Astrocytes in multiple sclerosis lack beta-2 adrenergic receptors.

BACKGROUND: In MS, T cells reactive to myelin proteins can cross the blood-brain barrier and release proinflammatory cytokines, such as interferon gamma. These can induce glial cells to express class II major histocompatibility complex (MHC) molecules, which are required to present myelin antigens to the T cells in order to mount a proper autoimmune response. Both microglia and astrocytes can function as antigen-presenting cells. In contrast to microglia, endogenous suppressors, including norepinephrine, regulate astrocytic class II MHC expression. The effects of norepinephrine are mediated through activation of P2 adrenergic receptors. OBJECTIVE: To investigate P, adrenergic receptors in astrocytes in MS. METHODS: Immunocytochemical techniques were applied in postmortem brain tissue from 10 patients with MS, three patients with a cerebral infarction, and six controls, and in spinal cord from three patients with ALS. RESULTS: beta2 adrenergic receptors were visualized on astrocytes in white matter of controls, and they were prominently expressed in reactive astrocytes at the boundary of cerebral infarctions and in the lateral corticospinal tract in ALS. In MS, beta2 adrenergic receptors could neither be visualized on astrocytes in normal-appearing white matter nor in reactive astrocytes in chronic active and inactive plaques, whereas they were normally present on neurons. MHC class II-positive astrocytes were only visualized in chronic active plaques. CONCLUSIONS: Because astrocytic beta2 adrenergic receptors are involved in suppressing inducibility of MHC class II molecules, we suggest that their lack of expression may play an important role in the induction or perpetuation of autoimmune reactions in MS.

Insulin-like growth factor binding protein-4 interacts with centrosomes and microtubules in primary astrocytes.

Insulin-like growth factor-1 and -2 (IGFs) are important for CNS development and have implications in pathological situations of the brain. Insulin-like growth factor binding protein-4 (IGFBP-4) regulates the biological effects of IGFs. We examined the expression of IGFBP-4 in primary rat and human astrocytes. IGFBP-4 mRNA was detectable by reverse transcription-polymerase chain reaction (RT-PCR) and protein expression was verified by Western blotting of cell lysates as well as conditioned culture medium. When astrocytes were immunostained for IGFBP-4 we detected an intracellular structure that did not derive from organelles involved in cellular trafficking. Ingestion of fluorescein isothiocyanate-labeled transferrin excluded detection of IGFBP-4 within vesicles of endosomal nature, in which the protein might have been incorporated. Double staining with gamma-tubulin demonstrated co-localization of IGFBP-4 with centrosomes of these cells. Treatment with nocodazole resulted in absence of IGFBP-4 signal on centrosomes, indicating a dependency on intact microtubules. Immunoelectron microscopy revealed IGFBP-4 localization not only at the centrioles but also a direct interaction with microtubules. There was no binding of IGFBP-4 to centrioles in primary rat oligodendrocytes, microglia or meningeal cells. The association of IGFBP-4 with centrioles and microtubules in astrocytes suggests an involvement of this molecule in microtubule functions of these cells.

Astrocytes in chronic active multiple sclerosis plaques express MHC class II molecules.

To initiate the inflammatory cascade leading to demyelination in multiple sclerosis (MS) T cells have to recognize their specific myelin antigen, which needs to be presented in the context of major histocompatibility (MHC) class II molecules expressed on antigen presenting cells. Whether astrocytes can express MHC class II molecules in vivo is a controversial issue. We performed double labeling immunohistochemistry in postmortem samples from nine patients with MS, three patients with a cerebral infarction and six controls. Astrocytes in controls, in normal appearing white matter in MS, and at the boundary of infarctions were MHC class II negative. In contrast, a subset of astrocytes in active chronic plaques immunostained for MHC class II, indicating potential antigen presenting interactions of astrocytes in MS.

Disappearance of beta2-adrenergic receptors on astrocytes in canine distemper encephalitis: possible implications for the pathogenesis of multiple sclerosis.

It has been reported that astrocytes in the white matter of patients with multiple sclerosis (MS) lack beta2-adrenergic receptors. This abnormality might explain why astrocytes in active MS plaques aberrantly express major histocompatibility (MHC) class II molecules, which play an important role in the immunological cascade leading to myelin destruction. Canine distemper (CD) virus primarily infects astrocytes and causes a demyelinating disease in dogs that closely resembles MS. In control dogs, including three dogs with another inflammatory disease, beta2-adrenergic receptor immunoreactivity was observed on both neurons and astrocytes. In dogs with CD encephalitis, beta2-adrenergic receptors were present on neurons, but were absent on astrocytes in acute lesions, demyelinated lesions, and normal-appearing white matter. Similar to MS, several astrocytes in demyelinated lesions expressed MHC class II. These findings suggest that MS and the demyelinating stages of CD encephalitis have a common pathogenetic factor, and that the loss of astrocytic beta2-adrenergic receptors in MS might be induced by a viral infection of astrocytes.

Insulin-like growth factor-I receptors in normal appearing white matter and chronic plaques in multiple sclerosis.

Preclinical studies suggest that insulin-like growth factor-I (IGF-I) plays an important role in oligodendrocyte survival and myelination. We used human recombinant [125I]IGF-I to study IGF-I receptors in post-mortem brain tissue from patients with multiple sclerosis (MS). In normal appearing white matter, we found that IGF-I receptor densities and binding characteristics were not different between MS patients and controls. In chronic plaques, histologically characterized by astrogliosis, we found densities of IGF-I receptors which were in the same range as those measured in the normal appearing white matter. In vitro studies have shown that IGF-I also acts as a mitogenic factor for astrocytes. Since MS lesions are rapidly invaded by reactive astrocytes, IGF-I may not only protect oligodendrocytes and stimulate remyelination but also enhance the astrogliosis that limits repair.

Insulin-like growth factor II receptors in human brain and their absence in astrogliotic plaques in multiple sclerosis.

Insulin-like growth factor (IGF) II receptors were studied in human adult brain by using autoradiography with [125I]IGF-II. Receptors were found to be widely distributed throughout all neuronal regions. The highest densities were found in plexus choroideus, granular layer of the cerebellar cortex, gyrus dendatus and pyramidal layer of the hippocampus, striatum, and cerebral cortex. White matter was devoid of IGF-II receptors. We also examined [125I]IGF-II binding in six plaques of multiple sclerosis, which were characterized by a dense network of astrocytes. We were unable to detect IGF-II receptors in any of the astrogliotic plaques, suggesting that IGF-II receptors in human brain are not involved in astrogliosis. The regional variations in neuronal distribution of IGF-II receptors suggest involvement of IGF-II in functions associated with specific neuronal pathways.

Autoradiographic distribution of D3-type dopamine receptors in human brain using [3H]7-hydroxy-N,N-di-n-propyl-2-aminotetralin.

The regional distribution of D3 dopamine receptors was studied in human brain by quantitative autoradiography with [3H]7-hydroxy-N,N-di-n-propyl-2-aminotetralin ([3H]7-OH-DPAT). The highest densities were found in the ventral striatum/nucleus accumbens, followed by the remainder of the neostriatum, cerebral cortex, and cerebellar cortex. Moderate amounts were found in the substantia nigra. Low densities of D3 dopamine receptors were visualized in pituitary gland (posterior lobe > anterior lobe), amygdala, and hippocampus. The globus pallidus and thalamus contained lower densities. The distribution pattern is much more widespread as detected by in situ hybridisation histochemistry for D3 mRNA in human and in rat brain. Our data confirm a predilection of D3 receptors in brain areas involved in cognitive and emotional functions. The presence of D3 receptors in non-limbic parts of the neostriatum and substantia nigra suggests that they also play a role in the dopaminergic control of motor processes. Its precise function in cerebellum and pituitary gland is at present uncertain.

Insulin-like growth factor-I receptor densities in human frontal cortex and white matter during aging, in Alzheimer's disease, and in Huntington's disease.

Using quantitative autoradiography we have investigated insulin-like growth factor (IGF)-I receptors in postmortem-obtained frontal cortex and white matter from 39 individuals without neurological disease, ranging in age from 0 to 95 years, and from 5 patients with Huntington's disease and 4 with Alzheimer's disease. IGF-I receptor densities in white matter were significantly higher in neonates than in adults; during adult life there was no further decline. The higher density of IGF-I receptors in white matter of neonates most likely reflects extensive formation of myelin. There was no significant decrease in IGF-I receptor densities in the cortical mantle with age, suggesting that the cells containing IGF-I receptors in frontal cortex are maintained during the entire life-span. There were no significant alterations in IGF-I receptor densities in frontal cortex and white matter from patients with Huntington's disease and Alzheimer's disease compared with controls from the same age groups.

[3H]dihydroalprenolol binding to beta adrenergic receptors in multiple sclerosis brain.

By using immunocytochemistry we previously reported the absence of beta(2) adrenergic receptors on astrocytes in multiple sclerosis (MS) white matter. Here, we measured beta(1) and beta(2) adrenergic receptor concentrations in postmortem brain sections of six MS patients and six controls by using quantitative autoradiography with [(3)H]dihydroalprenolol. White matter contained no beta(1) adrenergic receptors. In white matter of controls low levels of beta(2) adrenergic receptors were detected. In agreement with the immunohistochemical study, we were unable to detect beta(2) adrenergic receptors in both normal appearing white matter and astrogliotic plaques in MS. Concentrations of beta(1) and beta(2) adrenergic receptors in cerebral cortex were not different between controls and MS patients.

[3H]Clozapine is not a suitable radioligand for the labelling of D4 dopamine receptors in postmortem human brain.

Clozapine displays nanomolar affinity for cloned human D4-type dopamine receptors expressed in tissue culture cells. Therefore, [3H]clozapine has been introduced as a radioligand for the labelling of the D4 dopamine receptors. We found that, in membranes of postmortem human striatum, amygdala, frontal cortex and substantia nigra, neither dopamine nor the dopamine agonist apomorphine displaced the binding of [3H]clozapine (5-20 nM). [3H]Clozapine competition curves with clozapine and loxapine revealed the presence of two binding sites. The high-affinity site was displaced by atropine and pirenzepine with nanomolar affinity. The low-affinity site did not correspond to a serotonin, adrenergic, gamma-amino butyric acid, N-methyl-D-aspartate, benzodiazepine, histamine, sigma, imidazoline receptor-binding site, or catecholamine uptake site. Our results suggest that [3H]clozapine in postmortem human brain binds to M1-type muscarinic cholinergic receptors and to a low-affinity binding site but is not a suitable radioligand for investigating the D4 dopamine receptors.

What intracranial tissues in humans contain sumatriptan-sensitive serotonin 5-HT1-type receptors?

We investigated the presence of sumatriptan-sensitive serotonin (5-HT)1 receptors in different human tissues by using a radioligand-binding technique with [3H]5-HT. Sumatriptan displaced [3H]5-HT from frontal cortical and striatal membranes in a biphasic manner, with a high-affinity site corresponding to binding to the 5-HT1D receptor. In blood platelet membranes, sumatriptan displaced [3H]5-HT with a 100-fold lower affinity. Sumatriptan failed to displace [3H]5-HT in membranes from large cerebral arteries, pial vessels, coronary arteries and dura mater. These findings suggest that either there are no sumatriptan-sensitive 5-HT1 receptors on intracranial blood vessels or they are so small in number that they cannot be detected by the radioligand-binding technique. Other mechanisms, possibly centrally mediated, may be responsible for the antimigraine action of sumatriptan.

Insulin-like growth factor system in serum and cerebrospinal fluid in patients with multiple sclerosis.

The insulin-like growth factor (IGF) system influences oligodendrocyte survival, myelination, and immune functions. We examined whether alterations in the circulating IGF system occur in multiple sclerosis (MS), a chronic inflammatory demyelinating disease of the central nervous system. We measured concentrations of IGF-I, IGF-II, and insulin-like growth factor binding proteins -1, -2, and -3 in both serum and cerebrospinal fluid from MS patients and age- and sex-matched controls. IGFBP-1 was not detectable in cerebrospinal fluid. We found no significant differences in any of the other components between patients with MS and controls.

Dopamine agonists used in the treatment of Parkinson's disease and their selectivity for the D1, D2, and D3 dopamine receptors in human striatum.

1. It has been suggested that an ideal antiparkinsonian treatment requires stimulation of both D1 and D2 dopamine receptors. Bromocriptine and lisuride are regarded as pure D2 receptor agonists, whereas pergolide and apomorphine are thought to stimulate both D1 and D2 receptors. 2. The aim of this study was to compare the affinities of bromocriptine, lisuride, pergolide, and apomorphine for the D1, D2, and D3 receptors in postmortem human striatum. The dissociation constants (Ki values) of the dopamine agonists were determined from competition binding experiments with selective radioligands. 3. The Ki values of the orally administered agonists--bromocriptine, pergolide, and lisuride--for the D2 receptors were proportional to their optimal doses against parkinsonism. Ki(D1)/Ki(D2) ratios were 23 for lisuride, 67 for pergolide, 60 for bromocriptine, and 2.6 for apomorphine. Ki(D3)/Ki(D2) ratios were 0.4 for lisuride, 1 for pergolide, 5.4 for bromocriptine, and 21 for apomorphine. 4. The present results support the hypothesis that the antiparkinsonian effect of dopamine agonists is mediated primarily by D2 receptors. Apomorphine is a mixed D1/D2 agonist, but pergolide has no more D1 agonist properties than bromocriptine and lisuride. The role of the D3 receptors is unknown, but their activation might either be associated with the generation of psychiatric side-effects or dyskinesias, or alternatively add to antiparkinsonian activity.

GFAP and S100B in the acute phase of mild traumatic brain injury.

OBJECTIVE: The biomarkers glial fibrillary acid protein (GFAP) and S100B are increasingly used as prognostic tools in severe traumatic brain injury (TBI). Data for mild TBI are scarce. This study aims to analyze the predictive value of GFAP and S100B for outcome in mild TBI and the relation with imaging. METHODS: In 94 patients biomarkers were determined directly after admission. Collected data included injury severity, patient characteristics, admission CT, and MRI 3 months postinjury. Six months postinjury outcome was determined with Glasgow Outcome Scale Extended (GOSE) and return to work (RTW). RESULTS: Mean GFAP was 0.25 µg/L (SD 1.08) and S100B 0.54 µg/L (SD 1.18). In 63% GFAP was not discernible. GFAP was increased in patients with an abnormal CT (1.20 µg/L, SD 2.65) compared to normal CT (0.05 µg/L, SD 0.17, p < 0.05). Also in patients with axonal injury on MRI GFAP was higher (0.65 µg/L, SD 0.91 vs 0.07 µg/L, SD 0.2, p < 0.05). GFAP was increased in patients with incomplete RTW compared to complete RTW (0.69 µg/L, SD 2.11 vs 0.12 µg/L, SD 0.38, p < 0.05). S100B was not related to outcome or imaging studies. In multivariate analysis GFAP was not predictive for outcome determined by GOSE and RTW. CONCLUSIONS: A relation between GFAP with imaging studies and outcome (determined by RTW) was found in contrast to S100B. As the positive predictive value of GFAP is limited in this category of TBI patients, this biomarker is not suitable for prediction of individual patient outcome.

Involvement of morbilliviruses in the pathogenesis of demyelinating disease.

Two members of the morbillivirus genus of the family Paramyxoviridae, canine distemper virus (CDV) and measles virus (MV), are well-known for their ability to cause a chronic demyelinating disease of the CNS in their natural hosts, dogs and humans, respectively. Both viruses have been studied for their potential involvement in the neuropathogenesis of the human demyelinating disease multiple sclerosis (MS). Recently, three new members of the morbillivirus genus, phocine distemper virus (PDV), porpoise morbillivirus (PMV) and dolphin morbillivirus (DMV), have been discovered. These viruses have also been shown to induce multifocal demyelinating disease in infected animals. This review focuses on morbillivirus-induced neuropathologies with emphasis on aetiopathogenesis of CNS demyelination. The possible involvement of a morbillivirus in the pathogenesis of multiple sclerosis is discussed.

Serum levels of insulin-like growth factor-1 and insulin-like growth factor binding protein-3 in relapsing and primary progressive multiple sclerosis.

Using radioimmunoassay we measured serum levels of insulin-like growth factor (IGF)-1 and IGF binding protein (IGFBP)-3 in patients with relapsing multiple sclerosis (MS) and a benign course (Expanded Disability Status Scale (EDSS) < or =3 despite > 10 years disease duration), relapsing MS with cumulative disability leading to an EDSS score >4 within 10 years of disease duration, primary progressive MS and healthy controls. We found no differences in IGF-1 and IGFBP-3 serum levels, and the IGF-1/IGFBP-3 ratio between the four groups. However, there was a significant correlation (P =0.005) between IGFBP-3 serum levels and both the progression index of disability and the Multiple Sclerosis Severity Score in patients with primary progressive MS.

Insulin-like growth factor-I receptors in human brain and pituitary gland: an autoradiographic study.

Insulin-like growth factor (IGF)-I receptors were studied in adult human postmortem brain and pituitary gland using quantitative autoradiography with human recombinant [125I]IGF-I. The highest densities were found in the choroid plexus, pituitary gland--where IGF-I receptors were mainly concentrated in the anterior lobe, pineal gland, glomerular layer of the olfactory bulb, and the molecular layer of the cerebellar cortex. Moderate densities were present in cerebral cortex, caudate nucleus, putamen, accumbens, the CA1, CA2, CA3 fields and dentate gyrus of the hippocampus, the dentate nucleus of the cerebellum, amygdala, thalamus, pontine nuclei, and substantia nigra. All other brain areas, including white matter, contained low densities of IGF-I receptors. The finding that several well-defined brain structures are enriched with IGF-I receptors suggests a neurotrophic/survival or neuromodulatory role of insulin-like growth factors on specific neuronal systems. IGF-I receptors observed in the white matter may be associated with oligodendrocytes.