Visualizing septin and cell dynamics in mammalian brain slices

2016 ◽  
pp. 295-309 ◽  
Author(s):  
H. Ito ◽  
R. Morishita ◽  
H. Tabata ◽  
K. Nagata
Keyword(s):  
Brain Slices ◽  
Mammalian Brain ◽  
Cell Dynamics

scholarly journals Imaging Subthreshold Voltage Oscillation With Cellular Resolution in the Inferior Olive in vitro

2020 ◽  
Vol 14 ◽  
Author(s):  
Kevin Dorgans ◽  
Bernd Kuhn ◽  
Marylka Yoe Uusisaari
Keyword(s):  
Inferior Olive ◽  
Brain Slices ◽  
Brain Regions ◽  
Mammalian Brain ◽  
Wide Field ◽  
Voltage Imaging ◽  
Subthreshold Oscillations ◽  
Cellular Resolution

Voltage imaging with cellular resolution in mammalian brain slices is still a challenging task. Here, we describe and validate a method for delivery of the voltage-sensitive dye ANNINE-6plus (A6+) into tissue for voltage imaging that results in higher signal-to-noise ratio (SNR) than conventional bath application methods. The not fully dissolved dye was injected into the inferior olive (IO) 0, 1, or 7 days prior to acute slice preparation using stereotactic surgery. We find that the voltage imaging improves after an extended incubation period in vivo in terms of labeled volume, homogeneous neuropil labeling with saliently labeled somata, and SNR. Preparing acute slices 7 days after the dye injection, the SNR is high enough to allow single-trial recording of IO subthreshold oscillations using wide-field (network-level) as well as high-magnification (single-cell level) voltage imaging with a CMOS camera. This method is easily adaptable to other brain regions where genetically-encoded voltage sensors are prohibitively difficult to use and where an ultrafast, pure electrochromic sensor, like A6+, is required. Due to the long-lasting staining demonstrated here, the method can be combined, for example, with deep-brain imaging using implantable GRIN lenses.

Perpetual inhibitory activity in mammalian brain slices generated by spontaneous GABA release

1991 ◽  
Vol 545 (1-2) ◽  
pp. 142-150 ◽  
Author(s):  
Thomas S. Otis ◽  
Kevin J. Staley ◽  
Istvan Mody
Keyword(s):  
Inhibitory Activity ◽  
Brain Slices ◽  
Gaba Release ◽  
Mammalian Brain

scholarly journals Spatiotemporal transcriptome at single-cell resolution reveals key radial glial cell population in axolotl telencephalon development and regeneration

2021 ◽  
Author(s):  
Xiaoyu Wei ◽  
Sulei Fu ◽  
Hanbo Li ◽  
Yang Liu ◽  
Shuai Wang ◽  
...  
Keyword(s):  
Single Cell ◽  
Glial Cell ◽  
Cell Population ◽  
Cell Types ◽  
Mammalian Brain ◽  
Regeneration Ability ◽  
Radial Glial Cell ◽  
Cell Dynamics ◽  
Brain Regeneration ◽  
Radial Glial

Brain regeneration requires a precise coordination of complex responses in a time- and region-specific manner. Identifying key cell types and molecules that direct brain regeneration would provide potential targets for the advance of regenerative medicine. However, progress in the field has been hampered largely due to very limited regeneration capacity of the mammalian brain and understanding of the regeneration process at both cellular and molecular level. Here, using axolotl brain with astonishing regeneration ability upon injury, and the Stereo-seq (SpaTial Enhanced REsolution Omics-sequencing), we reconstruct the first architecture of axolotl telencephalon with gene expression profiling at single-cell resolution, and fine cell dynamics maps throughout development and regeneration. Intriguingly, we discover a marked heterogeneity of radial glial cell (RGC) types with distinct behaviors. Of note, one subtype of RGCs is activated since early regeneration stages and proliferates while other RGCs remain dormant. Such RGC subtype appears to be the major cell population involved in early wound healing response and gradually covers the injured area before presumably transformed into the lost neurons. Altogether, our work systematically decodes the complex cellular and molecular dynamics of axolotl telencephalon in development and regeneration, laying the foundation for studying the regulatory mechanism of brain regeneration in future.

Dual GABAergic Synaptic Response of Fast Excitation and Slow Inhibition in the Medial Habenula of Rat Epithalamus

2007 ◽  
Vol 98 (3) ◽  
pp. 1323-1332 ◽  
Author(s):  
Uhnoh Kim ◽  
Lee-yup Chung
Keyword(s):  
Brain Slices ◽  
Mammalian Brain ◽  
Dependent Manner ◽  
Tonic Firing ◽  
Medial Habenula ◽  
Acid Type ◽  
Gabaergic Synapses ◽  
Cell Firing ◽  
Fast Excitation ◽  
Dual Activation

We report here a novel action of GABAergic synapses in regulating tonic firing in the mammalian brain. By using gramicidin-perforated patch recording in rat brain slices, we show that cells of the medial habenula of the epithalamus generate tonic firing in basal conditions. The GABAergic input onto these cells at postnatal days 18–25 generates a combinatorial activation of fast excitation and slow inhibition. The fast excitation, mediated by γ-aminobutyric acid type A receptors (GABAARs), is alone capable of triggering robust action potentials to increase cell firing. This excitatory influence of GABAergic input results from the Cl− homeostasis that maintains intracellular Cl− at high levels. The GABAA excitation is often followed by a slow inhibition mediated by GABABRs that suppresses tonic firing. Interestingly, in a subpopulation of the cells, the GABAB inhibition exhibits a remarkably low threshold for synaptic activation in that low-strength GABAergic input often activates selectively the GABAB slow inhibition, whereas the GABAA excitation requires further increases in stimulus strength. Our study demonstrates that the dual activation of GABAergic excitation and inhibition through GABAARs and GABABRs generates distinct temporal patterns of cell firing that alter the cellular output in an activity-dependent manner.

scholarly journals Facilitatory actions of guanidine on synaptic transmission in mammalian brain slices

1980 ◽  
Vol 67 (1) ◽  
pp. 234-246 ◽  
Author(s):  
M. Galvan ◽  
P. Grafe ◽  
G. ten Bruggencate
Keyword(s):  
Synaptic Transmission ◽  
Brain Slices ◽  
Mammalian Brain

scholarly journals In Vivo Mammalian Brain Imaging Using One- and Two-Photon Fluorescence Microendoscopy

2004 ◽  
Vol 92 (5) ◽  
pp. 3121-3133 ◽  
Author(s):  
Juergen C. Jung ◽  
Amit D. Mehta ◽  
Emre Aksay ◽  
Raymond Stepnoski ◽  
Mark J. Schnitzer
Keyword(s):  
Laser Scanning ◽  
Frame Rate ◽  
Mammalian Brain ◽  
Cell Dynamics ◽  
Ca1 Neurons ◽  
Two Photon ◽  
Two Photon Fluorescence ◽  
Rats And Mice ◽  
Photon Fluorescence

One of the major limitations in the current set of techniques available to neuroscientists is a dearth of methods for imaging individual cells deep within the brains of live animals. To overcome this limitation, we developed two forms of minimally invasive fluorescence microendoscopy and tested their abilities to image cells in vivo. Both one- and two-photon fluorescence microendoscopy are based on compound gradient refractive index (GRIN) lenses that are 350–1,000 μm in diameter and provide micron-scale resolution. One-photon microendoscopy allows full-frame images to be viewed by eye or with a camera, and is well suited to fast frame-rate imaging. Two-photon microendoscopy is a laser-scanning modality that provides optical sectioning deep within tissue. Using in vivo microendoscopy we acquired video-rate movies of thalamic and CA1 hippocampal red blood cell dynamics and still-frame images of CA1 neurons and dendrites in anesthetized rats and mice. Microendoscopy will help meet the growing demand for in vivo cellular imaging created by the rapid emergence of new synthetic and genetically encoded fluorophores that can be used to label specific brain areas or cell classes.

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