THE FORMATION AND STRUCTURE OF MYELIN SHEATHS IN THE CENTRAL NERVOUS SYSTEM

The development and structure of myelin sheaths have been studied in the optic nerves of rats and of Xenopus laevis tadpoles. Both potassium permanganate- and osmium-fixed material was examined with the electron microscope. In the first stage of myelinogenesis the nerve fibre is surrounded by a cell process which envelops it and forms a mesaxon. The mesaxon then elongates into a loose spiral from which the cytoplasm is later excluded, so that compact myelin is formed. This process is similar to myelinogenesis in the peripheral nervous system, although in central fibres the cytoplasm on the outside of the myelin is confined in a tongue-like process to a fraction of the circumference, leaving the remainder of the sheath uncovered, so that contacts are possible between adjacent myelin sheaths. The structure of nodes in the central nervous system has been described and it is suggested that the oligodendrocytes may be the myelin-forming cells.     

THE GEOMETRY OF PERIPHERAL MYELIN SHEATHS DURING THEIR FORMATION AND GROWTH IN RAT SCIATIC NERVES

In rat sciatic nerves, a small bundle of fibers was identified in which myelin sheaths were absent at birth, appeared within 3 days, and grew rapidly for 2 wk. During this interval, nerves were removed from littermates and were sectioned serially in the transverse plane. Alternating sets of thin and thick sections were used to prepare electron micrograph montages in which single myelinating axons could be identified and traced distally. During the formation of the first spiral turn, the mesaxon's length and configuration varied when it was studied at different levels in the same Schwann cell. The position of the mesaxon's termination shifted while its origin, at the Schwann cell surface, remained relatively constant. Along myelin internodes composed of two to six spiral turns, there were many variations in the number of lamellae and their contour. Near the mesaxon's origin, longitudinal strips of cytoplasm separated the myelin layers. Thicker sheaths were larger in circumference, more circular in transverse sections, and more uniform at different levels. Irregularities were confined to the paranodal region, and separation of lamellae by cytoplasm occurred at Schmidt-Lantermann clefts. Approximate dimensions of the bundle, its largest fibers, and their myelin sheaths were measured and calculated. The myelin membrane's transverse length and area increased exponentially with time; the growth rate increased rapidly during the formation of the first four to six spiral layers and remained relatively constant during the subsequent enlargement of the compact sheath.     

FURTHER OBSERVATIONS ON THE STRUCTURE OF MYELIN SHEATHS IN THE CENTRAL NERVOUS SYSTEM

Direct evidence has been presented to confirm the existence of a spiral in the myelin sheaths of the central nervous system. An account of some of the variations in structure of central myelin sheaths has been given and it has been shown that the radial component of myelin sheaths has the form of a series of rod-like thickenings of the intraperiod line. These thickenings extend along the intraperiod line in a direction parallel to the length of the axon. The relative position of the internal mesaxon and external tongue of cytoplasm has been determined in a number of transverse sections of sheaths from the optic nerves of adult mice, adult rats, and young rats. In about 75 per cent of the mature sheaths examined, these two structures were found within the same quadrant of the sheath, so that the cytoplasm of the external tongue process tends to lie directly outside that associated with the internal mesaxon. The frequency with which the internal mesaxon and external tongue lie within the same quadrant of the sheath increases both with the age of the animal and with the number of lamellae present within a sheath. The possible significance of these findings is discussed.     

THE ROLE OF WATER IN THE STRUCTURE OF PERIPHERAL NERVE MYELIN

In the study of the drying kinetics of nerve fibres, at least five "phases" of water evaporation can be distinguished. A consideration of the accompanying changes in low-angle x-ray diffraction patterns permits a tentative identification of the "phases" and a quantitative interpretation of the data in terms of the water distribution in nerve fibres. These results suggest that the myelin sheath of frog sciatic nerve contains 40 to 50 per cent water, and it is suggested further that the greater part of this water is "organised" in relation to the hydrophilic groups of the lipide and protein components.     

麦冬花药绒毡层和乌氏体的细微结构

麦冬(Ophiopogon japonicus)的绒毡层发育为分泌型。在小孢子母细胞时期,绒毡层细胞达到了发育的高峰。此时,绒毡层细胞中细胞器非常丰富,具大量线粒体、高尔基体和质体,尤以肉质网含量最多;原乌氏体出现较早,在小孢子母细胞时期绒毡层细胞中就已出现;四分体时期,大量原乌氏体被排入内切向面的质膜和纤维素壁之间;到了小孢子早期,绒毡层细胞失去细胞壁,原乌氏体分布在质膜的凹陷处,孢粉素物质在其上沉积,发育为乌氏体,乌氏体有单个和复合两种类型;当花粉成熟时,绒毡层细胞完全解体。     

THE FINE STRUCTURAL ORGANIZATION OF NERVE FIBERS, SHEATHS, AND GLIAL CELLS IN THE PRAWN, PALAEMONETES VULGARIS

In view of reports that the nerve fibers of the sea prawn conduct impulses more rapidly than other invertebrate nerves and look like myelinated vertebrate nerves in the light microscope, prawn nerve fibers were studied with the electron microscope. Their sheaths are found to have a consistent and unique structure that is unlike vertebrate myelin in four respects: (1) The sheath is composed of 10 to 50 thin (200- to 1000-A) layers or laminae; each lamina is a cellular process that contains cytoplasm and wraps concentrically around the axon. The laminae do not connect to form a spiral; in fact, no cytoplasmic continuity has been demonstrated among them. (2) Nuclei of sheath cells occur only in the innermost lamina of the sheath; thus, they lie between the sheath and the axon rather than outside the sheath as in vertebrate myelinated fibers. (3) In regions in which the structural integrity of the sheath is most prominent, radially oriented stacks of desmosomes are formed between adjacent laminae. (4) An ~200-A extracellular gap occurs around the axon and between the innermost sheath laminae, but it is separated from surrounding extracellular spaces by gap closure between the outer sheath laminae, as the membranes of adjacent laminae adhere to form external compound membranes (ECM's). Sheaths are interrupted periodically to form nodes, analogous to vertebrate nodes of Ranvier, where a new type of glial cell called the "nodal cell" loosely enmeshes the axon and intermittently forms tight junctions (ECM's) with it. This nodal cell, in turn, forms tight junctions with other glial cells which ramify widely within the cord, suggesting the possibility of functional axon-glia interaction.     

THE FINE STRUCTURE OF THE PURKINJE CELL

This paper describes the fine structure of the Purkinje cell of the rat cerebellum after fixation by perfusion with 1 per cent buffered osmium tetroxide. Structures described include a large Golgi apparatus, abundant Nissl substance, mitochondria, multivesicular bodies, osmiophilic granules, axodendritic and axosomatic synapses, the nucleus, the nucleolus, and the nucleolar body. A new and possibly unique relationship between mitochondria and subsurface cisterns is described. Possible functional correlations are discussed.     

THE FINE STRUCTURE AND ELECTROPHYSIOLOGY OF HEART MUSCLE CELL INJURY

Injured frog heart cells electrically uncouple from their uninjured neighbors within 30 min after injury. This uncoupling process can be shown by the disappearance of an injury potential measured between such injured and uninjured cells. In the present study, the time course of the decline of injury potentials, and thus of electrical uncoupling, in bullfrog atrial trabeculae was determined. Tissue was fixed with glutaraldehyde and osmium tetroxide at various times after injury to determine the morphological changes which accompany this uncoupling process. In some cases, ruthenium red was included in the fixatives. Normal atrial cells are long and narrow, with intercellular junctions located along the lateral surfaces of the cells. Two types of intercellular junctions have been observed: cardiac adhesion plaques (CAPs), and close junctions. Close junctions occur only infrequently. Ruthenium red penetrates all around the cells, leaving only small areas within the CAPs unstained. After injury, the cells are very dense and the myofilaments disarranged. Both types of intercellular junction remain intact, and only slight changes within CAPs are observed. The results are discussed in relation to current concepts of intercellular communication.     

THE FINE STRUCTURE OF THE GRASS GUARD CELL

Brown, W. V., and Sr. C. Johnson. (U. Texas, Austin.) The fine structure of the grass guard cell. Amer. Jour, Bot. 49 (2): 110–115. Illus. 1962.—An electron microscopic study of 16 species of grasses classified in 10 tribes and 5 subfamilies has revealed some hitherto unknown facts about guard-cell structure. In species of 3 subfamilies, but not in the Festucoideae, there are membranes on the guard cells overarching the stoma. In the Festucoideae, the membrane is rudimentary or absent and is associated with a different cross-sectional shape of the guard cell. The central canal through the thick-walled region of the guard cell is structurally quite complex. The wall between the central canal and the subsidiary cell is thin and lacks plasmodesmata. There are plastids but no developed chloroplasts in grass guard cells. Mitochondria are abundant, but vacuoles are undetectable. At the ends of the guard-cell pair, the wall between them is incomplete and the protoplasts are confluent.     

REDUNDANT MYELIN SHEATHS AND OTHER ULTRASTRUCTURAL FEATURES OF THE TOAD CEREBELLUM

Some of the myelin sheaths in the cerebellum of normal adult toads exhibit extensive evaginations of their full thickness. These redundant flaps of myelin are collapsed; i.e., they contain no axon and have no lumen. They extend away from the parent axonal myelin sheaths and tend to enfold other myelinated fibers or granule cell perikarya, producing bizarre configurations of myelin and what appear to be partially or completely myelinated cell bodies. In some instances, only the redundant flap of myelin appears in the plane of section, and its attachment to an axonal myelin sheath in another plane is only inferred. Single lamellae of myelin also tend to invest cerebellar granule cells and other processes, and these too appear to fold on themselves producing two- or four-layered segments. It is suggested that there are two phases of myelinogenesis: an initial "wrapping" phase, followed by a prolonged second phase during which internodes of myelin increase in both length and girth by a process other than wrapping, and that the occurrence of redundant myelin sheaths may reflect overgrowth of myelin during the second phase. Observations on the general organization of the toad cerebellum and on the ultrastructural cytology of its layers are also presented.     

THE FINE STRUCTURE OF MITOSIS AND CELL DIVISION IN THE CHRYSOPHYCEAN ALGA OCHROMONAS DANICA1

At the ultrastructural level, cell division in Ochromonas danica exhibits several unusual features. During interphase, the basal bodies of the 2 flagella replicate and the chloroplast divides by constriction between its 2 lobes. The rhizoplast, which is a fibrous striated root attached to the basal body of the long flagellum, extends under the Golgi body to the surface of the nucleus in interphase cells. During proprophase, the Golgi body replicates, apparently by division, and a daughter rhizoplast, appears. During prophase, the 2 pairs of flagellar basal bodies, each with their accompanying rhizoplast and Golgi body, begin to separate. Three or 4 flagella are already present at this stage. At the same time, there is a proliferation of microtubules outside the nuclear envelope. Gaps then appear in the nuclear envelope, admitting the microtubules into the nucleus, where they form a spindle. A unique feature of mitosis in O. danica is that the 2 rhizoplasts form the poles of the spindle, spindle microtubules inserting directly onto the rhizoplasts. Some of the spindle microtubules extend from pole to pole; others appear to attach to the chromosomes. Kinetochores, however, are not present. The nuclear envelope breaks down, except, in the regions adjacent, to the chloroplasts; chloroplast ER remains intact throughout mitosis. At late anaphase the chromosomes come to lie against part of the chloroplast ER. This segment of the chloroplast ER appears to be incorporated as part of the reforming nuclear envelope, thus reestablishing the characteristic nuclear envelope—chloroplast ER association of the interphase cell.     

SOME OBSERVATIONS ON THE FINE STRUCTURE OF THE SYMPATHETIC GANGLION OF BULLFROG

Electron microscope observations of abdominal sympathetic ganglia of American bullfrogs, Rana catesbiana, have demonstrated the presence of specific areas of cytoplasm in the superficial zone of the perikaryon which are devoid of granulated endoplasmic reticulum. These areas are occupied almost exclusively by granules 200 to 400 A in diameter which can be stained intensely with lead hydroxide but faintly with uranyl acetate. Each granule shows subgranular internal structure after the lead staining. Granules of similar properties are found in synapses also, and may be glycogen. From the satellite cell there extends a number of leaf- or finger-like cytoplasmic projections around the root portion of the nerve process. Some of these projections directly cover the surface of the nerve process. Many others, however, are separated from the neuron by a fairly wide interspace. Multivesicular bodies of the neuron are occasionally observed in a configuration which suggests that they are being extruded from the root of the nerve process into the interspace. Filaments about 100 A in thickness are found in the satellite cell cytoplasm. They are arranged more or less parallel to each other and are especially well developed around synapses and nerve fibers.     

FINE STRUCTURE OF THE SYNAPTIC DISCS SEPARATED FROM THE GOLDFISH MEDULLA OBLONGATA

Synaptic discs are structures localized in the club ending synapses on the Mauthner cell lateral dendrite of the goldfish medulla oblongata. The synaptic discs present a hexagonal array of particles ~8.5 nm center-to-center when observed in en face view. This lattice covers the entire surface Divalent cations are important in the stabilization of this particular hexagonal array of particles When a synaptic disc-rich fraction is treated with chelating agents (EDTA or EGTA), definite changes occur in the hexagonal lattice. First, the synaptic membranes show zones without particles interspersed with zones covered with the hexagonal array of particles Second, the synaptic discs break down and a new structure characterized by two parallel dense bands (7 nm each), separated by a 4 nm gap, is observed. The negative stain fills the gap region showing striations spaced ~10 nm center-to-center crossing the gap, but it does not penetrate the dense bands This "double band" structure is interpreted as an edge on view of a fragment of the synaptic membrane complex. Further treatment of this fraction with a chelating agent plus 0.3% deoxycholate produces an increase in the number of double band structures. However, EDTA plus Triton X-100 (a treatment known to produce solubilization of membrane proteins) never shows such double band structure An ordered material was observed associated with the cytoplasmic leaflets of the double bands This material consists of rows of beads ~4 nm in diameter and spaced at intervals of ~7 nm. Each of these beads is joined to the band by a thin stalk.     

THE ULTRASTRUCTURE OF MAUTHNER CELL SYNAPSES AND NODES IN GOLDFISH BRAINS

An electron microscope study of goldfish Mauthner cells is reported.¹ The cell is covered by a synaptic bed ~ 5 µ thick containing unusual amounts of extracellular matrix material in which synapses and clear glia processes are implanted. The preterminal synaptic neurites are closely invested by an interwoven layer of filament-containing satellite cell processes. The axoplasm of the club endings contains oriented mitochondria, neurofilaments, neurotubules, and relatively few synaptic vesicles. That of the boutons terminaux contains many unoriented mitochondria and is packed with synaptic vesicles and some glycogen but no neurofilaments or neurotubules. The bare axons of club endings are surrounded by a moderately abundant layer of matrix material. The synaptic membrane complex (SMC) in cross-section shows segments of closure of the synaptic cleft ~ 0.2 to 0.5 µ long. These alternate with desmosome-like regions of about the same length in which the gap widens to ~ 150 A and contains a condensed central stratum of dense material. Here, there are also accumulations of dense material in pre- and postsynaptic neuroplasm. The boutons show no such differentiation and the extracellular matrix is largely excluded around them. The axon cap is a dense neuropil of interwoven neural and glial elements free of myelin. It is covered by a closely packed layer of glia cells. The findings are interpreted as suggestive of electrical transmission in the club endings.     

CHANGES IN THE OSMIOPHILIA OF MYELIN AND LIPID CONTENT IN THE KITTEN OPTIC NERVE

Myelin osmiophilia has been shown to develop significantly later than myelin staining by Luxol fast blue and Sudan black, in the developing kitten optic nerve. These histological changes are accompanied by alterations in the lipid composition of the optic nerve. Although myelination commences at about 10 days post partum in the nerve the appearance of cerebrosides is unexpectedly delayed. Changes in fatty acid chain length and lipid composition of optic nerve are consistent with the suggestion that ‘early’ myelin may be unchanged glial plasma membrane.     

STUDIES ON THE FINE STRUCTURE OF ULTRACENTRIFUGED SPINAL GANGLION CELLS

The following structures were observed in electron micrographs of the mouse spinal ganglion cells: Nissl bodies composed of both aggregated rough-type, largely oriented, membranes of the endoplasmic reticulum and discrete particles; short rodlike mitochondria with well-developed transverse, obliquely or longitudinally arranged cristae, and a relatively typical Golgi complex. The components of ultracentrifuged ganglion cells (400,000 times gravity for 20 minutes) are stratified, the layers appearing in the order of their decreasing density as follows: (1) A microsomal or ergastoplasmic layer which may be further divided into three sublayers without sharp boundaries, namely, a discrete particle layer, a layer of discrete particles and highly distorted membranes of the endoplasmic reticulum, and a layer composed of relatively intact, but stretched membranes of the endoplasmic reticulum and discrete particles. (2) Mitochondria constitute a relatively broad layer. They are sometimes stretched; however, they retain most of their fine structure. The stratified nucleus is found within the mitochondrial layer. (3) A relatively wide layer of tightly packed vesicles. (4) At the centripetal end, resting against the cell membrane, are a few lipid vacuoles. A comparison is made between the ultrastructure of the stratified layers in situ and those described by others in differentially ultracentrifuged homogenates.     

THE FINE STRUCTURE OF THE NUCLEOLUS DURING MITOSIS IN THE GRASSHOPPER NEUROBLAST CELL

The behavior of the nucleolus during mitosis was studied by electron microscopy in neuroblast cells of the grasshopper embryo, Chortophaga viridifasciata. Living neuroblast cells were observed in the light microscope, and their mitotic stages were identified and recorded. The cells were fixed and embedded; alternate thick and thin sections were made for light and electron microscopy. The interphase nucleolus consists of two fine structural components arranged in separate zones. Concentrations of 150 A granules form a dense peripheral zone, while the central regions are composed of a homogeneous background substance. Observations show that nucleolar dissolution in prophase occurs in two steps with a preliminary loss of the background substance followed by a dispersal of the granules. Nucleolar material reappears at anaphase as small clumps or layers at the chromosome surfaces. These later form into definite bodies, which disappear as the nucleolus grows in telophase. Evidence suggests both a collecting and a synthesizing role for the nucleolus-associated chromatin. The final, mature nucleolar form is produced by a rearrangement of the fine structural components and an increase in their mass.