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.     

A STRUCTURAL ANALYSIS OF THE MYELIN SHEATH IN THE CENTRAL NERVOUS SYSTEM

The cerebral white matter of rats subjected to a variety of noxious experimental conditions was examined in the electron microscope. Several unusual configurations of the myelin sheath are identified in addition to the usual configuration. These variations include the presence of (a) formed organelles within the inner and outer loops, (b) isolated islands of cytoplasm in unfused portions of the major dense lines, (c) apparently unconnected cell processes between the sheath and the axon, and (d) concentric, double myelin sheaths. A generalized model of the myelin sheath based on a hypothetical unrolling of the sheath is described. It consists of a shovel-shaped myelin sheet surrounded by a continuous thickened rim of cytoplasm. Most of the unusual myelin configurations are explained as simple variations on this basic theme. With the help of this model, an explanation of the formation of the myelin sheath is offered. This explanation involves the concept that myelin formation can occur at all cytoplasmic areas adjacent to the myelin proper and that adjacent myelin lamellae can move in relation to each other.     

ELECTRON MICROSCOPIC OBSERVATIONS OF THE CENTRAL NERVOUS SYSTEM

In order to establish criteria for the identification of the neural and glial cells of the central nervous system, sections of the brains and spinal cords of mice, rabbits, guinea pigs, and rats; and portions of tumors of the human brain have been examined by electron microscopy. Identification of neurons is made possible by the characteristic cytoplasmic picture, in which there is a distinct granular and less constant membranous ergastoplasmic pattern. In no other cell of the central nervous system is such a distinct granular component present in the ergastoplasm. The shape of the neuron in electron microscopic preparations is similar to that seen by light microscopy with several dendrites containing a similar cytoplasm arising from the perikaryon. Synapses are relatively common on the surface of the neuron and its dendrites. Microglial cells are relatively small and dense with few processes, and are arranged as perineuronal and perivascular satellites for the most part. Occasionally phagocytized material is present in their cytoplasm. The oligodendroglial cells are identifiable by their position as perineuronal satellites and in the white matter as cells arranged in rows. They have a uniformly round to ovoid nucleus with a pale cytoplasm, which has a sparse, finely granular component and a few small mitochondria. The processes are few and relatively straight when cut in longitudinal section. The predominant cellular type in an oligodendroglioma was similar, with a pale cytoplasm. The astrocytes are variable in appearance. Their nuclei are moderately large, irregularly ovoid, and the cytoplasm adjacent to the nucleus is finely granular and scant. In the protoplasmic astrocytes the cytoplasm has a complicated infolded arrangement with reduplication of the plasma membrane, numerous processes extending radially from the cell and rebranching. To a certain extent this same folded plasma membrane was noted in the fibrous astrocytes. However, their more distant processes were narrowed, relatively straight, and filled with numerous dense fibrils. The processes of the astrocyte often surrounded axons, and other cellular processes, and surrounded some vessels, while attaching to a part of the wall of another vessel. Proliferating cells in experimentally produced gliosis and in astrocytic neoplasms were similar in structure. The ependymal cells and the epithelium of the choroid plexus have a specialized surface with microvillous projections of the cytoplasm covered by the plasma membrane. Cilia in varying numbers are present in both epithelia.     

MYELIN PROTEINS FROM DIFFERENT REGIONS OF THE CENTRAL NERVOUS SYSTEM

—The protein composition of myelin prepared from specific anatomical regions of the bovine brain and spinal cord was studied by a modification of the method of Gonzalez -Sastre (1970). Spinal cord myelin contained lesser amounts of chloroform-methanol soluble protein and proteolipid protein and had a lower activity of the enzyme 2′,3′-cyclic nucleotide 3′-phosphohydrolase than did myelin from subcortical white matter. There was no difference, however, in the protein composition of myelin from the various levels of the spinal cord. The amino acid composition of both proteolipid and basic protein showed no significant regional differences. Myelin preparations from both brain and spinal cord contained DM-20 protein.     

ULTRAVIOLET ABSORPTION CHARACTERISTICS OF MYELIN FROM THE CENTRAL NERVOUS SYSTEM

—New data are presented and published data reviewed to show that the protein content of rat brain myelin must be close to 21 per cent. Ultraviolet absorption measurements in the 220 nm region, however, indicate an apparent protein content of 44·3 per cent which, after solubilization with lysophosphatidylcholine falls to 35·8 per cent. It is shown that this latter proportion can be accounted for in terms of u.v. absorption by myelin protein and lipids, contributions being made by sphingolipids, phospholipids and cholesterol. It is concluded that in the environment of the intact myelin structure, u.v. absorption due to some of the chromophores is enhanced and that this effect is relaxed by lysophosphatidylcholine solubilization. Supportive evidence is given from measurements in the 280 nm region.     

MYELIN IN THE CENTRAL NERVOUS SYSTEM AS OBSERVED IN EXPERIMENTALLY INDUCED EDEMA IN THE RAT

The compact arrangement of cells in the normal white matter of the brain makes an analysis of cellular architecture difficult. To overcome this difficulty, cerebral edema was induced in rats by means of the unilateral intracerebral implantation of silver nitrate. Within 48 hr, the brains were fixed by perfusion with glutaraldehyde followed by immersion in Dalton's chrome-osmium. Sections of the callosal radiations were studied in the electron microscope. The untreated hemisphere appeared entirely unaltered, whereas in the edematous hemisphere the edema fluid separated individual cell processes and small groups of them. The myelin sheaths and their relationships to the axons appeared essentially unaltered. In this material, analysis of cellular architecture was relatively easy, and the widely held theory of spiral wrapping could be confirmed. In addition, several other aspects of the myelin and myelin-forming cell relationships became apparent in the edematous tissue. Most of these were later confirmed by extensive and careful study of the nonedematous tissue. These included the presence of occasional isolated cytoplasmic areas in myelin and the presence of two complete sheaths around a single axon. Other observations, such as the appearance of mitochondria and dense bodies within the outer loop and the separation of myelin lamellae, are apparently limited to the edematous tissue.     

A CHOLESTEROL-ESTERIFYING ENZYME IN RAT CENTRAL NERVOUS SYSTEM MYELIN1

Aontrary to our earlier finding (Eto & Suzuki , 1971), the myelin fraction purified from young adult rat brain consistently showed cholesterol-esterifying activity. The specific activity in myelin was the highest among subcellular fractions. Extensive washing wiih various aqueous salt solutions failed to remove the activity from myelin. The enzyme was evenly distributed among the arbitrarily defined light, medium and heavy myelin subfractions. The myelin-localized activity showed the pH optimum and heat stability identical to the microsome-bound activity. Although there were minor differences in the effect of detergents or exogenous lipids added to the reaction mixture, no firm evidence was obtained to indicate that the myelin-bound cholesterol-esterifying enzyme is different from that in other subcellular fractions. On the other hand, the distribution among the myelin subfractions and heat stability of the myelin-bound cholesterol-esterifying activity were different from those of the myelin-specific cholesterol ester hydrolase. Therefore, the esterification does not appear to be a mere reverse reaction catalyzed by the previously known myelin-specific hydrolase. The rat brain myelin, therefore, is capable of both synthesizing and hydrolyzing cholesterol esters.     

THE OCCURRENCE OF TWO MYELIN BASIC PROTEINS IN THE CENTRAL NERVOUS SYSTEM OF RODENTS IN THE SUBORDERS MYOMORPHA AND SCIUROMORPHA

Extracts containing myelin basic proteins have been prepared from CNS tissue of representatives of the three suborders of Rodentia—Myomorpha, Hystricomorpha and Sciuro-morpha. Analyses of the extracts by electrophoresis at low pH showed that one type (L) of myelin basic protein is present in the CNS of all of the rodents examined (rat, mouse, hamster, guinea pig, chinchilla, prairie dog, woodchuck and squirrel). This protein is comparable in molecular size and charge to the CNS myelin basic proteins found in several other mammalian orders. In the CNS of the myomorphs (rat, mouse, hamster) and sciuro-morphs (prairie dog, woodchuck, squirrel) there is an additional type (S) of myelin basic protein of higher cathodic mobility and smaller molecular size. This additional protein is absent from the CNS of the hystricomorphs (guinea pig, chinchilla). These findings indicate that the presence of two myelin basic proteins originally reported in the CNS of the inbred rat is not an anomaly of inbreeding. These data further suggest that the presence of a single L-type CNS myelin basic protein might be a general characteristic of hystricomorphs, while the presence of both L- and S-type CNS myelin basic proteins might be a general characteristic of the myomorphs and sciuromorphs. Analyses by electrophoresis at low pH failed to reveal differences in mobility among either the L-type or the S-type CNS myelin basic proteins of the different species. In contrast, when electrophoresis was carried out cathodically at high pH, species-related differences in mobility were observed among the L- and S-type CNS myelin basic proteins.     

FURTHER OBSERVATIONS ON THE STRUCTURE OF SELAGINELLITES CRASSICINCTUS

Leisman , Gilbert A. (Kansas State Teachers Coll., Emporia.) Further observations on the structure of Selaginellites crassicinctus. Amer. Jour. Bot. 48(3): 224–229. Illus. 1961.—Specimens of Selaginellites crassicinctus Hoskins and Abbott in coal balls from southeastern Kansas have yielded new information concerning the anatomy of this strobilus. The axis contains an exarch protostele apparently suspended in an air-cavity by endodermal trabeculae. Sporophylls are attached in alternating whorls of 4 each. The strobilus is compared with modern Selaginella and a related compression fossil species.     

FURTHER OBSERVATIONS ON THE FINE STRUCTURE OF THE MACRONUCLEUS IN TOKOPHRYA INFUSIONUM

In a previous paper (8) an organized structure was described in the macronuclei of certain old organisms of Tokophrya infusionum. It was found that the same honeycomb structure appears in great abundance in the macronuclei of overfed organisms. This permitted a better three-dimensional reconstruction of the described structure. Since the defined structure may be experimentally induced, it offers an opportunity for further more detailed studies as to its nature and meaning.     

FORMATION OF MYELIN IN THE CENTRAL NERVOUS SYSTEM OF MICE AND RATS, AS STUDIED WITH THE ELECTRON MICROSCOPE

Sections of brain and spinal cord of mice and rats at 1, 3, 5, and 17 days after birth were examined with the electron microscope. In the early stages of myelinization only two to four lamellae surround the axon. These laminae are formed from the plasma membraces of glial processes, and in particular of oligodendroglial processes. In a later stage of myelinization a larger number of flattened glial processes surround the axon with enclosed cytoplasm trapped within some of the membranes. Multiple extensions of the membranes of the flattened glial processes to the lamellae of the myelinated sheath are evident at this stage, with a variable number of membranes within the sheath at various positions along the fiber. Newly formed myelinated sheaths are sometimes larger than their enclosed fibers, extending as a projection of sheath which does not surround axoplasm. Loci are present in which the myelinated sheath is incomplete or interrupted.     

SYNAPSES IN THE CENTRAL NERVOUS SYSTEM

A number of different synapses have been described in the medulla, cerebellar cortex, and cerebral cortex of the rat. All of these possess the same fundamental fine structure as follows: 1. Close apposition of the limiting membranes of presynaptic and postsynaptic cells without any protoplasmic continuity across the synapse. The two apposed membranes are separated by a cleft about 200 A wide, and display localized regions of thickening and increased density. 2. The presynaptic expansion of the axon, the end-foot or bouton terminal, contains a collection of mitochondria and clusters of small vesicles about 200 to 650 A in diameter. Although the significance of these structures in the physiology of the synapse is still unknown, two suggestions are made: that the mitochondria, by means of the relation between their enzymatic activity and ion transport, participate in the electrical phenomena about the synapse; and that the small synaptic vesicles provide the morphological representation of the prejunctional, subcellular units of neurohumoral discharge at the synapse demanded by physiological evidence.     

ERGOTHIONEINE IN THE CENTRAL NERVOUS SYSTEM

Abstract— Further investigations have been made into ergothioneine in the brains of several mammalian species, and the distribution of ergothioneine in the brain of the ox is described. It has not been possible to confirm many of the findings of earlier workers and the results do not appear to support their conclusion that ergothioneine is identical with the cerebellar factor.     

PROTEIN COMPOSITION OF MYELIN OF THE PERIPHERAL NERVOUS SYSTEM

Abstract— Myelin was purified from the peripheral nervous system (PNS) of several species. The protein composition of these preparations was examined by discontinuous polyacrylamide gel electrophoresis in buffers containing sodium lauryl sulphate. Proteins characteristic of all samples include, in order of increasing mobility: a series of high molecular weight proteins, the major peripheral nerve protein (P₀), two uncharacterized proteins, and two basic proteins (P₁ and P₂). Quantitative results, obtained by densitometry of gels stained with Fast Green showed differences in protein distribution, both between species, and from different types of nerves obtained from the same animal. The relative amounts of P₁ and P₂ proteins were the most variable; e.g. myelin from guinea-pig sciatic nerve had little or no P₂ protein, whereas 15 per cent of the myelin protein of beef posterior intradural root was Pz protein. P₀, P₁ and P₂ proteins from rabbit sciatic nerve and P₀ and P₂ proteins from beef dorsal and ventral intradural roots were purified and their amino acid compositions were determined. Our results indicated that the P₁ protein is very similar in size and amino acid composition to the basic protein of central nervous system myelin, whereas the P₀ and P₂ proteins are unique to the PNS.