Electron-dense Axon Research Articles

Persisting axonal degeneration in the hippocampus after transection of the fimbria-fornix

Degeneration within the hippocampus was examined at the light microscopic level using the Gallyas silver stain two, four or nine months after bilateral transection of the fimbria-fornix and commissural connections. At two or four months after the lesion the strata oriens and radiatum of the subicular end of the CA1 subfields were strongly argyrophilic as was the inner third of the molecular layer of the dentate gyrus. At nine months post-lesion argyrophilia diminished but clearly persisted in the same layers. Electron microscopic examination revealed a large number of electron-dense axon terminals in the argyrophilic areas, most of them making asymmetric synaptic contacts with dendritic spines. These findings suggest that at least a portion of the Schaffer collaterals of the CA3 pyramidal cells and associational collaterals of hilar neurons were in a process of acute degeneration at all time points after the initial surgical trauma. This persistent synaptic reorganization of intrahippocampal circuits may be related to abnormal electrical activity observed several months after fimbria-fornix transection.

Transganglionic degeneration in vibrissae innervating primary sensory neurons of the rat: a light and electron microscopic study.

Previous studies have shown that transection of peripheral branches of primary sensory neurons leads to light microscopical degeneration argyrophilia and ultrastructural changes in the central termination areas of these neurons. This type of degeneration has been termed transganglionic degeneration (TGD). In the present experiments TGD has been studied specifically in neurons innervating the rat vibrissae at the light and electron microscopic levels. Light microscopically, small amounts of degeneration argyrophilia are observed in the magnocellular zone of the trigeminal subnucleus caudalis at 8-14 days survival. At longer survival times there are substantial amounts of degeneration in this area. At the ultrastructural level the first signs of TGD are observed at 6 days survival, when some terminals show a small increase in electron density, loss of synaptic vesicles, and mitochondrial disintegration. Terminals showing a more advanced increase in electron density become common at 8 days survival, but few of them are still left at 14 days survival. Neurofilamentous terminals appear in small numbers 8-14 days postoperatively. Various forms of degeneration in myelinated axons are observed from 8 days survival and are common also at 80 days survival. Electron-dense axons are rather unfrequent, but more or less disrupted myelin sheaths containing disintegrated axoplasmic remnants and empty areas are common as well as extremely expanded myelin sheaths. Glial cells containing axonal and myelin debris are seen from 8 days survival and become a more common finding at longer survivals. A most striking finding 8-10 days postoperatively is a complex relationship between glial cells and less darkened terminals, indicating phagocytosis before reaching an entirely darkened state. The findings clearly show that peripheral nerve transection leads to severe central alterations in a population of mechanoreceptor neurons innervating the vibrissae of the adult rat.

The fate of axonal debris in Wallerian degeneration of rat optic and sciatic nerves. Electron microscopy and immunofluorescence studies with neurofilament antisera.

Immunofluorescence demonstrated that axonal debris reacting with neurofilament antisera persist up to 4 months in rat optic nerves undergoing Wallerian degeneration. Antisera used in this study allowed the isolation of the 72,000- and 150,000-dalton neurofilament polypeptides from rat spinal cord by immunoaffinity chromatography. After 2 weeks of degeneration, proteins co-migrating with these neurofilament polypeptides were no longer identifiable in rat optic nerves, which suggests that immunofluorescent structures persisting in the nerves after this period contained neurofilament degradation products of different molecular weight. Additional evidence as to the persistence of axonal debris in degenerated optic nerves was obtained by electron microscopy. Two distinct types of axonal degeneration were observed in rat optic nerves by this method, floccular swelling and increased electron density of the axoplasm. In both types of degeneration, axoplasmic filaments and tubules were not identifiable. Although floccular material disappeared after 2 weeks of degeneration, so that only empty myelin sheaths remained, electron-dense axons persisted longer and were probably phagocytosed together with their myelin sheaths. In sciatic nerves, cross-reaction with neurofilament antisera had almost completely disappeared 10 days after transection. The same was true for nerves which had been tightly ligated to prevent axonal growth and to squeezed nerves which showed vigorous regeneration. A few scattered, brightly immunofluorescent fragments which persisted in nerves up to 2 weeks after transection were exception to these findings.

The synaptic terminations of certain midbrain-olivary fibers in the opossum.

The nuclear origin and distribution of midbrain-olivary fibers has been described in a previous study utilizing axonal transport techniques (Linauts and Martin, '78a). The present report extends their results to the electron microscopic level and details the postsynaptic distribution of such fibers. Lesions within the ventral periaqueductal grey and adjacent tegmentum, the red nucleus or the nucleus subparafascicularis result in electron dense axon terminals within the olive at survival times of 48, 72 and 96 hours. At 72 hours, many degenerating presynaptic profiles shrink, become irregular in shape and are totally or partially surrounded by glial processes. The principal olivary nucleus contains the majority of these profiles. However, the subparafascicular terminals are more abundant in the rostral and intermediate parts of the medial accessory nucleus and the rubral terminals are concentrated within the dorsal lamella of the principal nucleus. The nuclear location of the degenerating terminals was determined by examination of 1 micrometer plastic sections cut in the transverse plane from each block face prior to thin sectioning. Degenerating terminals were counted in three cases, one from each of the three lesion sites described above. When taken together these cases show that just over 50% of the degenerating terminals are presynaptic to spiny appendages and are located within the synaptic clusters (glomeruli) described previously (King, '76). The percentage of degenerating terminals in the glomeruli increases to 70% when the lesion is in the ventral periaqueductal grey and adjacent tegmentum. The remaining degenerating terminals contact dendritic shafts outside the astrocytic boundaries of the synaptic clusters. The synpatic vesicle populations within the degenerating terminals vary with the location of the lesion. Lesions in the ventral periaqueductal grey and the adjacent tegmentum result in the degeneration of terminals with either clear spherical vesicles or endings with both clear spherical vesicles and a variable number of large dense core vesicles. In contrast, the primary degenerative changes that occur after destruction of the red nucleus or the nucleus subparafascicularis are in terminals with clear spherical vesicles. When the synaptic complex was present in the plane of section, regardless of the site of the lesion, the degenerating terminals could be classified as Gray's type I. Thus, we have demonstrated that afferents from the mesencephalon terminate within synpatic clusters located in the principal and medial accessory (part A) subnuclei of the inferior olive. Although the mesencephalic afferents have multiple origins (Linauts and Martin, '78a), many of their synaptic terminals contact spiny appendages within the synaptic clusters. This postsynaptic site also receives cerebellar terminals (King et al., '76). The origin of presynaptic profiles within the synaptic clusters that contain clear pleomorphlic vesicles is yet to be determined.

The direct spinal area of the inferior olivary nucleus: an electron microscopic study.

Identification of the direct spinal areas (portions of the dorsal and medial accessory nuclei) within the opossum inferior olivary complex was accomplished by mapping the location of the terminal degeneration by the Fink-Heimer technique subsequent to cervical cord lesions. Following similar lesions, sampling of these same regions for electron microscopic study was assured by examination of transversely oriented, 1 mu plastic sections prior to thin sectioning. The first evidence of electron dense axon terminals was found at a survival time of 24 hours. At survival times of 36, 48 and 72 hours, degenerating presynaptic profiles shrink, become irregular in shape and are totally or partially surrounded by glial processes. Spinal terminals average 1-2 mu in their greatest dimension, contain round, clear synaptic vesicles and generally contact small diameter (0.4--1.8 mu) dendritic shafts or occasional spiny appendages. The spiny dendritic appendages make up the central core of the olivary glomeruli and these juxtaposed dendritic processes exhibit gap junctions. At longer survival times (5, 7 and 9 days) many presynaptic profiles with either round or pleomorphic synaptic vesicles remain normal in appearance and contact dendritic shafts or the spiny appendages within glomeruli. Afferents from other sources (possible including intrinsic neurons) must terminate within the direct spinal portion of the nuclear complex to account for the numerous axon terminals which retain normal morphology after such long survival times.

The synaptic organization of lemniscal projections to the ventrobasal thalamus of the cat

The fine structure of degenerating synapses in VB following ablation of the dorsal column nuclei has been studied with the electron microscope. Within 24 h after the lesion, large synaptic knobs forming asymmetrical synapses are seen to fill with neurofilaments. The neurofilamentous degenerative changes become more marked 2–3 days following the lesion. The degenerating knobs exhibit an electrondense appearance after 3–5 days, and then are phagocytosed and most disappear after 5–10 days. The degenerating lemniscal synapses are found to synapse principally upon dendrites, and also upon other synaptic knobs or dendritic profiles giving rise to synapses. The fine structure of degeneration is compared to degeneration seen with the light microscope. Neurofilamentous synaptic degeneration corresponds to theformation of neurofibrillar rings stained by the Glees method. Electron-dense axons and synapses are correlated with staining of degeneration with Nauta methods. The morphological findings support physiological postulates concerning secure relations between lemniscal terminals and VB neurons, but are at variance with physiological models used to explain presynaptic inhibition.

AN ELECTRON MICROSCOPICAL STUDY OF DEVELOPING SYMPATHETIC NEURONS IN MAN.

An electron microscopic study has been made of the sympathetic ganglia of a 15 and a 17 week old male human fetus. The fetal sympathetic neurons were densely packed in a scanty connective tissue matrix which also contained blood vessels. The fetal sympathetic neurons had a large, electron-light nucleus with one or two nucleoli, and was of a somewhat mottled appearance due to irregularly dispersed aggregates of fine and coarse granules. The perikaryon usually formed a thin envelope around the nucleus and contained, except for large pigment granules, all intracytoplasmic structures which were also found in mature sympathetic neurons. Adjacent sympathetic cells were either in immediate contact, or slightly separated by a wedge of electron-light satellite expansions, or lined by primitive axons. The satellite cells were in the early state of development. Electron-dense axons either stood side by side with, or were slightly engulfed by light Schwann cell expansions and formed distinct bundles surrounded by a common basement membrane. There was practically no trace of myelin formation or Schwann cell wrapping characteristic for unmyelinated fibers as seen in the adult.