.ir
Bio-protocol author page: a4391
 and Mahmoudreza Hadjighassem
Mahmoudreza HadjighassemAffiliation 1: Department of Neuroscience, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
Affiliation 2: Brain and Spinal Cord Injury Research Center, Neuroscience Institute, Tehran University of Medical Sciences, Tehran, Iran
Bio-protocol author page: a4392
date: 4/20/2017, 72 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2241.

Brief version appeared in Mol Neurobiol, Oct 2016
Gliobalstoma multiforme (GBM) is a grade 4 astrocytoma tumor in central nervous system. Astrocytes can be isolated from human GBM. Study of astrocytes can provide insights about the formation, progression and recurrence of glioblastoma. For using isolated astrocytes, new studies can be designed in the fields of pharmacology, neuroscience and neurosurgery for glioblastoma treatment. This protocol describes the details for preparing high purity primary astrocytes from human GBM. Tumor tissue is disrupted using mechanical dissociation and chemical digestion in this protocol. 2 weeks after plating the cell suspension in culture, primary astrocytes are available for further subculturing and immunocytochemistry of S100-beta antigen.

Measuring Behavioral Individuality in the Acoustic Startle Behavior in Zebrafish

Featured protocol,  Authors: Carlos Pantoja*
Carlos PantojaAffiliation: Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, USA
Bio-protocol author page: a4294
Adam Hoagland*
Adam HoaglandAffiliation: Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, USA
Bio-protocol author page: a4295
Elizabeth Carroll
Elizabeth CarrollAffiliation: Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, USA
Bio-protocol author page: a4296
David Schoppik
David SchoppikAffiliation: Department of Molecular and Cellular Biology, Harvard University, Cambridge MA, USA
Present address: Departments of Otolaryngology, Neuroscience & Physiology, and the Neuroscience Institute New York University Langone School of Medicine, New York, USA
Bio-protocol author page: a4297
 and Ehud Y. Isacoff
Ehud Y. IsacoffAffiliation 1: Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, USA
Affiliation 2: Helen Wills Neuroscience Institute, University of California Berkeley, Berkeley, USA
Affiliation 3: Physical Bioscience Division, Lawrence Berkeley National Laboratory, Berkeley, USA
For correspondence: ehud@berkeley.edu
Bio-protocol author page: a4298
 (*contributed equally to this work) date: 4/5/2017, 128 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2200.

Brief version appeared in Neuron, Aug 2016
The objective of this protocol is to provide a detailed description for the construction and use of a behavioral apparatus, the zBox, for high-throughput behavioral measurements in larval zebrafish (Danio rerio). The zBox is used to measure behavior in multiple individuals simultaneously. Individual fish are housed in wells of multi-well plates and receive acoustic/vibration stimuli with simultaneous recording of behavior. Automated analysis of behavioral movies is performed with MATLAB scripts. This protocol was adapted from two of our previously published papers (Levitz et al., 2013; Pantoja et al., 2016). The zBox provides an easy to setup flexible platform for behavioral experiments in zebrafish larvae.

Testing Depression in Mice: a Chronic Social Defeat Stress Model

Featured protocol,  Authors: Hee-Dae Kim
Hee-Dae KimAffiliation: Department of Basic Medical Sciences, University of Arizona College of Medicine-Phoenix, Phoenix, USA
Bio-protocol author page: a4302
Tanessa Call
Tanessa CallAffiliation: Department of Basic Medical Sciences, University of Arizona College of Medicine-Phoenix, Phoenix, USA
Bio-protocol author page: a4303
Samantha Carotenuto
Samantha CarotenutoAffiliation: Department of Basic Medical Sciences, University of Arizona College of Medicine-Phoenix, Phoenix, USA
Bio-protocol author page: a4304
Ross Johnson
Ross JohnsonAffiliation: Department of Basic Medical Sciences, University of Arizona College of Medicine-Phoenix, Phoenix, USA
Bio-protocol author page: a4305
 and Deveroux Ferguson
Deveroux FergusonAffiliation: Department of Basic Medical Sciences, University of Arizona College of Medicine-Phoenix, Phoenix, USA
For correspondence: dferguson@email.arizona.edu
Bio-protocol author page: a4306
date: 4/5/2017, 147 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2203.

Brief version appeared in J Neurosci, Aug 2016
A vast challenge within neuropsychiatric research has been the development of animal models that accurately reflect symptoms associated with affective disorders. An ethologically valid model that has been shown to be effective in studying depression is the chronic social defeat stress model. In this model, C57BL/6J mice are subjected to chronic social defeat stress induced by CD-1 aggressor mice for 10 consecutive days. Discussed here is a protocol describing the screening process of the CD-1 aggressor mice, the confrontations between the C57BL/6J and CD-1 aggressor mice, and analysis of social avoidance scores as an indication of depression-like behaviors.

Evaluation of Muscle Performance in Mice by Treadmill Exhaustion Test and Whole-limb Grip Strength Assay

Authors: Beatriz Castro
Beatriz CastroAffiliation: Department of Animal Sciences, Purdue University, West Lafayette, USA
For correspondence: beacb@purdue.edu
Bio-protocol author page: a4386
 and Shihuan Kuang
Shihuan KuangAffiliation 1: Department of Animal Sciences, Purdue University, West Lafayette, USA
Affiliation 2: Center for Cancer Research, Purdue University, West Lafayette, USA
For correspondence: skuang@purdue.edu
Bio-protocol author page: a2572
date: 4/20/2017, 93 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2237.

[Abstract] In vivo muscle function testing has become of great interest as primary phenotypic analysis of muscle performance. This protocol provides detailed procedures to perform the treadmill exhaustion test and the whole-limb grip strength assay, two methods commonly used in the neuromuscular research field....

The Object Context-place-location Paradigm for Testing Spatial Memory in Mice

Authors: Edith Lesburguères
Edith LesburguèresAffiliation: Center for Neural Science, New York University, New York, USA
Bio-protocol author page: a4246
Panayiotis Tsokas
Panayiotis TsokasAffiliation 1: Department of Physiology and Pharmacology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Affiliation 2: Department of Anesthesiology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Bio-protocol author page: a4243
Todd Charlton Sacktor
Todd Charlton SacktorAffiliation 1: Department of Physiology and Pharmacology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Affiliation 2: Department of Neurology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
Bio-protocol author page: a4244
 and André Antonio Fenton
André Antonio FentonAffiliation 1: Center for Neural Science, New York University, New York, USA
Affiliation 2: Department of Physiology and Pharmacology, the Robert F. Furchgott Center for Neural and Behavioral Science, Downstate Medical Center, State University of New York, New York, USA
For correspondence: afenton@nyu.edu
Bio-protocol author page: a4245
date: 4/20/2017, 92 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2231.

[Abstract] This protocol was originally designed to examine long-term spatial memory in PKMζ knockout (i.e., PKMζ-null) mice (Tsokas et al., 2016). Our main goal was to test whether the ability of these animals to maintain previously acquired spatial information was sensitive to the type and complexity of the spatial information that needs to be remembered. Accordingly, ...

Preparation of Primary Astrocyte Culture Derived from Human Glioblastoma Multiforme Specimen

Authors: Mansoureh Hashemi
Mansoureh HashemiAffiliation: Functional Neurosurgery Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
For correspondence: mansoureh.hashemi@sbmu.ac.ir
Bio-protocol author page: a4391
 and Mahmoudreza Hadjighassem
Mahmoudreza HadjighassemAffiliation 1: Department of Neuroscience, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
Affiliation 2: Brain and Spinal Cord Injury Research Center, Neuroscience Institute, Tehran University of Medical Sciences, Tehran, Iran
Bio-protocol author page: a4392
date: 4/20/2017, 72 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2241.

[Abstract] Gliobalstoma multiforme (GBM) is a grade 4 astrocytoma tumor in central nervous system. Astrocytes can be isolated from human GBM. Study of astrocytes can provide insights about the formation, progression and recurrence of glioblastoma. For using isolated astrocytes, new studies can be designed in the fields of pharmacology, neuroscience and neurosurgery ...

Measuring Behavioral Individuality in the Acoustic Startle Behavior in Zebrafish

Authors: Carlos Pantoja*
Carlos PantojaAffiliation: Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, USA
Bio-protocol author page: a4294
Adam Hoagland*
Adam HoaglandAffiliation: Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, USA
Bio-protocol author page: a4295
Elizabeth Carroll
Elizabeth CarrollAffiliation: Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, USA
Bio-protocol author page: a4296
David Schoppik
David SchoppikAffiliation: Department of Molecular and Cellular Biology, Harvard University, Cambridge MA, USA
Present address: Departments of Otolaryngology, Neuroscience & Physiology, and the Neuroscience Institute New York University Langone School of Medicine, New York, USA
Bio-protocol author page: a4297
 and Ehud Y. Isacoff
Ehud Y. IsacoffAffiliation 1: Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, USA
Affiliation 2: Helen Wills Neuroscience Institute, University of California Berkeley, Berkeley, USA
Affiliation 3: Physical Bioscience Division, Lawrence Berkeley National Laboratory, Berkeley, USA
For correspondence: ehud@berkeley.edu
Bio-protocol author page: a4298
 (*contributed equally to this work) date: 4/5/2017, 128 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2200.

[Abstract] The objective of this protocol is to provide a detailed description for the construction and use of a behavioral apparatus, the zBox, for high-throughput behavioral measurements in larval zebrafish (Danio rerio). The zBox is used to measure behavior in multiple individuals simultaneously. Individual fish are housed in wells of multi-well plates and ...

Testing Depression in Mice: a Chronic Social Defeat Stress Model

Authors: Hee-Dae Kim
Hee-Dae KimAffiliation: Department of Basic Medical Sciences, University of Arizona College of Medicine-Phoenix, Phoenix, USA
Bio-protocol author page: a4302
Tanessa Call
Tanessa CallAffiliation: Department of Basic Medical Sciences, University of Arizona College of Medicine-Phoenix, Phoenix, USA
Bio-protocol author page: a4303
Samantha Carotenuto
Samantha CarotenutoAffiliation: Department of Basic Medical Sciences, University of Arizona College of Medicine-Phoenix, Phoenix, USA
Bio-protocol author page: a4304
Ross Johnson
Ross JohnsonAffiliation: Department of Basic Medical Sciences, University of Arizona College of Medicine-Phoenix, Phoenix, USA
Bio-protocol author page: a4305
 and Deveroux Ferguson
Deveroux FergusonAffiliation: Department of Basic Medical Sciences, University of Arizona College of Medicine-Phoenix, Phoenix, USA
For correspondence: dferguson@email.arizona.edu
Bio-protocol author page: a4306
date: 4/5/2017, 147 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2203.

[Abstract] A vast challenge within neuropsychiatric research has been the development of animal models that accurately reflect symptoms associated with affective disorders. An ethologically valid model that has been shown to be effective in studying depression is the chronic social defeat stress model. In this model, C57BL/6J mice are subjected to chronic social ...

Thinned-skulled Cranial Window Preparation (Mice)

Authors: Lifeng Zhang
Lifeng ZhangAffiliation: Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Baltimore, USA
For correspondence: lfzhang916@hotmail.com
Bio-protocol author page: a4186
Bo Liang
Bo LiangAffiliation: Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Baltimore, USA
Bio-protocol author page: a4187
Yun Li
Yun LiAffiliation: Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Baltimore, USA
Bio-protocol author page: a4188
 and Da-Ting Lin
Da-Ting LinAffiliation 1: Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Baltimore, USA
Affiliation 2: The Jackson Laboratory, Bar Harbor, USA
Affiliation 3: The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, USA
Bio-protocol author page: a4189
date: 3/5/2017, 301 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2158.

[Abstract] Imaging structural plasticity or activity of neurons in the brain circuit will facilitate understanding the neural mechanisms underlying animal behavior. Here we describe a modified procedure, the polished and reinforced thinned-skull cranial window preparation, by which we can image dendrites and spines in mouse layer I cortex for weeks (Zhang et ...

Olfactory Habituation-dishabituation Test (Mouse)

Authors: Hiroo Takahashi
Hiroo TakahashiAffiliation: Laboratory for Molecular Biology of Neural System, Advanced Medical Research Center, Nara Medical University , Kashihara, Nara, Japan
Bio-protocol author page: a4174
 and Akio Tsuboi
Akio TsuboiAffiliation: Laboratory for Molecular Biology of Neural System, Advanced Medical Research Center, Nara Medical University , Kashihara, Nara, Japan
For correspondence: atsuboi@naramed-u.ac.jp
Bio-protocol author page: a4175
date: 3/5/2017, 294 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2154.

[Abstract] Olfaction plays a fundamental role for the various behaviors such as feeding, mating, nursing, and avoidance in mice. Behavioral tests that characterize abilities of odor detection and recognition using genetically modified mice reveal the contribution of target genes to the olfactory processing. Here, we describe the olfactory habituation-dishabituation ...

Olfactory Avoidance Test (Mouse)

Authors: Hiroo Takahashi
Hiroo TakahashiAffiliation: Laboratory for Molecular Biology of Neural System, Advanced Medical Research Center, Nara Medical University , Kashihara, Nara, Japan
Bio-protocol author page: a4174
 and Akio Tsuboi
Akio TsuboiAffiliation: Laboratory for Molecular Biology of Neural System, Advanced Medical Research Center, Nara Medical University , Kashihara, Nara, Japan
For correspondence: atsuboi@naramed-u.ac.jp
Bio-protocol author page: a4175
date: 3/5/2017, 295 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2153.

[Abstract] In mice, olfaction plays a pivotal role for the various behaviors, such as feeding, mating, nursing and avoidance. Behavioral tests that analyze abilities of odor detection and recognition using genetically modified mice reveal the contribution of target genes to the olfactory processing. Here, we describe the olfactory avoidance test to investigate ...

Axonal Conduction Velocity Measurement

Authors: Margaret Louise DeMaegd
Margaret Louise DeMaegdAffiliation: School of Biological Sciences, Illinois State University, Normal, IL, USA
Bio-protocol author page: a4177
Carola Städele
Carola StädeleAffiliation: School of Biological Sciences, Illinois State University, Normal, IL, USA
For correspondence: carola@neurobiologie.de
Bio-protocol author page: a4150
 and Wolfgang Stein
Wolfgang SteinAffiliation: School of Biological Sciences, Illinois State University, Normal, IL, USA
For correspondence: wstein@neurobiologie.de
Bio-protocol author page: a4151
date: 3/5/2017, 333 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2152.

[Abstract] Action potential conduction velocity is the speed at which an action potential (AP) propagates along an axon. Measuring AP conduction velocity is instrumental in determining neuron health, function, and computational capability, as well as in determining short-term dynamics of neuronal communication and AP initiation (Ballo and Bucher, 2009; Bullock, ...

Extracellular Axon Stimulation

Authors: Carola Städele
Carola StädeleAffiliation: School of Biological Sciences, Illinois State University, Normal, IL, USA
For correspondence: carola@neurobiologie.de
Bio-protocol author page: a4150
Margaret Louise DeMaegd
Margaret Louise DeMaegdAffiliation: School of Biological Sciences, Illinois State University, Normal, IL, USA
Bio-protocol author page: a4177
 and Wolfgang Stein
Wolfgang SteinAffiliation: School of Biological Sciences, Illinois State University, Normal, IL, USA
For correspondence: wstein@neurobiologie.de
Bio-protocol author page: a4151
date: 3/5/2017, 339 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2151.

[Abstract] This is a detailed protocol explaining how to perform extracellular axon stimulations as described in Städele and Stein, 2016. The ability to stimulate and record action potentials is essential to electrophysiological examinations of neuronal function. Extracellular stimulation of axons traveling in fiber bundles (nerves) is a classical technique in ...
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Immunofluorescence Staining on Mouse Embryonic Brain Sections

Author: Xuecai Ge
Xuecai GeAffiliation 1: Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology (MIT), Cambridge, USA
Affiliation 2: , Howard Hughes Medical Institute, Cambridge, USA
For correspondence: xuecaige@stanford.edu
Bio-protocol author page: a46
date: 6/5/2012, 11299 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.192.

[Abstract] This protocol comprises the entire process of immunofluorescence staining on mouse embryonic brains, starting from tissue preparation to mounting of the tissue sections....

Stereotaxic Injection of LPS into Mouse Substantia Nigra

Author: Huiming Gao
Huiming GaoAffiliation: National Institute of Environmental Health Sciences, Research Triangle Park, NC, USA
For correspondence: gao2@niehs.nih.gov
Bio-protocol author page: a17
date: 4/20/2012, 11256 views, 4 Q&A
DOI: https://doi.org/10.21769/BioProtoc.153.

[Abstract] Stereotaxic injection is an attractive approach for studying genetic, cellular and circuit functions in the brain. Injection of anatomical tracers, site-targeted lesions and gene delivery by recombinant adeno-associated viruses and lentiviruses in mice are powerful tools to study nervous system development ...

Mouse Cochlear Whole Mount Immunofluorescence

Authors: Omar Akil
Omar AkilAffiliation: Department Of Otolaryngology-HNS, University of California, San Francisco, USA
For correspondence: oakil@ohns.ucsf.edu
Bio-protocol author page: a238
 and Lawrence R. Lustig
Lawrence R. LustigAffiliation: Department Of Otolaryngology-HNS, University of California, San Francisco, USA
Bio-protocol author page: a239
date: 3/5/2013, 10956 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.332.

[Abstract] This protocol comprises the entire process of immunofluorescence staining on mouse cochlea whole mount, starting from tissue preparation to the mounting of the tissue. This technique provides “three-dimensional” views of the stained components in order to determine the localization of a protein of interest ...

c-Fos and Arc Immunohistochemistry on Rat Cerebellum

Author: Soyun Kim
Soyun KimAffiliation: Neuroscience Program, University of Southern California, Los Angeles, USA
For correspondence: soyunkimucsd@gmail.com
Bio-protocol author page: a45
date: 5/20/2012, 10793 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.191.

[Abstract] This protocol aims to introduce methods for sacrificing rats by transcardial perfusion and extracting the brain, and introduce methods for staining the rat brain tissue with c-Fos and Arc antibodies. Please note the expression of the proteins is very sensitive to behavioral paradigm that triggers neural ...

[Bio101] Microglia Cultures and Mixed Glial Culture

Author: Huiming Gao
Huiming GaoAffiliation: Neuropharmacology Section, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC, USA
For correspondence: gao2@niehs.nih.gov
Bio-protocol author page: a17
date: 11/5/2011, 10766 views, 3 Q&A
DOI: https://doi.org/10.21769/BioProtoc.149.

[Abstract] Primary rodent microglia-enriched cultures are the most popular model to study microglial biology in vitro and to explore immune signaling pathways. Mixed glial cultures that contain microglia and astroglia are very useful for investigating the precise mechanisms of microglia-astroglia interaction during ...

Novel Object Recognition for Studying Memory in Mice

Authors: Tzyy-Nan Huang
Tzyy-Nan HuangAffiliation: Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan
For correspondence: eugene02@gate.sinica.edu.tw
Bio-protocol author page: a1680
 and Yi-Ping Hsueh
Yi-Ping HsuehAffiliation: Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan
For correspondence: yph@gate.sinica.edu.tw
Bio-protocol author page: a1681
date: 10/5/2014, 10594 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1249.

[Abstract] Memory tests are important indexes of the brain functions for rodents behavior assay. Many memory tasks require external forces (e.g. electric shocks) or intrinsic forces (e.g. hunger and thirsty) to trigger the responses. Under those conditions, rodents are under stresses, such as pain, tired, malnutrition ...

In utero Electroporation of Mouse Embryo Brains

Author: Xuecai Ge
Xuecai GeAffiliation 1: Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology (MIT), Cambridge, USA
Affiliation 2: , Howard Hughes Medical Institute, Cambridge, USA
For correspondence: xuecaige@stanford.edu
Bio-protocol author page: a46
date: 7/20/2012, 10235 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.231.

[Abstract] This is a non-invasive technique to introduce transgenes into developing brains. In this technique, DNA is injected into the lateral ventricle of the embryonic brains, and is incorporated into the cells through electroporation. Embryos then continue their development in normal conditions in vivo. The ...

A Protocol for Electrophoretic Mobility Shift Assay (EMSA) from Primary Neuron

Author: Jiali Li
Jiali LiAffiliation: Department of Cell Biology and Neuroscience, Nelson Biological Laboratories, Rutgers University, Piscataway, NJ, USA
For correspondence: jli@dls.rutgers.edu
Bio-protocol author page: a179
date: 12/5/2012, 10198 views, 3 Q&A
DOI: https://doi.org/10.21769/BioProtoc.300.

[Abstract] The interaction of transcriptional or co-transcriptional factors with DNA is crucial for changes of neuronal gene expression during normal brain development as well as neurodegeneration. The electrophoretic mobility shift assay (EMSA) is a very powerful technique for studying changes of neuronal gene ...

Optical Clearing Using SeeDB

Authors: Meng-Tsen Ke
Meng-Tsen KeAffiliation: Laboratory for Sensory Circuit Formation, RIKEN Center for Developmental Biology, Kobe, Japan
Bio-protocol author page: a1144
Satoshi Fujimoto
Satoshi FujimotoAffiliation: Laboratory for Sensory Circuit Formation, RIKEN Center for Developmental Biology, Kobe, Japan
Bio-protocol author page: a1145
 and Takeshi Imai
Takeshi ImaiAffiliation: Laboratory for Sensory Circuit Formation, RIKEN Center for Developmental Biology, Kobe, Japan
For correspondence: imai@cdb.riken.jp
Bio-protocol author page: a1146
date: 2/5/2014, 9571 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1042.

[Abstract] We describe a water-based optical clearing agent, SeeDB (See Deep Brain), which clears fixed brain samples in a few days without quenching many types of fluorescent dyes, including fluorescent proteins and lipophilic neuronal tracers. SeeDB is a saturated solution of fructose (80.2% w/w) in water with ...

Hippocampal Neuron Dissociation Transfection and Culture in Microfluidics Chambers

Author: Yang Geng
Yang GengAffiliation: Department of Pediatrics and Bioengineering, Stanford University School of Medicine, Stanford, USA
For correspondence: yanggeng@stanford.edu
Bio-protocol author page: a64
date: 7/20/2012, 7538 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.235.

[Abstract] Microfluidics chamber is an ideal tool to study local events that occurring in neuronal projections by perfectly compartmentalizing the cell soma from certain branches. It is very well suited for live cell imaging or immunohistochemistry staining. This protocol has been carefully modified in detail ...
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