scholarly journals Two-Photon in vivo Imaging of Dendritic Spines in the Mouse Cortex Using a Thinned-skull Preparation

10.3791/51520 ◽  
2014 ◽  
Author(s):  
Xinzhu Yu ◽  
Yi Zuo
Keyword(s):  
Dendritic Spines ◽  
Two Photon ◽  
Mouse Cortex

scholarly journals Skull optical clearing window for in vivo imaging of the mouse cortex at synaptic resolution

2017 ◽  
Vol 7 (2) ◽  
pp. 17153-17153 ◽  
Author(s):  
Yan-Jie Zhao ◽  
Ting-Ting Yu ◽  
Chao Zhang ◽  
Zhao Li ◽  
Qing-Ming Luo ◽  
...  
Keyword(s):  
Dendritic Spines ◽  
In Vivo Imaging ◽  
Great Promise ◽  
Critical Periods ◽  
Optical Clearing ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Neuroscience Research ◽  
Mouse Cortex

Abstract Imaging cells and microvasculature in the living brain is crucial to understanding an array of neurobiological phenomena. Here, we introduce a skull optical clearing window for imaging cortical structures at synaptic resolution. Combined with two-photon microscopy, this technique allowed us to repeatedly image neurons, microglia and microvasculature of mice. We applied it to study the plasticity of dendritic spines in critical periods and to visualize dendrites and microglia after laser ablation. Given its easy handling and safety, this method holds great promise for application in neuroscience research.

scholarly journals Microglia motility depends on neuronal activity and promotes structural plasticity in the hippocampus

2019 ◽  
Author(s):  
Felix C. Nebeling ◽  
Stefanie Poll ◽  
Lena C. Schmid ◽  
Manuel Mittag ◽  
Julia Steffen ◽  
...  
Keyword(s):  
Neuronal Activity ◽  
Dendritic Spines ◽  
Synapse Formation ◽  
Brain Parenchyma ◽  
Two Photon ◽  
Contact Rates ◽  
Health And Disease ◽  
The Impact ◽  
The Brain

AbstractMicroglia, the resident immune cells of the brain, play a complex role in health and disease. They actively survey the brain parenchyma by physically interacting with other cells and structurally shaping the brain. Yet, the mechanisms underlying microglia motility and their significance for synapse stability, especially during adulthood, remain widely unresolved. Here we investigated the impact of neuronal activity on microglia motility and its implication for synapse formation and survival. We used repetitive two-photon in vivo imaging in the hippocampus of awake mice to simultaneously study microglia motility and their interaction with synapses. We found that microglia process motility depended on neuronal activity. Simultaneously, more dendritic spines emerged in awake compared to anesthetized mice. Interestingly, microglia contact rates with individual dendritic spines were associated with their stability. These results suggest that microglia are not only sensing neuronal activity, but participate in synaptic rewiring of the hippocampus during adulthood, which has profound relevance for learning and memory processes.

scholarly journals Bidirectional in vivo structural dendritic spine plasticity revealed by two-photon glutamate uncaging in the mouse neocortex

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Jun Noguchi ◽  
Akira Nagaoka ◽  
Tatsuya Hayama ◽  
Hasan Ucar ◽  
Sho Yagishita ◽  
...  
Keyword(s):  
Dendritic Spines ◽  
Time Course ◽  
Hippocampal Slices ◽  
Low Frequency ◽  
Structural Plasticity ◽  
Experimental Conditions ◽  
Two Photon ◽  
Spine Plasticity

Abstract Most excitatory synapses in the brain form on dendritic spines. Two-photon uncaging of glutamate is widely utilized to characterize the structural plasticity of dendritic spines in brain slice preparations in vitro. In the present study, glutamate uncaging was used to investigate spine plasticity, for the first time, in vivo. A caged glutamate compound was applied to the surface of the mouse visual cortex in vivo, revealing the successful induction of spine enlargement by repetitive two-photon uncaging in a magnesium free solution. Notably, this induction occurred in a smaller fraction of spines in the neocortex in vivo (22%) than in hippocampal slices (95%). Once induced, the time course and mean long-term enlargement amplitudes were similar to those found in hippocampal slices. However, low-frequency (1–2 Hz) glutamate uncaging in the presence of magnesium caused spine shrinkage in a similar fraction (35%) of spines as in hippocampal slices, though spread to neighboring spines occurred less frequently than it did in hippocampal slices. Thus, the structural plasticity may occur similarly in the neocortex in vivo as in hippocampal slices, although it happened less frequently in our experimental conditions.

scholarly journals In Vivo Two-Photon Imaging of Dendritic Spines in Marmoset Neocortex

eNeuro ◽  
2015 ◽  
Vol 2 (4) ◽  
pp. ENEURO.0019-15.2015 ◽  
Author(s):  
Osamu Sadakane ◽  
Akiya Watakabe ◽  
Masanari Ohtsuka ◽  
Masafumi Takaji ◽  
Tetsuya Sasaki ◽  
...  
Keyword(s):  
Dendritic Spines ◽  
Two Photon ◽  
Photon Imaging ◽  
Two Photon Imaging

In vivo two-photon uncaging of glutamate revealing the structure–function relationships of dendritic spines in the neocortex of adult mice

2011 ◽  
Vol 71 ◽  
pp. e206
Author(s):  
Jun Noguchi ◽  
Akira Nagaoka ◽  
Satoshi Watanabe ◽  
Graham C.R. Ellis-Davies ◽  
Kazuo Kitamura ◽  
...  
Keyword(s):  
Structure Function ◽  
Dendritic Spines ◽  
Two Photon ◽  
Adult Mice

scholarly journals Transient Effects of Anesthetics on Dendritic Spines and Filopodia in the Living Mouse Cortex

2011 ◽  
Vol 115 (4) ◽  
pp. 718-726 ◽  
Author(s):  
Guang Yang ◽  
Paul C. Chang ◽  
Alex Bekker ◽  
Thomas J.J. Blanck ◽  
Wen-Biao Gan
Keyword(s):  
Dendritic Spines ◽  
Pyramidal Neurons ◽  
Excitatory Synapses ◽  
Transient Effects ◽  
Yellow Fluorescence Protein ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Mouse Cortex ◽  
Living Mouse ◽  
Fluorescence Protein

Background Anesthetics are widely used to induce unconsciousness, pain relief, and immobility during surgery. It remains unclear whether the use of anesthetics has significant and long-lasting effects on synapse development and plasticity in the brain. To address this question, the authors examined the formation and elimination of dendritic spines, postsynaptic sites of excitatory synapses, in the developing mouse cortex during and after anesthetics exposure. Methods Transgenic mice expressing yellow fluorescence protein in layer 5 pyramidal neurons were used in this study. Mice at 1 month of age underwent ketamine-xylazine and isoflurane anesthesia over a period of hours. The elimination and formation rates of dendritic spines and filopodia, the precursors of spines, were followed over hours to days in the primary somatosensory cortex using transcranial two-photon microscopy. Four to five animals were examined under each experimental condition. Student t test and Mann-Whitney U test were used to analyze the data. Results Administration of either ketamine-xylazine or isoflurane rapidly altered dendritic filopodial dynamics but had no significant effects on spine dynamics. Ketamine-xylazine increased filopodial formation whereas isoflurane decreased filopodial elimination during 4 h of anesthesia. Both effects were transient and disappeared within a day after the animals woke up. Conclusion Studies suggest that exposure to anesthetics transiently affects the dynamics of dendritic filopodia but has no significant effect on dendritic spine development and plasticity in the cortex of 1-month-old mice.

scholarly journals In vivo Two-Photon Imaging of Anesthesia-Specific Alterations in Microglial Surveillance and Photodamage-Directed Motility in Mouse Cortex

2019 ◽  
Vol 13 ◽  
Author(s):  
Weilun Sun ◽  
Kunimichi Suzuki ◽  
Dmytro Toptunov ◽  
Stoyan Stoyanov ◽  
Michisuke Yuzaki ◽  
...  
Keyword(s):  
Two Photon ◽  
Mouse Cortex ◽  
Directed Motility ◽  
Photon Imaging ◽  
Two Photon Imaging
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