小鼠脊髓血管-神經(jīng)干細(xì)胞龕與神經(jīng)細(xì)胞的增殖和分化

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1、萬方數(shù)據(jù)第45卷第2期2014年4月解剖學(xué)報(bào)ACTAANATOMICASINICAV01．45，No．2Apr．2014’155’小鼠脊髓血管-神經(jīng)干細(xì)胞龕與神經(jīng)細(xì)胞的增殖和分化席艷1’2薛帥1李莉莉1(1．河南大學(xué)神經(jīng)生物學(xué)研究所，河南開封475004；2．聶夢月1鄧錦波h河南大學(xué)醫(yī)學(xué)院病理學(xué)教研室，河南開封475004)[摘要]目的探討小鼠脊髓發(fā)育過程中血管一千細(xì)胞龕對(duì)神經(jīng)干細(xì)胞分化、增殖和遷移的影響；探討龕中的各種細(xì)胞、血管在發(fā)育中的變化及相互作用。方法發(fā)育過程中的胚胎鼠或生后小鼠150只，取脊髓腰膨大處的橫斷面切片。采用5一溴脫氧尿嘧啶核苷(BrdU)免疫熒光標(biāo)記神

2、經(jīng)干細(xì)胞；用尼氏(Nissl)染色觀察神經(jīng)元的發(fā)育和形態(tài)變化；利用血管墨汁灌注結(jié)合免疫熒光三重標(biāo)記的方法檢測神經(jīng)干細(xì)胞、神經(jīng)元和神經(jīng)膠質(zhì)細(xì)胞在小鼠胚胎期及生后的形態(tài)分布變化；同時(shí)對(duì)脊髓內(nèi)血管發(fā)育的動(dòng)態(tài)變化進(jìn)行形態(tài)學(xué)觀察和體視學(xué)處理。結(jié)果妊娠14d(E14)小鼠脊髓內(nèi)即有BrdU標(biāo)記的陽性細(xì)胞，均勻分布。新生的神經(jīng)元自神經(jīng)管側(cè)腦室下帶遷移至中間層附近，逐漸分化為神經(jīng)元，并集中于灰質(zhì)呈現(xiàn)典型的“H”型。統(tǒng)計(jì)學(xué)分析表明，BrdU陽性細(xì)胞計(jì)數(shù)在發(fā)育中呈現(xiàn)拋物線，高峰值位于出生3d(P3)左右。此外，神經(jīng)膠質(zhì)細(xì)胞的祖細(xì)胞最早集中于邊緣白質(zhì)，隨后又向中央的灰質(zhì)回遷。血管發(fā)育經(jīng)歷了早期均

3、勻分布，成年后集中在灰質(zhì)分布的過程。血管、神經(jīng)干細(xì)胞、神經(jīng)元和神經(jīng)膠質(zhì)細(xì)胞構(gòu)成了所謂的血管一千細(xì)胞龕。血管．干細(xì)胞龕主要位于側(cè)腦室下帶和灰質(zhì)附近，其中的血管、神經(jīng)干細(xì)胞、神經(jīng)元和神經(jīng)膠質(zhì)細(xì)胞互相交織、緊密聯(lián)系。結(jié)論血管一神經(jīng)干細(xì)胞龕與脊髓發(fā)育緊密聯(lián)系，同時(shí)影響神經(jīng)元、神經(jīng)膠質(zhì)細(xì)胞和血管的相互作用。血管一神經(jīng)干細(xì)胞龕為脊髓發(fā)育及神經(jīng)元和神經(jīng)膠質(zhì)細(xì)胞分化提供了適宜的微環(huán)境。[關(guān)鍵詞]脊髓發(fā)育；神經(jīng)干細(xì)胞；血管一神經(jīng)干細(xì)胞龕；5一溴脫氧尿嘧啶核苷免疫熒光標(biāo)記；血管墨汁灌注法；小鼠[中圖分類號(hào)]R322．81[文獻(xiàn)標(biāo)志碼]A[DOI]10．3969／j．issn．0529-1356

4、．2014．02．002Vascularandstem-cellularnicheandtheneuralproliferationanddifferentiationinspinalcordofthemouseXIYanl一，XUEShuail，LILi．．．1i，NIEMeng—yuel，DENGJin．b01+(，．InstituteofNeurobiology，He’nanUniversity，He’nanKa洳ng475004，China2．MedicalCollege，He’nanUniversity，He’nanKa治ng475004，China)[Abstr

5、act]ObjectiveTostudyhowthevascularandstem—cellularnicheaffecttheneuralproliferation，differentiationandmigrationinthemousespinalcordandtoinvestigatetheinteractionamongneurons，neurogliaandvasculatureinthedevelopingspinalcord．MethodsAtotalof150micefromE17toP180wereusedinthisproject．BrdUassay，

6、Nissl’Sstainingandimmunofluorescentlabeling(NeuNandGFAP)methodswereapplied．Bloodvesselsandneuralstemcellsweremeasuredwithstereologiealmethods．ResultsBrdUpositiveneuralstemcellsappearedevenlyintheembryonicspinalcordatasearlyasE14，andthenewbornneuronsmigratedfromthesubventicularzoneoftheneur

7、altubetotheintermediatelayeraroundit．Withageincreasing，theneuralstemcellsdifferentiatedintoneuronsgradually，andtheneuronsmainlyconcentratedintheputativegraymatterasa“H”shape．Accordingtothestatisticalanalysis．theBrdUpositivestemcellsdevelopedasparaboliccurvewit