中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2012年
9期
781-788
,共8页
尚艳峰%张静%宫华青%陈霞
尚豔峰%張靜%宮華青%陳霞
상염봉%장정%궁화청%진하
眼震,先天性%斜视%动眼肌%神经末稍%神经纤维%腱%肌纤维
眼震,先天性%斜視%動眼肌%神經末稍%神經纖維%腱%肌纖維
안진,선천성%사시%동안기%신경말초%신경섬유%건%기섬유
Nystagmus,congenital%Strabismus%Oculomotor muscles%Nerve endings%Nerve fibers%Tendons%Muscle fibers
目的 研究人眼外肌栅栏状神经终末(PE)结构的形态、分布和功能,观察其在先天性眼球震颤(CN)中发生的改变,探讨CN的病因和发病机制,并为指导临床治疗提供理论依据.方法 眼肌手术时留取眼外肌远端的肌-腱连接区部分,标本分为CN组和对照组(共同性斜视患者),两组再按相近年龄大小(差别＜3岁)、相同取材部位配对分组并进行下述各项检测.(1)超微结构观察:标本经4％戊二醛前固定后,自附着点处向后连续每1 mm切取1或2块横断的1 mm ×2 mm×3 mm组织,常规电子显微镜标本处理,透射电子显微镜下观察两组眼外肌及PE的超微结构.(2)眼外肌肌-腱连接区整体经4％多聚甲醛固定、胆碱乙酰转移酶抗体标记后,行25 μm厚的纵、横断面冰冻切片,激光扫描共聚焦显微镜下观察PE的胆碱乙酰转移酶表达情况、形态及分布.(3)两组选取相同部位的肌-腱连接区组织,提取组织蛋白,进行胆碱乙酰转移酶的免疫印迹检测.采用两个独立样本t检验,对CN组与对照组内直肌横断面PE的数量、肌-腱连接区胆碱乙酰转移酶阳性表达量进行比较.结果 与对照组相比,透射电子显微镜下CN组PE的结构水肿,电子密度增高呈明显的嗜锇酸样改变；肌细胞内线粒体肿胀,肌浆网扩张.人眼外肌PE的胆碱乙酰转移酶呈阳性表达.眼外肌纵断面切片可见供应PE的神经纤维起自肌肉,延伸入腱后返回,发出栅栏状分支,分布于肌纤维末端.横断面切片可见内直肌在附着点后约3～4 mm处PE的分布最为集中,对照组和CN组栅栏状终末分布数量分别为26.800±3.421、39.200±3.564,差异有统计学意义(t=-5.613,P＜0.05).CN组的ChAT表达条带的灰度值(1.503±0.203)高于对照组(1.000±0.113),差异有统计学意义(t=-3.730,P＜0.05).结论 人眼外肌PE为胆碱能神经,分布于眼外肌远端的肌-腱连接处,可能与眼外肌的收缩有关.CN患者的眼外肌PE结构发生了明显改变,可能导致异常的眼球运动.
目的 研究人眼外肌柵欄狀神經終末(PE)結構的形態、分佈和功能,觀察其在先天性眼毬震顫(CN)中髮生的改變,探討CN的病因和髮病機製,併為指導臨床治療提供理論依據.方法 眼肌手術時留取眼外肌遠耑的肌-腱連接區部分,標本分為CN組和對照組(共同性斜視患者),兩組再按相近年齡大小(差彆＜3歲)、相同取材部位配對分組併進行下述各項檢測.(1)超微結構觀察:標本經4％戊二醛前固定後,自附著點處嚮後連續每1 mm切取1或2塊橫斷的1 mm ×2 mm×3 mm組織,常規電子顯微鏡標本處理,透射電子顯微鏡下觀察兩組眼外肌及PE的超微結構.(2)眼外肌肌-腱連接區整體經4％多聚甲醛固定、膽堿乙酰轉移酶抗體標記後,行25 μm厚的縱、橫斷麵冰凍切片,激光掃描共聚焦顯微鏡下觀察PE的膽堿乙酰轉移酶錶達情況、形態及分佈.(3)兩組選取相同部位的肌-腱連接區組織,提取組織蛋白,進行膽堿乙酰轉移酶的免疫印跡檢測.採用兩箇獨立樣本t檢驗,對CN組與對照組內直肌橫斷麵PE的數量、肌-腱連接區膽堿乙酰轉移酶暘性錶達量進行比較.結果 與對照組相比,透射電子顯微鏡下CN組PE的結構水腫,電子密度增高呈明顯的嗜鋨痠樣改變；肌細胞內線粒體腫脹,肌漿網擴張.人眼外肌PE的膽堿乙酰轉移酶呈暘性錶達.眼外肌縱斷麵切片可見供應PE的神經纖維起自肌肉,延伸入腱後返迴,髮齣柵欄狀分支,分佈于肌纖維末耑.橫斷麵切片可見內直肌在附著點後約3～4 mm處PE的分佈最為集中,對照組和CN組柵欄狀終末分佈數量分彆為26.800±3.421、39.200±3.564,差異有統計學意義(t=-5.613,P＜0.05).CN組的ChAT錶達條帶的灰度值(1.503±0.203)高于對照組(1.000±0.113),差異有統計學意義(t=-3.730,P＜0.05).結論 人眼外肌PE為膽堿能神經,分佈于眼外肌遠耑的肌-腱連接處,可能與眼外肌的收縮有關.CN患者的眼外肌PE結構髮生瞭明顯改變,可能導緻異常的眼毬運動.
목적 연구인안외기책란상신경종말(PE)결구적형태、분포화공능,관찰기재선천성안구진전(CN)중발생적개변,탐토CN적병인화발병궤제,병위지도림상치료제공이론의거.방법 안기수술시류취안외기원단적기-건련접구부분,표본분위CN조화대조조(공동성사시환자),량조재안상근년령대소(차별＜3세)、상동취재부위배대분조병진행하술각항검측.(1)초미결구관찰:표본경4％무이철전고정후,자부착점처향후련속매1 mm절취1혹2괴횡단적1 mm ×2 mm×3 mm조직,상규전자현미경표본처리,투사전자현미경하관찰량조안외기급PE적초미결구.(2)안외기기-건련접구정체경4％다취갑철고정、담감을선전이매항체표기후,행25 μm후적종、횡단면빙동절편,격광소묘공취초현미경하관찰PE적담감을선전이매표체정황、형태급분포.(3)량조선취상동부위적기-건련접구조직,제취조직단백,진행담감을선전이매적면역인적검측.채용량개독립양본t검험,대CN조여대조조내직기횡단면PE적수량、기-건련접구담감을선전이매양성표체량진행비교.결과 여대조조상비,투사전자현미경하CN조PE적결구수종,전자밀도증고정명현적기철산양개변；기세포내선립체종창,기장망확장.인안외기PE적담감을선전이매정양성표체.안외기종단면절편가견공응PE적신경섬유기자기육,연신입건후반회,발출책란상분지,분포우기섬유말단.횡단면절편가견내직기재부착점후약3～4 mm처PE적분포최위집중,대조조화CN조책란상종말분포수량분별위26.800±3.421、39.200±3.564,차이유통계학의의(t=-5.613,P＜0.05).CN조적ChAT표체조대적회도치(1.503±0.203)고우대조조(1.000±0.113),차이유통계학의의(t=-3.730,P＜0.05).결론 인안외기PE위담감능신경,분포우안외기원단적기-건련접처,가능여안외기적수축유관.CN환자적안외기PE결구발생료명현개변,가능도치이상적안구운동.
Objective To evaluate the morphology,distribution and function of palisade endings (PE) in human extraocular muscles (EOM),and observe the alterations in eyes with congenital nystagmus (CN).The etiology and pathogenesis of CN were also investigated.Methods It was a experimental study.The distal myotendinous junctions of the EOM were obtained during operation for CN ( CN group) and concomitant strabismus ( control group). The samples from patients with similar age and same extraction sites in the two groups were compared.The muscles cut during operation were immediately put into 4％ glutaraldehyde fixative solution.And 1 - 2 transverse bands of tissue were cut every 1 mm from tendon insertion for specimens processing.The ultrastructure of EOM and PE in the two groups was observed by transmission electron microscopy. The distal parts of EOM cut during operation were put into 4％ paraformaldehyde promptly.Myotendinous junction region whole mounts were labeled with antibodies against choline acetyltransferase (ChAT).Muscle fibers were counterstained with phalloidin.And longitudinal and transverse cryostat serial sections were cut at 25 μm and analyzed by confocal laser scanning microscopy.The ChAT expression,morphology and distribution of PE were observed.The same fragment of myotendinous junction in the two groups was selected.After the total protein was extracted,ChAT was detected by western blot.The expression level of ChAT was analyzed.Results Compared with the controls,the ultrastructure in the CN group had considerable variations.The axon of PE was swelled and deformed partly.The electron density was increased and presented as addicted to osmic acid. In the muscle cells,mitochondria was swelled,and sarcoplasmic reticulum was dilated.All PE exhibited ChAT immunoreactivity in human EOM.In the longitudinal section,nerve fibers extended from the muscle into the tendon,looped back and divided into several terminal arborizations (palisade endings) around the muscle fiber tip.The PE of medial rectus were richest at the location 3 - 4 mm from tendon insertion.In the cross section,the amount of PE in the CN group was higher than the control group ( t =- 5.613,P ＜ 0.05).The expression level of ChAT in the CN group was higher than the control group ( t =- 3.730,P ＜ 0.05 ).Conclusions Palisade endings in myotendinous junction of human EOM are cholinergic nerves,which might innervate the contraction of EOM.Significant changes of palisade endings in the EOM of the CN subjects may affect eve movement.
