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  • 1
    ISSN: 1460-9568
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: A major reason for the insufficient recovery of function after motor nerve injury are the numerous axonal branches which often re-innervate muscles with completely different functions. We hypothesized that a neutralization of diffusable neurotrophic factors at the lesion site in rats could reduce the branching of transected axons. Following analysis of local protein expression by immunocytochemistry and by in situ hybridization, we transected the facial nerve trunk of adult rats and inserted both ends into a silicon tube containing (i) collagen gel with neutralizing concentrations of antibodies to NGF, BDNF, bFGF, IGF-I, CNTF and GDNF; (ii) five-fold higher concentrations of the antibodies and (iii) combination of antibodies. Two months later, retrograde labelling was used to estimate the portion of motoneurons the axons of which had branched and projected into three major branches of the facial trunk. After control entubulation in collagen gel containing non-immune mouse IgG 85% of all motoneurons projecting along the zygomatic branch sprouted and sent at least one twin axon to the buccal and/or marginal-mandibular branches of the facial nerve. Neutralizing concentrations of anti-NGF, anti-BDNF and anti-IGF-I significantly reduced sprouting. The most pronounced effect was achieved after application of anti-BDNF, which reduced the portion of branched neurons to 18%. All effects after a single application of antibodies were concentration-dependent and superior to those observed after combined treatment. This first report on improved quality of reinnervation by antibody-therapy implies that, in rats, the post-transectional collateral axonal branching can be reduced without obvious harmful effects on neuronal survival and axonal elongation.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1433-0458
    Keywords: Schlüsselwörter Reinnervation ; Nervennaht ; Ratte ; N. facialis ; FluoroGold ; FastBlue ; Markierung ; Key words Rat facial nerve ; Axonal regrowth ; Retrograde fluorescent tracer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Introduction: In recent studies we identified two morphological phenomena in the facial nucleus of the brainstem that play an important role in the recovery of facial movement: hyperinnervation and misdirected reinnervation. While the hyperinnervation could easily be quantified by cell counting, the extent of misdirected reinnervation could not be estimated accurately. In the present study we developed a method for accurate quantification of this misdirected reinnervation. Material and methods: In 6 rats we injected the fluorescent tracer FluoroGold into the whiskerpad. After 4 days the facial nerve was transected and a facial-facial anastomosis (FFA) performed. Eight weeks later the fluorescent tracer Fast Blue was injected in the same site of the whiskerpad, as done before with the FluoroGold. Following sacrifice, the brainstems of the animals were removed and labeled motoneurons in the facial nucleus counted. Results: Three different types of labeled motoneurons could be identified: (a) white fluorescent motoneurons (labeled preoperatively only with FluoroGold), (b) blue fluorescent motoneurons (labeled only postoperatively with Fast Blue) and (c) green-grey fluorescent motoneurons double-labeled pre- and postoperatively. The double-labeled motoneurons were seen to project to the same sites in the whiskerpad pre- and postoperatively, demonstating no misdirected reinnervation. In our experiment we counted 478±45 green-grey labeled neurons from a total number of 1446±131 postoperatively labeled cells (double-labeled and single FB-labeled). These findings show that 33% of the regenerating motoneurons were correctly redirected after FFA in our animal model and 67% were misdirected.
    Notes: Zusammenfassung Hintergrund: Nach nervenplastischen Eingriffen des N. facialis kommt es regelmäßig zu Defektheilungszuständen, deren morphologisches Korrelat die fehlgeleitete Reinnervation und die Hyperinnervation ist. Fragestellung: Die fehlgeleitete Reinnervation ließ sich bisher im Gegensatz zur Hyperinnervation nicht exakt quantifizieren. Wir entwickelten eine Methode zur exakten Bestimmung dieses Parameters. Material and Methode: Präoperativ wurde der Fluoreszenzmarker FluoroGold in die Bartplatte der Ratte injiziert. 4 Tage später wurde eine Fazialis-Fazialis-Anastomose durchgeführt und nach Abwarten der Reinnervation der Fluoreszenzmarker Fast Blue exakt an gleicher Stelle in die Bartplatte injiziert. Die gewonnen Gehirnschnitte wurden bildanalytisch ausgewertet. Ergebnisse: 3 verschieden gefärbte Zelltypen konnten entdeckt werden: weiße, nur präoperativ mit FluoroGold gefärbte Motoneurone, blaue nur postoperativ mit Fast Blue gefärbte Motoneurone und doppelt markierte grau-grüne Motoneurone, die sowohl prä- als auch postoperativ markiert wurden. Die doppelt markierten Neurone, deren Anteil lediglich 33% betrug, entsprechen den Zellen, die sowohl prä- als auch postoperativ dieselbe Zielmuskulatur innervierten.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-7381
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Neuron-specific enolase as an enzyme of the glycolytic pathway is localized in the cytoplasm of nerve cells, but not in the cell nucleus. We have applied immunocytochemistry with 1∶64 000 polyclonal anti-rat neuron-specific enolase to the brainstem of male and female adult Wistar rats following: (a) transection of the facial nerve with immediate microsurgical nerve suture (facial-facial anastomosis), (b) transection of the hypoglossal nerve with immediate suture (hypoglossal-hypoglossal anastomosis) and (c) transection of the facial and hypoglossal nerve with immediate suture of the proximal hypoglossal to the distal facial nerve stump (hypoglossal-facial anastomosis). Studying the intracellular immunolocalization of neuron-specific enolase in neurons of the facial and hypoglossal nucleus we detected that (1) in normal rats about 20% of all facial and hypoglossal neurons display not only cytoplasmic, but also intranuclear neuron-specific enolase-like immunoreactivity and (2) following any axotomy of the facial or hypoglossal peripheral nerve, the perikarya of all injured motoneurons react by an outstanding increase of neuron-specific enolase-like immunoreactivity in the karyoplasm. Similar findings were obtained in experiments on non-fixed cultured Neuro-2a cells that had been lesioned with hydrogen peroxide. Counting the absolute numbers of normal and reactive neurons at 1–365 days post axotomy revealed that the increase of neuron-specific enolase in neuronal cell nuclei is temporary and reversible. It is first detected at 2 days post axotomy, reaches its maximum at 10–18 days post axotomy and is no longer evident 56 days following surgery. These findings suggest that the intranuclear neuron-specific enolase-like immunoreactive material may serve a regulatory function on the genome.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-7381
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Facial and hypoglossal nerves were resected unilaterally in a total of 108 rats. Rats were divided into two groups; one group received standard food pellets (placebo), the other received food pellets containing the Ca2+-blocking agent nimodipine. The expression of glial fibrillary acidic protein was examined in paraffin sections of the brainstem using light microscopical immunocytochemistry, and the degree of glial process ensheathment of the surviving neuronal perikarya in the hypoglossal and facial nuclei quantified on electron micrographs. Up to 28 days post-axotomy no differences in glial fibrillary acidic protein-immunoreactivity were observed between placebo and nimodipine-treated animals. By 42–days, glial fibrillary acid protein-immunoreactivity was stronger in the nimodipine treated animals and by 112 days, glial fibrillary acid protein-immunoreactive astrocytes occured only in nimodipine-treated animals. Thin astrocytic processes were seen to ensheath neurons in both placebo and nimodipine-treated animals. By 28 days post axotomy, lesioned neurons in nimodipine treated animals were covered by a mean of 2.6 (hypoglossal) and 2.9 (facial nucleus) astrocytic lamellae, compared with 1.7 lamellae in the placebo group. This relatively greater ensheathment of hypoglossal and facial neurons was maintained up to 112 days post-lesion, but reduced in the placebo-treated group to ∼ 1.4 lamellae. It is concluded that nimodipine enhances the formation of astrocytic lamellae on lesioned neurons and that this process may be associated with a protective role for activated astrocytes directed towards motoneurons suffering from permanent target-deprivation.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 257 (1989), S. 207-216 
    ISSN: 1432-0878
    Keywords: Morphogenesis ; Meninges ; Mesenchyme ; Ultrastructure ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The meninges of albino Wistar rat embryos, aged between the 11th embryonic day (ED) and birth, were sectioned using a specially constructed device. This technique permits optimal microanatomical preservation of all tissues covering the convexity of the brain: skin, muscle, cartilage or bone, and the meninges. At ED11, the zone situated between the epidermis and the brain is occupied by a mesenchymal network. At ED12, part of this delicate network develops as a dense outer cellular layer, while the remainder retains its reticular appearance, thus forming an inner layer (the future meningeal tissue). At ED13, the dura mater starts to differentiate. At ED14, the bony anlage of the skull can be identified, and along with the proceeding maturation of dura mater some fibrillar structures resembling skeletal muscle fibers appear in the developing arachnoid space. At ED15–17, a primitive interface zone — dura mater/ arachnoid — is formed, comprised by an outer electronlucent and an inner electron-dense layer marking the outer aspect of the arachnoidal space. At ED18–19, the innermost cellular row of the inner durai layer transforms into neurothelium, which is separated from the darker arachnoidal cells by an electron-dense band. The arachnoidal trabecular zone with the leptomeningeal cells is formed at ED19. By the end of the prenatal period (ED20–21), its innermost part organizes into an inner arachnoidal layer and an outer and inner pial layer. The results from this study indicate (i) that dura mater and leptomeninges develop from an embryonic network of connective tissue-forming cells, and (ii) that the formation of cerebrospinal fluid (CSF)-containing spaces accompanies the differentiation of the meningeal cellular layers.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0878
    Keywords: Dura mater ; Arachnoid ; Alkaline phosphatase ; Adenosine triphosphatase ; Ultrastructural histochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of the activity of alkaline phosphatase and Mg-dependent adenosine triphosphatase was studied in the encephalic dura mater-arachnoid borderline (interface) zone of albino Wistar rats. Intense clustering of electron-dense granules that indicated alkaline phosphatase activity was observed in the inner dural cells, the neurothelial cells, the outermost row of the outer arachnoidal cells and in the intercellular cleft between the latter two (the so-called electron-dense band). The remainder of the outer arachnoidal cells contained almost no reaction product. Mg-adenosine triphosphatase activity was distributed differently; a lack of reaction product was observed not only in the outer arachnoidal cells, but also in the zone occupied by the electron-dense band. The data confirm histochemically the barrier properties of the dura mater-arachnoid interface zone.
    Type of Medium: Electronic Resource
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