Evaluation of preferential motor reinnervation by measuring choline acetyltransferase activity and by Karnovsky staining

通过测量胆碱乙酰转移酶活性和卡诺夫斯基染色评估优先运动神经支配

• 批准号: 08457394
• 负责人: KANAYA Fuminori
• 金额: $ 4.29万
• 依托单位: University of the Ryukyus
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457394/
• 关键词: selective motor reinnervation; choline acethltransferase activity; Karnovsky staining; misdirection

A hundred-eight Wister rats were used to examine whether preferential motor fiber regeneration exists. Rat femoral nerve bifurcates into motor branch (quadriceps branch) and sensory branch (saphenous nerve). Femoral nerve was sectioned just proximal to the bifurcation and both nerve stumps were inserted into silicone tube leaving 2mm gap (2mm gap group) and 4mm gap (4mm gap group) between stumps. Right femoral nerve was repaired without rotation (no rotation group) and left femoral nerve was repaired with 180 degrees rotation (rotation group). At 4,8,16,24,36,48 weeks after surgery, nerve specimens were harvested from both motor and sensory branches 5mm distal from distal nerve stumps. Choline acetyltransferase (CAT) activity of these specimens and those from normal nerves were measured. Also frozen sections of these specimens were stained with Karnovsky staining in order to see the localization of acetylcholine esterase, which is specific to motor axon, at 4,8,12,24 weeks after surgery. Regenerated motor axons were counted.At 2mm gap group, CAT activity in motor branch became significantly higher than that in sensory branch at 4,8,16,24 weeks in no rotation group and at 8 weeks in rotation group (p<0.05). At 4mm gap group, CAT activity in motor branch became significantly higher than that in sensory branch at 4,8,16,24,36 weeks in no rotation group and at 8,16,36weeks in rotation group (p<0.05). Histrogic examination showed no specific motor exon staining was seen 4 weeks after surgery. Regenerated motor axons started to show staining in both motor and sensory branches after 8 weeks in each group. Number of regenerated exons increased with time in motor and sensory branches in no rotation and rotation groups at 2mm gap group, and was not different between motor and sensory branches. This result and significant higher CAT activity in motor branch suggests that motor axons regenerated into motor branch matured more than those regenerated into sensory branch.

108只Wister大鼠被用来检查是否存在优先运动纤维再生。大鼠股神经分为运动分支（股四头肌分支）和感觉分支（隐神经）。将股神经在分叉处近端切断，将两个神经残端插入硅胶管中，在残端之间留出2mm间隙（2mm间隙组）和4mm间隙（4mm间隙组）。右侧股神经不旋转修复（不旋转组），左侧股神经旋转180 °修复（旋转组）。分别于术后4、8、16、24、36、48周，取距远端神经残端5mm处的运动支和感觉支神经标本。测定了这些标本和正常神经的胆碱乙酰转移酶（CAT）活性。术后4、8、12、24周取标本冰冻切片行Karnovsky染色，观察运动轴突特异性乙酰胆碱酯酶的定位。2mm间隙时，无旋转组运动分支CAT活性在4、8、16、24周时显著高于感觉支，旋转组在8周时显著高于感觉支（p<0.05）。在4mm间隙下，无旋转组在4、8、16、24、36周时运动分支CAT活性显著高于感觉分支，旋转组在8、16、36周时运动分支CAT活性显著高于感觉分支（p<0.05）。术后4周组织学检查未见特异性运动外显子染色。8周后，各组再生的运动和感觉分支的运动轴突开始染色。在无旋转组和旋转2mm间隙组，运动支和感觉支的再生外显子数随时间增加而增加，运动支和感觉支之间无差异。该结果和运动分支中显著较高的CAT活性表明，再生到运动分支中的运动轴突比再生到感觉分支中的运动轴突更成熟。

项目成果

普天間 朝上: "酵素活性(CAT活性)を用いた神経線維の選択的再生能の検討-シリコンユ-ブ架橋(2mm間隙)神経修復のCAT活性-" 日本手の外科学会誌. 11・3. 434-437 (1994)

浅上普天间：“利用酶活性（CAT活性）检查神经纤维的选择性再生能力——硅管交联（2mm间隙）神经修复中的CAT活性”日本手外科学会杂志11·3。 434-437 (1994)

普天間 朝上: "再生運動神経と知覚神経の識別法に関する実験的検討" 日本マイクロサージャリー学会誌. 7・1. 28-33 (1994)

浅上普天间：“再生运动神经和感觉神经的区分方法的实验研究”日本显微外科杂志7・1（1994年）。

Chojo Futenma: "Differentation of Regenerated Motor and Sensory Fiber by Karnovsky Staining and CAT Activity" J.Jpn.S.R.M. vol.7-No.1. 28-33 (1994)

Chojo Futenma：“通过卡诺夫斯基染色和 CAT 活性区分再生运动和感觉纤维”J.Jpn.S.R.M.

Chojo Futenma: "Evaluation of Selective Motor Reinnervation by Measuring Choline Acetyltransferase Activity" J.Jpn.Soc.S urg.Hand. vol.11-No.3. 434-437 (1994)

Chojo Futenma：“通过测量胆碱乙酰转移酶活性评估选择性运动神经支配”J.Jpn.Soc.S urg.Hand。

Chojo Futenma: "Evaluation of Selective Motor Reinnervation by Measuring Choline Acetyltransferase Activity" J.Jpn.Soc.S urg.Hand. vol.12-No.3. 455-458 (1995)

Chojo Futenma：“通过测量胆碱乙酰转移酶活性评估选择性运动神经支配”J.Jpn.Soc.S urg.Hand。