Synapses

 

Synapses are specialised interneuronal contacts which can be electrical or chemical. Electrical synapses function by the propagation of electrical impulses and do not require elaborate structural organisation of the plasma membrane and cyto­plasmic organelles. These synapses, which are rare in matnmals but common in lower vertebrates, are formed by the close apposition of the cell membranes at a gap junction. Chemical synapses, in contrast, utilise various substances-neuro­transmitters and neuropeptides-for intercellular communication:  this  chemical  transmission requires a sophisticated subcellular mechanism well demonstrated by electron microscopy. By light microscopy, using silver impregnation tech­niques, only the profile of the end-bulb or bouton terminal of the axon abutting onto the surface of the perikaryon or dendrite can be seen. A synapse is composed of three constituents: the presynaptic element, the synaptic cleft and the postsynaptic component (Fig. 1.5). Both the pre- and the postsynaptic membranes display densities along the speci~ised stretch of cell membrane and these, together with the intervening cleft, are referred to as the synaptic junction.

 

Originally two types of synapse were distin­guished on ultrastructural examination of the pyramidal cells of the cerebral cortex. The type I junction is usually extensive, the postsynaptic density prominent, rendering the synapse asym­metrical, and the wide synaptic cleft contains a dense plaque. In the type Ii synapse, in contrast, the postsynaptic density is less readily identifiable and the narrower cleft does not contain a dense plaque; the overall configuration is symmetrical.15 The presynaptic element in most cases is an axon, either at its end (bouton terminal) or along its course (bouton en passant): the axonal terminal can form a synapse with any part of another neuron and thus axo-dendritic, axo-somatic and axo­axonal synapses are distinguished. The presyn­aptic element, however, does not need to be an axonal terminal and synapses can be formed between any part of two neurons, providing a wide variety of synapses.

 

The presynaptic element contains the synaptic vesicles which, in turn, are filled with neuro­transmitters (Fig. 1.5). The size, shape and content of the vesicles vary, depending upon the type of synapse. The most frequently Occurring vesicles are apparently clear, spherical and 40-50 nm in diameter. Elongated or flattened vesicles with clear centres are 20 nm wide and 50 nm long. There have been many attempts to correlate vesicular shape with functional activity: spherical vesicles occur in excitatory, type I synapses, whilst flattened vesicles are associated with inhibitory, type II synapses. Larger vesicles of up to 60nm in diameter with a dense core are encountered in areas of catecholamine activity and contain noradrenaline, dopamine or 5-hydroxy-tryptamine. Another, larger dense-cored vesicle, up to lSOnm with a spherical dense core of 50-70 nm in diameter, is most frequently seen in the presynaptic terminals of autonomic ganglia, but also occurs in various parts of the central nervous system intermingled with clear vesicles. The presynaptic element, particularly the axonal terminal, also contains other organelles, including mitochondria, neurofilaments,  microtubules, smooth endoplasmic reticulum and glycogen.

 

The synaptic cleft, separating the pre- and postsynaptic membranes, is 20-3Onm wide and usually contains a dense plaque of intercellular material. In addition, filaments appear to traverse the cleft and contribute to the formation of the plaque.

 

The  postsynaptic  membrane  is  rendered conspicuous by an accumulation of dense material on its cytoplasmic surface (Fig. 1.5). This post-synaptic density, composed of granular material and an occasional filamentous structure, is more pronounced in asymmetrical, type I, synapses. The existence of postsynaptic density is dependent upon the presence of the presynaptic element: if the  presynaptic  structures  degenerate,  the postsynaptic density will eventually disappear. The postsynaptic element ~so contains cisternac of the smooth endoplasmic reticulum and the spine apparatus, which is composed of 2-3 flattened cisternac, separated by plaques of dense material.

 

The morphological appearances of synapses reflect  the  functional  dynamics  of neuro­transmission. Accordingly, in chemical synapses the vesicles contain quanta of the neurotransmitter which is discharged into the synaptic cleft after the vesicle has fused with the presynaptic membrane. Thus, the uptake of calcium into the nerve terminal triggers off a chain of events leading to exocytosis: vesicular apposition, membrane fusion and fission.16 Although this vesicular or exocytotic hypothesis of neurotransmission'7 provides a plausible explanation for the quantal nature of  transmitter release, an increasing body of evidence now suggests that this mav not be the only mechanism by which chemical signals in the central nervous system are propagated. Chemical substances may be released from non-synaptic axon terminals and even from regions of nerve cells other than presynaptic axon.

 

A parasynaptic system is envisaged in which so-called neuroactive or informational substances reach specific target cell receptors by diffusion from release points through the extracellular space. The mechanism of exocytosis itself remains poorly understood not only morpho­logically, but also neurochemically, and various alternative biochemical mechanisms have been considered to explain the release of neuro­transmitters.

 

Neuronal processes

 

AXONS

 

The transitional zone between the neuronal perikaryon and the axon is the cone-shaped axon hillock. In larger, multipolar neurons the axon hillock lacks Nissl substance, and it is in this region that cytoskeletal elements, the microtubules and neurofilaments, converge to become parallel as they enter the axon. The microtubules then form a fascicle in the initial segment of the axon, a feature 'vhich enables axons to be distinguished from dendrites, in which microtubules are more evenly distributed. The more distal portion of the axon may contain various organelles, but granular endoplasmic reticulum and polyribosomes are usually absent. The internal structure of the axon has recently been visualised as a three-dimensional lattice: the longitudinally orientated neurofila­ments and microtubules are extensively cross-linked to each other and to the plasma membrane by thin filaments. Similar bridges also connect membrane-bound organelles with the components of the cytoskeleton and with each other .There is axonal transport of material from the perikaryon towards the periphery (anterograde transport) and to a lesser extent in the opposite direction (retrograde transport). Since the axon terminal, the most active site of neurotrans­mission, can be far removed from the perikaryon in which proteins are synthesised, this axonal transport is vital for the proper functioning of the nerve cell. Axonal flow has a slow and a fast component, 0~2-8~0mm and 50-500 mm per day respectively. Fast axoplasmic flow carries organ­elles, including vesicles and mitochondria, and membrane-bound substances like proteins and neurotransmitters  materials which are essential for synaptic activity. The fast axonal flow is effected by microtubules, but the possibility that some membrane-bound proteins are transported by the smooth endoplasmic reticulum Or another tubulo-vesicular system cannot be excluded. Slow flow, in contrast, transports high molecular weight and soluble materials which are involved in the growth and maintenance of the axon. The structural basis of slow transport is controversial, but the movement of the cytoplasmic matrix itself may represent the prime force.23 Mechanisms of anterograde and retrograde transport are similar: both need metabolic energy, have similar ionic requirements and are sensitive to the same drugs.23 Both anterograde and retrograde transport can be blocked by low temperatures and suspended by colchicine and vinblastine, agents which disrupt microtubules. The factors which determine the direction of axonal transport remain conjectural: alternatives envisage either two, oppositely polar­ised transport systems or a single mechanism in which the direction of movement is determined by the nature of the material or organelle to be trans­ported. Investigation of the molecular mechan­isms of axonal transport should contribute to our further understanding of this vital neuronal function.

 

Dendrites

 

There is wide variation in the number and organisation of these cell processes in the central nervous system. The character of the dendritic tree reflects the afferent neuronal connections and consequently indicates the functional activity of the cell. The complex configuration of dendrites is best appreciated in Golgi preparations; these have allowed the classification of neurons into three groups based on their dendritic arborisations (Fig. 1.1). Isodendritic neurons issue straight dendrites which run in all directions, whilst allodendritic cells are distinguished by shorter, branched den­drites which are restricted in their wavy course. Idiodendritic neurons have a unique dendritic tree characteristic  of and  determined  by  their location.27 A further, more detailed classification of neurons has been achieved by the dendritic branching pattern.


Although clear differences exist, it may not be always easy to distinguish a dendrite from an axon. Dendrites  have  irregular  contours,  taper gradually, branch at relatively acute angles, are unmyelinated and contain Nissi substance. Axons, in contrast, display a smoother contour, have a relatively even diameter along their course, branch at obtuse or at right angles, can be myelinated or unmyclinated and lack Nissl substance. The smaller axons and dendrites become, the more difficult is their positive identification; the presence of clustered ribosomes, particularly in association with cisternac of rough-surfaced endoplasmic reticulum, remains the most reliable identifying feature of small dendritic processes.

 

The dendritic tree represents the most sub­stantial receptive area of the nerve cells Synapses are formed either on the dendritic trunks them­selves or on the dendritic spines, the specialised structures projecting from them. The spines are composed of a stalk or neck which connects the dendrite with the ovoid bulb or head. There is a considerable variation in their configuration, but their overall length appears to be relatively con­stant at 2 um. Spines can also be present on the perikaryon and they are responsible for 43()() of the total surface area of the dendrites and the cell body: there are 4000 spines on a single pyramidal neuron.~ The function of the dendritic spines is still largely unknown. Although they synapse with axon terminals, their role is not restricted to increasing receptive surfaces; they may regulate the excitatory input to a neuron.1

 

 

The neuropil and the extracellular space

 

The neuropil is composed of the complex and intricate network of neuronal cell processes. This intermingling and interconnection of myelinated and unmyelinated axons and dendrites may appear random, but these processes form the neuronal circuitry of a particular area. Cell processes of astrocytes, oligodendrocytes and microglial cells further add to the variety of structures in the neuropil (Fig. 1.3).

Cell processes and perikarya, bounded by typical unit membranes, are separated by an extra-cellular space with an average width of 15-20 run. Electron-dense markers, electrical impedance and the distribution of radiotracers have conclusivelv demonstrated the existence of this extracellular space, which constitutes 18~25o() of the total gre~' and white matter.29

 

Myelin

 

Myelin in the central nervous system is produced by oligodendrocytes (see p. 22-24). The myclin sheath is formed by the spiral wrapping of the cytoplasmic processes of the myelinating cells around axons, and the subsequent extrusion of the cytoplasm leads to a compact, tightly spiralled, multi-layered envelope.  Electron microscopy reveals the characteristic structure of the myelin sheath: major dense lines alternate with thinner intraperiod lines to form the repeating units. The major dense line results from the fusion of the thicker, inner leaflet of the oligodendrocytic plasma membrane, whereas the intraperiod line is formed by the apposition of the thinner, outer leaflet of this membrane. Unlike Schwann cells (the myelinating cells of the peripheral nervous system which are concerned with the myelination of a single axon), each oligodendrocyte may provide myelin for many axons. From this it follows that the motion by which the sheath is produced by the myelinating cell may be different in the central and peripheral nervous Systems. Whilst Schwann cells can, in principle, lay down myelin by rotating around a single axon, the oligodendrocyte, being in connection with many axons, cannot perform a revolving motion. Con­sequently, ensheathment takes place by the progressive lengthening of the oligodendrocytic process which encircles the axon completely and an internal mesaxon is formed by apposition of the free edges of the myclinating process. The outer­most lamella of the sheath encloses the outer tongue and this, together with the internal mesaxon, represents the tenuous connections which remain between the fully formed myclin and the myelinating cell.

 

Nodes of Ranvier occur regularly along the course of central myclinated fibres; the segment between two nodes is referred to as internodal myclin, and the region where the lamellae terminate is the paranode. The thickeness of the myelin sheath is related to axonal diameter: larger axons are surrounded by thicker myelin sheaths. Evidence from experimental animals has con­vincingly shown that the formation of myelin is preceded by the proliferation of glial precursors which develop into young oligodendrocytes. The processes of these active oligodendrocytes are frequently wrapped around axons, but this direct connection is difficult to demonstrate in later life.' Biologically, the formation of myclin is a two-step process: first, the oligodendrocyte relates to the axon in response to an axonal myelinogenic stimulus and, second, it produces myelin, the volume of which is determined by the internodal axonal surface area. 30

Mvelination in man progresses slowly and generally proceeds centripetally: it commences peripherally and whilst many axons, particularly in the spinal cord) are myclinated at birth, the more central fibres in the frontal and parietal lobes remain unmyclinated well into posmatal life. Completion of myelin formation is achieved largely during the first two years after birth.31 Immunohistochemistry for myelin basic protein confirms earlier findings of the chronological sequence of myelination: phylogenetically older regions acquire myelin first.32 The sequential nature of myelination reflects physiological demand: the time and rapidity of myelination are related to the relative significance of a fibre system at various periods of cerebral development.

 

Biochemically, myelin is composed of altern­ating layers of proteins and lipids. Of the myelin proteins, a proteolipid protein constitutes approximately 50 mu, basic protein (the antigenic agent capable of inducing experimental allergic encephalomyclitis) 30%, and an acidic proteolipid protein 200/0 .3' Myelin-associated glycoprotein is an acidic, concanavalin A binding minor com­ponent which is also present in the cytoplasm of oligodendrocytes before and during myelination. The various lipids to be found in the myelin include cholesterol, phosphatidylethanolamine, phosphatidylserine  and  phosphatidylcholine, sphingomyelin   and   glycolipids.   Galacto­cerebroside is the main glycolipid component and the cell membrane of the myelinating cell contains a high percentage of this compound. This suggests that galactocerebroside may play an important role in the successive layering of cell membranes, a

process unique to myelination.