Bert Csillik

Fine structure of growth cones in the upper dorsal horn of the adult primate spinal cord in the course of reactive synapto-neogenesis

SummaryFollowing transganglionic degenerative atrophy of primary afferent terminals induced by a crush-injury of the sciatic nerve, a regenerative process takes places in the upper dorsal horn of the lumbar spinal cord in the primate Macacus rhesus. Axonal growth cones are characterized by cisterns of axoplasmic reticulum; filopodia emanating from growth cones are electron-optically translucent sheet-like expansions, often containing growth-cone vesicles. Axoplasmic reticulum appears also in preterminal portions of regenerating axons. Dendritic growth cones contain a fine, filamentous matrix; electron-dense membrane specializations can be seen in well-defined areas of their surfaces. Immature synapses are formed between filopodia of axonal growth cones and dendritic growth cones. Electron-microscopic structures of this unique CNS regeneration are similar to those seen in the course of embryonic development of the spinal cord.

Alterations in glial fibrillary acidic protein immunoreactivity in the upper dorsal horn of the rat spinal cord in the course of transganglionic degenerative atrophy and regenerative proliferation

Transection of a peripheral nerve induces marked increase in the glial fibrillary acidic protein (GFAP) immunoreactivity in the ipsilateral, segmentally related upper dorsal horn. Increase of GFAP immunoreaction is similar to, but not identical with, that observed after dorsal rhizotomy. If the peripheral nerve succeeds in regenerating, GFAP immunoreactivity in the upper dorsal horn returns to normal. It is concluded that the amount and distribution of GFAP is determined by transganglionic degenerative atrophy. Wallerian degeneration and regenerative proliferation of dorsal root axon terminals, respectively.

Treatment of chronic pain syndromes with iontophoresis of vinca alkaloids to the skin of patients

Abstract Repeated iontophoretic administration of the microtubule inhibitors vinblastic or vincristine to the segmentally related dermatomes of patients suffering from postherpetic, trigeminal and other neuralgias permanently alleviates chronic, autochthonoue pain. The beneficial effect of this therapy is probably due to transganglionic degenerative atrophy of primary central sensory terminals in the Rolando substance by blockade of retrograde axoplasmic transport in sensory nerves.

Iontophoretically applied microtubule inhibitors induce transganglionic degenerative atrophy of primary central nociceptive terminals and abolish chronic autochtonous pain

Transcutaneous iontophoresis of microtubule inhibitors (Vinblastin®, Vincristin®, Formyl‐Leurosin® in rats induces depletion of fluoride‐resistant acid phosphatase (FRAP) and transganglionic degenerative atrophy (trggl. deg. atr.) of the central terminals of primary nociceptive neurons, probably via blockade of axoplasmic transport in the peripheral sensory nerves. Radiochemical experiments prove that about 0.2% of the microtubule inhibitors applied iontophoretically at the skin reach the level of nociceptive axon terminals. 40 out of 48 patients suffering from chronic intractable pain of diverse etiology (postherpetic, paresthetic, ischaemic and trigeminal neuralgia, alcoholic and diabetic polyneuropathy, meralgia, brachialgia, discopathia, arthropathia and terminal pain) were successfully treated with Vinblastin® or Vincristin® iontophoresis. Iontophoretically applied microtubule inhibitors do not affect the blood cell count, have no side‐effects and do not impair the skin at the site of application.

Kynurenine aminotransferase in the supratentorial dura mater of the rat: Effect of stimulation of the trigeminal ganglion

Electrical stimulation of the trigeminal ganglion has been widely used as a model of nociception, characterizing migraine. This treatment is known to evoke release of neuropeptides and neurotransmitters from nerve fibers of the dura mater. On the basis of immunocytochemical investigations, we found that under normal conditions, surface membranes of Schwann cells surrounding nerve fibers in the supratentorial dura mater display kynurenine aminotransferase-immunoreaction (KAT-IR); also KAT-IR are the granules of mast cells and the cytoplasms of macrophages (histiocytes). In consequence of stimulation of the trigeminal ganglion, Schwann cells in the dura mater became conspicuously swollen while their KAT-IR decreased considerably; also KAT-IR of mast cells and macrophages decreased significantly. At the same time, nitric oxide synthase (NOS)-IR of nerve fibers in the dura mater increased, suggesting release of nitric oxide (NO), this is known to be involved in NMDA receptor activation leading to vasodilation followed by neurogenic inflammation. Because kynurenic acid (KYNA) is an antagonist of NMDA receptors, we hypothesize that KYNA and its synthesizing enzyme, KAT, may play a role in the prevention of migraine attacks.

A case for transmitter plasticity at the molecular level: Axotomy-induced VIP increase in the upper spinal dorsal horn is related to blockade of retrograde axoplasmic transport of nerve growth factor in the peripheral nerve

Blockade of retrograde axoplasmic transport in peripheral nerves, by means of perineurally applied microtubule inhibitors, results in an increased vasoactive intestinal polypeptide (VIP) reaction of the segmentally related, ipsilateral upper dorsal horn. Similar effect is elicited by the perineural application of an anti-Nerve Growth Factor (anti-NGF) serum. At the same time, both treatments result in depletion of Substance P from the same region of the spinal cord. It is assumed that this striking example of transmitter plasticity, obviously taking place at the molecular level, is due to a stimulating effect of NGF upon the perikaryal Substance P-synthesizing mechanism in dorsal root ganglion cells, and the inhibitory effect of NGF upon the VIP synthesizing machinery in these same nerve cells.

Transneuronal degeneration in the Rolando substance of the primate spinal cord evoked by axotomy-induced transganglionic degenerative atrophy of central primary sensory terminals

SummaryTransection of the sciatic nerve in Rhesus monkeys and the consequent transganglionic degenerative atrophy (TDA) of central terminals of primary afferents result in transneuronal degeneration of substantia gelatinosa (SG) cells. Severe degeneration is characterized by an increased electron density of the nucleus and by conspicuous shrinkage of the cytoplasm, mitochondrial swelling, dilation of cisterns of the rough-surfaced endoplasmic reticulum, accumulation of free ribosomes and an electron-dense material in the cytoplasm. In the mild form, dilation of cisternal elements of the endoplasmic reticulum, swollen mitochondria and accumulation of free ribosomes takes place. About 10% of SG cells in segment L5 undergo the severe form whereas the rest shows signs of the mild form. Cytoplasmic alterations that occur during transneuronal degeneration seem to start at the level of subsurface cisterns. Dendrites and axons of transneuronally degenerating SG cells also show a conspicuous electron density. By analyzing the synaptic relationships of such “darkened” dendrites, connections in the upper dorsal horn can be deciphered. Modular units of the primary nociceptive analyzer that evaluate noxious and innocuous inputs on the basis of thin versus thick (AδC/Aβ) afferent activity and subjecting them to descending control appear to be recruited from structurally dispersed elements of synaptic glomeruli. These are arranged alongside dendritic processes of large antenna cells which relay impulses to projection cells of the spinothalamic tract.

Fine structural correlates of VIP-like immunoreactivity in the upper spinal dorsal horn after peripheral axotomy: Possibilities of a neoro-glial translocation of a neuropeptide

After transection of the peripheral nerve, VIP-like immunoreactivity (VIPLI) increases markedly in the ipsilateral upper dorsal horn of the spinal cord. Immunoreactivity has been studied by means of light- and electron microscopic immunocytochemistry. Under normal conditions, there is little VIPLI present in the superficial dorsal horn, confined to small dot-like elements corresponding to axonal and glial profiles. At the electron microscopic level, immunostaining was found mainly in preterminal and, partly, also in en passant terminal swellings or varicosities. The reaction was confined to the axoplasm and, to a lesser extent, to large dense core vesicles. VIPLI is also present in several astroglial processes. 13, 19, and 25 d after transection of the sciatic nerve, increased immunoreactivity was present in the medial 2/3 of the superficial dorsal horn. Electron microscopically, VIPLI was seen mainly in preterminal axons and in many astroglial processes surrounding axon terminals while VIPLI in the en passant axon terminals themselves decreases. 2 months after peripheral axotomy, the amount of axonally localized VIPLI decreases considerably and most of the immunocytochemically detectable VIPLI is found in expansions and processes of astroglial cells. Perikarya of glial cells rarely exhibit VIPLI. VIPLI also increased after crushing the related peripheral nerve; however, as soon as the nerve fibers regenerate, VIPLI decreases again to normal levels. It appears that blockade of the retrograde axoplasmic transport induces a switch in the neuropeptide synthesizing machinery of dorsal root ganglion cells which results in the expression of VIP instead of substance P, somatostatin and CGRP. It is proposed that VIP is released from axon terminals affected by transganglionic degenerative atrophy. Subsequently, astroglial cells equipped with receptors for VIP, might bind and internalize the released VIP.

Mapping the distribution of thiamine monophosphatase, fluoride-resistant acid phosphatase, and substance P in the spinal cord with a personal computer compatible program

Distribution of the marker enzymes fluoride-resistant acid phosphatase (FRAP) and thiamine monophosphatase (TMPase) as well as that of the marker neuropeptide substance P (SP) has been mapped by means of the semi-automatic computer program MUDH, in various experimental conditions, including degeneration, regeneration, and anti-Nerve Growth Hormone treatment of the sciatic nerve. The program is written in standard Pascal language; it is applicable to Commodore 64 and IBM compatible personal computers. Graphical illustration of values obtained in serial sections is a powerful and highly valuable contribution in the analysis of various biodynamic reactions related to transganglionic regulation of primary sensory neurons.

Structural, Functional and Cytochemical Plasticity of Primary Afferent Terminals in the Upper Dorsal Horn

Structural and functional plasticity of central terminals of primary sensory neurons is regulated by nerve growth factor supplied by retrograde axoplasmic transport to dorsal root ganglion cells. Two important aspects of this regulatory system: transganglionic degenerative atrophy and regenerative synaptoneogenesis are reviewed in view of electron microscopic histochemical and immunocytochemical studies.