Preparation of Acute Brain Slices Using an Optimized &lt;em&gt;N&lt;/em&gt;-Methyl-D-glucamine Protective Recovery Method

Ting, Jonathan T.; Lee, Brian; Chong, Peter Han Joo; Soler-Llavina, Gilberto; Cobbs, Charles; Koch, Christof; Zeng, Hongkui; Lein, Ed

doi:10.3791/53825

Cited by 238 publications

(216 citation statements)

References 56 publications

Supporting

Mentioning

201

Contrasting

Order By: Relevance

“…Preparation of acute brain slices. Preparation followed a published protocol 62 and was performed as follows. Animals were anesthetized using intraperitoneal administration of Nembutal (50 mg/kg).…”

Section: Methodsmentioning

confidence: 99%

Identification of region-specific astrocyte subtypes at single cell resolution

et al. 2020

View full text Add to dashboard Cite

Astrocytes, a major cell type found throughout the central nervous system, have general roles in the modulation of synapse formation and synaptic transmission, blood-brain barrier formation, and regulation of blood flow, as well as metabolic support of other brain resident cells. Crucially, emerging evidence shows specific adaptations and astrocyte-encoded functions in regions, such as the spinal cord and cerebellum. To investigate the true extent of astrocyte molecular diversity across forebrain regions, we used single-cell RNA sequencing. Our analysis identifies five transcriptomically distinct astrocyte subtypes in adult mouse cortex and hippocampus. Validation of our data in situ reveals distinct spatial positioning of defined subtypes, reflecting the distribution of morphologically and physiologically distinct astrocyte populations. Our findings are evidence for specialized astrocyte subtypes between and within brain regions. The data are available through an online database (https://holt-sc. glialab.org/), providing a resource on which to base explorations of local astrocyte diversity and function in the brain.

show abstract

Section: Methodsmentioning

confidence: 99%

Identification of region-specific astrocyte subtypes at single cell resolution

et al. 2020

View full text Add to dashboard Cite

show abstract

“…LLH reaches full amplitude and duration 3-5 weeks postnatal. The use of primarily younger animals for acute slice preparations (Ting et al, 2018) may be one of the main reasons why LLH has been overlooked in electrophysiological studies. However, slow inhibition has been extensively studied in other hippocampal cells and GCs (Schwartzkroin, 1975;Alger and Nicoll, 1982;Rausche et al, 1989;Miles, 1990;Buhl et al, 1996;De Koninck and Mody, 1997;Thomson and Destexhe, 1999;Pouille and Scanziani, 2004;Lovett-Barron et al, 2012).…”

Section: Delayed Postnatal Maturation Of Inhibitory Circuitsmentioning

confidence: 99%

Interplay of Entorhinal Input and Local Inhibitory Network in the Hippocampus at the Origin of Slow Inhibition in Granule Cells

2019

View full text Add to dashboard Cite

Neuronal activity from the entorhinal cortex propagates through the perforant path (PP) to the molecular layer of the dentate gyrus (DG) where information is filtered and converted into sparse hippocampal code. Nearly simultaneous signaling to both granule cells (GC) and local interneurons (INs) engages network interactions that will modulate input integration and output generation. When triggered, GABA release from interneurons counteracts the glutamatergic signals of PP terminals, scaling down the overall DG activation. Inhibition occurs at fast or slow timescales depending on the activation of ionotropic GABA A -R or metabotropic GABA B -R. Although postsynaptic GABA A and GABA B -R differ in their location at the synapse, mixed GABA A/B -R IPSPs can also occur. Here we describe a slow inhibition mechanism in mouse GCs recorded from either sex, mediated by GABA A/B -R in combination with metabotropic glutamate receptors. Short burst PP stimulation in the gamma frequency range lead to a long-lasting hyperpolarization (LLH) of the GCs with a duration that exceeds GABA B -R IPSPs. As a result, LLH alters GC firing patterns and the responses to concomitant excitatory signals are also affected. Synaptic recruitment of feedforward inhibition and subsequent GABA release from interneurons, also successfully trigger mixed GABA responses in GCs. Together these results suggest that slow inhibition through LLH leads to reduced excitability of GCs during entorhinal input integration. The implication of LLH in regulation of neuronal excitability suggests it also contributes to the sparse population coding in DG. Significance StatementOur study describes a long-lasting hyperpolarization (LLH) in hippocampal granule cells. We used whole-cell patch-clamp recordings and an optogenetic approach to characterize this event. LLH is a slow inhibitory mechanism that occurs following the stimulation of the perforant pathway in the molecular layer of the dentate gyrus. We found that it is mediated via postsynaptic ionotropic and metabotropic GABA and metabotropic glutamate receptors. The duration of LLH exceeds previously described IPSPs mediated by any of these receptors. The activation of LLH requires presynaptic gamma frequency bursts and recruitment of the local feedforward inhibition. LLH defines prolonged periods of low excitability of GCs and a restrained neuronal discharge. Our results suggest that LLH can contribute to sparse activation of GCs.

show abstract

“…Slices from older animals (>1 month of age) may benefit from lower temperatures (ß32°C; see, e.g., Ting et al, 2018).…”

Section: Protocol Stepsmentioning

confidence: 99%

Neocortical Microdissection at Columnar and Laminar Resolution for Molecular Interrogation

Kole

Celikel

2018

CP Neuroscience

View full text Add to dashboard Cite

The heterogeneous organization of the mammalian neocortex poses a challenge for elucidating the molecular mechanisms underlying its physiological processes. Although high‐throughput molecular methods are increasingly deployed in neuroscience, their anatomical specificity is often lacking. In this unit, we introduce a targeted microdissection technique that enables extraction of high‐quality RNA and proteins at high anatomical resolution from acutely prepared brain slices. We exemplify its utility by isolating single cortical columns and laminae from the mouse primary somatosensory (barrel) cortex. Tissues can be isolated from living slices in minutes, and the extracted RNA and protein are of sufficient quantity and quality to be used for RNA sequencing and mass spectrometry. This technique will help to increase the anatomical specificity of molecular studies of the neocortex, and the brain in general, as it is applicable to any brain structure that can be identified using optical landmarks in living slices. © 2018 by John Wiley & Sons, Inc.

show abstract

Preparation of Acute Brain Slices Using an Optimized <em>N</em>-Methyl-D-glucamine Protective Recovery Method

Cited by 238 publications

References 56 publications

Identification of region-specific astrocyte subtypes at single cell resolution

Identification of region-specific astrocyte subtypes at single cell resolution

Interplay of Entorhinal Input and Local Inhibitory Network in the Hippocampus at the Origin of Slow Inhibition in Granule Cells

Neocortical Microdissection at Columnar and Laminar Resolution for Molecular Interrogation

Contact Info

Product

Resources

About