LIGHT AND ELECTRON MICROSCOPE STUDIES OF THE PRIMARY XYLEM OF RICINUS COMMUNIS

Scott , Flora Murray , Virginia Sjaholm, and Edwin Bowler (U. California, Los Angeles.) Light and electron microscope studies of the primary xylem of Ricinus communis. Amer. Jour. Bot. 47(3) : 162-173. Illus. 1960.–The development of annular and spiral vessels in Ricinus communis has been examined under light and electron miscroscopes. Under the light microscope it is seen that spiral elements make up the bulk of the primary xylem. Pits and plasmodesmata are ubiquitous and are demonstrable in vertical and end walls. Plasmodesmata are evident in spiral thickenings. During tissue growth, intercellular spaces are formed between surrounding cells and developing vessels. These circum-vessel spaces are first lined with and later occluded by suberinlike substances. Traces of a material similar in microchemical reaction are laid down in the middle lamella. A suberin-like lining, termed in this paper the lipid lining, stainable with dimethylaminoazobenzene, occurs in mature living vessel elements. Innumerable minute fat-like droplets, refringent, and stainable with Sudan III, Sudan Black and also with osmic acid, occur in the outer cytoplasm and appear to be attached to the vessel lining by fine protoplasmic strands. They presumably are the source of the wall deposits. After the death of the protoplast, the vessel walls appear completely suberized. When contiguous cells are removed by treatment with I₂ki-H₂SO₄, their site and the site of the intercellular spaces remain marked by linear suberized ridges on the vessel wall. Annular and spiral thickenings arise as cellulose strands and begin to lignify only when the vessel reaches maximum diameter. In transverse section, the broken end of an extracted spiral thickening appears stratified. Under the electron microscope, pits and plasmodesmata are evident in procambial and in differentiating xylem elements in all walls. Annular and spiral thickening are distinguishable first as closely woven microfibrillar cellulose bands. As lignin is deposited in the microfibrillar mesh, the thickenings become dense to the electron beam. Irradiation with the full strength of the electron beam distinguishes between spiral thickenings in younger and older vessels. Older spirals remain apparently unchanged. Younger spirals instantly swell, volatilize in part, and assume a moniliform outline. The bead-like swellings consist of a matrix partly transparent to the electron beam and an internal framework of a material comparatively dense to the electron beam. Similar intense irradiation differentiates between younger and older vessel linings. Older linings appear unchanged, while the younger react violently, volatilize in part and stabilize as an irregular coagulum set in a basic mesh. The volatilized substances appear as granules on lining surface or on substrate. The changing microfibrillar pattern of the cell wall is observed from the procambial stage to the final deposition of the lipid vessel lining.     

A STUDY OF THE STRUCTURE OF SPLENIC SINUSES IN MAN AND IN THE ALBINO RAT WITH THE LIGHT MICROSCOPE AND THE ELECTRON MICROSCOPE

The splenic sinuses in the spleens of 5 human beings and 7 albino rats have been studied in the light microscope and electron microscope after fixation in Dalton's fluid and Palade's fluid and embedding in n-butyl methacrylate. Splenic sinuses are tortuous vascular channels of large but variable diameter which represent the first venous vessels in the spleen and make up almost the entire red pulp in man and in rats. These vessels are composed of reticulo-endothelial cells flattened to endothelial form and sheathed by a netted reticulum. The luminal surface of the endothelium is made highly irregular by delicate and variable cytoplasmic protrusions, slender corridors separating adjacent endothelial cells, anastomotic openings to other sinuses, bulgings of entire cells, and even thrusts of endothelium spanning the sinai lumen. The supporting reticulum presents a well developed latticed appearance in tangential sections of sinuses, but in most cuts is punctate or linear. The reticulum is composed of strands without limiting membranes, which, in substance, are amorphous and resemble basement membrane. Material identical in appearance to the substance of the reticulum may be present in the endothelium, suggesting that the reticulum is formed by endothelial cells. The endothelium also contains deposits of presumed ferritin and hemosiderin. The extreme luminal bulgings of endothelium suggest production of circulating monocytes or lymphocytes by detachment of endothelial cells. Sinuses are patent and collapsed to varying degrees. Patent sinuses are separated by collapsed sinuses and these collapsed sinuses appear to constitute splenic (Billroth) cords.     

LIGHT AND ELECTRON MICROSCOPE STUDIES OF THE CELL WALL STRUCTURE OF THE ROOT HAIRS OF RAPHANUS SATIVUS

Dawes , Clinton J., and Edwin Bowler . (U. of California, Los Angeles.) Light and electron microscope studies of the cell wall structure of the root hairs of Raphanus sativus. Amer. Jour. Bot. 46(8): 561–565. Illus. 1959.—The structure and development of the cell wall of the root hair of Raphanus sativus were studied under the light and electron microscopes. The outer layer of the root hair consists of mucilage which covers the entire hair and forms a thick cap at the tip. Beneath the mucilage a thin cuticle covers the inner layers of the cell wall. These layers consist of cellulose microfibrils, varying in pattern, in a granular matrix, presumably pectic in nature. The microfibrils of the outer layer, apparently laid down at the tip, are reticulate in arrangement. In mature regions of the root hair, the wall is thickened by an inner layer of parallel and longitudinally orientated microfibrils. Pores in the cellulose wall are evident and increase in number and size near the base of the hair.     

LIGHT AND ELECTRON MICROSCOPE STUDIES OF MORPHOLOGICAL CHANGES OF MITOCHONDRIA DURING SPERMATOGENESIS IN THE GRASSHOPPER

1. Observations on the morphological changes of mitochondria preparatory to the formation of the nebenkern, as well as changes within the nebenkern, are reported. 2. Mitochondria enlarge and divide during the meiotic divisions. 3. The mitochondria of the spermatid elongate, become filamentous, form a crescent, and partially encircle the nucleus. 4. Nodes which develop on either end of the crescent become entwined as they move toward each other. 5. The two nodes coalesce to form a filamentous or early type nebenkern which is described by others as chromophilic. 6. Internal rearrangement and partial dissolution of the filaments result in the development of the late or chromophobic nebenkern which separates into two distinct bodies. 7. The nebenkern moieties send out processes toward the centrosome, and after making contact, elongate and occupy part of the space between the tail filaments and sheath of the spermatozoon.     

疥螨消化系统的显微和超微结构观察

疥螨消化系统由咽、食道、中肠、１对侧囊、结肠、直肠、肛门以及唾腺组成。应用透射电镜,可将中肠和侧囊壁上皮细胞分为鳞状细胞期、柱状细胞期、核变性圆细胞期和全变性细胞期等４种不同生理功能状态。可将唾腺细胞分为Ｉ期、Ⅱ期和Ⅲ期等３种不同生理功能时期。食道和中肠及侧囊内含物为絮状物。直肠前段肠壁具有较多微管。     

钉螺体内日本血吸虫尾蚴发育期的形态及其扫描电镜观察

本文通过活体标本、连续切片、全封标本及扫描电镜观察了大陆品系日本血吸虫Schistosomajaponicum子胞蚴体内尾蚴发育期的形态。日本血吸虫尾蚴发育分为五期,即胚细胞期、胚球期、尾蚴雏体期、成熟前期和成熟期。与曼氏血吸虫尾蚴发育进行比较,日本血吸虫尾蚴在胚球期无极化现象;尾蚴雏体期的尾芽形态多非圆球形,尾叉形成较早。钻腺及焰细胞均在此期发生,较曼氏血吸虫尾蚴早。除了上述头器、头腺、钻腺、焰细胞及消化器官进一步发育外,体内散在胚细胞结集为生殖始基,体表感觉乳突分化与体尾肌细胞的发育,尾蚴从成熟前期过渡到成熟期。 电镜首次原位观察尾蚴发育在子胞蚴育腔内的自然状态。尽管偶尔可见到胞蚴体壁与胚元之间某些结构联系,但大部份的胚元各自独立混杂在育腔之中。此外尚注意到尾蚴体棘发生在成熟前期,但其密度较成熟尾蚴小,而其大小却比成熟尾蚴的大。尾蚴头器上感觉乳突和围褶可能先于内部腺体细胞的分化。     

LIGHT AND ELECTRON MICROSCOPE OBSERVATIONS OF ALGIROSPHAERA ROBUSTA (PRYMNESIOPHYCEAE)1

The coccolithophore Algirosphaera robusta (Lohmann) R. E. Norris was isolated into laboratory culture for the first time. This species is of particular interest as the first deep photic coccolithophore to be cultured, the only member of the Rhabdosphaeraceae to have been successfully isolated, and the first coccolithophore with a coccolith structure including a complex disjunct central area to have been studied in detail. Observations on the culture strain supported the previous inference that the commonly recognized species A. robusta, A. oryza Schlauder, and A. quadricornu (J. Schiller) R. E. Norris are conspecific. However, A. meteora (Müller) R. E. Norris and A. cucullata (Lecal‐Schlauder) J. R. Young, Probert et Kleijne were recognized as discrete species. Coccolith rim formation in A. robusta follows the pattern of biomineralization documented in other heterococcoliths and was suggested to be universal. However, the prominent central hood had a unique ultrastructure and appeared to be formed by a distinctively different biomineralization mode. We suggest that this can provide a key to reinterpreting homology in coccolith structure and that this species is a promising target for comparative biochemical and genomic studies of biomineralization. In terms of cell ultrastructure, A. robusta exhibited marked similarities to Syracosphaera pulchra Lohmann, and a close evolutionary relationship between the families Rhabdosphaeraceae and Syracosphaeraceae is suggested.     

A CORRELATED LIGHT AND ELECTRON MICROSCOPE STUDY OF THE NUCLEOLAR MATERIAL DURING MITOSIS IN VICIA FABA

Root meristematic cells of Vicia faba were examined, with both light and electron microscopes, in order to study the behaviour of the nucleolar material during the mitotic process. Under light microscopy, the preprophase nucleolus is seen to consist of a densely stained material in which are embedded several unstained vacuole-like structures of varying size. The electron microscope reveals that the dense nucleolar material is formed of two structurally distinct components, each segregated into irregularly shaped zones blending with one another. One of these components is represented by 150 A granules which, in places, are arranged into thread-like structures approximately 0.1 µ in diameter; the other component apparently consists of fibrils 60 to 100 A in diameter. The large and medium sized intranucleolar vacuoles contain loosely scattered granules and fibrils similar to those just described. The granular and fibrillar components of the denser portion of the nucleolus persist as such during prophase and disperse throughout the nuclear cavity at the time of nucleolar disintegration. After nuclear membrane breakdown, these granules and fibrils, as well as those of the nucleoplasm, mix freely with similar elements already present within the forming spindle. No evidence has been obtained that, during or after nucleolar disintegration, the structural components of the nucleolus become associated as such with the chromosomes to form an external or internal matrix. Our observations suggest the existence, of a matrix substance within late prophase, metaphase, and anaphase chromosomes, the fine structure of which bears strong resemblance to that of their constituent coiled chromonemata. Data are presented, moreover, that indicate that part of this matrix substance, presumably formed at some time during prophase, is released from the chromosomes during their anaphasic movement. A number of observations indicate that the main bulk of the next nucleolus is derived from a prenucleolar fibrillogranular material, arranged into thread-like structures some 0.1 µ in diameter, which collect in the interchromosomal spaces during early and midtelophase. Finally, our data would seem to favour the view that most of this prenucleolar material results from a resumption of the synthetic activity of the early and midtelophase chromosomes rather than from a mere shedding of a preexisting matrix substance.     

ENZYME-LABELED ANTIBODIES FOR THE LIGHT AND ELECTRON MICROSCOPIC LOCALIZATION OF TISSUE ANTIGENS

Enzymes, either acid phosphatase or horseradish peroxidase, were conjugated to antibodies with bifunctional reagents. The conjugates, enzymatically and immunologically active, were employed in the immunohistochemical localization of tissue antigens utilizing the reaction product of the enzymatic reaction as the marker. Tissues reacted with acid phosphatase-labeled antibodies directed against basement membrane were stained for the enzyme with Gomori's method, and those reacted with peroxidase-labeled antibody were stained with Karnovsky's method. The reaction products of the enzymes localized in the basement membrane. Unlike the preparations of the fluorescent antibody technique, enzyme-labeled antibody preparations were permanent, could be observed with an ordinary microscope, and could be examined with the electron microscope. In the latter, specific localization of antibody occurred in the basement membrane and in the endoplasmic reticulum of cells known to synthesize basement membrane antigens. The method is sensitive because of the amplifying effect of the enzymatic activity. The ultrastructural preservation and localization were better with acid phosphatase-labeled antibody than with peroxidase-labeled antibody, but acid phosphatase conjugated antibody was unstable and difficult to prepare. Peroxidase-antibody conjugates were stable and could be stored for several months at 4°C, or indefiniely in a frozen state.     

云南树鼩肝脏结构的光,电镜观察

本文报告了树肺脏的一般结构和超微结构。与人和灵长目相似，其肺实质也是由导气部和呼吸部构成。但不同的是其细支气管粘膜形成很高的皱襞。在电镜下Ｃｌａｒａ细胞电子密度高，顶部胞质中含有大量膜包颗粒，这些结构与大白鼠和家兔的结构相似。许多毛细血管外方都包绕着基膜和肺泡Ⅰ型上皮细胞的胞质。气血屏障由肺泡上皮细胞、融合的基膜和内皮细胞胞质构成。说明树肺脏不但是呼吸器官，也是一些激素和介质产生及代谢的重要器官。本文为研究树的正常生理功能及分类提供形态学资料。     

A LIGHT AND ELECTRON MICROSCOPE STUDY OF INTERLAMELLAR POLYGLUCOSIDE BODIES IN OSCILLATORIA CHALYBIA

A light and electron microscopic study of interlamellar granules in Oscillatoria chalybia was made to determine their physiological nature. Oscillatoria chalybia was cultured under continuous light in media of high nitrogen content, moderate nitrogen content and low nitrogen content. Cultures growing vigorously in a medium of moderate nitrogen content were placed in darkness for an additional 96 hr. Periodic acid-Schiffs reagent tests were made on specimens from these 4 cultural conditions. Electron microscopic studies of interlamellar granules were correlated with the cytochemical tests. It is shown that diastase digestion will eliminate the PAS-positive substance and the interlamellar granules. Conclusions are that the interlamellar granules are polyglucoside in nature and that they vary in number and size with available nitrogen, light intensity and age of culture in such a way as to indicate that they are food reserves.     

ELECTRON MICROSCOPE OBSERVATIONS ON THE STRUCTURE AND DISCHARGE OF THE STENOTELE OF HYDRA

Sections of the stenotele type of nematocyst of Chlorohydra hadleyi have revealed that the stenotele, upon firing, completely everts its stylets and spines and the long, thin tubule, much as the eversion of the tubule of the nematocyst of the jewel anemone (Picken, 1953; Robson, 1953). Alternative mechanisms for supplying the energy necessary to forcefully discharge the stenotele contents are discussed as well as the possible significance of several regions containing highly ordered periodic structure. The origin of nematocysts as kinetosomal derivatives is discussed as a possibility suggested by the symmetry of the stenotele contents and the structure, location, and function of the cnidocil.     

LIGHT AND ELECTRON MICROSCOPE OBSERVATIONS ON THE TUBE-DWELLING DIATOM AMPHIPLEURA RUTILANS (TRENTEPOHL) CLEVE

The tubular sheath of Amphipleura rutilans is composed of small fibrils 20–25 A in diameter. The endotubular environment is further partitioned by individual cell sheath formation. The tube boundary seems to be an effective barrier against damage from extreme salinity changes, but does not prevent passage of EDTA, neutral red, surfactants, or caffeine. Terminal swellings develop on growing tips during extremely cold periods. These give rise to 2 or more separate growing tips which proceed to form regular tubes. There was no evidence for sexual reproduction or auxospore formation within the terminal swellings, even though adjacent swellings fused. Bacterial, algal, and fungal epiphytes were abundant on the extratubular surface. Hollow lipid bodies occurred in many cells. A rimring strand of presumed DNA occurs in the chloroplast. It is proposed that the tube material serves not only as a protective boundary but also functions as an active nutrient trap.