Impairment of mitochondria dynamics by human A53T α-synuclein and rescue by NAP (davunetide) in a cell model for Parkinson's disease.

The formation of oligomers and aggregates of overexpressed or mutant α-synuclein play a role in the degeneration of dopaminergic neurons in Parkinson's disease by causing dysfunction of mitochondria, reflected in their disturbed mobility and production of ROS. The mode of action and mechanisms underlying this mitochondrial impairment is still unclear. We have induced stable expression of wild-type, A30P or A53T α-synuclein in neuronally differentiated SH-SY5Y neuroblastoma cells and studied anterograde and retrograde mitochondrial trafficking in this cell model for Parkinson's disease.

Early transient presence of implanted bone marrow stem cells reduces lesion size after cerebral ischaemia in adult rats.

Aims: Previous studies on the therapeutic time window for intravascular administration of bone marrow stem cells (BMSCs) after stroke have shown that early intervention (from 3 h after onset) in the middle cerebral artery occlusion (MCAO) rat model is the most effective approach to reduce ischaemic lesion size. We have confirmed these observations but noticed that 2 weeks after transplantation, almost none of the grafted BMSCs could be detected in or around the lesion. The present experiments aimed to assess the fate and kinetics of intravascularly injected BMSCs shortly after administration in correlation to the development of the ischaemic lesion after MCAO.

p75NTR independent oligodendrocyte death in cuprizone-induced demyelination in C57BL/6 mice.

Feeding C57Bl/6 J mice the copper chelator cuprizone leads to selective apoptosis of mature oligodendrocytes and concomitant demyelination predominantly in the corpus callosum. The process of oligodendrocyte apoptosis in this animal model for multiple sclerosis (MS) involves early microglial activation, but no infiltration of T-lymphocytes. Therefore, this model could mimic early stages of oligodendrocyte degeneration Affected oligodendrocytes express the common neurotrophin receptor, p75(NTR), a 'stress-receptor' which under certain circumstances can induce apoptosis.

Functionally deficient neuronal differentiation of mouse embryonic neural stem cells in vitro.

Embryonic mouse neural stem cells (NSCs) were isolated from E14 mice, multiplied in medium containing epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) and plated in laminin-coated wells in basic serum-free neurobasal medium. After 7 days in vitro, approximately 20% of the embryonic mouse NSCs developed into morphologically and biochemically fully maturated neurons, with extensive dendrites and multiple synaptic contacts. However, even after 22 days of culture, none of these neurons developed voltage-dependent sodium-channels characteristic for a functional neuron.

Reduced p75NTR expression delays disease onset only in female mice of a transgenic model of familial amyotrophic lateral sclerosis.

hSOD1 (G93A) transgenic mice develop pathological changes similar to those in patients with familial amyotrophic lateral sclerosis (FALS). In particular, the progressive degeneration of motoneurons is charactered in this mouse model. One feature of stressed motoneurons in ALS and the hSOD1 mice is the induction of the p75 neurotrophin receptor, which is thought, under certain circumstances, to be a death-signaling molecule.

Expression of the low affinity neurotrophin receptor p75 in spinal motoneurons in a transgenic mouse model for amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis is a lethal neurodegenerative disorder involving motoneuron loss in the cortex, brainstem and spinal cord, resulting in progressive paralysis. Aberrant neurotrophin signalling via the low affinity neurotrophin receptor p75 has been suggested to be involved in the motoneuron death by the activation of apoptotic pathways. In order to investigate the involvement of neurotrophin receptor p75 in the amyotrophic lateral sclerosis related motoneuron degeneration process, we have studied the expression of this receptor in the spinal cord of transgenic mice carrying a mutated human Cu, Zn superoxide dismutase gene.

Elevated levels of neurotrophins in human biceps brachii tissue of amyotrophic lateral sclerosis.

Previous studies suggest that neurotrophins support regeneration and survival of injured motoneurons. Based on these findings, brain-derived neurotrophic factor (BDNF) has been clinically investigated for its therapeutic potential in amyotrophic lateral sclerosis (ALS), a rapidly progressing and fatal motoneuronal disease. We questioned whether imbalances of neurotrophic levels are indeed involved in the pathology of ALS.

Ultrastructural localisation of intramuscular expression of BDNF mRNA by silver-gold intensified non-radioactive in situ hybridisation.

A non-radioactive in situ hybridisation method is described for the detection of low intramuscular levels of brain-derived neurotrophic factor (BDNF) mRNA at the electron microscope level. Application of high-grade silver-gold intensification of the diaminobenzidine end product of in situ hybridisation revealed that in adult rat muscle the constitutive expression of muscular BDNF is confined to the myofibres; satellite cells, Schwann cells, endothelial cells, fibroblasts or axons do not appear to contribute to BDNF production in normal muscle. Although muscular BDNF is a neurotrophic factor for innervating motoneurons and supposedly released only at the motor endplates, the production of BDNF mRNA appears to occur along the entire length of the myofibres and is not confined to nuclei in the postsynaptic regions.

Expression of interleukin-1 beta in rat dorsal root ganglia.

The expression of interleukin-1beta was examined in dorsal root ganglion (DRG) neurons from adult rats using non-radioactive in situ hybridization and immunocytochemistry. At all spinal levels, approximately 70% of the DRG neurons appeared to express IL-1beta mRNA; about 80% of these DRG neurons actually appeared to produce the IL-1beta protein at markedly varying levels. The expression of IL-1beta was found in large as well as in intermediate diameter sensory neurons but only sporadically in the population of small sensory neurons.

Fetal porcine ventral mesencephalon graft. Determination of the optimal gestational age for implantation in parkinsonian patients.

Human fetal ventral mesencephalon tissue has been used as dopaminergic striatal implants in Parkinsonian patients, so far with variable effects. Fetuses from animals that breed in large litters, e.g.

Neurotrophic requirements of rat embryonic catecholaminergic neurons from the rostral ventrolateral medulla.

The factors that regulate the ontogeny and differentiation of C1 adrenergic neurons located in the rostral ventrolateral medulla (RVLM) are completely unknown. In the present study, we have investigated the effects of a number of neurotrophic factors on the survival of E18-19 rat C1 adrenergic neurons in culture. Immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) were used to study the expression of tyrosine hydroxylase (TH), an enzyme present in all catecholaminergic neurons, and of phenylethanolamine N-methyltransferase (PNMT), the final enzyme in the synthesis of adrenalin, as markers for the C1 RVLM neurons.

Discharge properties of motoneurones: how are they matched to the properties and use of their muscle units?

A general survey is given of old as well as more recent findings concerning matches between electrophysiological properties of motoneurones and contractile properties of their muscle fibres. Mechanisms for creating and maintaining such matches are discussed. It is pointed out that it is not sufficient to describe the variation of functional motoneurone characteristics simply in terms of 'fast' or 'slow': all properties seem continuously graded and there is cytochemical evidence for several, seemingly independent parameters of functional specialisation.

Neurotrophin-3 mRNA expression in rat intrafusal muscle fibres after denervation and reinnervation.

We have studied the regulation of the expression of neurotrophin-3 (NT-3) mRNA in neonatal and adult rat muscle spindles after denervation and after denervation followed by reinnervation. Denervation of the intrafusal fibres did not result in an upregulation of the NT-3 mRNA expression but decreased this expression below the detection limit of the in situ hybridization method. Reinnervation of intrafusal fibres restored the NT-3 mRNA expression.

Dopamine-immunoreactivity in the rat mesencephalic trigeminal nucleus: an ultrastructural analysis.

The ultrastructure and distribution of dopaminergic boutons within the rat mesencephalic trigeminal (Me5) nucleus was examined with the use of electronmicroscopic immunocytochemistry. A total of 5102 boutons, comprising axosomatic and axodendritic synaptic terminals as well as non-synaptic boutons (or varicosities), located in the ventrocaudal portion of Me5 was analysed. Approximately 20% of these boutons were dopamine-immunoreactive.

Calreticulin expression in spinal motoneurons of the rat.

We have examined the expression of calreticulin in rat spinal motoneurons in order to reveal the occurrence and distribution of Ca2(+)-storage organelles in these neurons. Calreticulin, the non-muscle equivalent of calsequestrin, is the low-affinity, high-capacity calcium-binding protein responsible for intracompartmental Ca2(+)-storage in a number of different cell types. The results of the present immunohistochemical study show that all spinal motoneurons express calreticulin at approximately the same level; no significant differences in cytoplasmic immunostaining intensity were observed between different motoneuron pools or between small and large spinal motoneurons.

Immunogold localization of serotonin within synaptic terminals in the rat mesencephalic trigeminal nucleus.

With the use of postembedding electron-microscopic immunogold cytochemistry, the vesicular distribution of serotonin within serotonergic synaptic terminals in the mesencephalic trigeminal nucleus was determined in order to obtain further insight into the mechanisms and functional significance of serotonin release to these jaw muscle spindle afferent neurons. Immunogold labelling was restricted to the previously described type I and type II terminals. The distribution of immunogold particles over the synaptic terminals indicated that serotonin was stored in small round or pleomorphic (RSV) vesicles and in large granular dense-cored (DSV) vesicles.

Fetal porcine ventral mesencephalon grafts: dissection procedure and cellular characterization in culture.

The objective of this study was to develop an optimal dissection procedure for fetal porcine ventral mesencephalon (VM) grafts and to characterize the cellular composition of such an explant, in particular with respect to the dopaminergic and GABAergic components. We have used a monolayer cell culture system to study and identify the various VM cell types. The in vitro development of the fetal VM cells and the effect of the addition of brain-derived neurotrophic factor (BDNF) was investigated during a culture period of 5 days.

Selective expression of neurotrophin-3 messenger RNA in muscle spindles of the rat.

The expression of neurotrophin-3 messenger RNA was studied by in situ hybridization in rat muscle spindles from the first embryonic stages of their formation until their mature appearance in adult animals. The first expression of neurotrophin-3 messenger RNA in developing muscles was observed at E19 in the firstly formed intrafusal fiber, the nuclear bag2 fiber. High levels of neurotrophin messenger RNA were found in the equatorial region of these intrafusal fibers in thin lines of cytoplasma around and between the packed-up nuclei.

Expression of calcium-binding proteins in the neurotrophin-3-dependent subpopulation of rat embryonic dorsal root ganglion cells in culture.

In this study we have examined the calcium-binding protein expression in rat embryonic (E16) dorsal root ganglia (DRG) neurons in vitro in the presence of neurotrophin-3 (NT-3). A comparison was made with the expression of calcium-binding proteins in DRG subpopulations that depended in vitro on nerve growth factor (NGF) or brain-derived neurotrophic factor (BDNF). Our results show that NT-3 promotes the survival of a DRG subpopulation of which over 75% expresses parvalbumin (PV).

Outstations of the Golgi complex are present in the processes of cultured rat oligodendrocytes.

Primary cultures of rat oligodendrocytes were incubated with a fluorescent sphingolipid precursor, 6-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]hexanoylceramide+ ++ (C6-NBD-ceramide). This compound is known to stain the Golgi complex specifically. Within 30 min of incubation at 37 degrees C most of the C6-NBD-ceramide was incorporated into the perinuclear Golgi system, as revealed by conventional and confocal laser fluorescence microscopy.

Survival and neurite formation of mesencephalic trigeminal neurones of the rat in vitro.

In order to study the development and functional properties of single, isolated, rat mesencephalic trigeminal neurones, a cell-culture procedure was developed for these specific primary sensory neurones. Mesencephalic trigeminal neurones were isolated from the brainstem of 16-day-old rat embryos. Various factors thought to promote the survival and growth of these neurones in vitro were examined.

GABA-ergic component of rat embryonic ventral mesencephalic grafts: an in-vitro study.

In order to establish the number, the viability and the developmental potential of GABAergic neurons present in dopaminergic ventral mesencephalic (VM) grafts from embryonic rat, we have studied the survival and development of these neurons in culture. The GABAergic fraction demonstrated a highly disproportionate survival in culture in relation to other VM neurons resulting in a drastic change in the neuronal composition of the dissociated VM grafts. The occurrence of a similar gradual dominance of GABAergic neurons at the site of intracerebral implantation, may affect the development of grafted dopaminergic VM neurons and their interaction with host striatal cells.

Serotonin-immunoreactive terminals in the mesencephalic trigeminal nucleus of the rat: an electron microscopic immunocytochemical study.

The morphological characteristics and distribution of serotonin-immunoreactive terminals within the rat mesencephalic trigeminal nucleus (Me5) were studied using immunocytochemical electron microscopy. Fibers, immunostained by specific antibodies raised against serotonin, were distributed throughout the entire rostrocaudal portion of the Me5. Examination of 355 serotonergic terminals in the caudal part of the Me5 revealed that 93% formed synaptic contacts with dendritic shafts: 68% on small (< 1.

Distribution of synaptic boutons in the mesencephalic trigeminal nucleus of the rat--a quantitative electron-microscopical study.

The distribution of synapses and synaptic bouton types in the mesencephalic trigeminal (Me5) nucleus was examined in a quantitative electron-microscopical study. Of 588 terminal boutons that were counted in the compact caudal part of the Me5 nucleus, less than 8% formed synapses on the somata of the predominantly unipolar Me5 neurons. About 79% formed synapses on fibres located between the Me5 somata, while about 13% of the vesicle-containing terminals had no clear synaptic specialization.