两种夜间活动壁虎的视网膜光学显微镜和电子显微镜观察

自然界大多数壁虎在夜间活动。它们的视网膜感受细胞呈杆状,通常称为杆细胞。但是这些杆细胞仍有一些锥细胞的形态特征。Walls(1934;1942)从种系发生学上研究了许多爬行动物的形态,指出现在的夜行壁虎的杆细胞是从其祖先白昼蜥蜴的锥细胞演变来的。Walls演变学说提出后,不仅得到了形态学研究的支持(Underwood 1951;Tansley1959;1964;Pedler,1964;Dunn,1966),他们所描述的夜行壁虎感受细胞的这个特性,更吸引了视觉研究工作者的注意。近十多年来,关于大壁虎视色素(Liebman,1972;     

锰离子和门冬酸对夜行壁虎视细胞终端摄入HRP的影响

我们研究了锰离子和门冬酸对一种夜行壁虎(Gekko japonicus)视细胞终端摄入HRP的影响。(1)暗适应网膜的视细胞摄入许多HRP,被HRP标记的突触小泡约为14％。光照压抑HRP摄入,标记小泡数降至2％左右。(2)5mM Mn~( )抑制视细胞摄入HRP,为HRP标记的突触泡小水平与明适应时相似。(3)100mM门冬酸促进视细胞摄入HRP,特别是为HRP标记的囊泡显著增多。(4)ERG记录表明,Mn~( )的作用在多数例子中显示出对b波有较a波为强的选择性抑制作用。看来Mn~( )对摄入HRP的抑制效应主要是由于阻断突触传递,Mn~( )是否对视细胞也存在某种直接作用尚不能肯定。(5)本工作支持目前普遍所认为的关于脊椎动物视细胞信息传递模型学说。     

大壁虎不同地理居群的遗传变异与分化

以线粒体细胞色素b(Cytb)基因作为分子标记,对中国广西12个地区,以及越南和老挝大壁虎(Gekko gecko)进行序列测定,获得Cytb基因424bp的序列片段,共有7个单倍型。以白脊壁虎和沙虎为外群,用邻接法和最大简约法构建了大壁虎不同地理种群的系统发育关系,其结果显示中国广西4个不同单倍型黑大壁虎之间的平均遗传距离为0.20%—1.20%,越南红大壁虎与老挝红大壁虎之间的平均遗传距离为0.50%,广西宁明红大壁虎与越南红大壁虎和老挝红大壁虎之间平均遗传距离分别为1.70%和2.20%。广西黑大壁虎种群与红大壁虎种群之间的平均遗传距离为8.60%—9.50%,达到了亚种或种分化的差异。     

大壁虎不同地理居群的遗传变异与分化

以线粒体细胞色素b (Cyt b) 基因作为分子标记，对中国广西12个地区，以及越南和老挝大壁虎(Gekko gecko) 进行序列测定，获得Cyt b基因424 bp的序列片段，共有7个单倍型。以白脊壁虎和沙虎为外群，用邻接法和最大简约法构建了大壁虎不同地理种群的系统发育关系，其结果显示中国广西4个不同单倍型黑大壁虎之间的平均遗传距离为0.20%—1.20%，越南红大壁虎与老挝红大壁虎之间的平均遗传距离为0.50%，广西宁明红大壁虎与越南红大壁虎和老挝红大壁虎之间平均遗传距离分别为1.70%和2.20%。广西黑大壁虎种群与红大壁虎种群之间的平均遗传距离为8.60%—9.50%，达到了亚种或种分化的差异。     

蛤蚧的视网膜电图(ERG)及其明、暗适应特性

蛤蚧是一种昼伏夜出的大壁虎,盛产于我国的广西、云南一带。牠的眼睛无眼睑,而由眼罩(Spectacle)所代替。瞳孔呈垂直窄缝形,收缩时在窄缝上可见四个小孔。其视网膜没有明显的中央凹,视细胞在与瞳孔垂直方向上排列成行。感受细胞层由纯视杆细胞构成,这些细胞在形态学上既具有视杆型的特征,又具有视锥型的特征。     

长时间暗适应和弱背景光对鲤鱼视网膜视锥水平细胞的影响——形态和生理的相关研究

本工作利用电子显微镜及细胞内记录技术研究了在暗适应和弱背景光下鲤鱼视锥与水平细胞间突触部位超微结构及视锥水平细胞光反应的变化。在长时间暗适应(>2h)后,视锥水平细胞对光反应受到强烈压抑,其突起末端则外形光滑、圆钝。在施加弱背景光(15min)后,这些细胞的反应显著增大,其末端出现大量深陷于视锥小足内部的刺形结构,这种刺形结构增加了视锥与水平细胞间的突触传递效率,可能是视锥水平细胞光反应性增高的形态学基础。     

视网膜中央凹视锥细胞椭球体光学作用分析

用光波导理论分析了视网膜中央凹视锥细胞椭球体的光学作用,发现中央凹视锥细胞椭球体能够有效减小中央凹视锥细胞内段的肌样体和外段之间的耦合损耗;能够匹配中央凹视锥细胞内段的肌样体和外段的模场,具有模场尺寸转换器的作用;椭球体的存在是中央凹视锥细胞实现高灵敏度和高分辨率的选择。     

壁虎药用价值的研究进展

壁虎是对蜥蜴亚目壁虎科所有蜥蜴的通称,其作为中药材使用具有悠久的历史。结合近年来国内外对壁虎药用价值的研究成果,综 述了壁虎在抗肿瘤、调节机体免疫力和促进再生等方面的活性及相关机制研究进展,旨在为壁虎药用价值的深入探索提供参考。     

果蝇视锥细胞聚集形态形成机理的研究

果蝇视锥细胞聚集形态与液态泡沫一致, 视锥细胞N-钙黏附分子间的黏附力可以类比为泡沫的表面张力. 把果蝇视锥细胞排布近似为两维由等同气泡组成的约束边界液态泡沫, 采用两维液态泡沫动力学模型模拟了2~6个视锥细胞稳定聚集形态的形成过程, 提出了在视锥细胞面积恒定的情况下周长达到极小值时(即表面能量极小)细胞即达到稳定聚集的假设, 在此假设基础上确定的排布方式与文献[4]报道的实验观测完全一致, 并预报了一个更稳定的6细胞聚集形态; 计算表明, 果蝇视锥细胞稳定聚集时, 表面能与细胞个数成正比     

AMPA对视网膜L型水平细胞所接收的红、绿敏视锥信号的不同调制作用

在离体灌流的鲫鱼视网膜,应用细胞内记录技术,研究了谷氨酸受体激动剂——AMPA对L型水平细胞(LHC)的作用.AMPA压抑LHC由红敏视锥驱动的反应,却增强由绿敏视锥驱动的反应.此效应可为AMPA受体特异的拮抗剂(GYKI53655)所逆转,表明仅有AMPA受体参与其中.印防己毒素(PTX)或双氢红藻氨酸(DHK)存在时,依然可以观察到这种效应,提示自LHC向视锥的负反馈和视锥谷氨酸转运体在此效应中不起显著作用.AMPA的不同调制作用提示,LHC上可能存在不同特性的AMPA受体亚型介导红敏和绿敏视锥的信号传递.     

THE RENEWAL OF ROD AND CONE OUTER SEGMENTS IN THE RHESUS MONKEY

The renewal of retinal rod and cone outer segments has been studied by radioautography in rhesus monkeys examined 2 and 4 days after injection of leucine-³H. The cell outer segment consists of a stack of photosensitive, membranous discs. In both rods and cones some of the newly formed (radioactive) protein became distributed throughout the outer segment. Furthermore, in rods (but not in cones), there was a transverse band of concentrated radioactive protein slightly above the outer segment base 2 days after injection. This was due to the formation of new discs, into which labeled protein had been incorporated. At 4 days, these radioactive discs were located farther from the outer segment base. Repeated assembly of new discs had displaced them away from the basal assembly site and along the outer segment. Measurements of the displacement rate indicated that each retinal rod produces 80–90 discs per day, and that the entire complement of outer segment discs is replaced every 9–13 days. To compensate for the continual formation of new discs, groups of old discs are intermittently shed from the apical end of the cell and phagocytized by the pigment epithelium. Each pigment epithelial cell engulfs and destroys about 2000–4000 rod outer segment discs daily. The similarity between visual cells in the rhesus monkey and those in man suggests that the same renewal processes occur in the human retina.     

真鲷视网膜结构及其视觉特性研究：Ⅱ视网膜光感受细胞超微结构研究

对真鲷光感受细胞的超微结构进行观察,结果表明：视杆外段膜盘为游离膜盘,视锥外段膜盘则为连续的膜结构,视锥和视杆均含有连接纤毛和辅助外段。花萼状突起起源于内段。椭体内充满线粒体,无球状小体。双锥椭圆体并生膜为六层,视锥内段无鳍状突起,视锥突触带,在明适应视网膜中数量增多,在暗适应视网中数量减少,视杆突触带在这两种适应网膜中数量不变,每一杆小球只有一个突触带,而锥小足有４－６个突触带。     

THE RENEWAL OF PHOTORECEPTOR CELL OUTER SEGMENTS

The utilization of methionine-³H by retinal photoreceptor cells has been studied by radioautographic technique in the rat, mouse, and frog. In all three species, the labeled amino acid is concentrated initially in the inner segment of the cell. Within 24 hr, the radioactive material is displaced to the base of the outer segment, where it accumulates as a distinct reaction band. The reaction band then gradually moves along the outer segment and ultimately disappears at the apex of the cell, which is in contact with the retinal pigment epithelium. These findings are interpreted to indicate that the photoreceptor cell outer segment is continually renewed, by the repeated lamellar apposition of material (membranous discs) at the base of the outer segment, in conjunction with a balanced removal of material at its apex. The outer segment renewal rate is accelerated in frogs when ambient temperature is raised, and is elevated in both frogs and rats when the intensity of retinal illumination is increased.     

Differential effects of protease digestion on photoreceptor lectin binding sites

The use of lectin cytochemistry together with proteolytic digestion techniques to partially characterize lectin binding sites of several intracellular compartments in frog photoreceptors was studied. Uniform access of reagents to all intracellular compartments was obtained by performing the experiments directly on semithin sections of retinal tissue embedded in a hydrophilic plastic resin. Protease pretreatment of sections of Xenopus laevis eyecup leads to a loss of wheat germ agglutinin (WGA) binding sites from most of the rod outer segment. Under experimental conditions used here, cone outer segment WGA binding sites are resistant to proteolytic digestion. Another major difference between rod and cone under segments is that rod outer segments are heavily labeled with succinylated WGA, whereas cone outer segments are barely labeled except for a region of intense staining thought to be at the connecting cilium. WGA binding sites in the shed outer segment tip (phagosome) are also relatively resistant to proteolytic digestion, as is the tip region of a few rod outer segments. This difference in lectin binding properties between the bulk of the outer segment membrane and the shed outer segment membrane is the only distinction we have observed between the two compartments in terms of their glycoconjugates. These results may be useful in terms of designing experiments to isolate cone and rod outer segments separately. They indicate that a change in outer segment glycoconjugates may accompany the shedding and phagocytosis events, as previously suggested, but this change does not necessarily involve the addition of saccharides to outer segment glycoproteins.     

中国大鲵视网膜的光镜和扫描电镜研究

用光镜和扫描电镜观察了大鲵视网膜各类细胞的形态及分布, 对视细胞和节细胞进行计数。视网腊中三个核层及两个网状层分布均匀,无中央凹。每张视网膜的视细胞总数约130000,节细胞约8000,视杆与视锥之比为8.5：1。扫描电镜下,视杆外节表面的小叶间沟清晰;视杆视锥外节均有从内节伸出的20-30条萼状突起;核周体表面亦有20-30条细胞质突起。文中还报道了幼体视细胞的形态及密度。讨论了上述结构的机能。     

Localization of peripherin/rds in the disk membranes of cone and rod photoreceptors: relationship to disk membrane morphogenesis and retinal degeneration

The outer segments of vertebrate rod photoreceptor cells consist of an ordered stack of membrane disks, which, except for a few nascent disks at the base of the outer segment, is surrounded by a separate plasma membrane. Previous studies indicate that the protein, peripherin or peripherin/rds, is localized along the rim of mature disks of rod outer segments. A mutation in the gene for this protein has been reported to be responsible for retinal degeneration in the rds mouse. In the present study, we have shown by immunogold labeling of rat and ground squirrel retinas that peripherin/rds is present in the disk rims of cone outer segments as well as rod outer segments. Additionally, in the basal regions of rod and cone outer segments, where disk morphogenesis occurs, we have found that the distribution of peripherin/rds is restricted to a region that is adjacent to the cilium. Extension of its distribution from the cilium coincides with the formation of the disk rim. These results support the model of disk membrane morphogenesis that predicts rim formation to be a second stage of growth, after the first stage in which the ciliary plasma membrane evaginates to form open nascent disks. The results also indicate how the proteins of the outer segment plasma membrane and the disk membranes are sorted into their separate domains: different sets of proteins may be incorporated into membrane outgrowths during different growth stages of disk morphogenesis. Finally, the presence of peripherin/rds protein in both cone and rod outer segment disks, together with the phenotype of the rds mouse, which is characterized by the failure of both rod and cone outer segment formation, suggest that the same rds gene is expressed in both types of photoreceptor cells.     

Correlation of rhodopsin biogenesis with ultrastructural morphogenesis in the chick retina

The developing chick retina from stages 39-45 has been examined by biochemical and electron microscope techniques. The levels of rhodopsin contained in the maturing chick retina were evaluated by detergent extraction and correlated with rod outer segment formation. It was found that the appearance of rhodopsin in significant levels preceded outer segment formation by at least 2 days, thus implying that rhodopsin is synthesized in the receptor cell inner segment and translocated to the outer limb when disk membrane biogenesis occurs. The level of rhodopsin continues to rise as the rod outer segment develops. Development of both rods and cones originates and proceeds most rapidly in the fundus or central region and proceeds toward the periphery. In general, rod outer segments were noted to develop far more rapidly than cone outer segments.     

Evolution of visual pigments in geckos

Most geckos are nocturnal forms and possess rod retinas, but some diurnal genera have pure-cone retinas. We isolated cDNAs encoding the diurnal gecko opsins, dg1 and dg2, similar to nocturnal gecko P521 and P467, respectively. Despite the large morphological differences between the diurnal and nocturnal gecko photoreceptor types, they express phylogenetically closely related opsins. These results provide molecular evidence for the reverse transmutation, that is, rods of an ancestral nocturnal gecko have backed into cones of diurnal geckos. The amino acid substitution rates of dgl and dg2 are higher than those of P521 and P467, respectively. Changes of behavior regarding photic environment may have contributed to acceleration of amino acid substitutions in the diurnal gecko opsins.     

Proteomics of photoreceptor outer segments identifies a subset of SNARE and Rab proteins implicated in membrane vesicle trafficking and fusion

The outer segment is a specialized compartment of vertebrate rod and cone photoreceptor cells where phototransduction takes place. In rod cells it consists of an organized stack of disks enclosed by a separate plasma membrane. Although most proteins involved in phototransduction have been identified and characterized, little is known about the proteins that are responsible for outer segment structure and renewal. In this study we used a tandem mass spectrometry-based proteomics approach to identify proteins in rod outer segment preparations as an initial step in defining their roles in photoreceptor structure, function, renewal, and degeneration. Five hundred and sixteen proteins were identified including 41 proteins that function in rod and cone phototransduction and the visual cycle and most proteins previously shown to be involved in outer segment structure and metabolic pathways. In addition, numerous proteins were detected that have not been previously reported to be present in outer segments including a subset of Rab and SNARE proteins implicated in vesicle trafficking and membrane fusion. Western blotting and immunofluorescence microscopy confirmed the presence of Rab 11b, Rab 18, Rab 1b, and Rab GDP dissociation inhibitor in outer segments. The SNARE proteins, VAMP2/3, syntaxin 3, N-ethylmaleimide-sensitive factor, and Munc 18 detected in outer segment preparations by mass spectrometry and Western blotting were also observed in outer segments by immunofluorescence microscopy. Syntaxin 3 and N-ethylmaleimide- sensitive factor had a restricted localization at the base of the outer segments, whereas VAMP2/3 and Munc 18 were distributed throughout the outer segments. These results suggest that Rab and SNARE proteins play a role in vesicle trafficking and membrane fusion as part of the outer segment renewal process. The data set generated in this study is a valuable resource for further analysis of photoreceptor outer segment structure and function.     

MORPHOGENESIS OF THE RETINAL RODS : AN ELECTRON MICROSCOPE STUDY

The morphogenesis of the retinal rods has been studied with the electron microscope in white mice from birth up to the 16th day of age. Observations have been mainly concentrated on specimens of the 8th and 12th days and on the differentiation of the inner and outer segments of the retinal rods. In the morphogenesis of the outer segment three main stages have been considered. The first stage consists in the development of a primitive cilium projecting from a bulge of protoplasm which constitutes the primordium of the inner segment. A basal body, nine pairs of peripheral filaments, a surface membrane, and a matrix filled with a fine vesicular material have been recognized as components of the primitive cilium. The vesicles are called "morphogenetic material" because it is believed that they represent the macromolecular primordium of the rod sacs of the future outer segment. The second stage corresponds to the great enlargment of the apical region of the primitive cilium due to the rapid building up of the lamellar material of the rod sacs. The primitive rod sacs appear to be connected with the ciliary filaments. The basal portion of the primitive cilium remains undifferentiated and constitutes the connecting cilium of the adult rod (1). The third stage consists in the remodelling and reorientation of the rod sacs into their permanent transverse disposition. This process starts in the middle portion of the outer segments and proceeds towards both extremities which can be considered as zones of growth of the outer segment. The inner segment is at the beginning a bulge of protoplasm containing unoriented mitochondria, a basal body, a small Golgi zone, and numerous dense particles. Then this region becomes elongated and the different components assume the stratified disposition found in the adult (1). The demonstration that the entire outer segment of the rod cell is the result of the differentiation of a primitive cilium is discussed in view of the conflicting interpretations found in the literature. The possible macromolecular mechanisms that may be involved in the submicroscopic morphogenesis of the rod sacs are discussed and the possible role of the morphogenetic material is considered. The results described in this paper confirm and extend the interpretation of the submicroscopic morphology of the adult rod cell as presented in a previous paper (1).