Biomedical subjects

M J Politis

Publications and source records attributed to M J Politis.

33 records · Page 2Linked to original sources

PubMed1985

The role of post-traumatic mitosis in elevation of anaerobic metabolism enzyme (lactic acid dehydrogenase) activity in degenerating central and peripheral nerve.

Glial and Schwann cells undergo marked biochemical and morphological alterations following axonal injury. In the present experiments, the extent of enzyme activity associated with anaerobic (LDH, lactic dehydrogenase) vs aerobic (SDH, succinic dehydrogenase) respiration was assessed distal to the site of nerve fiber injury. Studies were performed in rat central (optic) and peripheral (sciatic) nerves at 2, 7 and 14 days postoperatively (d.p.o.). In sciatic nerves, LDH activity rose 3-fold in traumatized (vs unoperated control) nerve tissue between 2 and 7 d.p.o. and remained elevated at 14 d.p.o. SDH activity in traumatized nerve was equal to that in unoperated nerve at 7 d.p.o., but decreased at 14 d.p.o. LDH activity in optic nerve at 2 d.p.o. was equivalent to that in control nerve, but rose approximately two-fold by 7 d.p.o. However, unlike peripheral nerve, activity in traumatized optic nerve decreased to control levels at 14 d.p.o. SDH activity in traumatized optic nerve remained unchanged at any timepoint examined. Taken in concert, these data are consistent with the hypothesis that there is an overall shift in CNS glial and Schwann cell metabolism from aerobic to anaerobic respiration following nerve injury. Additional studies were performed to determine if this shift requires prior Schwann or glial cell mitosis. Administration of mitotic inhibitor (AraC, cytosine arabinofuranoside) inhibited post-traumatic elevations in LDH activity in optic, but not peripheral nerve. No significant effect of the drug on axonal degeneration (as assessed by saxitoxin binding) was observed.

Animals↗

PubMed1985

The roles of reactive gliosis and mitosis on tropic factor production in traumatized nervous system tissue.

Proximal stumps of rat sciatic nerves were attached to inlet ends of Y-shaped silastic implants and offered a 'choice' of growing toward an Elvax pellet containing homogenate from previously crushed optic nerve which had been exposed to saline or cytosine arabinofuranoside (AraC). Previous studies indicate that AraC administration inhibits reactive gliosis in crushed optic nerve. Preferential or exclusive growth of axons occurred in implant forks attached to pellets containing saline- (vs drug-) exposed optic nerve homogenates. In contrast, inhibition of Schwann cell mitosis had no discernible effect on tropic factor production in distal stumps of transected sciatic nerves. Tropic activity of homogenates from cultures containing reactive-like astrocytes was nearly 4 times higher than homogenates not containing these cells. Results suggest a possible link between formation of hypertrophic reactive astrocytes and production of neurotropic factor.

Animals↗

PubMed1985

Specificity in mammalian peripheral nerve regeneration at the level of the nerve trunk.

Previous studies indicate that distal stumps of transected rat peripheral nerves secrete 'tropic' factors which can attract/support axonal regeneration over distances of several mm in vivo. The present study was undertaken in order to determine if there is specificity of neurotropic interaction at the level of the nerve trunk. Proximal stumps of transected peroneal or tibial nerves were inserted into the single inlet end of Y-shaped Silastic implants and offered alternative 'lures' at the paired outlet ends (specifically, grafts of peroneal vs tibial distal stump tissue). Several weeks later, the overwhelming majority of preparations showed exclusive growth of nerve fibers in implant forks attached to 'native' (originally associated) nerve stumps. Inversion of the distal stump grafts (such that the proximal stump was facing an analogous native distal stump, but a different region of it) diminished the frequency and extent of native preference. Taken together, data suggest the possibility that there can be a specificity of nerve regeneration at the level of the nerve trunk.

Animals↗

PubMed1985

Tropic factors in reactive mammalian central nervous system tissue.

Previous studies suggest that distal stumps of transected peripheral nerves contain diffusible factors which can attract/support axonal regeneration over distances of several mm in vivo. The present experiments were undertaken to determine if this is so for distal regions of traumatized central (i.e., optic) nerves. Proximal stumps of transected rat sciatic nerves were inserted into the single inlet ends of 6 mm long Y-shaped Silastic implants. Alternative 'lures' were attached to the paired outlets, the ability of these lures to attract/support regeneration of nerve fibers in their associated forks assessed 3.5 weeks postoperatively. Exclusive or preferential growth of nerve fibers occurred in implant forks associated with optic nerve grafts, of Elvax pellets containing homogenate obtained from previously crushed (reactive) optic nerves. Grafts of tendon, as well as homogenate from unoperated optic nerve had no effect. Results suggest that, with respect to the assay used, degenerating optic nerve tissue contains factor(s) which can attract/support regenerating nerve fibers.

Animals↗

PubMed1985

Effect of cytosine arabinofuranoside (AraC) on reactive gliosis in vivo. An immunohistochemical and morphometric study.

Glial reactivity is believed to contribute to the lack of functional recovery after injury to the mammalian central nervous system. The role of glial mitosis in the progression of events associated with reactive gliosis has received little attention. In the present study, the expression of reactive gliosis distal to the site of crush in rat optic nerves was assessed in the presence and absence of a chronically administered mitotic inhibitor, cytosine arabinofuranoside (AraC). Right optic nerves were crushed and animals sacrificed 1, 2 or 3 weeks later. Parameters assessed were (1) glial hypertrophy, (2) degradation of myelin sheaths and (3) ability of tissue to stain with antisera raised against glial filament protein (GFA), actin and vimentin. In saline treated animals, greater than 90% of the myelin sheaths distal to the site of axonal injury had degraded within 7 days postoperatively. Glial hypertrophy was evident by the second week after after crush and increased progressively. The number of GFA-positive profiles (i.e., cells) increased between 1 and 3 weeks postoperatively. Vimentin staining increased 4-fold between 1 and 2 weeks after injury and subsequently showed no change. Actin staining rose 3-fold between 1 and 2 weeks after injury, but decreased by the third postoperative week. In AraC treated animals, almost 50% of the myelin sheaths distal to the injury site were preserved a week after surgery. A delay in myelin degradation continued until the second postoperative week. Glial hypertrophy was evident at the 2 and 3 week time points. However, the extent of hypertrophy was substantially lower in drug (vs saline) treated animals. Vimentin staining never rose above minimal levels in AraC treated animals. Actin staining in AraC rats at 2 weeks postoperatively was equivalent to that in saline injected animals, but in contrast to the results in the latter group, increased (3-fold) between 2 and 3 weeks after crush. Results indicate a delay in the expression of reactive gliosis with chronic administration of AraC. It is proposed that this might be due to a delay in the appearance of 'signals' (e.g., myelin debris) which initiate the process of reactive gliosis.

Actins↗

PubMed1985

Electromyographic evaluation of a novel surgical preparation to enhance nerve-muscle specificity that follows mammalian peripheral nerve trunk transection.

Previous studies indicated that axons from proximal stumps of transected peripheral nerves "prefer" to grow through Silastic tubes attached to their native (originally associated) rather than foreign (not originally associated) distal stumps. We determined whether or not this specificity is expressed at the level of the neuromuscular junction. Proximal stumps of transected rat sciatic nerves (peroneal and tibial branches) were attached to single inlet ends of 6-mm-long, Y-shape Silastic implants. One outlet was attached to the distal peroneal and the other to the distal tibial stump. Ten weeks later, innervation of the anterior tibialis and interosseous muscles (normally innervated predominantly by peroneal and tibial nerve fibers, respectively) was assessed by measuring compound muscle action potential amplitudes and latencies that follow supramaximal peroneal and tibial nerve stimulation. Results showed higher amplitudes in anterior tibialis muscle, induced by "native" peroneal (vs. tibial) stimulation in four of five animals, and higher amplitudes in interosseous muscles after "native" tibial (vs. peroneal) stimulation in all cases examined. Preparations in which bridges between proximal and distal nerve stumps were bridged with unbranched tubes showed random patterns of muscle innervation. The results suggest that if allowed to express "specificity" at the level of nerve trunk transection, regenerating mammalian peripheral axons can grow into, and form functional connection with, native (vs. foreign) muscle groups. This finding has possible clinical significance.

Action Potentials↗

PubMed1983

An in vivo assay of neurotropic activity.

Previous studies in this laboratory indicate that diffusible factors from distal stumps of transected peripheral nerves exert a neurotropic effect on regenerating nerve fibers in vivo. The present study was performed (a) to strengthen this hypothesis, and (b) to begin to determine the distribution and identity of putative neurotropic factors in peripheral nerves. Rat sciatic nerves were transected and inserted into the inlet end of a hollow 6 mm-long Y-shaped implant made of medical-grade Silastic tubing. To one of the outlet ends was attached an Elvax pellet impregnated with whole homogenate from a single sciatic nerve. The other outlet was attached to a pellet not containing homogenate. Homogenates assayed were obtained from distal stumps of transected nerves derived 14 days postoperatively or from unoperated sciatic nerves. After 3.5 weeks, nerve fiber bundles consisting of myelinated regenerating axons were only present in implant forks attached to pellets containing denervated nerve homogenate. This activity was heat- and trypsin-sensitive. These data (a) strengthen the hypothesis that diffusible factors from distal nerve stumps of transected nerves can support nerve fiber regeneration, (b) indicate that those factors are protein and/or protein dependent, and (c) suggest that unoperated peripheral nerve (at the levels assayed) either do not contain such factors or contain substances which inhibit such factors.

Animals↗

PubMed1983

Reactive glial protein synthesis and early disappearance of saxitoxin binding in degenerating rat optic nerve.

The biochemistry of gliotic CNS tissue was assessed by monotoring changes in de novo protein synthesis distal to site of crush in the rat optic nerve between 3 and 20 days post-operatively. Radioactivity profiles on 12% polyacrylamide SDS gels showed reproducible peaks associated with protein(s) with dissociated molecular weights of 57K, 51K, 42K, 40K, 37K and 23K. Differences between crush and control nerves were observed with respect to the latter two peaks. De novo synthesis of 23K protein (comigrant with myelin proteolipid protein) was evident in control but not crushed nerves. Synthesis of 37K protein (identity unknown) was evident at 7, 10 and (to a lesser extent) 20 days post-operatively in crushed nerves, but not in crushed nerves at 3 days post-operatively or in unoperated nerves at any time point. The appearance of the synthesis of the 37K protein coincides with a drop in the level of functional axolemma (assessed by [3H]saxitoxin binding) in crushed nerves from 78% to 36% of control levels between 3 and 7 days post-operatively.

Animals↗

PubMed1983

Rapid reorganization of the axonal cytoskeleton induced by a gamma diketone.

Microtubules and neurofilaments are the major components of nerve fiber axoplasm and are evenly distributed longitudinally. Rearrangement of this distribution can be induced by a number of chemicals, including 2,5-hexanedione (2,5-HD). Utilizing 2,5-HD, this study explores the earliest time points of observable rearrangement of the cytoskeletal elements. Intrafascicular injections into the sciatic nerve shows central accumulation of microtubules and peripheral relocation of neurofilaments within 1 min of injection, suggesting a direct effect of the toxin on these components.

Animals↗

PubMed1982

Tropism in nerve regeneration in vivo. Attraction of regenerating axons by diffusible factors derived from cells in distal nerve stumps of transected peripheral nerves.

We re-examined the hypothesis of Cajal3, later refuted by Weiss and Taylor20, that cells in distal stumps of transected peripheral nerves exert an attractive (tropic) effect on regenerating axons. This question was re-assessed in vivo using surgical materials and assay procedures not available to those workers. Proximal stumps of transected rat sciatic or cat peroneal nerves were inserted into the single inlet end of a hollow, Y-shaped Silastic implant. Regenerating axons were provided with alternative targets consisting of a vacant arm vs one occupied by a sciatic nerve graft (rat), or a tibial (Tout) vs peroneal (Pout) distal nerve stump (cat). In some cases Pout was rendered metabolically compromised relative to Tout by exposing the former to dry ice and inhibitors of DNA and RNA synthesis. At 4.5 or 6 weeks postoperatively, the number of regenerating axons in each fork of the implant was assessed by morphometric analysis (total number of non-myelinated and myelinated axons greater than 1 micron in diameter at 4.5 weeks, and total number of myelinated axons at 6 weeks postoperatively), or by quantification of an axonally transported label. Rat sciatic nerve fibers exclusively regenerated toward the nerve graft, suggesting the existence of a neurotropic lure. In cats, morphometric analysis revealed a 10-(4.5 week) and 6-fold (6 week) greater number of axons growing towards untreated Tout vs treated Pout. When both distal stumps were untreated, more axons were seen in forks leading to Pout. Analysis of transported label confirmed the preferential growth of axons towards untreated Tout vs treated Pout for both motor and sensory axons. In separate experiments, Nuclepore filters (0.2 microns, pore size) were inserted between distal nerve stumps and outlet ends of Silastic implants. Preferential regeneration toward untreated stumps was observed if the distance between proximal and distal nerve stumps was equal to but not greater than 4-5 mm. These results suggest that peripheral nerve fiber regeneration in vivo can be directed by cells in distal stumps of transected nerves, and that this effect can be mediated over distances of several millimeters via diffusible factors.

Animals↗

PubMed1982

Studies on the control of myelinogenesis. IV. Neuronal induction of Schwann cell myelin-specific protein synthesis during nerve fiber regeneration.

The temporal sequence of axon-Schwann cell interaction during regeneration is examined in cat tibial nerves surgically denervated for 8 weeks and, subsequently, coated to a freshly severed peroneal nerve for 3 weeks. Prior to association with regenerating axons, Schwann cells resident in denervated and reinnervated stumps failed to synthesize proteins co-migrant with P0, P1, and P2 myelin proteins in contrast to normal nerves. Axonal association with Schwann cells stimulated synthesis of amino acid-labeled proteins co-migrating with myelin-specific proteins prior to elaboration of myelin lamellae. Radioactivity from these peaks was precipitated by antibodies raised against myelin-specific proteins. The synthesis of P1 and P2 proteins was evident before P0 synthesis in reinnervated stumps. Immunocytochemical staining with antibody to P0, P1, P2, and myelin-associated glycoprotein (MAG) appeared only after myelin lamellae had been formed. These data suggest that Schwann cells: (a) synthesize proteins co-migrant with P1, P2, P0, and MAG in normal cat nerves; (b) cease detectable manufacture of these proteins after axonal loss; (c) regain their capacity to synthesize these proteins upon re-establishment of axonal association; and (d) during regeneration, express the synthesis of P1 and P2 before that of P0.

Animals↗

PubMed1981

A method to separate spatially the temporal sequence of axon-Schwann cell interaction during nerve regeneration.

This paper describes a surgical method of manipulating feline peripheral nerve regeneration to separate spatially the temporal sequence of events of axon-Schwann cell interaction during nerve fibre formation. The method allows regenerating axons from the peroneal nerve to reinnervate the distal stump of a axon- and myelin-free, Schwann cell-enriched, chronically denervated tibial nerve distal stump. Three zones can be morphologically delineated in the tibial nerve stump after three weeks of reinnervation: (1) a proximal myelinated zone, (2) a more distal, non-myelinated, axon-Schwann cell contact zone, and (3) a distal axon-free Schwann cell non-contact zone. The distal limit of the second zone can be determined accurately by the front of an axonally transported label. The large volume of available tissue makes this method suitable for interdisciplinary studies to elucidate the control of axon elongation, axon growth, and axon-Schwann cell inter-relationships.

Animals↗

PubMed1980

Ultrastructural studies of the dying-back process. V. Axonal neurofilaments accumulate at sites of 2,5-hexanedione application: evidence for nerve fibre dysfunction in experimental hexacarbon neuropathy.

This study examined the thesis that 2,5-dexanedione (2,5-HD) produces distal (dying-back) axonopathy by direct toxic action on nerve fibers. Single or repeated application of undiluted or 10% 2,5-HD to exposed rat sciatic nerves caused some fibres to develop focal axonal swellings filled with abnormally large numbers of 10 nm neurofilaments. Such changes occurred within four days of application and were especially prominent in equivalently treated nerve segments obtained from animals orally exposed to 0.5% 2,5-HD before surgery. Nerve fibers along the perimeter of tibial nerve fascicles exposed to 2,5-HD or 2,4-hexanedione (2,4-HD), a compound unable to produce systemic neuropathy, underwent non-specific breakdown and Schwann cell necrosis. Nerve fibres located in the centre of such fascicles only developed an hypertrophied paranuclear Schwann cell cytoplasm and did not proliferate intermediate filaments. Saline, hydrochloric acid and 1,6-hexanediol, a water-soluble hexacarbon also lacking systemic neurotoxic properties, produced no intra-fascicular changes when locally applied to the sciatic nerve. It is concluded (1) that 2,5-HD causes a giant axonal swellings by direct toxic action on the nerve fibre, (2) 2,5-HD does not induce a generalized disorder of cytoplasmic intermediate filaments and (3) primary Schwann cell changes produced by locally applied 2,5-HD or 2,4-HD are non-specific and unrelated to the formation of the giant axonal swellings.

Animals↗

PubMed1979

Axonal transport of nucleosides, nucleotides and 4S RNA in the neonatal rat visual system.

The axonal migration of RNA, the nucleoside uridine and its nucleotide derivates (NS/NT) were compared in neonatal and young adult rat optic axons. Tritiated uridine was injected into right eyes of developing (1- or 4-day-old) and young adult (40-day-old) rats which were sacrificed at times after injection ranging from 6 h to 20 days. Right and left lateral geniculates were removed and assayed for trichloroacetic acid soluble (NS/NT) and RNA radioactivity. Left minus right geniculate (L-RLG) radioactivity was used as an index of axonally migrating radioactivity. Results showed that uridine and its phosphorylated derivatives were transported along both neonatal and young adult rat optic axons. Greater than 90% of right geniculate (blood-borne) TCA soluble radioactivity was metabolized to volatile substances (probably 3H2O) by three days after injection, leaving approximately 3% of the neonatal and approximately 10% of the adult activity as [3H]NS/NT. In left geniculate fractions (containing transported material) approximately 15% and 40% of total TCA soluble radioactivity was present as [3H]NS/NT in neonates and adults, respectively. Thus, axonal NS/NT appears to be relatively protected from degradation when compared with blood-borne NS/NT. The amount of L-RLG [3H]RNA in the neonates was 10 times higher than in young adults. Peaks of neonatal [3H]RNA occurred at 5 and 10 days after birth, whether injections were made at 1 or 4 days of age indicating that this [3H]RNA may be linked to developmental events. Gel electrophoretic analysis of neonatal geniculate RNA indicated that a small portion of the [3H]RNA in the first peak represented axonally transported 4S RNA. The remainder of the L-RLG [3H]RNA in the neonates was probably due to a rapid and efficient incorporation of axonally transported [3H]NS/NT into extraaxonal geniculate RNA. In contrast, little or no axonal RNA transport could be demonstrated in the young adults.

Age Factors↗

PubMed1979

Axonal transport of taurine along neonatal and young adult rat optic axons.

Studies in this laboratory have indicated that taurine is axonally transported along goldfish optic nerves. In the present experiments the axonal transport of taurine was examined in neonatal and young adult rat optic axons. [35S]taurine was injected into the vitreous humor of right eyes of developing (1--15-day-old) or young adult (40-day-old) rats. At various times after injection ranging from 3 h to 7 days, right retinae and left and right geniculates were removed and assayed for radioactivity, left minus right lateral geniculate (L-RLG) radioactivity being used as an index of axonally transported [35S]taurine. Results indicated that taurine was rapidly transported along both neonatal and young optic axons, in contrast to other amino acids (i.e., leucine and proline) which are not axonally transported in this system. Significant developmental variations were seen in both L-RLG and right retinal [35S]taurine activity 24 h after injection. The amounts of L-RLG [35S]taurine corrected for retinal ganglion cell uptake in animals injected at 1,4,7 and 11 days after birth (prior to and during the major period of synaptogenesis in the geniculates) were 4.5, 3.1, 2.3 and 2.6 times higher, respectively, than those in the young adults. In contrast, the amount of corrected L-RLG [35S]taurine in animals injected at 15 days after birth (after synaptogenesis) were not significantly different from that in the young adult.

Age Factors↗