Targeted Expression of Channelrhodopsin-2 to the Axon Initial Segment Alters the Temporal Firing Properties of Retinal Ganglion Cells

Zhang, Zhifei; Feng, Jiaxuan; Wu, Cunkai; Lu, Qi; Pan, Zhuo Hua

doi:10.1371/journal.pone.0142052

Cited by 17 publications

(14 citation statements)

References 25 publications

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“…Animals in the NpHR group received a 3-mL injection of AAV-hSyn-eNpHR3.0-EYFP targeted to the ipsilateral ACC. The concentration of the viral vectors was 2.3 × 10 13 genome copies (GC)/mL 20. We made an incision in the skin over the skull and drilled a hole in the skull to inject the AAV plasmids or PBS into the ACC using a Hamilton syringe and an automated microsyringe pump (Harvard Apparatus, Holliston, MA, USA) at a flow rate of 0.5 mL/min 21.…”

Section: Methodsmentioning

confidence: 99%

Optical inactivation of the anterior cingulate cortex modulate descending pain pathway in a rat model of trigeminal neuropathic pain created via chronic constriction injury of the infraorbital nerve

Moon¹,

Heo²,

Kim

et al. 2017

JPR

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PurposeThe anterior cingulate cortex (ACC) plays a critical role in the initiation, development, and maintenance of neuropathic pain. Recently, the effects of optical stimulation on pain have been investigated, but the therapeutic effects of optical stimulation on trigeminal neuralgia (TN) have not been clearly shown. Here, we investigated the effects of optical inhibition of the ACC on TN lesions to determine whether the alleviation of pain affects behavior performance and thalamic neuron signaling.Materials and methodsTN lesions were established in animals by generating a chronic constriction injury of the infraorbital nerve, and the animals received injections of AAV-hSyn-eNpHR3.0-EYFP or a vehicle (phosphate-buffered saline [PBS]) in the ACC. The optical fiber was fixed into the ipsilateral ACC after the injection of adeno-associated virus plasmids or vehicle. Behavioral testing, consisting of responses to an air puff and cold allodynia, was performed, and thalamic neuronal activity was monitored following optical stimulation in vivo. Optical stimulation experiments were executed in three steps: during pre-light-off, stimulation-light-on, and post-light-off states. The role of the optical modulation of the ACC in response to pain was shown using a combination of optical stimulation and electrophysiological recordings in vivo.ResultsMechanical thresholds and facial cold allodynia scores were significantly improved in the TN lesion group during optical stimulation compared to those in the control group. Thalamic neuronal activity, consisting of the firing rate (spikes/s) and burst rate (bursts/s), was also decreased during optical stimulation.ConclusionReciprocal optical inhibition of the ACC can alleviate pain-associated behavior and decrease abnormal thalamic sensory neuron activity in the trigeminal neuropathic rat model. The descending pain pathway can modulate thalamic neurons from the ACC following optical stimulation.

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Section: Methodsmentioning

confidence: 99%

Optical inactivation of the anterior cingulate cortex modulate descending pain pathway in a rat model of trigeminal neuropathic pain created via chronic constriction injury of the infraorbital nerve

Moon¹,

Heo²,

Kim

et al. 2017

JPR

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“…The ultimate goal for this approach is to enable transcriptomic cell type identification of specific cells with clear connectomic results. We demonstrate an example of this desired outcome by first injecting a virus encoding Cre-dependent ChrimsonR-EYFP-Kv2.1 21,22 into the brain of an Sst-IRES-Cre mouse. The channelrhodopsin and reporter is sparsely expressed in Sst-expressing cells.…”

Section: B -Filled Cell (Orange Marker) Shows Expression Of Slc17a7 Amentioning

confidence: 99%

“…In other experiments (Figure 4), ~P28 Sst-IRES-Cre driver mice were anesthetized with an isoflurane/oxygen mixture and 200nL of adeno associated virus (AAV) serotype 1 (~5×10 12 genome copy units/mL) was injected through a small craniotomy at 0.3 mm and 0.6 mm beneath the surface of primary visual cortex (3.16mm anterior and 2.75mm lateral to bregma). The AAV delivered a bicistronic construct consisting of ChrimsonR fused to a Kv2.1 somatic localization motif 21,22 that has been shown to enhance resolution and sensitivity for two photon optogenetic mapping experiments 15…”

Section: Animals and Transgene Expressionmentioning

confidence: 99%

“…Figure 4 shows an example of this desired outcome. A slice expressing a channelrhodopsin (via injection of virus encoding Cre-depenent ChrimsonR-Kv2.1 21,22 ) in Sst-IRES-Cre+ cells probed in the two photon synaptic physiology pipeline demonstrates a connection between an optically-targeted cell ( Fig 4A and 4B, red) and a patched cell. Neighboring cells show no detected connection ( Fig 4A and B, blue and yellow) to the patched cell.…”

Section: Figure 3 -Mfish Light Sheet On a Two-photon Synaptic Physiolmentioning

confidence: 99%

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Multimodal cell type correspondence by intersectional mFISH in intact tissues

Nicovich

Taormina

et al. 2019

Preprint

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Defining a complete set of cell types within the cortex requires reconciling disparate results achieved through diverging methodologies. To address this correspondence problem, multiple methodologies must be applied to the same cells in multiple single-cell experiments. Here we present an approach in which spatial transcriptomics using multiplexed fluorescence in situ hybridization (mFISH) on tissue previously interrogated through two-photon optogenetic mapping of synaptic connectivity can resolve the anatomical, transcriptomic, connectomic, electrophysiological, and morphological characteristics of single cells within the mouse cortex. MainThe parable of the Blind Men and the Elephant tells the story of a group of blind men, naïve to the nature of an elephant, presented with such a creature to try to conceptualize while observing only through touch ( Figure 1A, upper). The men are each concerned with a single area and draw a conclusion for the whole based on their limited perspective. As scientists, we are similarly a group of observers each indirectly observing one portion of a large, hidden, and complex problem. The parable's moral is that these challenging concepts may have many facets individually revealed to those with diverging perspectives. To strive towards a comprehensive description, we must be able to synthesize these unique perspectives into a consistent whole.This challenge of differing perspectives leading to seemingly conflicting outcomes is exemplified in attempts to understand the number and features of cell types within the mouse cortex (Fig 1A, lower). Recent studies report self-consistent outcomes based on anatomy 1 , transcriptomics 2,3 , electrophysiology 4 , morphology 4 , long-range 5 and local connectomics 6 , and in vivo recordings 7 . However, these studies markedly disagree on the number of underlying cell types. Even if they did agree on the number of types, how results from one study map to those of another is not obvious. To reconcile this problem of cell type correspondence, single-cells must traverse multiple modalities.Each of these modalities, save transcriptomics, explicitly or implicitly carries spatial information within the specimen at single-cell resolution. Multiplexed fluorescence in situ hybridization (mFISH) enables a transcriptomic readout with this same single-cell spatial resolution. We have extended the hairpin

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“…The concentration of the viral vectors was 2.3 × 10 13 genome copies (GC)/mL. 20 We made an incision in the skin over the skull and drilled a hole in the skull to inject the AAV plasmids or PBS into the ACC using a Hamilton syringe and an automated microsyringe pump (Harvard Apparatus, Holliston, MA, USA) at a flow rate of 0.5 mL/min. 21 After the injection was complete, the syringe was maintained in place for 10 minutes to allow the solution to absorb.…”

Section: Surgical Preparation For Tn Modelsmentioning

confidence: 99%

Optical inactivation of the anterior cingulate cortex modulate descending pain pathway in a rat model of trigeminal neuropathic pain created via chronic constriction injury of the infraorbital nerve

et al. 2019

View full text Add to dashboard Cite

Purpose: The anterior cingulate cortex (ACC) plays a critical role in the initiation, development, and maintenance of neuropathic pain. Recently, the effects of optical stimulation on pain have been investigated, but the therapeutic effects of optical stimulation on trigeminal neuralgia (TN) have not been clearly shown. Here, we investigated the effects of optical inhibition of the ACC on TN lesions to determine whether the alleviation of pain affects behavior performance and thalamic neuron signaling. Materials and methods: TN lesions were established in animals by generating a chronic constriction injury of the infraorbital nerve, and the animals received injections of AAV-hSyn-eNpHR3.0-EYFP or a vehicle (phosphate-buffered saline [PBS]) in the ACC. The optical fiber was fixed into the ipsilateral ACC after the injection of adeno-associated virus plasmids or vehicle. Behavioral testing, consisting of responses to an air puff and cold allodynia, was performed, and thalamic neuronal activity was monitored following optical stimulation in vivo. Optical stimulation experiments were executed in three steps: during pre-light-off, stimulation-light-on, and post-light-off states. The role of the optical modulation of the ACC in response to pain was shown using a combination of optical stimulation and electrophysiological recordings in vivo. Results: Mechanical thresholds and facial cold allodynia scores were significantly improved in the TN lesion group during optical stimulation compared to those in the control group. Thalamic neuronal activity, consisting of the firing rate (spikes/s) and burst rate (bursts/s), was also decreased during optical stimulation. Conclusion: Reciprocal optical inhibition of the ACC can alleviate pain-associated behavior and decrease abnormal thalamic sensory neuron activity in the trigeminal neuropathic rat model. The descending pain pathway can modulate thalamic neurons from the ACC following optical stimulation.

show abstract

Targeted Expression of Channelrhodopsin-2 to the Axon Initial Segment Alters the Temporal Firing Properties of Retinal Ganglion Cells

Cited by 17 publications

References 25 publications

Optical inactivation of the anterior cingulate cortex modulate descending pain pathway in a rat model of trigeminal neuropathic pain created via chronic constriction injury of the infraorbital nerve

Optical inactivation of the anterior cingulate cortex modulate descending pain pathway in a rat model of trigeminal neuropathic pain created via chronic constriction injury of the infraorbital nerve

Multimodal cell type correspondence by intersectional mFISH in intact tissues

Optical inactivation of the anterior cingulate cortex modulate descending pain pathway in a rat model of trigeminal neuropathic pain created via chronic constriction injury of the infraorbital nerve

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