An inhalation anaesthesia approach for neonatal mice allowing streamlined stereotactic injection in the brain

Ho, Hinze; Fowle, Adam; Coetzee, Marisa; Greger, Ingo H.; Watson, Jake F.

doi:10.1016/j.jneumeth.2020.108824

Cited by 12 publications

(9 citation statements)

References 15 publications

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“…In vivo, the viral injection was performed according to 76 . P1 neonatal mice were anaesthetised using isoflurane, bilaterally injected using a pulled glass needle in the hippocampal area with 0.5 μL AAV-hSyn-Cre-EGFP at 3 × 10 12 GC ml −1 (Addgene #105540, virus produced by U -Penn Vector Core), and returned to the parent cage until P7, when tissue was harvested for organotypic culture.…”

Section: Methodsmentioning

confidence: 99%

AMPA receptor anchoring at CA1 synapses is determined by N-terminal domain and TARP γ8 interactions

Watson

Striessnig

et al. 2021

Nat Commun

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AMPA receptor (AMPAR) abundance and positioning at excitatory synapses regulates the strength of transmission. Changes in AMPAR localisation can enact synaptic plasticity, allowing long-term information storage, and is therefore tightly controlled. Multiple mechanisms regulating AMPAR synaptic anchoring have been described, but with limited coherence or comparison between reports, our understanding of this process is unclear. Here, combining synaptic recordings from mouse hippocampal slices and super-resolution imaging in dissociated cultures, we compare the contributions of three AMPAR interaction domains controlling transmission at hippocampal CA1 synapses. We show that the AMPAR C-termini play only a modulatory role, whereas the extracellular N-terminal domain (NTD) and PDZ interactions of the auxiliary subunit TARP γ8 are both crucial, and each is sufficient to maintain transmission. Our data support a model in which γ8 accumulates AMPARs at the postsynaptic density, where the NTD further tunes their positioning. This interplay between cytosolic (TARP γ8) and synaptic cleft (NTD) interactions provides versatility to regulate synaptic transmission and plasticity.

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Section: Methodsmentioning

confidence: 99%

AMPA receptor anchoring at CA1 synapses is determined by N-terminal domain and TARP γ8 interactions

Watson

Striessnig

et al. 2021

Nat Commun

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“…Adenovirus 9 (AAV9)‐based overexpression vector of TGIF2 (AAV9‐TGIF2) and the empty vector (AAV9‐NC) were procured from Genechem Co., Ltd (Shanghai, China). The AAV9‐based vectors were injected into the newborn mice (P1) in the VPA group intracerebroventricularly according to a previous report (Ho et al., 2020 ). The injection was performed using a small animal stereotactic frame, and the virus was delivered using Hamilton syringes and microinjection adaptors.…”

Section: Methodsmentioning

confidence: 99%

Downregulation of TGIF2 is possibly correlated with neuronal apoptosis and autism‐like symptoms in mice

Lei

Deng

2022

Brain and Behavior

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Background TGFB‐induced factor homeobox 2 (TGIF2) has been reported to exert essential functions in brain development. This study aimed to elucidate the correlation of TGIF2 with autism, a neurodevelopmental condition which presents with severe communication problems. Methods An autism‐related gene expression dataset GSE36315 was used to analyze aberrantly expressed genes in autistic brain tissues. Maternal mice were treated with valproate (VPA), and their offspring were selected as model mice with autism. The functions of TGIF2 in autism‐like symptoms in mice were examined by behavioral tests and histological examination of their hippocampal tissues. Mouse hippocampal neurons were extracted for in vitro studies. A gene set enrichment analysis was performed to analyze the signaling pathways involved, and the upstream factors influencing TGIF2 expression were explored in the ENCODE database and validated by ChIP‐qPCR assays. Results TGIF2 was poorly expressed in autistic patients in the GSE36315 dataset as well as in the temporal cortex tissues of autistic mice. Adenovirus‐mediated overexpression of TGIF2 suppressed autism‐like symptoms and neuronal apoptosis in autistic mice. TGIF2 activated the Wnt/β‐catenin signaling pathway. TGIF2 could be regulated by monomethylation of histone H3 Lys4 (H3K4me1). The histone demethylase LSD1 was highly expressed in the tissues of autistic mice and bound to TGIF2 promoter, which was possibly responsible for TGIF2 downregulation. Conclusion This research suggests that the downregulation of TGIF2, possibly regulated by LSD1/H3K4me1, is correlated with neuronal apoptosis and development of autism in mice through the inactivation of the Wnt/β‐catenin pathway.

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“…Wait for loss of motor response. Once anesthetized, transfer the pup to the heating pad and place the snout into a 3D-printed whole-body mouse mold (we use a mold from a previously published protocol 7 ). Keep the mouse under anesthesia (oxygen 1 L/min, isoflurane 4%).…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“…2022 1 Addgene Cat#191213 Software and algorithms Matlab 2019a or later versions MathWorks https://www.mathworks.com/ RStudio 1.2.1335 RStudio Team http://www.rstudio.com/ Custom-made Matlab script This study https://doi.org/10.5281/zenodo.6980507 Other 1 L Flasks N/A N/A 3D printed whole-body mouse mold Ho et al. 7 N/A Bunsen burner FALC INSTRUMENTS Cat#140205010 Cover slips Menzel-Gläser Cat#BB02400600A113MNZ0 ECM 399 Electroporation System BTX Cat#45-0000 Eppendorf PCR clean tubes Eppendorf Cat#0030 123.328 Falcon® 14 mL Round Bottom Tubes Falcon Cat#352059 Heating pad N/A N/A Insulin syringe (30G × 1/2″) B Braun Cat#9151125S Isoflurane vaporizer with oxygen extractor/bottle Provet N/A Magnetic Stirrer N/A N/A Microgrinder NARISHIGE EG-45 Micropipette Puller Sutter Instrument Flaming/Brown Model P-87 Microscope with ocular micrometer N/A N/A Nail polish N/A N/A Nanodrop Thermo Scientific Nanodrop 1000 Nipper N/A N/A PCR glass micropipettes Drummond ...…”

Section: Key Resources Tablementioning

confidence: 99%

Sparse postnatal labeling and quantification of superficial cortical cell synapses in the mouse neocortex

Gesuita¹,

Argunşah²,

Karayannis³

2022

STAR Protocols

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An inhalation anaesthesia approach for neonatal mice allowing streamlined stereotactic injection in the brain

Cited by 12 publications

References 15 publications

AMPA receptor anchoring at CA1 synapses is determined by N-terminal domain and TARP γ8 interactions

AMPA receptor anchoring at CA1 synapses is determined by N-terminal domain and TARP γ8 interactions

Downregulation of TGIF2 is possibly correlated with neuronal apoptosis and autism‐like symptoms in mice

Sparse postnatal labeling and quantification of superficial cortical cell synapses in the mouse neocortex

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