Cellular control over spicule formation in sea urchin embryos: A structural approach

The spicules of the sea urchin embryo form in intracellular membrane-delineated compartments. Each spicule is composed of a single crystal of calcite and amorphous calcium carbonate. The latter transforms with time into calcite by overgrowth of the preexisting crystal. Relationships between the membrane surrounding the spiculogenic compartment and the spicule mineral phase were studied in the transmission electron microscope (TEM) using freeze-fracture. In all the replicas observed the spicules were tightly surrounded by the membrane. Furthermore, a variety of structures that are related to the material exchange process across the membrane were observed. The spiculogenic cells were separated from other cell types of the embryo, frozen, and freeze-dried on the TEM grids. The contents of electron-dense granules in the spiculogenic cells were shown by electron diffraction to be composed of amorphous calcium carbonate. These observations are consistent with the notion that the amorphous calcium carbonate-containing granules contain the precursor mineral phase for spicule formation and that the membrane surrounding the forming spicule is involved both in transport of material and in controlling spicule mineralization.     

Multiscale structure of calcite fibres of the shell of the brachiopod Terebratulina retusa

The shells of rhynchonelliform brachiopods have an outer (primary) layer of acicular calcite and an inner (secondary) layer of calcite fibres which are parallel to the shell exterior. Atomic force microscopy (AFM) reveals that these fibres are composed of large triangular nanogranules of about 600-650 nm along their long axis. The nanogranules are composites of organic and inorganic components. As the shell grows, the fibres elongate with the calcite c-axis perpendicular to the fibre axis as demonstrated by electron backscatter diffraction (EBSD). Thus, despite being a composite structure comprising granules that are themselves composites, each fibre is effectively a single crystal. The combination of AFM and EBSD reveals the details of the structure and crystallography of these fibres. This knowledge serves to identify those aspects of biological control that must be understood to enable comprehension of the biological control exerted on the construction of these exquisite biomineral structures.     

Electron microscopic observation of cell wall structure during appressorium formation in Colletotrichum lagenarium

Summary The formation of cell walls during the appressorium formation inColletotrichum lagenarium was observed by electron microscope on the materials prepared by replicas and sectioning. The outer layer of conidia cell walls ruptured at the time of germination and the inner layer bulged out to form a germ tube. The germ tubes and primordia of appressoria had smooth surface and were consisted of one-layered cell wall. However, as the appressorium matured, the electron dense materials appeared on the outer surface of the cell wall which grew into granules. These granules are believed to form the outer layer of appressoria. The under side of the appressorium in contact with the glass surface showed a round pore.Contribution No. 191.     

CRYSTAL PROPERTY OF THE LARVAL SEA URCHIN SPICULE*

A pair of pluteus skeletal spicules arises from a pair of calcareous granules via the triradiate form. In polarized light, each spicule behaves as though carved out of a single crystal of magnesian calcite. The optic axis lies perpendicular to the plane of the triradiate and parallel to the body rod of the pluteus. However, in the scanning electron microscope, the spicule surface appeared smooth or somewhat spongy and manifested no crystal faces. Neither etching nor fracturing revealed underlying crystalline texture. Nevertheless, rhombohedral calcite crystals could be grown epitaxially onto isolated spicules immersed in a medium containing CaCl₂ and NaHCO₃. The optic axes of all crystals coincided with the optic axis of the spicule on which they were grown. Corresponding faces of the crystals were all aligned parallel to each other despite the complex shape of each spicule. Where the left and right spicules joined, two mutually tilted sets of crystals were observed but not crystals of intermediate orientation. Thus, the sea urchin larval spicule is built from a stack of molecularly contiguous microcrystals but its overall shape is generated by the mesenchyme cells independent of the magnesian calcite crystal habit.     

Surface Ultrastructure of Some Tests of Recent Foraminiferida from the Dry Tortugas, Florida

SYNOPSIS. The surface ultrastructure of 10 species of benthic Foraminiferida from the Dry Tortugas, Florida, has been investigated using an electron microscope. Tests of miliolid Foraminiferida appear to be made up of at least 2 distinct layers. An outermost pavement-like layer, consisting of a single layer of calcite rhombohedrons, and an underlying "randomly oriented layer" comprise most of the wall thickness. It was possible to differentiate each miliolid species on the basis of a statistical analysis of calcite rhombs making up the pavement-like layer. Pseudopores were found on the tests of Miliolinella subrotunda (Montagu) and Archaias angulatus (Fichtel and Moll). None of these pseudopores completely penetrated the wall, and all floored in the "randomly oriented layer." It is believed that these structures are formed at time of calcification, rather than secondarily at some later time.
Calcification in Foraminiferida appears to be a biphasic process similar to that found in most other organisms. Groups of crystals were discernible on surfaces of most porcelaneous and "radiate" calcareous species. This suggests that parallel growth of crystals may be an important process in calcification of many Foraminiferida.
A sieve-plate was found in a canal paralleling the test's surface in Cymbaloporetta squammosa (d'Orbigny).     

The fine structure and development of calcified skeletal elements in the body wall of holothurian echinoderms

The calcareous ossicles and iron-rich calcified granules in the integument of sea cucumbers (Echinodermata: Holothuroida) have been examined by light and electron microscopy. Most ossicles are perforated, platelike structures that measure 100–500 μm long and consist of magnesium-rich calcite. The formation of ossicles occurs within multinucleated syncytia of sclerocytes that are situated in the dermal layer of the body wall. Sclerocytes possess fairly large numbers of mitochondria and are characteristically enveloped by an external lamina. Each ossicle arises as a minute rodlike spicule that branches and develops into a fenestrated skeletal element. When viewed by SEM, fully developed ossicles appear non-crystalline. Following decoration with synthetically grown calcite seeds, however, each ossicle reveals an ordered array of crystallites that seem to be aligned perpendicular to the ossicle's original plane of growth. Examinations of sectioned ossicles indicate that the mineral phase in each skeletal element lacks a well developed matrix of EDTA-insoluble organic substances. Ossicles first arise in the ciliated, lecithotrophic larvae of Eupentacta quinquesimita at about 1 week postfertilization. Two-week-old specimens adopt an epibenthic existence and subsequently become enveloped by imbricated ossicles that are secreted by sclerocytes resembling those found in adults. In molpadiid holothurians, the adult body wall contains numerous reddish-brown granules that are chiefly composed of iron-rich subunits. The dermal granules differ from calcitic ossicles in developing extracellularly in association with finely filamentous material.     

Fine structure of the dorsal epithelium of the tongue of the freshwater turtle,Geoclemys reevesii (Chelonia,Emydinae)

Scanning electron microscopy shows that lingual papillae occur all over the dorsal surface of the tongue of the freshwater turtle, Geoclemys reevesii. The surface of each papilla is composed of compactly distributed hemispherical bulges, each composed of a single cell. Microvilli are widely distributed over the surface of cells. Histological examination reveals that the connective tissue penetrates deep into the center of papillae and that the epithelium is stratified columnar. Under the transmission electron microscope, the cells of the basal and the deep intermediate layers of the epithelium appear rounded. A large nucleus lies in the central area of each cell. The cytoplasm contains mitochondria, endoplasmic reticulum and free ribosomes. The cell membrane form numerous processes. The shallow intermediate layer contains two types of cell. The cytoplasm of the first has numerous fine granules, in addition to mitochondria, ribosomes, and endoplasmic reticulum. The other type of cell contains highly electron-dense granules. The surface layer shows two cell types. One type consists of typical mucous cells. The other type of cell contains fine, electron-lucent granules. The latter cells lie on the free-surface side, covering the mucous cells, and have microvilli on their free surfaces.     

水鳖科9属15种植物花粉形态的研究

应用光学显微镜、扫描电镜和透射电镜对水鳖科Hydrocharitaceae 9属15种植物的花粉形态进行 了观察。水鳖科植物花粉为圆球形至近椭球形,无萌发孔或偶为单沟萌发孔,外壁纹饰通常为小刺状纹 饰,刺密集或稀疏,花粉表面具瘤状、疣状、颗粒状、皱波状突起或光滑。外壁由覆盖层、柱状层和基层组 成。覆盖层厚或较薄,柱状层小柱发育不明显,基层薄。水鳖科植物在花粉大小、纹饰类型、刺的长短、 密度、形态、萌发孔的有无以及花粉壁的结构等方面表现出了较为明显的差异,这些特征对探讨类群间 关系具有较重要意义。由于黑藻属Hydrilla和Stratiotes属花粉较为特殊,支持将它们各自作为一个独立 的族处理。水鳖科植物花粉外壁纹饰和结构特点表明该科与水雍科Aponogetonaceae、泽泻科Alismataceae 和花蔺科Butomaceae等近缘,而该科植物花粉大多无萌发孔等则反应了该科与茨藻目Najadales植物有密切联系。     

Cortical granules remaining after fertilization in Xenopus laevis

Eggs of Xenopus laevis were examined in an electron microscope at unfertilized egg, 1-cell, 2-cell, 32-cell, and blastula stages. Granules closely resembling cortical granules were observed within the “germinal plasm” as well as in the peripheral cytoplasm of all the eggs examined. A staining method was developed that makes it easier to count cortical granules in thick Epon sections. Light and electron microscope examinations revealed that granules remaining after fertilization possessed morphological characteristics wholly consistent with those of cortical granules of unfertilized eggs. These granules were confirmed to be true cortical granules.     

美乐多（Melodorum fruticosum Lour．）花粉形态观察

利用扫描电子显微镜（SEM）和透射电子显微镜（TEM）观察了美乐多（Melodorum fruticosum）的花粉形态特征。美乐多花粉为球形或扁圆形的单粒花粉,外壁纹饰为微褶皱状,有点状凹陷,无任何萌发孔或萌发沟。花粉外壁由外壁外层包括覆盖层（连续）、覆盖下层、基足层（1～3层薄片层结构,偶断裂或扭曲至6～10层）和外壁内层（连续）组成。其中,覆盖下层,其厚度为整个花粉外壁厚度的1／2,为混合型结构,即小柱状和颗粒状同时存在,但以颗粒状为主。花粉内壁分为内壁外层和内壁内层,其厚度逐渐变薄。美乐多的花粉特征（单粒、无萌发孔或沟、覆盖层连续、基足层为薄片层结构、花粉外壁内层薄等）与紫玉盘族其他类群一致。     

THE INFLUENCE OF ENVIRONMENT ON THE SHELL STRUCTURE OF STARCH GRANULES

It is well known from light microscope studies of potato starch that the granules formed in a constant environment (of light and temperature) have a ring formation indistinguishable from that of granules formed under field conditions. Electron microscope studies have confirmed that normal potato starch granules have a fine shell structure not usually resolved by the light microscope, and also that shells do not develop in barley granules grown in a constant environment. The paper presented here reports a further study of the dependence of shell formation on environment. Potatoes were grown in a constant environment and starch granules from the newly formed tubers were examined in the light microscope, and in the electron microscope after corrosion by acid. No difference between these granules and normal granules was observed; both wide (light microscope) rings and fine lamellae developed in both granules. Parallel studies were made on wheat starch granules. In this case, shells were not differentiated in granules that developed in a constant environment, but they could be produced at will by imposing a dark period. Thus, shell formation in potato granules must be controlled by an endogenous rhythm, whereas in wheat granules it must be controlled by external environment.     

STRUCTURE OF THE CONCHIOLIN CASES OF THE PRISMS IN MYTILUS EDULIS LINNE

The prisms in the shell of Mytilus edulis Linné are calcite needles. Their small size and their thin conchiolin cases distinguish them from the prisms of many other species of mollusks. These Mytilus prisms have been studied with the electron microscope. The material consisted of positive replicas of surfaces of the prismatic layer, etched with chelating agents, and of preparations of tubular cases from decalcified prisms which were compared with the conchiolin from decalcified mother-of-pearl of the same species. In the replicas, the cases appear as thin pellicles in the intervals between the prism crystals. Both the prism cases and the nacreous conchiolin, disintegrated by exposure to ultrasonic waves and sedimented on supporting films, appear in the form of tightly meshed, reticulated sheets, described as "tight pelecypod pattern" in former studies on nacreous conchiolin of Mytilus. The results show that in the shell of this species the same conchiolin structure is associated with aragonite in mother-of-pearl and with calcite in the prismatic layer.     

ELECTRON MICROSCOPY OF BASOPHILIC STRUCTURES OF SOME INVERTEBRATE OOCYTES : II. FINE STRUCTURE OF THE YOLK NUCLEI

RNA containing yolk nuclei from the surf clam Spisula solidissima have been studied with the light microscope and with the electron microscope. A variety of structures can be seen with both and a correlation can be made between bodies studied with the electron microscope and those studied with the light microscope. The electron microscope shows many of these structures to be composed of double walled lamellae arranged in space, in various ways. The variety of patterns seen with the electron microscope can be satisfactorily explained on the basis of a hypothetical model. The presence of yolk platelets within the yolk nuclei lends support to classical observations on the synthesis of yolk within yolk nuclei or yolk nuclei derivatives. Small granules (about 100 A) are included within the double walled lamellae and their presence probably accounts for the basophilic nature of the entire body. The presence of small granules attached to electron-dense layers relates the yolk nuclei described here to ergastoplasm discussed by others.     

Ultrastructural observations on the biogenic amines in the carotid and aortic-abdominal bodies of the human fetus

Summary The carotid and aortic-abdominal bodies of two human fetus at fifth month of pregnancy have been studied with the light and electron microscope. A personal variation of the glutaraldehyde ammoniacal silver (GA/S) method has been used, which consists in performing the silver reaction on the ultramicrotomical sections of the tissues, first fixed by glutaraldehyde perfusion and then included in Epikote 812.In the light microscope, the GA/S reaction is certainly positive in the aortic-abdominal bodies and it is negative or dubious in the carotid bodies. — In the electron microscope, however, the reaction is positive both in the aortic-abdominal and in the carotid bodies. — In the aortic-abdominal bodies, the silver precipitate accumulates into thick cytoplasmic granules, which have been shown to be NA-storing granules. — In the carotid bodies however, the silver precipitate accumulates into much smaller cytoplasmic granules, which are interpreted as 5-HT-storing granules.     

Electron microscopy of basophilic structures of some invertebrate oocytes. II. Fine structure of the yolk nuclei

RNA containing yolk nuclei from the surf clam Spisula solidissima have been studied with the light microscope and with the electron microscope. A variety of structures can be seen with both and a correlation can be made between bodies studied with the electron microscope and those studied with the light microscope. The electron microscope shows many of these structures to be composed of double walled lamellae arranged in space, in various ways. The variety of patterns seen with the electron microscope can be satisfactorily explained on the basis of a hypothetical model. The presence of yolk platelets within the yolk nuclei lends support to classical observations on the synthesis of yolk within yolk nuclei or yolk nuclei derivatives. Small granules (about 100 A) are included within the double walled lamellae and their presence probably accounts for the basophilic nature of the entire body. The presence of small granules attached to electron-dense layers relates the yolk nuclei described here to ergastoplasm discussed by others.     

Biochemical study of the adepidermal granules of Urodela and Anura—Especially on lipids

Summary The lipids of the adepidermal granules of Hynobius tokyoensis and Rana japonica were analysed qualitatively. To extract the lipids from the adepidermal granules, the epidermis of the larvae of both animals were removed from the body. Then the granules were extracted from the surface of the basal lamina by means of wiping with the cotton balls containing the solvent. It was ascertained with the electron microscope whether the adepidermal granules were extracted by the cotton balls or not.By the analysis which was made with thin layer chromatography, the same kind of phospholipids or neutral lipids were detected from the extracts of both R. Japonica and H. tokyoensis. Moreover, four spots which seemed to contain some sialic acids were detected from the both extracts.     

The Test Structure and Composition of the Foraminifer Rosalina floridana*

SYNOPSIS. The composition of the test of Rosalina floridana (Cushman) was examined histochemically, and its structure was studied with the electron microscope by means of thin sections and carbon replicas. The test is composed of a thick organic lining overlain by one or more calcite layers bounded above and below by thin membranes. The membranes are fused to organic pore processes composed of coarse fibers that penetrate the calcite layers. The ***lining, consisting of coarse fibers matted into a laminated sheet, is considered a strengthening element of the test. The membranes covering each calcite layer are composed of fine, headed fibrils which in aggregate have a striated pattern; they are thought to be the crystal-nucleating agent during calcification and to form a protective covering for the previously deposited calcite layers. The pore processes, which are devoid of an internal entrance for cytoplasm, are considered to be points of attachment for the membranes; they tie the organic test components into a unified whole. The calcite layers and the chambers lack this unity, being separated from each other and from the preceding chambers by membranes so that there are no calcite-to-calcite boundaries between them. An organic, sievelike structure of undetermined function has been found in the foramina of chambers near the prolocular region of the test. Histochemical methods show that the lining contains proteins, polysaccharides, and unidentified substances; the membranes and the pore processes stain as a protein-polysaccharide complex free of other substances.     

Ultrastructure of the acinar cells in the submandibular gland of the nine-banded armadillo

There are two discrete lobes comprising the armadillo subman-dibular gland. These two lobes can be defined grossly, histochemically and morphologically with the light and electron microscope. The minor lobe stains more intensely with PAS and AB. When viewed in the electron microscope, the secretory granules of the acinar cells within this lobe appear mucous-like. The granules of the demilune cells are slightly different in appearance. The secretory granules of the acinar cells in the major lobe contain many dense foci embedded in a fibrillar matrix, a substructure not described previously. The demilune cells of this lobe contain secretory granules with a mucous-like structure which is consistent throughout the entire lobe. As in the minor lobe, these demilune cells stain very intensely with PAS and AB.     

Structure and composition of cyanophycin granules in the cyanobacterium Aphanocapsa 6308

Cyanophycin granules in the unicellular cyanobacterium Aphanocapsa 6308 were examined with the electron microscope in both thin section and by freeze-fracture techniques. Purified granules were examined with the electron microscope, by arginine determinations, by chromatography, and by elemental analysis. They are similar in ultrastructure and composition to those isolated from the nitrogen-fixing cyanobacterium Anabaena cylindrica, consisting of equal molar quantities of L-arginine and L-aspartic acid.     

Bacteria-Induced Cementation in Sandy Soils

Bacteria-induced calcite precipitation (BICP) is a promising technique that utilizes bacteria to form calcite precipitates throughout the soil matrix, leading to an increase in soil strength and stiffness. This research investigated BICP in two types of sands under sterile and nonsterile conditions. Bacteria formation and BICP in the sterilized sand specimens are higher than those in the nonsterilized sand specimens. The development of calcite with time is initially greater for the sand specimens containing calcite. Scanning electron microscope imaging allowed the detection of cementation from calcite precipitation on the surface and pores of the sand matrix. The effects of injecting nutrient mediums and bacteria into the specimens, as well as pH of soil samples on BICP were investigated. The bearing capacity of biologically treated vs. untreated sand specimens were determined especially by laboratory foundation loading tests.