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J R Wujek

Publications and source records attributed to J R Wujek.

15 recordsLinked to original sources
PubMed2003

Axonal loss in the pathology of MS: consequences for understanding the progressive phase of the disease.

Axonal degeneration has been identified as the major determinant of irreversible neurological disability in patients with multiple sclerosis (MS). Axonal injury begins at disease onset and correlates with the degree of inflammation within lesions, indicating that inflammatory demyelination influences axon pathology during relapsing-remitting MS (RR-MS). This axonal loss remains clinically silent for many years, and irreversible neurological disability develops when a threshold of axonal loss is reached and compensatory CNS resources are exhausted. Experimental support for this view-the axonal hypothesis-is provided by data from various animal models with primary myelin or axonal pathology, and from pathological or magnetic resonance studies on MS patients. In mice with experimental autoimmune encephalomyelitis (EAE), 15-30% of spinal cord axons can be lost before permanent ambulatory impairment occurs. During secondary progressive MS (SP-MS), chronically demyelinated axons may degenerate due to lack of myelin-derived trophic support. In addition, we hypothesize that reduced trophic support from damaged targets or degeneration of efferent fibers may trigger preprogrammed neurodegenerative mechanisms. The concept of MS as an inflammatory neurodegenerative disease has important clinical implications regarding therapeutic approaches, monitoring of patients, and the development of neuroprotective treatment strategies.

Animals↗
PubMed1997

Reduction of peridural fibrosis after lumbar laminotomy and discectomy in dogs by a resorbable gel (ADCON-L).

STUDY DESIGN: A canine lumbar laminotomy and discectomy model was used to evaluate the effects of a resorbable gel, ADCON-L (Gliatech Inc., Cleveland, OH), in reducing peridural scar formation and to assess the healing progress of immediately adjacent normal structures. OBJECTIVES: To compare peridural scar formation and anulus fibrosis healing with and without the use of ADCON-L in an animal model that closely replicates the common human surgical procedure. SUMMARY OF BACKGROUND DATA: ADCON-L has been shown to reduce the development of peridural scars in rat, rabbit, and canine laminectomy models in previous investigations; discectomies were not performed in these previous studies, however, and thus anular healing could not be assessed. METHODS: Seven adult mongrel dogs underwent a three-level unilateral lumbar hemilaminotomy and anular fenestration. In each dog, the ADCON-L was applied to two randomly assigned sites around the discectomy, the nerve roots, and the hemilaminotomy. The third site underwent surgery, but was left untreated (sham surgery only). The canines were killed 8 weeks after surgery, and a gross anatomic assessment of scar formation was done using microdissection by an observer blinded to treatment. A numerical rating system was developed to assess the relative amount and tenacity of the anterior scarring (discectomy site) and posterior scarring (hemilaminotomy site). The healing of the anular defect was assessed histologically. RESULTS: ADCON-L significantly reduced peridural fibrosis in this lumbar discectomy model compared with the sham treated sites (p < 0.05). Further, the superficial layers, the posterior longitudinal ligament, and the anulus fibrosus healed well in the ADCON-L treated sites. CONCLUSIONS: ADCON-L reduced local peridural fibrosis after lumbar laminotomy and discectomy without impacting the healing of the surrounding tissues.

Animals↗
PubMed1993

beta-Amyloid of Alzheimer's disease induces reactive gliosis that inhibits axonal outgrowth.

Pathological lesions in the brains of patients with Alzheimer's disease (AD) are characterized by dense deposits of the protein beta-amyloid. The link between the deposition of beta-amyloid in senile plaques and AD-associated pathology is, at present, controversial since there have been conflicting reports on whether the 39-43 amino acid beta-amyloid sequence is toxic or trophic to neurons. In this report, we show that beta-amyloid peptide when presented as an insoluble substrate which mimics its conformation in vivo can induce cortical glial cells in vitro and in vivo to locally deposit chondroitin sulfate containing proteoglycan. In vitro the proteoglycan-containing matrix deposited by glia on beta-amyloid blocks the usual ability of the peptide to allow cortical neurons to adhere and grow. Chondroitin sulfate-containing proteoglycan was also found in senile plaques of human AD tissue. We suggest that an additional effect of beta-amyloid in the brain, which compounds the direct effects of beta-amyloid on neurons, is mediated by the stimulation of astroglia to become reactive. Once in the reactive state, glial cells deposit large amounts of growth-inhibitory molecules within the neuropil which could impair neuronal process survival and regeneration leading to neurite retraction and/or dystrophy around senile plaques in AD.

Alzheimer Disease↗
PubMed1993

The reduction of postlaminectomy peridural fibrosis in rabbits by a carbohydrate polymer.

Spinal peridural fibrosis following total laminectomy in New Zealand White rabbits was significantly decreased by the intraoperative application of GT1587, a semi-synthetic carbohydrate polymer. The application of a similar polymer, GT1043, or phosphate-buffered saline (PBS) was not as effective. Laminectomies were performed at L-2 and L-4 in 25 rabbits. Absorbable gelatin sponge soaked with GT1043, GT1587, or PBS was applied in a blinded fashion to the operative sites, with untreated (sham) laminectomy sites serving as controls. Animals were sacrificed after 2 or 4 weeks. The extent of peridural fibrosis was evaluated by gross microdissection and histological analysis. Dense scar formation and dural adhesions were evident at both time intervals in the sham- and PBS-treated laminectomy sites. The sites treated with GT1587 showed significantly decreased peridural scar formation and dural adhesions, whereas GT1043 treatment caused modest reduction of scar formation at only the 2-week examination. The healing of skin and lumbosacral fascia was not affected by treatment of the laminectomy site with GT1587. These results suggest that GT1587 may prove beneficial in preventing postlaminectomy dural adhesions and peridural fibrosis in humans.

Animals↗
PubMed1991

A carbohydrate polymer that effectively prevents epidural fibrosis at laminectomy sites in the rat.

We demonstrate that a carbohydrate polymer, designated GL402, effectively inhibits epidural fibrosis in a rat laminectomy model. A total laminectomy in Lewis rats was performed at lumbar vertebrae 3 and 5. GL402 or phosphate buffer solutions in gelatin sponges were applied to the laminectomy sites. Epidural fibrosis was measured, using a double-blind protocol, 2 weeks postoperatively either by gross anatomical evaluation (blunt dissection) or by histological evaluation. Local application of GL402 produced nearly complete inhibition of epidural fibrosis, whereas extensive scar formation and bone growth occurred after local application of buffer or other purported anti-fibrotics. In laminectomy sites treated with GL402 the dura mater was essentially free of adhering fibrosis and bone growth was dramatically decreased. With reduction of postlaminectomy fibrosis, the spinal nerve roots are more mobile and therefore may be less prone to recurrent nerve root compression. The dramatic reduction of epidural fibrosis by GL402 will make reoperative disc surgery safer due to greater accessibility of the laminectomy site. This compound may be useful in preventing surgical adhesions in other sites as well.

Animals↗
PubMed1990

Evidence that the B2 chain of laminin is responsible for the neurite outgrowth-promoting activity of astrocyte extracellular matrix.

Extracellular matrix (ECM) derived from cerebral cortical astrocytes stimulates neurite outgrowth from pheochromocytoma (PC12) cells in the absence of the classical nerve growth factor (NGF). We have shown here that astrocyte ECM can also stimulate neurite outgrowth from primary cultures of central nervous system (CNS) neurons. Using PC12 cells for a quantitative assay, we also demonstrated that the neurite growth-promoting activity increased as the astrocytes matured in vitro: ECM from older astrocytes (3-12 weeks in vitro) exhibited two-fold more neurite growth-promoting activity than ECM for younger astrocytes (5 days to 2 weeks in vitro). We applied various antibodies to identify the neurite growth-promoting factor of astrocyte ECM and found that anti-laminin inhibited neurite outgrowth by 50%, whereas anti-fibronectin and anti-NGF had no effect. Immunoblots, using laminin chain-specific antibodies, and cDNA hybridization of laminin mRNA demonstrated that cultured astrocytes synthesize only the B2 chain of laminin. This suggests that the B2 chain of laminin suffices to stimulate neurite outgrowth.

Animals↗
PubMed1988

Smooth muscle cells transiently express NCAM.

NCAM (neural cell adhesion molecule) polypeptides were first detected on neuronal cells and were subsequently found to be expressed at least transiently by a number of other cell types including skeletal and cardiac but not smooth muscle. We report here that rat smooth muscle expresses NCAM in vitro and transiently in vivo. Using a monoclonal antibody 3F4 which reacts with most rat NCAM polypeptides, NCAM was found on the surface of cultured rat aortic smooth muscle lines A10 and A7r5 and mouse smooth muscle like line BC3H1 in abundances equal to or greater than those of cardiac muscle, skeletal muscle, and neuronal cell lines. The major NCAM polypeptide of muscle cells was Mr = 140 kDa with lesser amounts of the 120 kDa form. Consistent with these results, a major NCAM mRNA of 6.7 kb was detected in Northern analyses with lesser amounts of the 4.3 and 2.9 kb mRNA size classes. The relative abundance of NCAM mRNA was similar in RNA prepared from smooth muscle A7r5 cells, L6 skeletal muscle cells, and 9-day-old rat brain. NCAM was distributed across the entire surface of cultured smooth muscle cells in a highly punctate manner. Cryostat sections of rat aorta, intestine and bladder were examined by immunofluorescence to determine if NCAM is also expressed on smooth muscle in vivo. In each case NCAM was found to be transiently expressed by the smooth muscle cells of these tissues. Highest NCAM levels were observed at embryonic day 17 which then declined to undetectable levels in tissues from adults. These results extend previous observations to indicate all muscle types transiently express NCAM in development.

Animals↗
PubMed1987

Extracellular matrix derived from astrocytes stimulates neuritic outgrowth from PC12 cells in vitro.

The ability of astrocyte extracellular matrix to stimulate axonal elongation was examined using an in vitro model system. Extracellular matrix (ECM) was derived from primary cultures of rat astrocytes or meningeal cells, or from a cell line of bovine aortic endothelial cells. The cells were grown in 35-mm tissue culture dishes for 24 h and then removed non-enzymatically, leaving ECM attached to the surface of the culture dishes. Subsequently, PC12 pheochromocytoma cells were seeded onto the ECM and de novo neurite outgrowth was measured. Within 24 h, the PC12 cells exhibited profuse neuritic outgrowth on ECM derived from astrocytes and endothelial cells, without addition of exogenous nerve growth factor. Over a period of 4 days, the neurites continued to elongate and branched extensively. Little or no neuritic outgrowth was observed from PC12 cells grown on uncoated culture dishes or on culture dishes treated with astrocyte-conditioned medium. Only a slight stimulation of neurite outgrowth was observed on meningeal cell-derived ECM. These results indicate that astrocyte ECM, as well as endothelial cell ECM, possesses one or more molecular factors that can stimulate and maintain de novo axonal elongation from PC12 cells. It is suggested that immature astrocytes secrete neurite-promoting factors as a component of the ECM which act to stimulate and possibly guide the growth of axons during in vivo development.

Animals↗
PubMed1986

Intraspinal transplantation of embryonic spinal cord tissue in neonatal and adult rats.

Fetal rat spinal cord tissue was obtained on gestational day 14 (E14) and transplanted into 2-4-mm-long intraspinal cavities produced by partial spinal cord lesions in adult and neonatal rats. At regular post-transplantation intervals, light and electron microscopy, autoradiographic demonstration of tritiated thymidine labelling, and immunocytochemical localization of glial fibrillary acidic protein (GFAP) were used to identify surviving donor tissues and to study their differentiation and extent of fusion with recipient spinal cords. In some experiments, wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) was also employed to examine whether neurons within the grafts projected axons into the host spinal cord and vice versa. Lastly, immunocytochemistry was used to determine whether any supraspinal serotoninergic (5-HT) axons from the host extended into the transplants. Over 80% of the grafts survived in lesions of both the neonatal and adult rat spinal cord for periods of 1-16 months (duration of experiment), and considerable maturation of donor tissue was evidenced, which even included the appearance of some topographical features of the normal spinal cord. Many of the transplants extended the entire length of the lesion, and were often closely apposed to the injured surfaces of the recipient spinal cords without an intervening dense glial scar. At post-transplantation intervals of 2-4 months, injection of WGA-HRP into the host spinal cord (5 mm from the transplant in adult animals or as much as 20 mm in neonatal recipients) demonstrated retrogradely labelled neurons and anterogradely labelled axons in the grafts. Likewise, injecting WGA-HRP into transplants in adult recipients resulted in labelling of neurons in adjacent segments of the host spinal cord; some labelled axons, derived from donor neurons, were also present in neighboring spinal gray matter. Finally, immunocytochemistry revealed 5-HT-like immunoreactive fibers in transplants that had been prelabelled with tritiated thymidine. These observations demonstrate the potential of embryonic spinal cord transplants to replace damaged intraspinal neuronal populations and to restore some degree of anatomical continuity between the isolated rostral and caudal stumps of the injured mammalian spinal cord.

Age Factors↗
PubMed1986

The amount of slow axonal transport is proportional to the radial dimensions of the axon.

Axons are fundamentally cylindrical and their geometry is defined by two basic parameters, i.e. diameter and length. The average cross-sectional diameter of an axon is determined primarily by the number and density of cytoskeletal structures (i.e. microtubules and neurofilaments) in the axon. The proteins that constitute these structures are synthesized in the nerve cell body and are conveyed through the axon by slow axonal transport. In particular, slow component a (SCa) supplies all of the axonal neurofilament proteins and most of the microtubule proteins to the axon. To study the relationship between slow axonal transport and axonal diameter, the slowly transported proteins were radiolabelled in rat dorsal root ganglion (DRG) cells. The amount of radiolabelled SCa proteins transported in individual unmyelinated and myelinated DRG axons was measured by the electron microscopic autoradiographic method. We found that the amount of SCa transported in the axons is proportional to axonal cross-sectional area. These results indicate that slow axonal transport of microtubules and neurofilaments is a primary determinant of axonal diameter.

Animals↗
PubMed1984

Astrocytic membrane morphology: differences between mammalian and amphibian astrocytes after axotomy.

Previous studies have shown that astrocytes in some nonmammalian species provide a favorable environment for axonal elongation, whereas mammalian astrocytes are thought to inhibit fiber outgrowth. The present study was performed to determine whether any plasma membrane differences exist between these glial elements which could account for their contrasting effects upon axonal outgrowth. Astrocytic scars were formed in optic nerves of rats, newts, and frogs by enucleation. Subsequently, the astrocytic membranes were examined with the freeze-fracture technique. Orthogonal arrays of small intramembranous particles (IMPs) are a prominent component of the plasma membranes of normal mammalian astrocytes; these arrays are most numerous in astrocytic membranes that form an interface between the CNS and nonneural tissue. Astrocytic membranes within the normal CNS parenchyma, however, possess much lower densities of arrays. Following axotomy and Wallerian degeneration, the density of arrays increased threefold within the parenchyma of the optic nerve, while remaining constant at the glia limitans. In striking contrast, only a few aggregates of IMPs that resembled orthogonal arrays could be found in normal and reactive astrocytes of amphibians, although the cytology of these glial cells and density of the scars are otherwise similar to those of their mammalian counterparts. These findings suggest (1) that a proliferation of orthogonal arrays in astrocytic plasma membranes is a prominent feature of gliosis in the mammalian CNS and (2) that differences in the composition of reactive mammalian and amphibian astrocytic membranes may account for variations in axonal-glial interactions within the injured CNS.

Aging↗
PubMed1983

Correlation of axonal regeneration and slow component B in two branches of a single axon.

We investigated the relationship between slow axonal transport and axonal regeneration in the rat dorsal root ganglion (DRG) cell. The DRG cell sends out a single axon which bifurcates within the ganglion; one axon proceeds centrally into the spinal cord and the other proceeds peripherally. The rate of axonal regeneration is approximately 2 times faster for the peripheral processes (4.6 +/- 0.9 mm/day) than for the central processes (2.1 +/- 0.5 mm/day). The peripheral and central processes regenerate through dissimilar environments (sciatic nerve and dorsal root, respectively); thus, environmental factors may account for the differences in regeneration rates. We tested this possibility by measuring the regeneration of motoneuron axons within the ventral root (histologically similar to the dorsal root). The motoneuron regeneration rate within the ventral root is similar to the motoneuron regeneration rate within the sciatic nerve, suggesting that factors within the DRG cell produce the differences in regeneration rate. Slow axonal transport is classified into two distinct components: slow component a (SCa), corresponding to the microtubule/neurofilament network of the axonal cytoskeleton, and slow component b (SCb), corresponding to the microfilament complex/axoplasmic matrix. The transport rate of SCa and SCb in the peripheral sensory axons is approximately 2 times faster than their counterparts in the central sensory axons. SCa moves at 1.0 to 3.0 mm/day in the peripheral processes and 0.5 to 1.0 mm/day in the central processes; SCb moves at 3.5 to 6.5 mm/day in the peripheral processes and 2.0 to 3.5 mm/day in the central processes. In each branch of the DRG cell, the rate of axonal regeneration is similar to the rate of SCb transport. These results support the hypothesis that SCb is a rate-limiting factor in axonal regeneration because of its role in providing the cytoskeletal elements which are directly involved in the motility of the growth cone and elongation of the axon.

Animals↗
PubMedDate not supplied

Deposits of A beta fibrils are not toxic to cortical and hippocampal neurons in vitro.

Amyloid beta peptide (A beta), which is deposited as insoluble fibrils in senile plaques, is thought to play a role in the neuropathology of Alzheimer's disease. We have developed a model in which rat embryonic cerebral cortical or hippocampal neurons are seeded onto culture dishes containing deposits of substrate-bound, fibrillar A beta. The neurons attached rapidly to A beta 1-40 and A beta 1-42 substrates and extended long, branching neurites. Quantitative assessment demonstrated that survival of neurons on the A beta matrices was equivalent to or better than on control substrates of poly L-lysine or poly L-ornithine. In contrast, preparations of A beta fibrils added directly to the culture medium caused neuronal death as previously reported in the literature. These results reveal that the response of neurons to deposited A beta 1-40 and A beta 1-42 is substantially different from that observed with suspensions of the amyloid peptides, with the former serving as growth-promoting substrates for cortical and hippocampal neurons. This may thus imply that fibrillar A beta of senile plaques is not sufficient by itself to cause the plaque-associated neuronal degeneration characteristic of AD.

Alzheimer Disease↗
PubMedDate not supplied

Evidence for glial-mediated inflammation in aged APP(SW) transgenic mice.

Chronic expression of inflammatory cytokines, including interleukin-1beta, tumor necrosis factor alpha, and interleukin-6, by glia may underlie the neurodegenerative events that occur within the brains of patients with Alzheimer's disease (AD). The present study determined whether these markers of inflammation could be observed within the brains of Tg(HuAPP695.K670N/M671L)2576 transgenic mice (Tg2576) that have recently been shown to mimic many features of AD. Interleukin-1beta- and tumor necrosis factor alpha-immunopositive microglia were localized with thioflavine-positive (fibrillar) Abeta deposits. Moreover, interleukin-6 immunoreactive astrocytes surrounded fibrillar Abeta deposits. These findings provide evidence that Tg2576 mice exhibit features of the inflammatory pathology seen in AD and suggest that these mice are a useful animal model for studying the role inflammation may play in this disease.

Aging↗