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Current Protocols in Neuroscience

Takako Kikkawa, Masanori Takahashi, Noriko Osumi
This unit describes basic methods for mammalian whole embryo culture (WEC) using embryonic day 10.5 mouse embryos, including the preparation of high-quality immediately centrifuged (IC) rat serum that is commonly used for WEC and is essential for normal growth and development of cultured mouse and rat embryos in vitro. An alternative protocol for different stages of rodent embryos is also introduced. Since embryos for WEC are dissected out of the uterus and manipulated under the microscope, one can overcome many of the difficulties of gene delivery encountered using in utero electroporation...
January 3, 2017: Current Protocols in Neuroscience
Cara L Wellman
This unit outlines a protocol for Golgi staining, which has been used extensively to reliably and quantitatively assess alterations in dendritic arborization and spine density as a result of a variety of factors, including chronic administration of glucocorticoids, chronic stress, and pharmacological manipulations. The method stains neurons in their entirety, allowing for sophisticated analyses of branch lengths and numbers as well as patterns of dendritic branching. Advantages of the technique include its usefulness in multisite collaborations and its utility in visualizing neurons in multiple regions within the same brain...
January 3, 2017: Current Protocols in Neuroscience
Marie-Theres Weil, Torben Ruhwedel, Wiebke Möbius, Mikael Simons
Intracerebral injections are an invasive method to bypass the blood brain barrier and are widely used to study molecular and cellular mechanisms of the central nervous system. The administered substances are injected directly at the site of interest, executing their effect locally. By combining injections in the rat brain with state-of-the-art electron microscopy, subtle changes in ultrastructure of the nervous tissue can be detected prior to overt damage or disease. The protocol presented here involves stereotactic injection into the corpus callosum of Lewis rats and the cryopreparation of freshly dissected tissue for electron microscopy...
January 3, 2017: Current Protocols in Neuroscience
Justyna Papciak, Rafal Rygula
An active-choice, operant, ambiguous-cue interpretation (ACI) paradigm is described that can be used for measuring cognitive judgement bias in rats. In this behavioral test, animals in an operant conditioning chamber are trained to press a lever to receive a food reward when a specific tone is presented, and to press another lever in response to a different tone to avoid punishment by an electric foot-shock. The tones, which serve as discriminative stimuli, acquire a positive or negative valence, and the training continues until the rats demonstrate a stable, correct discrimination between these two stimuli...
January 3, 2017: Current Protocols in Neuroscience
Zackary A Cope, Jared W Young
Deficits in the domains of attention and response inhibition are central to many psychiatric disorders. As such, animal models of disorders purporting to replicate these behavioral deficits first require tests that can accurately assess the behaviors with high fidelity. The gold-standard clinical test of attention and response inhibition is the continuous performance test (CPT). Although there are a number of CPTs, all share the premise of responding to target stimuli and inhibiting from responding to non-target stimuli...
January 3, 2017: Current Protocols in Neuroscience
Emilie Pacary, François Guillemot
Brain electroporation is a rapid and powerful approach to study neuronal development. In particular, this technique has become a method of choice for studying the process of radial migration of projection neurons in the embryonic cerebral cortex. This method has considerably helped to describe in detail the different steps of radial migration and to characterize the molecular mechanisms controlling this process. Delineating the complexities of neuronal migration is critical to our understanding not only of normal cerebral cortex formation but also of neurodevelopmental disorders resulting from neuronal migration defects...
October 3, 2016: Current Protocols in Neuroscience
Candice A Gellner, Daisy D Reynaga, Frances M Leslie
Animal models are used to study many human diseases, one of which is tobacco addiction. Most preclinical models use nicotine alone, although there are >7000 constituents present in tobacco smoke. The clinical literature suggests that cigarettes have a strong addictive potential, which is not paralleled in preclinical studies using nicotine alone. In order to address the gap between clinical and preclinical literature on tobacco dependence, cigarette smoke extracts containing tobacco constituents have been developed...
October 3, 2016: Current Protocols in Neuroscience
Anne Marie Brady
Animal models are crucial to the study of the neurobiological bases of psychiatric disorders, but schizophrenia is a particularly challenging disorder to model given the complexity and heavily verbal nature of its symptoms. This unit describes a developmental surgical rodent model of schizophrenia, the neonatal ventral hippocampal lesion (NVHL) model. This widely used model produces reliable behavioral abnormalities that are comparable to those observed in patients, as well as anatomical and neurophysiological disruptions in forebrain areas that are also implicated in schizophrenia...
October 3, 2016: Current Protocols in Neuroscience
Dara L Dickstein, Daniel R Dickstein, William G M Janssen, Patrick R Hof, Jacob R Glaser, Alfredo Rodriguez, Nate O'Connor, Paul Angstman, Susan J Tappan
Determining the density and morphology of dendritic spines is of high biological significance given the role of spines in synaptic plasticity and in neurodegenerative and neuropsychiatric disorders. Precise quantification of spines in three dimensions (3D) is essential for understanding the structural determinants of normal and pathological neuronal function. However, this quantification has been restricted to time- and labor-intensive methods such as electron microscopy and manual counting, which have limited throughput and are impractical for studies of large samples...
October 3, 2016: Current Protocols in Neuroscience
T Gregg Davis
The ability to assess the potential for gastrointestinal adverse events in a preclinical setting is a challenge in the development of new drugs, as the vast majority of in vivo research is conducted in rodent species lacking a vomiting reflex. The use of higher species capable of emesis is often limited by cost, technical experience, and relevant efficacy models to define a therapeutic index. Additionally, investigators should be mindful of ethical considerations when using more sentient species when an alternative in lower species is available...
October 3, 2016: Current Protocols in Neuroscience
Laurent Mouchiroud, Vincenzo Sorrentino, Evan G Williams, Matteo Cornaglia, Michael V Frochaux, Tao Lin, Amandine A Nicolet-Dit-Félix, Gopal Krishnamani, Tarik Ouhmad, Martin A M Gijs, Bart Deplancke, Johan Auwerx
Phenotyping strategies in simple model organisms such as D. melanogaster and C. elegans are often broadly limited to growth, aging, and fitness. Recently, a number of physical setups and video tracking software suites have been developed to allow for accurate, quantitative, and high-throughput analysis of movement in flies and worms. However, many of these systems require precise experimental setups and/or fixed recording formats. We report here an update to the Parallel Worm Tracker software, which we termed the Movement Tracker...
October 3, 2016: Current Protocols in Neuroscience
Angela M Yarnell, Erin S Barry, Andrea Mountney, Deborah Shear, Frank Tortella, Neil E Grunberg
Motor and sensory deficits are common following traumatic brain injury (TBI). Although rodent models provide valuable insight into the biological and functional outcomes of TBI, the success of translational research is critically dependent upon proper selection of sensitive, reliable, and reproducible assessments. Published literature includes various observational scales designed to evaluate post-injury functionality; however, the heterogeneity in TBI location, severity, and symptomology can complicate behavioral assessments...
April 8, 2016: Current Protocols in Neuroscience
Markus Wöhr, Dominik Seffer, Rainer K W Schwarting
Rats are able to produce ultrasonic vocalizations (USVs). Such USVs are an important component of the rat social behavior repertoire and serve distinct communicative functions as socio-affective signals. Depending on the emotional valence of the situation, juvenile and adult rats utter (1) aversive 22-kHz USVs conveying an appeasing and/or alarming function; or (2) appetitive 50-kHz USVs, which act as social contact calls, amongst others. A 50-kHz USV radial maze playback paradigm that allows assessment of the behavioral responses displayed by the recipients in a highly standardized manner has been developed...
April 8, 2016: Current Protocols in Neuroscience
Rita Báldi, Dipanwita Ghosh, Brad A Grueter, Sachin Patel
Endocannabinoids (eCBs) are a class of bioactive lipids that mediate retrograde synaptic modulation at central and peripheral synapses. The highly lipophilic nature of eCBs and the pharmacological tools available to interrogate this system require unique methodological consideration, especially when applied to ex vivo systems such as electrophysiological analysis in acute brain slices. This unit provides protocols for measuring cannabinoid and eCB-mediated synaptic signaling in mouse brain slices, including analysis of short-term, long-term, and tonic eCB signaling modes, and the unique considerations for working with eCBs and TRPV1/cannabinoid ligands in acute brain slices...
April 8, 2016: Current Protocols in Neuroscience
W Scott Young, June Song, Éva Mezey
Expression of genes is manifested by the production of RNA transcripts within cells. Hybridization histochemistry (or in situ hybridization) permits localization of these transcripts with cellular resolution or better. Furthermore, the relative amounts of transcripts detected in different tissues or in the same tissues in different states (e.g., physiological or developmental) may be quantified. This unit describes hybridization histochemical techniques using either oligodeoxynucleotide probes (see Basic Protocols 1 and 2, Alternate Protocol 1) or RNA probes (riboprobes; see Basic Protocols 3 and 5)...
April 8, 2016: Current Protocols in Neuroscience
Christopher J Heath, Benjamin U Phillips, Timothy J Bussey, Lisa M Saksida
This unit is designed to facilitate implementation of the fixed and progressive ratio paradigms and the effort-related choice task in the rodent touchscreen apparatus to permit direct measurement of motivation and reward-related decision making in this equipment. These protocols have been optimized for use in the mouse and reliably yield stable performance levels that can be enhanced or suppressed by systemic pharmacological manipulation. Instructions are also provided for the adjustment of task parameters to permit use in mouse models of neurodegenerative disease...
January 4, 2016: Current Protocols in Neuroscience
Kevin T Beier, Nathan A Mundell, Y Albert Pan, Constance L Cepko
Viruses have been used as transsynaptic tracers, allowing one to map the inputs and outputs of neuronal populations, due to their ability to replicate in neurons and transmit in vivo only across synaptically connected cells. To date, their use has been largely restricted to mammals. In order to explore the use of such viruses in an expanded host range, we tested the transsynaptic tracing ability of recombinant vesicular stomatitis virus (rVSV) vectors in a variety of organisms. Successful infection and gene expression were achieved in a wide range of organisms, including vertebrate and invertebrate model organisms...
January 4, 2016: Current Protocols in Neuroscience
Eisuke Koya, Gabriella Margetts-Smith, Bruce T Hope
Learned associations about salient experiences (e.g., drug exposure, stress) and their associated environmental stimuli are mediated by a minority of sparsely distributed, behaviorally activated neurons coined 'neuronal ensembles.' For many years, it was not known whether these neuronal ensembles played causal roles in mediating learned behaviors. However, in the last several years the 'Daun02 inactivation technique' in Fos-lacZ transgenic rats has proved very useful in establishing causal links between neuronal ensembles that express the activity-regulated protein Fos and learned behaviors...
2016: Current Protocols in Neuroscience
Hassiba Beldjoud, Fany Messanvi, Nael Nadif Kasri, Benno Roozendaal
Histone posttranslational modifications (PTMs), by their action on the chromatin state, play a central role in the regulation of gene expression. The discovery that some PTMs in the brain are dynamically regulated by experience and environmental factors makes them an important subject for the study of plasticity changes in learning and memory, addiction, and psychiatric disorders. Current histone isolation protocols, however, require large amounts of tissue, which limits their application for analyzing small tissue samples from a specific brain region...
2016: Current Protocols in Neuroscience
Gloria E Hoffman, Kelley J Murphy, Luciane V Sita
When using immunocytochemistry, investigators may not know how to optimize staining or how to troubleshoot the method when staining fails. Lacking are guides for comparing techniques and applying information derived from one staining method to another. Newer methods amplify signal detection, but will not necessarily work at the same primary antibody concentrations used for less sensitive reactions. Recommendations of optimal titers are often not accurate and are not usually accompanied by information on the method used to test those antibodies or the specifics of the assay...
2016: Current Protocols in Neuroscience
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