ラット視神経への注入のための低侵襲テクニック
Summary
Direct injection into the rat optic nerve is useful for regenerative research. We demonstrate a minimally-invasive technique for direct injection into a rat optic nerve that does not involve opening the skull. Using this method, surgical complications are minimized and recovery is more rapid.
Abstract
The rat optic nerve is a useful model for stem cell regeneration research. Direct injection into the rat optic nerve allows delivery into the central nervous system in a minimally-invasive surgery without bone removal. This technique describes an approach to visualization and direct injection of the optic nerve following minor fascial dissection from the orbital ridge, using a conjunctival traction suture to gently pull the eye down and out. Representative examples of an injected optic nerve show successful injection of dyed beads.
Introduction
視神経は、中枢神経系（CNS）は、視神経炎、緑内障および外傷などの眼科条件を含む再生研究のための理想的なロケーションを提供します。幹細胞の種々の注射は、いずれかの軸索の数を増加させ、および/ または変性疾患を予防効果を示したか、失われたミエリンの取り付けの見込みを示している。1,2
人間の視神経が約3.0〜3.5ミリメートルの直径を有する視交叉に網膜から走行約120万平行軸索が含まれています。3、実験室でヒト疾患をモデル化するために、ラットは、頻繁に使用されています。成体ラットの視神経は、約0.5mmの直径内に約10万軸索を含んでいます。CNS再生研究の主な制限の4つは、直接骨なしアクセスです。頭蓋骨や脊椎骨が除去されたときに、動物に、合併症、手術リスクが高くなっています。の利点と同様に、頭蓋骨のオファー減少合併症、より迅速な回復を開かずに脊椎における低侵襲アプローチ、5直接視神経注射。
この技術は、以前の研究で使用されてきた。6本稿および付随ビデオでは、我々はラット視神経に幹細胞を注入するための低侵襲性の手順を示します。
Protocol
Representative Results
Discussion
Direct injection into the optic nerve of stem cells or other products intended to facilitate regeneration provides a convenient model compared to other means of injections into the CNS. This technique takes less time, requires less total anesthesia, avoids drilling or removing skull or bone tissue, reduces complications rates and allows for more rapid recovery following surgery.
The most critical steps in this protocol include: 1. Adequate hemostasis in the surgical field to allow clear visua…
Disclosures
The authors have nothing to disclose.
Acknowledgements
This study was supported by NeuralStem, Inc., and Johns Hopkins Project RESTORE.
Materials
| Name of Material/ Equipment | Company | Catalog Number | Comments/Description |
| Lewis rat | Charles River | 4 | Any rat strain will work. |
| Anesthesia machine | Surgivet | CDS9000 | CDS 9000 Small Animal Anesthesia Machine – Pole Mount |
| Infusion pump | Stoelting | 53129 | |
| Dissection microscope | National Optical | 409-411-1105 | |
| Fiber-optic light source | Fisher Scientific | 12-562-21 | |
| Dissection and Stereotaxic Instrument | Stoelting | 51400 | |
| Pipette Puller | Kopf | 750 | |
| Pipettes | World Precision Instruments | 18150-6 | |
| Disposable scalpel blades | Harvard Apparatus | 810-15-021 | |
| Iridectomy scissors | Electron Microscopy Sciences | Uniband LA-4XF |
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