Biomedical subjects

J R Wolff

Publications and source records attributed to J R Wolff.

At least 19 recordsLinked to original sources

PubMed2001

Rapid morphological changes in astrocytes are accompanied by redistribution but not by quantitative changes of cytoskeletal proteins.

Astrocytes have the potential to acquire very different morphologies, depending on their regional location in the CNS and on their functional interactions with other cell types. Morphological changes between a flat or a fibroblast-like and a stellate or process-bearing appearance, and vice versa, can occur rapidly, but very little is known as to whether morphological transformations are based on quantitative changes of cytoskeletal proteins in microfilaments, intermediate filaments, and/or microtubules. Using a cell culture of selective type 1 astrocytes, we compared the distribution and protein amounts of a number of cytoskeletal proteins both during primary process growth induced by specific media conditions and after secondary transformations induced by dBcAMP. Our data presented in this report support the idea that astrocytes can undergo dramatic changes in their morphology requiring subcellular redistribution of most cytoskeletal proteins but no quantitative modifications of the amount of the respective proteins. After pharmacological treatment with lysophosphatic acid and genistein we show that astrocytes can acquire intermediate morphologies reminiscent of both fibroblast and stellate-like cells. These experiments demonstrate that the recently described RhoA-mediated signaling cascade between the cell surface and cytoskeletal proteins is only one of several signaling pathways acting on the astrocytic cytoskeleton.

Actinin↗

PubMed2000

Modulation of the truncated GAD25 by estrogen in the olfactory bulb of adult rats.

The aim of our study was to investigate the influence of gonadal steroids on the expression of different GAD isoforms. Here we show that, in addition to the adult GAD forms, the two embryonic splice variants of GAD67 mRNA and the truncated GAD25 are present in the adult rat olfactory bulb, a brain region with high synaptic plasticity, which has preserved some features of the developing brain. By Western blot analysis, we could demonstrate that the expression of the embryonic GAD25 is cyclic in females: its quantity is higher on estrus day. Furthermore, in ovariectomized animals 17-beta-estradiol treatment induced an increase of GAD25 within 3 h, reaching a maximum at 9-12 h. Our data are compatible with the interpretation that the embryonic GAD isoforms may play a role in the neuroplastic changes induced by sexual steroids.

Alternative Splicing↗

PubMed2000

Facial nerve injury-induced disinhibition in the primary motor cortices of both hemispheres.

Unilateral facial nerve transection induces plastic reorganization of the somatotopic order in the primary motor cortex area (MI). This process is biphasic and starts with a transient disinhibition of connections between cortical areas in both hemispheres. Little is known about the underlying mechanisms. Here, cortical excitability has been studied by paired pulse electrical stimulation, applied either within the MI or peripherally to the trigeminal nerve, while the responses were recorded bilaterally in the MI. The ratios between the amplitudes of the second and first evoked potentials (EPs or fEPSPs) were taken as measures of the inhibitory capacity in the MI ipsilateral or contralateral to the nerve injury. A skin wound or unilateral facial nerve exposure immediately caused a transient facilitation, which was followed by a reset to some level of inhibition in the MI on both sides. After facial nerve transection, the first relatively mild reduction of inhibition started shortly (within 10 min) after denervation. This was followed by a second step, involving a stronger decrease in inhibition, 40-45 min later. Previous publications have proved that sensory nerve injury (deafferentation) induces disinhibition in corresponding areas of the sensory cortex. It is now demonstrated that sham operation and, to an even greater extent, unilateral transection of the purely motoric facial nerve (deefferentation), each induce extended disinhibition in the MIs on both sides.

Animals↗

PubMed2000

Transient changes in cortical distribution of S100 proteins during reorganization of somatotopy in the primary motor cortex induced by facial nerve transection in adult rats.

In adult rats, the primary motor cortex (MI) comprises a somatotopic map of muscle representations. This somatotopy is modified after transection of the facial nerve (N7x). Mapping with cortical stimulation revealed that the underlying cortical reorganization is biphasic. Primary changes cause a transient disinhibition of long cortico-cortical connections in both hemispheres. While the first reaction vanishes within a few hours, short intra-areal connections are disinhibited within MI contralateral to N7x. The resulting co-operation between adjacent parts of MI persists as long as peripheral reinnervation is prevented. Cellular mechanisms underlying this cortical reorganization are largely unknown. Here, we utilized changes in immunoreactivity of S100 proteins (S100-IR) known as a sensitive indicator of astroglial reactions during plastic reactions in the central nervous system. Within 1 h of N7x, zones with enhanced S100-IR appeared in both hemispheres. Between 3. 5 and 18 h, reaction patterns with changing topography were transiently prominent in many cortical areas including parts of MI which surrounded the facial muscle representation fields. After 24 h, the facial muscle representation contralateral to N7x became labelled while S100-IR enhancement disappeared in most of the cortex. S100-IR-enhancement vanished completely during the next day of survival. Data presented suggest that (i) enhancement of S100-IR labels cortical tissue during the functional reorganization that is induced by N7x, (ii) large parts of the cerebral cortex participate in the reorganization, before it is finally focused on the representation field of MI that corresponds with contralateral N7x, and (iii) temporo-spatial patterns of astrocytic reactions apparently play a role in the underlying plasticity reaction.

Animals↗

PubMed1999

Facial nerve injury produces a latent somatosensory input through recruitment of the motor cortex in the rat.

Short-latency effects of unilateral facial nerve transection were studied on neuronal activation evoked in the primary motor cortex (MI) on both sides by vibrissa stimulation in adult rats. In the controls, unilateral trigeminal stimulation evoked activity in the whisker representation of both the contralateral somatosensory cortex (SI) and MI, but never in the ipsilateral MI. Unilateral transection of the facial motoric nerve facilitated evoked responses in the contralateral MI, and induced further neuronal activation (gross potentials and unit activity) in the MI ipsilateral to the stimulation. Since these changes appeared rapidly and could be mimicked by picrotoxin application onto the SI contralateral to the stimulation, they are considered to be based on the disinhibition of preexisting associative and commissural connections, which are unmasked by facial nerve transection.

Animals↗

PubMed1999

Activation of the primary motor cortex by somatosensory stimulation in adult rats is mediated mainly by associational connections from the somatosensory cortex.

In anaesthetized adult rats, facial nerve injury causes a disinhibition of the interhemispheric connections between homotopic representation fields in the primary motor cortex with a latency of 4 min (Toldi et al., 1996, Neurosci Lett. 203, 179-182). One possible explanation for the induction of such rapid changes is an alteration of the somatosensory input to the motor cortex. To test this hypothesis, unit activity in primary motor cortex was recorded during electrical stimulation of trigeminal afferents in the contralateral whisker-pad. About one-third of all recorded primary motor cortex neurons responded with latencies shorter than in the ventrolateral and posterior nuclei of the thalamus. Responses failed at stimulation frequencies > or = 10 Hz and after elimination or inactivation of the somatosensory cortex. Within primary motor cortex, the activatable neurons displayed a bilaminar distribution and were identified as pyramidal neurons by neurobiotin labelling. The results suggest that trigeminal afferents participate in modulation of the activity of primary motor cortex output neurons via primary somatosensory cortex-to-primary motor cortex associational connections, even under anaesthesia.

Afferent Pathways↗

PubMed1998

Autocellular coupling by gap junctions in cultured astrocytes: a new view on cellular autoregulation during process formation.

Neocortical astrocytes make two types of gap junctions, intercellular ones create a functional syncytium, while reflexive gap junctions mediate autocellular coupling and serve unknown functions (Rohlmann and Wolff, 1996). Here, the question is addressed whether solitary astrocytes in vitro express connexin43 (Cx43) and establish gap junctions in the absence of intercellular contacts. In all media conditions tested, immunocytochemistry visualized Cx43-expression and gap junctions irrespective of the presence or absence of intercellular contacts. Reflexive gap junctions were associated with mechanical junctions (adherent spots and fascia adherens) connecting surface membranes and cytoskelal components, respectively. Both were characteristically located along incompletely separated borders between developing processes and/or branches. In addition, Cx43-immunoreactivity was found on some non-junctional membranes: i) intracellular vesicle clusters sited to forming processes and at the basis of filopodia; ii) the surface membrane of filopodial subpopulations usually appearing in bunches. Results suggest changes in the resumptive role of Cx43 in cultivated astrocytes: 1) Cx43 is not confined to intercellular gap junctions, it may even selectively compose reflexive ones; 2) from intracellular stores (vesicle aggregates), Cx43 may be incorporated into the surface membrane of filopodia; 3) by contacting other parts of the same cell surface (or neighboring cells), filopodia and membrane patches carrying Cx43-half channels may be essential in initial steps of gap junction formation; 4) the distribution of reflexive gap junctions is compatible with the hypothesis that autocellular coupling serves reorganization of cytoskeleton during the formation of cell processes and branches; 5) in general, gap junctions may be important for coordinating the cytoskeleton across intercellular contacts and within cells with complex shape.

Animals↗

PubMed1997

Distribution of astroglia in glomeruli of the rat main olfactory bulb: exclusion from the sensory subcompartment of neuropil.

During an entire lifetime, sensory axons of regenerating olfactory receptor neurons can enter glomeruli in the olfactory bulb and establish synaptic junctions with central neurons. The role played by astrocytes in this unique permissiveness is still unclear. Glomerular astrocytes have been identified by immunocytochemistry for glial fibrillary acidic protein and S100 proteins at the light and electron microscopic levels. The latter labeling included submicroscopic lamellar and filopodial extensions of astroglial processes. Cell bodies and processes accumulate along the border between juxtaglomerular walls and glomerular neuropil. Within glomeruli, a network of astroglial processes encloses mesh-like neuropil zones devoid of astroglia. Electron microscopy confirmed the division into subcompartments of glomerular neuropil: 1) The "sensory-synaptic subcompartment" includes all sensory axon terminals and terminal dendritic branches receiving sensory input, whereas astroglia are excluded; 2) in the "central-synaptic subcompartment," astroglial processes are intermingled with other neuropil components: dendrites of relay cells and interneurons, dendrodendritic synapses, centrifugal (cholinergic and serotonergic) axons, their axodendritic synapses, and blood vessels. Unevenly distributed astroglial processes in this subcompartment are attached to vascular basal laminae, stem dendrites, and subpopulations of dendrodendritic synapses, especially those colocalized with centrifugal projections ("triadic synapses"). Astroglia-free parts of the "central" subcompartment contain segments of dendrites and subpopulations of dendrodendritic synapses. Because of the subdivision of the glomerular neuropil into portions with and without glial components, glia do not completely demarcate the border between the "sensory" and the "central" subcompartments. Interdigitation between the subcompartments varies among glomeruli and even within a single glomerulus. The mesh width of astroglial networks covaries with numerical relations between sensory and dendrodendritic synapses. This distribution pattern of astrocytes suggests that these glial cells monitor brain-derived effects on olfactory glomerular neuropil rather than olfactory input and that astroglial processes are (re-)arranged accordingly.

Animals↗

PubMed1997

Rapid astroglial reactions in the motor cortex of adult rats following peripheral facial nerve lesions.

We report on changes in the motor cortex of adult rats that rapidly and transiently followed various types of facial nerve lesions. These reactions led to enhanced immunoreactivities of various astroglial markers: S-100 protein (a Ca2+- and Zn2+-binding protein predominantly located in the cytosol of astrocytes), glial fibrillary acidic protein (a cytoskeletal protein) and connexin 43 (the astroglial gap junction protein). Reactions could be visualized 1 h after the facial nerve lesion and disappeared within about 5 days after surgery. Combined lesions of the facial and trigeminal nerves modified the spatial pattern of the astroglial reaction, similar to intramuscular injections of botulinum toxin, which inhibits the release of acetylcholine in motor endplates. Data presented suggest that peripheral interference with muscular functions rapidly induces modifications in the motor cortex.

Acetylcholine↗

PubMed1997

Localization and biochemical characterization of acid phosphatase isoforms in the olfactory system of adult rats.

Localization of acid phosphatases was studied with the use of beta-glycerophosphate and p-nitrophenyl phosphate as substrates in the brain with special emphasis on the olfactory system of adult rat at light and electron microscopic level. With the use of beta-glycerophosphate, a selective substrate for the lysosomal acid phosphatase, lead-containing reaction product was found in primary and secondary lysosomes of neurons, glial cells and perivascular macrophages as well as in the cytoplasm of olfactory sensory axons. Incubation with p-nitrophenyl phosphate as substrate additionally revealed a cytoplasmic isoform of acid phosphatase, which could not be inhibited by tartrate or fluoride and was predominantly located in dendrites. Acid phosphatase isoforms were biochemically characterized in samples prepared separately from the olfactory mucosa, olfactory nerve layer, olfactory bulb and its dendrodendritic synaptosomes isolated by subcellular fractionation. In the olfactory mucosa and olfactory nerve layer the lysosomal type (high molecular weight form) was the most prominent acid phosphatase form, whereas the isoform located in dendrites corresponded to the tartrate-resistant extralysosomal, cytosolic type (low molecular weight form). The functional significance of different isoforms of acid phosphatase in the olfactory sensory axons and dendritic elements is discussed.

Acid Phosphatase↗

PubMed1997

Non-conventional role of lysosomal acid phosphatase in olfactory receptor axons: co-localization with growth-associated phosphoprotein-43.

Olfactory receptor neurons undergo a continuous turnover in adult mammals. It is largely unknown how their axons invade the olfactory bulb and induce synaptic re-organization in glomeruli. Here, the cytochemical localization of lysosomal acid phosphatase has been studied in olfactory bulbs of adult rats and mice. The enzyme has been identified by specific substrate, inhibitors and absence in lysosomal acid phosphatase-knockout mice. Lysosomal acid phosphatase is located in primary and secondary lysosomes, which are unevenly distributed in the olfactory nerve layer and among olfactory glomeruli. In consecutive sections of glomeruli, the intensity of lysosomal acid phosphatase immunoreactivity co-varied with that of growth-associated phosphoprotein. Electron microscopically, differential lysosomal acid phosphatase staining in glomeruli corresponded to different proportions of labelled and unlabelled axons. Quantification revealed that lysosomal acid phosphatase labelling was strongest in non-synaptic profiles of terminal axons, while it was weak in or even missing from most synaptic profiles. Hence, growing olfactory axons apparently carry more lysosomal acid phosphatase than those which have established synaptic contacts. Following olfactory deafferentation both lysosomal acid phosphatase activity and growth-associated phosphoprotein-43 are lost from glomeruli, suggesting that both proteins are expressed in olfactory sensory axons during growth, while lysosomal acid phosphatase is apparently not a marker of anterograde terminal degeneration.

Acid Phosphatase↗

PubMed1997

Beta-galactosidase-labelled relay neurons of homotopic olfactory bulb transplants establish proper afferent and efferent synaptic connections with host neurons.

The vertebrate olfactory system has long been an attractive model for studying neuronal regeneration and adaptive plasticity due to the continuous neurogenesis and synaptic remodelling throughout adult life in primary and secondary olfactory centres, its precisely ordered synaptic network and accessibility for manipulation. After homotopic transplantation of fetal olfactory bulbs in bulbectomized neonatal rodents, newly regenerated olfactory neurons form glomeruli within the graft, and the efferent mitral/tufted cells of the transplant innervate the host brain, terminating in higher olfactory centres. However, the synaptic connections of the transplanted relay neurons within the graft and/or host's olfactory centres could not be characterized mainly because of lack of suitable cell-specific markers for these neurons. In this study, we have used olfactory bulbs from transgenic fetuses, in which the majority of the mitral/tufted cells express the bacterial enzyme beta-galactosidase, for homotopic olfactory bulb transplantation following complete unilateral bulbectomy. In the transplants, the cell bodies and terminals of the donor mitral/tufted cells were identified by beta-galactosidase histochemistry and immunocytochemistry at both light and electron microscope levels. We demonstrate that transplanted relay neurons re-establish specific synaptic connections with host neurons of the periphery, source of the primary signal and central nervous system, thereby providing the basis for a functional recovery in the lesioned olfactory system.

Afferent Pathways↗

PubMed1997

Visualization of beta-galactosidase by enzyme and immunohistochemistry in the olfactory bulb of transgenic mice carrying the LacZ transgene.

In the olfactory bulb (OB) of a transgenic mouse line that carries the bacterial LacZ gene under the control of the 5'-regulatory region of the GAD67 gene, expression of the beta-galactosidase was confined almost exclusively to the non-GABAergic mitral and tufted cells. By light microscopy, enzyme histochemistry showed strong staining in the cell bodies and faint diffuse staining in the axons and dendrites. With immunohistochemistry for beta-galactosidase the entire cytoplasm, including the axons and dendrites, was strongly stained. By electron microscopy, beta-galactosidase enzyme histochemistry resulted in a submicroscopic reaction product that was diffusely distributed in the cytoplasm of neurons. In addition, large deposits of the reaction product were also seen attached to the cytoplasmic side of the membranes. In contrast, when the intracellular localization of beta-galactosidase was determined by immunohistochemistry, homogeneous cytoplasmic staining was obtained that filled the entire cytoplasm including the terminal dendrites and fine axons. Therefore, synaptic contacts of the beta-galactosidase-positive output neurons with other beta-galactosidase-negative neuronal cells were readily recognized in the OB. As we demonstrated, transgenic mouse lines expressing the LacZ reporter gene in a well-defined neuronal subpopulation can be used to follow beta-galactosidase-positive neurons and to directly identify their synaptic connections.

Animals↗

PubMed1996

Biphasic reorganization of somatotopy in the primary motor cortex follows facial nerve lesions in adult rats.

Effects of facial nerve transection were studied on muscle responses evoked by electrical stimulation in the primary motor cortex (MI) of adult rats. In intact animals, activated muscles varied according to the somatotopic representation map, and responses were restricted to the contralateral side. Unilateral transection of the facial nerve extinguished contralateral vibrissal responses, while ipsilateral vibrissae began to respond within 4 min. This abnormal response (primary change) was transient and gradually disappeared within hours to days. Instead, contralateral movements of forepaw and eye/eyelid muscles could be evoked from increasing portions of the former vibrissal field (secondary change), in which many points became unresponsive. After 4 days, the former vibrissal field had shrunk to a small central part, where ipsilateral vibrissae responsiveness remained. The secondary modification was stable for at least 2 weeks. Since the primary change is rapid, transient and may be mimicked by picrotoxin, it may be based on disinhibition of commissural connections, while the secondary change is longlasting and therefore may include some form of reorganization of associational synapses.

Animals↗

PubMed1996

Changes in glial fibrillary acidic protein immunoreactivity in the rat facial nucleus following various types of nerve lesions.

We report about changes on astrocytes in the facial nucleus of the rat following various types of peripheral nerve lesions. Astrocyte-specific glial fibrillary acidic protein (GFAP) was labeled by immunohistochemistry and served as a marker for these changes. Increased GFAP immunoreactivity was found in the facial nucleus on the lesioned side within 2-3 days after axotomy. This change lasted longer (up to 1 year) when axon regeneration was prevented or delayed by placing a metal clip on the proximal nerve stump. Lesion of the trigeminal nerve prior to axotomy reduced the degree of GFAP immunoreactivity. No side differences were observed after botulinum toxin application.

Animals↗

PubMed1996

Complex synaptic arrangements in the rat suprachiasmatic nucleus: a possible basis for the "Zeitgeber" and non-synaptic synchronization of neuronal activity.

A special type of complex synaptic arrangement occurs in the ventro-lateral portion of the rat suprachiasmatic nucleus. These arrangements are polycentric, with about equal numbers of pre- and postsynaptic elements. Because of an incomplete astroglial covering, these synaptic complexes are connected with each other and form a continuous reticulum or sponge-like system throughout the ventro-lateral region of the nucleus. In two partially reconstructed complex synaptic arrangements, boutons from retinal afferents could be seen to make up the majority of presynaptic elements. They form asymmetric and symmetric synaptic appositions with dendritic elements. Non-optic axo-dendritic synapses of unknown origin with asymmetric and symmetric appositions and dendro-dendritic synapses with symmetric appositions are also seen in complex synaptic arrangements. Within complex synaptic arrangements, dendrites often run in bundles, with some dendrites spiralling around others. Membranes of neighbouring dendrites are closely apposed. These interdendritic appositions are possibly ephapses and may, together with intersomatic contacts, mediate non-synaptic synchronization of neuronal activity in the suprachiasmatic nucleus, as described by other authors. The activity of optic and non-optic synapses in complex synaptic arrangements over a 24 h period may also produce an integrated response that influences the circadian rhythm of neuronal activity in this nucleus.

Animals↗

PubMed1996

Neuronal plasticity induced by neonatal monocular (and binocular) enucleation.

Monocular (ME) and binocular enucleation has become a useful experimental tool for analyzing the mechanisms of neural plasticity. ME when performed during an early postnatal period (up to 15 days after birth) initiates a series of adaptive reactions in the visual (and other sensory) system(s) which tend to compensate for the lost sensory capacity. Extirpation of one eye (usually the right) destroys afferents to both lateral geniculate bodies dorsal nucleus (CGLd) and superior colliculi (CS), being severely impaired by the degeneration of retino-geniculate and collicular synapses. The sprouting of retinogeniculate fibers coming from the remaining eye replaces these synapses in both CGLds. Ipsilateral representation of the remaining eye (usually of minor significance) becomes extended in the left CGLd and consequently in the left visual area, just as in the superior colliculi. A similar but somewhat smaller extension takes place in the contralateral CGLd and visual cortex. The strengthening of commissural connections results in a remarkable extension of callosally connected stripes and patches in both hemispheres. After ME in the critical period, the control over behavior is taken over by the remaining eye. Its power of resolution is improved because of the higher survival of (mainly ipsilaterally projecting) ganglion cells. Therefore, both hemispheres are still available for storing visual information. In ME rats the learning of visual tasks requires both hemispheres, but relearning is still possible after extirpation of the contralateral one. The possible two main mechanisms of adaptive plastic changes are: (i) replacement of degenerated synapses by sprouting collaterals of ingrowing foreign fibers, and (ii) neurons having morphologically intact but inactive synapses establishing connections with afferent fibers other than the usual. The same mechanism is seen operating in cross-modal adaptive reactions as well.

Animals↗

PubMed1996

The structural localization of galanin, and its function in modulating acetylcholine release in the olfactory bulb of adult rat.

The localization of galanin immunoreactivity was analyzed within the olfactory bulb of adult rats. Galanin-positive neurons were differentially distributed among the bulb layers. The density of stained neurons was highest in the glomerular and external plexiform layers. According to morphology, size, location and arrangement, a large proportion of galanin-immunoreactive neurons corresponds to external tufted cells and short-axon neurons in the superficial part of the external plexiform and glomerular layers. A smaller number were middle tufted cells and short-axons neurons while only a few short-axon neurons were labeled in the granule cell layer. Galanin-stained nerve fibers had different structures (thick fibers with or without varicosities, and thin fibers with or without varicosities). Among them were afferent immunoreactive nerve fibers entering the bulb through the olfactory nerve layer, but penetrating superficial layers. Correspondingly, a large number of galanin-positive axons (with or without varicosities) were observed in the olfactory nerve layer. A number of galanin-positive nerve fibers was also present in the glomerular and internal plexiform layers, while these fibers were scarce in the granule cell layer, their density was lowest in the external plexiform layer. These results suggest that galanin-positive axons present in the olfactory bulb originate from at least four different sources. From the periphery axon bundles enter the bulb together with olfactory nerve fibers from the rostral direction and with a fiber bundle from the ventral posterior surface, i.e. at the border between the olfactory tract and the main olfactory bulb along a large blood vessel. Central sources are local interneurons in the olfactory bulb and some extrabulbar brain regions. Double-labeling experiments combining acetylcholinesterase histochemistry with galanin immunocytochemistry did not show any co-localization of acetylcholinesterase and galanin in nerve cell perikarya or nerve fibers. Synthetic porcine galanin (1-29) promoted acetylcholine release in olfactory bulb tissue slices, suggesting that galanin can effectively modulate cholinergic transmission and perhaps other forms of neuronal transmission. It is concluded that galanin may be significantly involved in olfactory processing at cellular and synaptic levels.

Acetylcholine↗