褶纹冠蚌外套膜组织培养的分泌物的偏光显微镜观察

以淡水育珠贝中珍珠形成较快的褶纹冠蚌为材料,用相差显微镜观察组织培养的外套膜的分泌物的形成和变化,用偏光显微镜观察分泌物的双折射现象,并与活体外套膜的分泌物、贝壳的角质层、棱柱层、珍珠层的双折射现象进行比较。结果表明;离体培养的外套膜细胞不仅能产生活体细胞相同的分泌物,而且分泌物还能在培养过程中形成结晶,并逐渐生长。发现外套膜的不同部位分区培养所形成的分泌物的性状与结晶性质和活体有一致性,表明组织培养的外套膜小片具有贝体原来的组织结构、分化特征和分泌功能。     

褶纹冠蚌外套膜组织培养的分泌物的偏光显微镜…

以淡水育珠贝中珍珠形成较快的褶纹冠蚌为材料,用相差显微镜观察组织培养的外套膜的分泌物的形成和变化,用偏光显微镜观察分泌物的双折射现象,并与活体外套膜的分泌物、贝壳的角质层、棱柱层、珍珠层的双折射现象进行比较。结果表明：离体培养的外套膜细胞不仅能产生活体细胞相同的分泌物,而且分泌物还能在培养过程中形成结晶,并逐渐生长。发现外套膜的不同部位分区培养所形成的分泌物的性状与结晶性质和活体有一致性,表明组织     

淡水育珠蚌外套膜表皮细胞分泌方式的研究

用光学显微镜、相差显微镜、透射电镜、扫描电镜对３种淡水育珠蚌外套膜表皮细胞的分泌方式进行了观察研究。几种显微镜观察对比研究的结果表明,外套膜细胞的分泌方式主要有小泡式的局部分泌;液流式、缺口式的顶浆分泌和巨浆分泌;分泌物排出时连同整个细胞一起排出和形成复层表皮后外层脱落的全浆分泌。内表皮细胞以小泡式和缺口式分泌为主;外表皮细胞多为液流式分泌和全浆分泌。内表皮分泌活动表现为持续性和连续性的特点,观察相邻细胞同时可见到各种不同的分泌时相;外表皮分泌活动呈节律性和区段性的特点,分泌活动旺盛的细胞区段和无分泌活动的区段相间出现。以上结果表明内、外表皮细胞在分泌方式及分泌物性质上均有显著差异,这种差异反映了内、外表皮功能上的不同。     

河蚌外套膜一新结构及分泌物的初步研究

30年代日本小林等对珍珠贝外套膜组织结构及珍珠囊形成组织学作过观察,指出了外套膜、外表皮(壳侧表皮)与珍珠囊表皮细胞形态上存在着明显差异及细胞分泌特点;后经许多珍珠学者不断拓展,逐步形成了目前已基本上定论的珍珠分泌组织学及分泌机制1-3.作者观察到几种淡水河蚌外套膜组织及一些尚未被描述的分泌结构:这些物质与外表皮分--泌物一致,能够解释贝类组织学及珍珠(贝壳)形成机制上的一些重要问题,且在某些见解上与上述文献报道有较多出入.因此,有必要对实验结果作分析报道,供今后进一步研究作参考.       

合浦珠母贝外套膜细胞engrailed、BMP2和Smad3的mRNA表达水平的相关性

软体动物engrailed蛋白和骨形成相关蛋白对胚胎贝壳区域边界形成可能具有重要作用,engrailed还被推测为调节基质蛋白在外套膜组织区域化表达的重要调控因子．因此,弄清调控engrailed在软体动物中特征表达的分子机制有着重要的研究意义．但是,由于贝类基因组测序尚不完整,目前也没有建立获得贝类细胞系,以致于许多预测可能参与调控的基因需要通过克隆来鉴定,而且经典的研究细胞信号通路的方法也很难得到应用．目前,在中国南海广泛养殖的合浦珠母贝中,已获知其BMP2和Smad3的cDNA全长,以该贝的基因组为模板,PCR扩增获得了一段engrailed编码区片段．经软件分析,该片段含有EH4结构域,且与其他物种engrailed蛋白具有很高的同源性．研究的贝中,特别是外套膜组织中,engrailed、BMP2和Smad3三者表达之间的相关性,将有助于我们理解贝壳形成的分子机制．贝壳缺刻后半定量PCR试验结果表明,三者均参与贝壳修复,且在贝壳缺刻后的修复过程中,engrailed和Smad3的mRNA表达变化规律非常相似,提示它们之间可能存在相互影响的联系．用地塞米松(DXM)和过氧化氢(H₂O₂)分别处理原代培养的贝外套膜组织迁出细胞,实时相对定量PCR检测engrailed、BMP2和Smad3的mRNA表达水平,统计分析结果表明,三者具有显著的相关性．上述所有结果为进一步研究贝类生物矿化的发育和信号转导机制提供了新的思路和基础．     

三角帆蚌Nacrein基因克隆、蛋白提纯及其对珍珠晶体成型的影响

为研究三角帆蚌Nacrein蛋白对珍珠晶体成型的影响,通过巢式及3′-RACE PCR对三角帆蚌基质蛋白Nacrein基因3′端序列进行克隆,得到850bp碱基片段,分析表明其与合浦珠母贝同源基因序列相似度为56%,与马氏珠母贝相似度为50%。试验通过水提法初步分离出珍珠质水溶性基质蛋白,并采用快速蛋白液相色谱(fast protein liquid chromatography,FPLC)分离出分子量约为60kD的Nacrein蛋白。此外,采用配对试验设计,对试验组外套膜组织培养样品加入Nacrein蛋白,研究Nacrein蛋白对晶体成型的影响,结果显示,加入Nacrein蛋白的试验组结晶成棱形状;而未加Nacrein蛋白的对照组结晶成雪花状,而且在晶体形成的时间上,试验组也较之对照组要早。研究表明,Nacrein蛋白能加速外套膜组织分泌的钙质形成棱状结晶,从而对晶体成型产生影响。本研究为进一步研究基质蛋白对生物矿化的作用提供试验依据,对珍珠培育生产有一定指导意义。     

离体培养嗅鞘细胞促进新生大鼠螺旋神经节细胞的存活

本文旨在探索离体实验中的嗅鞘细胞(olfactory ensheathing cells,OECs)有无促进耳蜗听觉传入神经元——螺旋神经节细胞(spiral ganglion cells,SGCs)存活作用及其可能机制。取成年大鼠嗅球和新生大鼠蜗轴组织块进行OECs与SGCs的培养,采用差速贴壁法纯化培养OECs。实验分OECs与SGCs共培养组和SGCs单独培养组。倒置相差显微镜下观察OECs和SGCs生长状态,神经营养因子受体p75免疫组织化学法鉴定OECs,神经元特异性标志物βIII-tubulin标记SGCs。为了研究OECs与SGCs共培养体系中,前者促进后者存活的可能机制,共培养组中分别加入脑源性神经生长因子(BDNF,500pg/mL)和BDNF抗体(IgY型,50μg/mL),对照组为未加任何处理的共培养组,然后检查各培养组中SGCs存活数量和存活时间。结果显示,OECs贴壁培养7d后形成一细胞单层,在OECs与SGCs共培养体系中,SGCs在OECs形成的细胞单层的表面生长,并伸出长突起,呈现典型的双极神经元形态;在培养的前6天内,随着培养时间的增加,两组中的SGCs都较接种前减少,但共培养组中SGCs存活数量明显高于SGCs单独培养组(P0.01);单独培养组的SGCs数量在培养的第6天出现大幅度减少,在培养的第9天几乎没有生长;共培养组的SGCs数量未见明显变化(P0.05);共培养中加入BDNF对OECs促进SGCs存活无明显影响,而加入BDNF抗体(IgY)后存活的SGCs减少(P0.01)。本研究结果提示,OECs与SGCs共培养能够促进新生大鼠SGCs存活和突起生长,延长存活时间,OECs分泌BDNF可能是促进SGCs存活的机制之一。     

大鼠输精管平滑肌细胞的体外培养及形态学特征(英文）

建立大鼠输精管平滑肌细胞的培养方法。取大鼠输精管,剥离外膜和内膜,用组织块法进行体外培养。用抗α-SMA(anti α-smooth muscle actin)免疫组化染色的方法鉴定培养的细胞。结果显示,在倒置显微镜下观察细胞形态多样,表现为长梭形或星形,细胞伸出突起互相接触,彼此融合,部分区域细胞多层重叠,部分区域细胞单层高低起伏,呈"峰-谷"状生长。免疫组化染色鉴定呈阳性反应,用该方法所分离、培养的输精管平滑肌细胞纯度达99%以上。应用组织块法培养大鼠输精管平滑肌细胞,操作简单,结果稳定。     

背瘤丽蚌(Lamprotula leai)的外套膜组织液对皮肤创伤愈合的初步探讨

珍贵药材之一的珍珠,根据记载(时逸人,1954;Jameson,1902;小串次郎,1938),系由贝类的外套膜上皮受刺激而分泌形成。中医用其为滋养强壮和补充组织亏损之药物。因此,推想在其组织的分泌液中可能存在有促进创伤愈合的化学物质,而初步实验结果也似乎证明了这一点。 材料与方法 外套膜组织液的制备,是在无菌条件下进行的。首先将新鲜背瘤丽蚌贝壳打开,去其积水,然后取出二侧之外套膜组织,经电动组织捣碎机搅成乳糜状,并离心15分钟,取出清液,去其沉淀,继用细菌漏     

文蛤花纹的形态及形成观察

通过研究辽宁、山东、浙江、江苏和广西沿海的文蛤(Meretrix meretrix)壳的颜色及花纹特征,发现这些地区的文蛤可以分为2种类型,即花纹明显型和不明显型.辽宁和山东海域的文蛤贝壳表面具有花纹,只有极少数文蛤个体贝壳表面没有花纹;其中1龄个体贝壳表面都具有花纹.江苏地区的文蛤群体中,绝大多数个体的贝壳表面没有花纹,仅有极少数个体贝壳表面具有花纹.浙江地区的文蛤外表与山东文蛤很相似.广西地区的文蛤绝大部分没有花纹分布.外套膜组织装片确定了文蛤外套膜中的黄色素带与贝壳的色泽及花纹有关.黑色素带则与文蛤的生长状态有关,生长速度快的文蛤黑色素带浓而宽.所以,随着文蛤年龄的增长,黑色素带的颜色逐渐减弱直至消失.     

小地老虎雄蛾中胚层生殖道和附腺的细胞结构和分泌功能

通过光镜、电镜及组织化学等方法,研究了小地老虎生殖前期雄蛾中胚层生殖道和附腺的腺细胞结构和分泌功能,以及与精子形态和数量变化的关系,结果表明：(1)以缢缩位置、解剖形态、细胞结构、分泌方式、精子形态变化和数量变动为依据,将中胚层生殖道划分为修精囊、输精管、贮精囊、精包腺1～5段等8个区段;(2)中胚层生殖道和附腺具有相同的组织层次,自内向外分为单细胞上皮层、底膜、肌肉层和围膜等4层,但缺少表皮质内膜;(3)中胚层生殖道和附腺的腺细胞具有旺盛的合成和分泌蛋白质的能力,主要有内质网型和液泡型两种,前者有发达的粗面内质网和高尔基体,后者具有致密的核糖体和分泌泡;至少有4种分泌方式：即颗粒顶泌、液泡顶泌、胞质局泌和胞间分泌;修精囊、贮精囊、雄性附腺、精包腺1段的顶泌物为糖蛋白性质(PAS阳性)、局泌物为非糖蛋白性质(PAS阴性)。     

The cytology of apocrine sweat glands

Summary The apocrine sweat glands of cat and monkey have been studied by light and electron microscopy. The apocrine secretory cells of the cat are columnar cells with prominent apical cytoplasmic caps extending into the gland lumen beyond the zone of terminal bars (zonulae occludentes). Many secretory vacuoles are present in the cytoplasm, and they contain acid mucopolysaccharide demonstrable by light microscopy. These secretory vacuoles arise from prosecretory vacuoles in the region of the Golgi apparatus and are liberated from the apical cell surface as in other merocrine cells. The apocrine duct is short and the cells have scant mitochondria. The apocrine secretory cells of the monkey have secretory vacuoles similar to those of the cat but are fewer in number. The monkey apocrine cells also contain unidentified bodies similar to those seen in Langerhans cells of the epidermis. These cells liberate secretory vacuoles in a merocrine manner. Apocrine or decapitation secretion is regarded as an artifact.This investigation was supported in part by United States Public Health Service research grants GM-03784 and GM-10102 from the Institute of General Medical Sciences.     

Morphology of the Harderian gland of the Gecko, Tarentola mauritanica

The Harderian gland of the gecko, Tarentola mauritanica, was studied at the histological, histochemical, and ultrastructural levels. It is a nonlobate compound acinar gland surrounded by a thin capsule of connective tissue. Numerous connective tissue-type mast cells, ultrastructurally similar to those described in other higher vertebrates, were identified in the interstitial tissue between the acini. Pyramidal or columnar-shaped secretory glandular cells were observed in the acini. In the glandular cells, two types of structures could be distinguished on the basis of their high or low electron density. Lipid droplets were found in the cytoplasm of the Harderian gland of both sexes. Histochemical tests showed that the Harderian gland of the gecko is a seromucous gland. The secretion is essentially merocrine, although an apocrine type of secretion is sometimes observed.     

小地老虎雄性附腺细微结构和功能及高温的影响

本文通过光镜、电镜和生化分析等方法,研究了小地老虎Agrotis ypsilon(Rottemberg)雄性附腺的细微结构和功能,结果表明：(1)雄性附腺是一对管状腺,基段粉红色,中段桔红色,端段乳白色。形态分化在蛹前期完成,分泌功能在羽化后4天进入旺盛期;(2)附腺细胞属蛋白质合成型,具有旺盛的合成蛋白质的能力,胞内含有致密的粗面内质网和游离核糖体颗粒,大量的分泌液泡均匀地分布在细胞质中;(3)顶浆分泌和局部分泌是腺体的二种主要分泌方式, 前者分泌的颗粒物为糖蛋白性质(Pas阳性),后者吩泌的网状物为非糖蛋白性质(Pas阴性),二者在腺腔内呈有规则的放射状排列“4”雄性附腺分泌物具有种的特异性,小地老虎、棉铃虫和粘虫等夜蛾科昆虫分泌物的蛋白电泳谱带存在明显的种间差异,高温(32℃)抑制了雄性附腺分泌某些蛋白质的能力,从而改变精液的成分。     

Quantitative evidence for merocrine secretion in insect midgut cells

The occurrence of apical extrusions from cells has been observed on a large number of occasions in a wide variety of histological investigations. But whether the extrusions observed are a normal physiological event or are artefacts of the experimental procedures adopted has been the subject of much controversy. Lehane (1976a) described the extrusion of apical portions of the opaque zone midgut cells of Stomoxys calcitrans which contained large numbers of storage granules (merocrine secretion). This process involved the cell in the loss of cytoplasm, mitochondria and membranes as well as secretory products. If merocrine secretion is a normal physiological event rather than a fixation artefact then large falls should occur in the quantities of these components in the cell following the secretion burst. Using stereological methods the opaque zone cells were quantitatively described before and after the period in which merocrine secretion occurs (using these timings any possibility of producing apical extrusions as fixation artefacts was avoided). The results are entirely consistent with merocrine secretion having occurred. The volume of the cytoplasm falls by 23.74%, mitochondria by 30.55% and the total area of membrane in the cell by 32.59%.     

Simultaneous apocrine and merocrine secretion in the rat coagulating gland

The coagulating gland of the rat synthesizes two prevalent secretory proteins (transglutaminase and 115 K) that are discharched in a different manner, one being secreted in an apocrine fashion (transglutaminase) and the other one in a merocrine way (115 K). Differences in the intra- cellular pathway and the release of either protein were studied using immunofluorescence on semithin sections, immunoelectron microscopy of preembedding-processed chopper sections and postembedding-processed ultrathin sections of rat coagulating gland. Immunohistochemical staining using an anti-transglutaminase antibody resulted in dense labeling of the cytoplasm of secretory cells and their apical blebs, whereas the cisternae of the rough endoplasmic reticulum and the Golgi apparatus were completely unlabeled. When, on the contrary, the anti-115 K antiserum was used, dense labeling of the cisternae of the rough endoplasmic reticulum, the Golgi apparatus, and the secretory granules was seen. Intraluminal secretion was also labeled, but the secretory blebs remained unlabeled. Our findings show that, in the coagulating gland of the male rat, the two secretory proteins studied are processed in parallel, but at completely different intracellular pathways. They are released via different extrusion mechanisms. Transglutaminase is synthesized outside the endoplasmic reticulum, reaches the apical cell pole by free flow in the cytoplasm, and is released via apocrine blebs, the membranes of which appear to be derived from the apical plasma membrane. The protein 115 K, on the other hand, follows the classic route, being synthesized within the cisternae of rough endoplasmic reticulum, subsequently glycosylated in the Golgi apparatus, and released in a merocrine fashion. The mutual exclusion of the two secretory pathways and the regulation of the alternative release mechanism are still unresolved issues.     

An autoradiographic,biochemical, and morphological study of the harderian gland of the mouse

Biochemical and morphological properties of the Harderian gland of the mouse were examined by combining autoradiographic, biochemical, and electron microscopic techniques. Autoradiographs show that the radioactive carbon from [U-¹⁴C]glucose injected into the abdominal cavity is completely incorporated into the acid-insoluble substances within 30 minutes. The results of chemical analysis show that the main components of this gland are glyceryl ether diesters and phospholipids. Scanning electron microscopy shows numerous lipid droplets in the secretory cells and alveolar lumina. Myoepithelial cells lie between the secretory cell base and the basement membrane and have a basket-like distribution of processes as confirmed by hydrochloric acid and collagenase digestions. Myofilaments are demonstrated in the cytoplasm. Two types of secretory cells (A and B) comprise the alveolar epithelium and can be differentiated under the electron microscope. The cytoplasm of both contains numerous vacuoles. The vacuoles are almost empty in A cells, which are a more numerous constituent of the alveolar epithelium than B cells. However, the vacuoles of the B cells contain densely osmiophilic material. In both, cell types show a merocrine mode of secretion. Unmyelinated nerve cell endings occur in the interstices of the connective tissue, and contain clear or cored vesicles.     

Morphology and innervation of the buccal glands of the Southern Hemisphere lamprey, Geotria australis.

The buccal glands of adults of the Southern Hemisphere lamprey Geotria australis consist of a pair of small, bean-shaped, hollow sacs, embedded within the basilaris muscle in the region below the eyes and to either side of the piston cartilage. Each gland, which is lined by a simple columnar epithelium and surrounded by an incomplete layer of skeletal muscle, discharges its contents into the oral cavity via a long, narrow duct. In downstream migrating young adults, the epithelial cells are low columnar, intermediate in electron density, and contain dark-staining inclusions and numerous lipid-like droplets. After saltwater acclimation, the epithelial cells become taller and the numbers of dark-staining inclusions increase whereas those of lipid-like droplets decline. By the end of the marine phase, the epithelium is more folded and now also contains dark and light cells. The ultrastructure of the epithelium shows the characteristics of both apocrine and merocrine secretion. Although intra-epithelial nerve endings were not observed, axons and occasional neurons are present in the lamina propria. Since the skeletal muscle capsule is also well innervated and contains neurons, a local feed-back mechanism may regulate the release of buccal gland fluid by monitoring the luminal pressure. Contractions of the skeletal muscle capsule and movements of the basilaris muscle during feeding would presumably assist the movement of secretion along the duct. The secretion possesses anticoagulating and haemolytic properties.