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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Establishment of Resiniferatoxin (RTX) Neuropathy
CAUTION:&#160;RTX is neurotoxic and hazardous. On contact, it acts as an irritant to the eyes, mucous membranes, and upper respiratory tract. Avoid inhalation and wear lab eyeglasses and coats during RTX preparation. Rinse with plenty of water in case of skin contact or after handling.
<ol>
	<li>Add 1 mg of RTX powder to a 200 &#181;L mixture of equal volume of Tween 80 and absolute ethanol (100 &#181;L for each solvent).</li>
	<li>Aliquot the RTX solution (12 &#181;L/vial) and store it at -20 &#176;C for up to 3 months. This constitutes the RTX stock; discard the remaining RTX solution at expiration.</li>
	<li>Dilute the RTX stock with normal saline to a final volume of 600 &#181;L. The final concentration of the RTX solution should be 0.01%, which equals 1 &#181;g RTX in 10 &#181;L vehicle solution.</li>
	<li>Use 8-week-old adult male ICR mice (35-40 g) as experimental animals and administer a single dose of RTX solution (dose: 200, 50, and 10 &#181;g/kg, respectively) intraperitoneally (i.p.) with a microinjection syringe to the mice.&#160;Mice were anesthetized by inhaler with 5 % isoflurane for deep anesthesia. If mice showed withdrawal action of extremities during RTX injection, mice should take longer inhalation of anesthesia.&#160;&#160; 
	Example: If the mouse weighs 40 g, then it will receive 20 &#181;L of the RTX solution, representing the dose of 50 &#181;g/kg.</li>
	<li>Give one group of mice an equal volume of vehicle (10% Tween 80 and 10% absolute ethanol in saline solution) as a control.</li>
	<li>After RTX injection, return the mice to a plastic cage on a 12-h light/12-h dark cycle and provide food and water&#160;ad libitum.</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Chemical reagent</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Resiniferatoxin</td>
			<td>Sigma</td>
			<td>R8756</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween 80</td>
			<td>Sigma</td>
			<td>P1754</td>
			<td></td>
		</tr>
	</tbody>
</table>

establishing a mouse model of small fiber neuropathy using resiniferatoxin

Source: Lee, Y. et al., <a href="https://app.jove.com/v/56651">Establishing a Mouse Model of a Pure Small Fiber Neuropathy with the Ultrapotent Agonist of Transient Receptor Potential Vanilloid Type 1.</a>&#160;J. Vis. Exp.&#160;(2018).
This video demonstrates a method for establishing a small fiber neuropathy model in mice using the TRPV1 agonist resiniferatoxin (RTX). It outlines the steps involved in administering RTX to mice and explains how RTX induces neuronal death in small-diameter sensory neurons, thereby establishing the neuropathy model.

Establishing a Mouse Model of Small Fiber Neuropathy Using Resiniferatoxin

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

Begin with a solution of resiniferatoxin (RTX), a high-affinity agonist of the TRPV1 receptor&#8212;pain-sensitive ion channels expressed on sensory neurons.
Take an anesthetized adult mouse and administer RTX intraperitoneally.
The drug enters systemic circulation and reaches the dorsal root ganglia (DRG), a cluster of sensory neuron cell bodies located near the spinal cord.
RTX crosses the blood-nerve barrier and interacts with small-diameter sensory neurons in the DRG that abundantly express TRPV1.
RTX binding to TRPV1 activates the receptors, inducing a massive calcium ion influx into the neurons.
Intracellular calcium activates enzymes that degrade cell membrane phospholipids, cellular proteins, and DNA, ultimately leading to neuronal death.
Subsequently, the intraepidermal nerve fibers (IENFs), the peripheral terminals of small-diameter sensory neurons that innervate the skin, begin to degenerate.
This degeneration of IENFs results in the loss of sensory input from the skin, thereby establishing a small fiber neuropathy model.

establishing-mouse-model-small-fiber-neuropathy-using

Research

JoVE Encyclopedia of Experiments

Neuroscience

Neuropathology

Neuromuscular Disorders

35 ビュー. 出典:Lee, Y. et al., 一過性受容体電位バニロイド1型の超強力なアゴニストによる純粋な小繊維ニューロパチーのマウスモデルの確立。J. Vis. Exp.(2018年)。 このビデオでは、TRPV1アゴニフェラトキシン(RTX)を使用してマウスの小繊維ニューロパチーモデルを確立する方法を示します。マウスへのRTX投与の手順を概説し、RTXが小径の感覚ニューロンでニューロン死を誘発し、それ?...

ビデオ: レシニフェラトキシンを用いた小繊維ニューロパチーのマウスモデルの確立 - 概念

Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history have&#160;remained poorly understood. Recently, we developed a reliable, reproducible animal model of PION and tested the treatment effect of some neurotrophic factors in this model1. The purpose of this video is to demonstrate our photochemically induced model of posterior ischemic optic neuropathy, and to evaluate its effects with retrograde labeling of retinal ganglion cells. Following surgical exposure of the posterior optic nerve, a photosensitizing dye, erythrosin B, is intravenously injected and a laser beam is focused onto the optic nerve surface. Photochemical interaction of erythrosin B and the laser during irradiation damages the vascular endothelium, prompting microvascular occlusion mediated by platelet thrombosis and edematous compression. The resulting ischemic injury yields a gradual but pronounced retinal ganglion cell dieback, owing to a loss of axonal input - a remote, injury-induced and clinically relevant outcome. Thus, this model provides a novel platform to study the pathophysiologic course of PION,&#160;and can be further optimized for testing therapeutic approaches for optic neuropathies as well as other CNS ischemic diseases.

The goal of this protocol is to photochemically induce ischemic injury to the posterior optic nerve in rat. This model is critical to studies of the pathophysiology of posterior ischemic optic neuropathy, and therapeutic approaches for this and other optic neuropathies, as well as of other CNS ischemic diseases.

光化学誘起後方虚血性視神経障害のラットモデル

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history ...

JoVE Journal

医学

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy (rNAION)

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their lifetime. NAION results in optic nerve edema, selective loss of the retinal ganglion cell neurons (RGCs) and atrophy of the optic nerve. A rodent model of NAION that expresses most NAION features and sequelae has been developed, which is applicable to both rats and mice. This model utilizes a focal laser application of 532 nm wavelength to illuminate a photoactive dye, Rose Bengal (RB), to cause capillary damage and leakage at the targeted anterior optic nerve (the laminar region). After rNAION induction, there is an early optic nerve ischemia, optic nerve edema, and intraneural inflammation, followed by selective RGC and axonal loss. Since the optic nerve is a CNS white matter tract, the rNAION model is applicable to mechanistic studies of selective white matter ischemia, as well as neuroprotective analyses and short and long-term mechanisms of glial and neuronal response to ischemia.

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy rNAION

The following report describes how to replicate the rodent model of nonarteritic anterior ischemic optic neuropathy (rNAION), using the appropriate dye, contact lens and laser parameters. We also reveal the appropriate steps for evaluating the rNAION lesion in vivo.

Nonarteritic前部虚血性視神経症の齧歯類モデル（rNAION）

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their ...

Modeling Neuronal Death and Degeneration in Mouse Primary Cerebellar Granule Neurons

Cerebellar granule neurons (CGNs) are a commonly used neuronal model, forming an abundant homogeneous population in the cerebellum. In light of their post-natal development, abundance, and accessibility, CGNs are an ideal model to study neuronal processes, including neuronal development, neuronal migration, and physiological neuronal activity stimulation. In addition, CGN cultures provide an excellent model for studying different modes of cell death including excitotoxicity and apoptosis. Within a week in culture, CGNs express N-methyl-D-aspartate (NMDA) receptors, a specific ionotropic glutamate receptor with many critical functions in neuronal health and disease. The addition of low concentrations of NMDA in conjunction with membrane depolarization to rodent primary CGN cultures has been used to model physiological neuronal activity stimulation while the addition of high concentrations of NMDA can be employed to model excitotoxic neuronal injury. Here, a method of isolation and culturing of CGNs from 6 day old pups as well as genetic manipulation of CGNs by adenoviruses and lentiviruses are described. We also present optimized protocols on how to stimulate NMDA-induced excitotoxicity, low-potassium-induced apoptosis, oxidative stress and DNA damage following transduction of these neurons.

This protocol describes a simple method for isolating and culturing primary mouse cerebral granule neurons (CGNs) from 6-7 day old pups, efficient transduction of CGNs for loss and gain of function studies, and modelling NMDA-induced neuronal excitotoxicity, low-potassium-induced cell death, DNA-damage, and oxidative stress using the same culture model.

神経細胞死とマウス プライマリ小脳顆粒ニューロンの変性をモデリング

Cerebellar granule neurons (CGNs) are a commonly used neuronal model, forming an abundant homogeneous population in the cerebellum. In light of their ...

Establishing a Mouse Model of Small Fiber Neuropathy Using Resiniferatoxin

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Establishing a Mouse Model of Small Fiber Neuropathy Using Resiniferatoxin