發(fā)布時(shí)間：2018-03-18 06:41

  本文選題：大鼠　切入點(diǎn)：眼水平同向運(yùn)動(dòng)　出處：《青島大學(xué)》2007年博士論文　論文類型：學(xué)位論文

【摘要】： 目的 探討大鼠腦干內(nèi)向動(dòng)眼神經(jīng)核內(nèi)直肌亞核傳入的運(yùn)動(dòng)前神經(jīng)元的定位和分布，建立眼水平同向運(yùn)動(dòng)相關(guān)的腦干中樞核團(tuán)與動(dòng)眼神經(jīng)核內(nèi)直肌亞核之間的神經(jīng)纖維通路。在此基礎(chǔ)上，進(jìn)一步檢測這些通路中可能存在的興奮性和抑制性的神經(jīng)遞質(zhì)，從而建立眼水平同向運(yùn)動(dòng)相關(guān)核團(tuán)向動(dòng)眼神經(jīng)核投射的抑制性和興奮性神經(jīng)遞質(zhì)通路，為眼水平同向運(yùn)動(dòng)障礙性疾病的發(fā)生和治療提供解剖學(xué)依據(jù)。 材料和方法 健康成年Wister大鼠30只，220～250g，隨機(jī)分成2組。第1組14只，大鼠麻醉后，置于腦立體定位儀上，將50％的辣根過氧化物酶(horseradish peroxidase，HRP)溶液0.1μl緩慢注入到大鼠腦的一側(cè)動(dòng)眼神經(jīng)核(oculomotor nucleus，OMN)內(nèi)直肌亞核(medial rectus subdivision)，存活48h后，灌注固定取腦，連續(xù)冠狀冰凍切片(25～40μm)分5套收集，經(jīng)TMB呈色反應(yīng)，1套切片用于觀察注射部位和腦干核團(tuán)中HRP逆行標(biāo)記細(xì)胞的分布。另外4套切片經(jīng)TMB反應(yīng)后分別進(jìn)行抗谷氨酸(glutamate，Glu)，γ-氨基丁酸(γ-amino-butyric acid，GABA)，谷氨酸脫羧酶(glutamic acid decarboxylase，GAD)和谷氨酸受體1型(Glutamate Receptor1，GluR1)血清的SABC免疫組織化學(xué)染色，檢測這些核團(tuán)中存在的神經(jīng)遞質(zhì)。第2組16只，大鼠麻醉后，置于腦立體定位儀上，，將3％的熒光金(fluorogold，F(xiàn)G)0.1μl緩慢注入大鼠腦一側(cè)動(dòng)眼神經(jīng)核內(nèi)直肌亞核。動(dòng)物存活5～7天，灌注固定取腦，連續(xù)冠狀冰凍切片(25μm)分5組收集。1套用于觀察注射部位和FG逆行標(biāo)記細(xì)胞在腦干核團(tuán)中的分布，另外4套分別進(jìn)行抗GABA、Glu、GluR1、GAD血清的免疫熒光組織化學(xué)反應(yīng)，觀察這些腦內(nèi)核團(tuán)中逆行標(biāo)記細(xì)胞、免疫熒光標(biāo)記細(xì)胞和雙標(biāo)記細(xì)胞的分布及其向動(dòng)眼神經(jīng)核投射的神經(jīng)遞質(zhì)聯(lián)系。 結(jié)果 (1)逆行標(biāo)記結(jié)果：將HRP或FG注入一側(cè)動(dòng)眼神經(jīng)核(OMN)內(nèi)直肌亞核，在對(duì)側(cè)展神經(jīng)核(abducens nucleus，ABN)、雙側(cè)前庭神經(jīng)內(nèi)側(cè)核(medial vestibular nucleus，MVN)、腦橋旁正中網(wǎng)狀結(jié)構(gòu)(pontine paramedian reticular formation，PPRF)及同側(cè)前庭上核(superior vestibular nucleus，SVN)和舌下前置核(nucleus of prepositus hypoglosal，PH)均可見逆行標(biāo)記細(xì)胞和纖維。 (2)GABA、GAD、Glu、GluR1的免疫組織(熒光)化學(xué)反應(yīng)的切片中分別可見逆行標(biāo)記細(xì)胞，對(duì)應(yīng)的一抗的免疫(熒光)陽性細(xì)胞，逆標(biāo)／免疫(熒光)雙標(biāo)細(xì)胞三種標(biāo)記細(xì)胞。在對(duì)側(cè)展神經(jīng)核和同側(cè)舌下前置核主要存在Glu免疫／逆標(biāo)和GluR1免疫／逆標(biāo)的雙標(biāo)記細(xì)胞，GABA免疫／逆標(biāo)、GAD免疫／逆標(biāo)的雙標(biāo)記細(xì)胞較少；在同側(cè)前庭神經(jīng)內(nèi)側(cè)核主要可見Glu免疫／逆標(biāo)和GluR1免疫／逆標(biāo)的雙標(biāo)記細(xì)胞，未見明顯的GABA免疫／逆標(biāo)、GAD免疫／逆標(biāo)的雙標(biāo)記細(xì)胞；而對(duì)側(cè)前庭內(nèi)側(cè)核中則四種免疫雙標(biāo)記細(xì)胞都有；腦橋旁正中網(wǎng)狀結(jié)構(gòu)中GABA免疫／逆標(biāo)、GAD免疫／逆標(biāo)、Glu免疫／逆標(biāo)、GluR1免疫／逆標(biāo)的雙標(biāo)記細(xì)胞分布較為均勻。 結(jié)論 (1)在對(duì)側(cè)展神經(jīng)核，雙側(cè)前庭神經(jīng)內(nèi)側(cè)核、同側(cè)前庭上核、舌下前置核以及腦橋旁正中網(wǎng)狀結(jié)構(gòu)中均有纖維投射到動(dòng)眼神經(jīng)核內(nèi)直肌亞核，并可能參與協(xié)調(diào)雙眼的水平同向運(yùn)動(dòng)。 (2)展神經(jīng)核核間神經(jīng)元向?qū)?cè)動(dòng)眼神經(jīng)核投射，其興奮性投射主要以Glu為神經(jīng)遞質(zhì)，部分通過GluR1型受體起作用；其抑制性投射中GABA可能不是起主要作用的神經(jīng)遞質(zhì)。舌下前置核向同側(cè)內(nèi)直肌運(yùn)動(dòng)神經(jīng)元投射，其興奮性投射和抑制性投射與展神經(jīng)核相似。 (3)前庭神經(jīng)核向同側(cè)動(dòng)眼神經(jīng)核內(nèi)直肌亞核的投射，主要是興奮性的，用谷氨酸作為神經(jīng)遞質(zhì)，部分通過GluR1受體起作用；而前庭神經(jīng)內(nèi)側(cè)核向?qū)?cè)內(nèi)直肌亞核的投射包含興奮性和抑制性的投射，分別用Glu和GABA作為相應(yīng)的神經(jīng)遞質(zhì)；腦橋旁正中網(wǎng)狀結(jié)構(gòu)向雙側(cè)動(dòng)眼神經(jīng)核內(nèi)直肌亞核的投射同樣存在興奮性和抑制性的投射，其中Glu和GABA分別起一定的作用，是否存在遞質(zhì)共存，需進(jìn)一步研究。
[Abstract]:objective
To investigate the localization and distribution of rat brain stem within the medial rectus subnucleus of the afferent premotor neurons, a conjugate horizontal eye movement related brainstem nuclei and medial rectus subnucleus between the nerve fiber pathway. On this basis, further testing of these pathways in possible excitatory and inhibitory nerve the transmitter, so as to establish the conjugate horizontal eye movements related nucleus to oculomotor nucleus of inhibitory and excitatory neurotransmitter pathways, as happened to the same level of eye movement disorders and treatment to provide the anatomical basis.
Materials and methods
30 healthy adult Wister rats, 220 ~ 250g, were randomly divided into 2 groups. The first group of 14 rats, rats were anesthetized and placed in a stereotaxic apparatus, 50% horseradish peroxidase (horseradish, peroxidase, HRP) solution 0.1 l slowly injected into the rat brain side of the oculomotor nucleus (oculomotor nucleus. OMN) medial rectus subnucleus (medial rectus subdivision), the survival of 48h after perfusion fixed brains, continuous coronal frozen sections (25~40 m) 5 sets collected by TMB color reaction, 1 sets of sections for observation of the injection site and brainstem nuclei in HRP retrograde labeled cells. The distribution of the other 4 sets section after the TMB reaction were anti glutamic acid (glutamate, Glu), GABA (gamma -amino-butyric acid GABA (glutamic acid), glutamic acid decarboxylase decarboxylase, GAD) and glutamate receptor type 1 (Glutamate Receptor1, GluR1 SABC) immunohistochemical staining to detect the serum. There are some neurotransmitters in the nuclei. The second group of 16 rats, rats were anesthetized and placed in a stereotaxic apparatus, fluorescent gold 3% (Fluorogold, FG) 0.1 l slowly injected into the rat brain side of medial rectus subnucleus. The animal survived for 5~7 days, take continuous coronary perfusion fixed brain frozen section (25 m) were divided into 5 groups to observe the distribution of.1 from the injection site and FG retrograde labeled neurons in the brainstem nuclei, the other 4 were anti GABA, Glu, GluR1, immunohistochemical reaction of GAD in serum, to observe the retrograde labeling of cells in these nuclei, the distribution of immunofluorescence the labeled cells and double labeled cells and projecting to the oculomotor nucleus of the neurotransmitter.
Result
Results: (1) retrogradely labeled HRP or FG into the unilateral oculomotor nucleus (OMN) medial rectus subnucleus, contralateral abducens nucleus (abducens, nucleus, ABN), bilateral medial vestibular nucleus (medial vestibular nucleus, MVN), PPRF (pontine paramedian reticular formation, PPRF) and the same the lateral superior vestibular nucleus (superior vestibular nucleus, SVN) and hypoglossal nucleus (nucleus of prepositus hypoglosal, PH) were retrogradely labeled cells and fibers.
(2) GABA, GAD, Glu, GluR1 immunohistochemistry (Ying Guang) chemical reaction sections were observed retrogradely labeled cells, a corresponding antibody (Ying Guang) positive cells labeled / immune (Ying Guang) three kinds of double labeled cells labeled cells in the contralateral abducens nucleus and ipsilateral hypoglossal pre nuclear are Glu / GluR1 and immune labeled immune / retrogradely labeled double labeled neurons and GABA labeled GAD / immune, immune / retrogradely labeled double labeled neurons in the ipsilateral vestibular less; medial nucleus mainly visible Glu and GluR1 retrogradely labeled immune / immune / retrogradely labeled double labeled cells, no obviously GABA / GAD retrogradely labeled immune, immune / retrogradely labeled double labeled neurons; while the contralateral medial vestibular nucleus in four immune cells have double labeling; PPRF GABA / GAD retrogradely labeled immune, immune / inverse standard, Glu / GluR1 retrogradely labeled immune, immune / inverse calibration the double labeled cells The distribution is more uniform.
conclusion
(1) in the lateral abducens nucleus, bilateral medial vestibular nucleus, ipsilateral superior vestibular nucleus, sublingual anterior nucleus and para Pontus medialis reticular structure, all fibers projected into the rectus subnucleus of the oculomotor nucleus, and may be involved in coordinating the horizontal horizontal movement of the eyes.
(2) the abducens nucleus nucleus neurons to the lateral projection of the oculomotor nucleus, the excitatory projection takes Glu as the main neurotransmitter part via GluR1 receptors; the inhibition of GABA projection may not mainneurotransmitter. Hypoglossal nucleus to the ipsilateral medial rectus motoneurons projecting, the the excitatory and inhibitory projection projection and abducens nucleus is similar.
(3) vestibular nuclei projecting to the ipsilateral medial rectus subnucleus, predominantly excitatory, use glutamate as neurotransmitter, partly mediated by GluR1 receptors; and the medial vestibular nucleus to the lateral projection of the medial rectus subnucleus contains excitatory and inhibitory projection, respectively Glu and GABA as the corresponding neurotransmitters; PPRF projecting to the bilateral medial rectus subnucleus have excitatory and inhibitory projections, which Glu and GABA play a role, whether there is coexistence of transmitters, to be further studied.

【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2007
【分類號(hào)】：R77;R33

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 遲煥芳,李國棟,王守彪;大鼠展神經(jīng)核的細(xì)胞構(gòu)筑及其向動(dòng)眼神經(jīng)核的投射[J];神經(jīng)解剖學(xué)雜志;2003年03期

本文編號(hào)：1628421