无尾两棲类心脏的决定与分化

内胚层在两栖类心脏发生过程中起着重要的作用。在有尾类神经期去除内胚层不能得到心脏的形成,心脏原基在尾芽期才能显示其自己分化的能力。在无尾类方面尚缺少系统的工作,本文用黑斑蛙Rana nigromaculata,泽蛙Rana limnocharis和狭口蛙 Callula borealis三种无尾类为材料,自原肠晚期至神经胚期进行下列三方面的实验研究。1.切除全部内胚层。2.分离心脏原基外植培养。3.取部分神经板与心脏原基一起外植培养。实验结果指出这三种无尾类在13期(早神经胚)心脏原基已开始显示其自己分化的能力,在缺少内胚层的情况下发育为分化程度不一的跳动的心脏结构。神经组织在早神经期并没有抑制心脏的发生,说明无尾类与有尾类不同,在早神经期的心脏原基已经充分决定,能按其预定意义分化。     

花背蟾蜍早期发育中内胚层细胞核的分化

以细胞核移植的方法,探讨花背蟾蜍早期发育中内胚层细胞核的分化。实验按供体胚胎的不同发育时期分7组,并以囊胚期动物极区细胞核的移植作对照。结果表明:原肠晚期以前的内胚层细胞核与囊胚期动物极区细胞核一样,是尚未分化的,它们仍具有发育的全能性。从神经胚期开始,细胞核有了明显的分化,但这种分化是渐进的,至摄食期的蝌蚪仍有部分细胞核保持着促使卵子发育的功能。     

半滑舌鳎早期胚胎性腺原基分化的组织学

采用连续组织切片对半滑舌鳎胚胎及仔鱼进行了观察研究,首次描述了半滑舌鳎胚胎发育过程中原始生殖细胞(primordial germ cells,PGCs)出现的部位及迁移特征,以及卵黄合胞体,鳔与性腺原基的发育分化.结果发现,PGCs出现于神经胚期的靠近卵黄囊的囊胚层.PGCs的特征为体积比周围细胞大,核大透亮,随后在肌肉期的脊索壁上出现.孵化前期的PGCs迁移到肠原基附近,肠系膜旁可见尚在移动的PGCs和10日龄的仔鱼中在肾管旁出现性腺原基,以后PGCs数量逐渐增多参与性腺的形成.本研究为半滑舌鳎的发育生物学以及养殖生产实践提供参考.     

日本沼虾胚胎复眼发生的研究

应用光镜技术和透射电镜技术,研究了日本沼虾胚胎复眼的发生.在原肠期,胚区前端的外胚层细胞增殖形成视叶原基.发育至后无节幼体期开始形成视神经节.在前蚤状幼体早期胚胎出现复眼色素，同时构成复眼的细胞开始分化。前蚤状幼体晚期，复眼色素区扩大到复眼直径的一半。复眼由个眼构成，每个个眼由角膜、角膜生成细胞、晶锥、小网膜细胞等组成。超微结构显示，胚胎的复眼结构与成体的相似，但感杆束直径比成体小。至胚胎孵化，眼柄尚未发育完善。     

''''晚籽银桂''''、''''多芽金桂''''花芽的形态分化

采用石蜡切片法观察了'晚籽银桂'、'多芽金桂'花芽形态分化过程.结果表明:'晚籽银桂'花芽分化从6月下旬苞片原基分化至9月初雌蕊原基形成历时近3个月,其过程可分为苞片分化期、花序原基分化期、小花原基分化期、顶花花萼分化期、花瓣分化期、雄蕊分化期和雌蕊分化期7个时期.其中雄蕊分化期历时长,分化较慢,其他6个时期历时较短.聚伞花序的中间顶花先分化,侧花后分化,各小花几乎同时完成花芽分化进程.'多芽金桂'的花芽分化除雌蕊发育与'晚籽银桂'不同外,其余基本一致.     

分棘四棱线虫的生活史研究

本文报道了分棘四棱线虫在中间宿主和终末宿主体内的发育情况。在16—22℃下,虫卵被异壳介虫吞下后数小时,第1期幼虫即孵化出来,进入宿主组织中寄生。第8天后进入第2期,消化道逐渐形成。第13天后进入第3期,尾端的棘刺排成1圈,圈中央为1个棘刺。生殖原基呈团状,雄虫的生殖原基靠近肠前部附近;雌虫的生殖原基在肠的后部靠近直肠。感染雏鸭后,幼虫生殖原基即开始迅速发育。至感染后第3天,幼虫蜕皮进入第4期,雄虫的交合刺与泄殖腔、雌虫的储精囊形成。感染后第7天,幼虫最后一次蜕皮,雌虫进入拉培根窝内寄生,虫体开始显著膨大。感染后13天雌雄虫分别出现精子与虫卵。感染后15天,出现发育成熟的虫卵。     

鸡胚胎干细胞的分离、培养与鉴定的初步研究

采用饲养层培养法,以高糖DM EM为基础培养基,同时添加胎牛血清以及5 ng/m l SCF,l0 ng/m l bFGF和1 000 IU/m l mL IF等细胞因子与鸡胚浸出液等,对鸡的X期胚盘细胞进行离体培养,并利用形态学鉴定、AKP染色、体外分化实验等方法对所获得的细胞克隆进行鉴定。实验结果表明,本实验建立的培养体系培养出了具有多分化潜能的类胚胎干细胞。     

南京椴花芽分化研究

利用石蜡切片法观察南京椴花芽分化过程.结果表明:南京椴越冬休眠芽为混合芽,萌发后叶腋处形成花芽.花芽分化时期为当年3月底至5月初,历时35～40 d,具有分化时间短、速度快的特点.花芽分化包括花序分化和小花分化两个过程,可划分为花芽分化始期、花序原基分化期、小花原基分化期、花萼原基分化期、花瓣原基分化期、雄蕊原基分化期、雌蕊原基分化期等7个时期.     

内耳干细胞研究取得突破进展

复旦大学附属眼耳鼻喉科医院李华伟副教授与美国哈佛大学眼耳医院Heller Stefan博士合作,经过艰苦努力,在成年鼠内耳前庭分离出胚胎样具有多向分化潜能干细胞,并将此干细胞经过体外细胞培养后移植到鸡胚胎并在发育的鸡胚胎内耳听囊中成功分化出毛细胞。此项研究成果揭示了内耳毛细胞可能再生的根     

鸡胚胸腺、法氏囊组织免疫功能发育研究

实验选取12-19胚龄的鸡胚和1日龄雏鸡的胸腺、法氏囊,石蜡切片HE染色后,对组织结构发育进行动态连续观察.通过MTT法检测胸腺T淋巴细胞和法氏囊B细胞的增殖率,了解不同胚龄T细胞、B细胞的免疫功能发育的状态.结果表明:随着胚龄的增加,鸡的胸腺和法氏囊发育逐渐完善,免疫功能随之增强;T淋巴细胞、B淋巴细胞的增殖率不断增加.     

Astrocytes induce blood-brain barrier properties in endothelial cells

The highly impermeable tight junctions between endothelial cells forming the capillaries and venules in the central nervous system (CNS) of higher vertebrates are thought to be responsible for the blood-brain barrier that impedes the passive diffusion of solutes from the blood into the extracellular space of the CNS. The ability of CNS endothelial cells to form a blood-brain barrier is not intrinsic to these cells but instead is induced by the CNS environment: Stewart and Wiley demonstrated that when avascular tissue from 3-day-old quail brain is transplanted into the coelomic cavity of chick embryos, the chick endothelial cells that vascularize the quail brain grafts form a competent blood-brain barrier; on the other hand, when avascular embryonic quail coelomic grafts are transplanted into embryonic chick brain, the chick endothelial cells that invade the mesenchymal tissue grafts form leaky capillaries and venules. It is, however, not known which cells in the CNS are responsible for inducing endothelial cells to form the tight junctions characteristic of the blood-brain barrier. Astrocytes are the most likely candidates since their processes form endfeet that collectively surround CNS microvessels. In this report we provide direct evidence that astrocytes are capable of inducing blood-brain barrier properties in non-neural endothelial cells in vivo.     

三疣梭子蟹消化系统的组织学研究

从组织学、组织化学二个方面研究了三疣梭子蟹的消化系统．消化系统主要包括消化管和消化腺．消化管的组织结构都由基本的四层构成，即粘膜层、粘膜下层、肌层和外膜．粘膜层是由单层柱状上皮和基膜构成的．除中肠外，其余部分管壁上皮表面分别被有弹性蛋白、胶原蛋白、粘蛋白和几丁质骨片等结构．在食道和后肠壁结缔组织中分别有食道腺和后肠腺，其分泌物为酸性粘多糖．肌层均为横纹肌．消化管的外膜均主要由疏构结缔组织构成．消化腺主要是肝胰腺，由无数的肝小管组成，每一小管均由位于基膜上的一层细胞构成．根据形态和功能，可把这些细胞分为四种类型：即分泌细胞（Ｂ细胞）、吸收细胞（Ｆ细胞）、储存细胞（Ｒ细胞）和胚细胞（Ｅ细胞）．     

The behaviour of Drosophila adult hindgut stem cells is controlled by Wnt and Hh signalling

The intestinal tract maintains proper function by replacing aged cells with freshly produced cells that arise from a population of self-renewing intestinal stem cells (ISCs). In the mammalian intestine, ISC self renewal, amplification and differentiation take place along the crypt-villus axis, and are controlled by the Wnt and hedgehog (Hh) signalling pathways. However, little is known about the mechanisms that specify ISCs within the developing intestinal epithelium, or about the signalling centres that help maintain them in their self-renewing stem cell state. Here we show that in adult Drosophila melanogaster, ISCs of the posterior intestine (hindgut) are confined to an anterior narrow segment, which we name the hindgut proliferation zone (HPZ). Within the HPZ, self renewal of ISCs, as well as subsequent proliferation and differentiation of ISC descendants, are controlled by locally emanating Wingless (Wg, a Drosophila Wnt homologue) and Hh signals. The anteriorly restricted expression of Wg in the HPZ acts as a niche signal that maintains cells in a slow-cycling, self-renewing mode. As cells divide and move posteriorly away from the Wg source, they enter a phase of rapid proliferation. During this phase, Hh signal is required for exiting the cell cycle and the onset of differentiation. The HPZ, with its characteristic proliferation dynamics and signalling properties, is set up during the embryonic phase and becomes active in the larva, where it generates all adult hindgut cells including ISCs. The mechanism and genetic control of cell renewal in the Drosophila HPZ exhibits a large degree of similarity with what is seen in the mammalian intestine. Our analysis of the Drosophila HPZ provides an insight into the specification and control of stem cells, highlighting the way in which the spatial pattern of signals that promote self renewal, growth and differentiation is set up within a genetically tractable model system.     

小鼠表皮生长因子对鸡胚角膜上皮组织的增殖效应

鸡胚角膜上皮组织离体培养试验表明，小鼠表皮生长因子对１０日龄鸡胚角膜上皮组织有显著的增殖效应。培养２４ｈ的实验组角膜上皮厚度较正常胚胎角膜上皮厚度增殖９．４３倍，提示鸡胚角膜上皮组织离体培养可作为评价表皮生长因子制剂活性的生物检定法。     

室内饲养松墨天牛幼虫不同肠段细菌的群落结构及功能分析

【目的】了解室内饲养松墨天牛(Monochamus alternatus)幼虫前、中、后肠段细菌的群落结构,比较不同肠段之间菌群多样性和优势菌群的差异,为揭示肠道细菌在松墨天牛获取营养、克服寄主植物化学防御的机理提供参考。【方法】分别提取室内饲养的松墨天牛4龄幼虫前、中、后肠各3组样本(每组5个前肠、5个中肠、5个后肠)的肠道DNA。利用Illumina HiSeq技术对松墨天牛幼虫肠道细菌的16S rDNA V3-V4区序列进行文库构建和高通量测序。原始序列使用Trimmomatic软件和FLASH软件分别进行质控和拼接。利用USEARCH软件对序列进行操作分类单元(OTUs)聚类,统计OTUs数量并绘制Venn图。在门和属分类水平上统计各样本的群落组成及物种丰度情况。通过Alpha多样性和Beta多样性分析反映不同样本的菌群多样性和相似性。采用PICRUSt软件预测松墨天牛幼虫肠道细菌映射到KEGG数据库上的功能,探究不同肠段细菌群落发挥的潜在功能。【结果】共获得643 404条高质量序列,在97% 相似度下将其聚类为1 614个操作分类单元(OTUs),共注释到35门、63纲、137目、250科、554属和844种。前肠OTUs最少,后肠OTUs最多,每个肠段的OTUs组成上既有相似性,也存在差异性。变形菌门(Proteobacteria)为3个肠段中最优势门;葡糖杆菌属(Gluconobacter)为前肠中最优势属,沙雷氏菌属(Serratia)为中肠的最优势属,葡糖杆菌属和沙雷氏菌属同为后肠的最优势属。Alpha多样性显示中、后肠群落多样性更丰富;Beta多样性分析表明,3个肠段的细菌群落组成存在差异,但中肠与后肠的群落组成较相近。功能预测表明,整个肠道菌群中代谢功能丰度最高,其中以糖类代谢和氨基酸代谢为主,这些功能集中在中、后肠。【结论】本实验中菌群功能是基于PICRUSt软件预测的结果,松墨天牛幼虫室内种群的前、中、后肠的细菌群落结构及不同肠段细菌的潜在功能存在差异,是由于不同肠段内的理化性质差异及其在消化中发挥的不同功能所致。肠道细菌与松墨天牛幼虫形成一个共生功能体,菌群在协助幼虫代谢物质、获取营养以及克服寄主植物化学防御方面发挥着重要作用。     

鸟类原生殖细胞的实验观察

本文系统地报导了鸟类原生殖细胞(PGC_s)的实验观察。初步提出了关于使用简易方法制备鸟类PGC_s标本的一系列技术(包括取胚、取胚血、制作胚血涂片、活胚标本、整胚装片和胚组织切片等)。实验观察表明,鸟类PGC_s可以在光镜下借助普通细胞学和组织化学相结合的方法进行鉴别。     

家兔B细胞的最早来源部位

采用光镜和电镜技术对不同发育时期的家兔肝脏、骨髓和阑尾中的Ｂ细胞及其动态变化进行了系统观察，结果表明，家兔的胚肝可能是Ｂ细胞最早发生和分化的场所，可能具有与鸟类法氏囊类似的功能．家兔阑尾的淋巴组织在出生后逐渐发育形成，与鸟类法氏囊滤泡髓质部的形成不同，因此，阑尾可能不是Ｂ细胞的最早来源部位，而是外周淋巴器官．     

A neuronal clone derived from a Rous sarcoma virus-transformed quail embryo neuroretina established culture

Neuroretina (NR) is an evagination of the central nervous system (CNS) which is composed of photoreceptors, glial (Müller) cells and horizontal, bipolar, amacrine and ganglion neuronal cells. We describe here the usefulness of Rous sarcoma virus (RSV) in the establishment of a neuronal clone from quail embryo neuroretina. When primary cultures of chick and quail embryo neuroretina cells are transformed by RSV, neuronal markers such as ribbon synapses, choline acetyltransferase (CAT) and glutamic acid decarboxylase (GAD) specific activity are present. These RSV-transformed primary cultures can be established into permanent cell lines from which neuronal clones have been isolated. One of them, clone QNR/D, can generate tetrodotoxin(TTX)-inhibitable action potentials on electrical stimulation, has a high GAD activity and binds monoclonal antibodies raised against chick embryo neuroretina. The presence of these neuronal markers suggests that the QNR/D clone is derived from cells of the amacrine or ganglionic lineage. This is the first time that a neuronal cell clone of defined origin has been obtained from the CNS. The neuronal markers of the QNR/D clone are expressed at both the permissive and the non-permissive temperatures for transformation.     

On the growth and form of the gut

The developing vertebrate gut tube forms a reproducible looped pattern as it grows into the body cavity. Here we use developmental experiments to eliminate alternative models and show that gut looping morphogenesis is driven by the homogeneous and isotropic forces that arise from the relative growth between the gut tube and the anchoring dorsal mesenteric sheet, tissues that grow at different rates. A simple physical mimic, using a differentially strained composite of a pliable rubber tube and a soft latex sheet is consistent with this mechanism and produces similar patterns. We devise a mathematical theory and a computational model for the number, size and shape of intestinal loops based solely on the measurable geometry, elasticity and relative growth of the tissues. The predictions of our theory are quantitatively consistent with observations of intestinal loops at different stages of development in the chick embryo. Our model also accounts for the qualitative and quantitative variation in the distinct gut looping patterns seen in a variety of species including quail, finch and mouse, illuminating how the simple macroscopic mechanics of differential growth drives the morphology of the developing gut.