Search

Aims: The NADPH oxidases constitute a major source of superoxide anion (·O2−) in hypertension. Several studies suggest an important role of NADPH oxidases in different effects mediated by transforming growth factor-β1 (TGF-β1). We investigated whether a chronic treatment with P144, a peptide synthesized from type III TGF-β1 receptor, inhibited NADPH oxidases in the renal cortex of spontaneously hypertensive rats (SHR). Results: Here, we show that chronic administration of P144 significantly reduced the NADPH oxidase expression and activity as well as the oxidative stress observed in control vehicle-treated SHR (V-SHR). In addition, P144 was also able to reduce the significant increase in the renal fibrosis and in mRNA expression of different components of collagen metabolism, as well as in the levels of connective tissue growth factor observed in V-SHR. Finally, TGF-β1-stimulated NRK52E exhibited a significant increase in NADPH oxidase expression and activity as well as a TGF-β1-dependent intrace...

Axonal collateralization is a major organizational feature of basal ganglia circuits. Single-axon tracing techniques reveal the full collateralization patterns of individual axons, providing key connectional information about target diversity. Previous studies from our group have analyzed the axonal branching patterns of two output nuclei of the basal ganglia in the rat, the ventral pallidum (VP) and the substantia nigra pars reticulata (SNr). In this review we compare individual VP and SNr neurons, highlighting common features and unique characteristics, which may contribute to a deeper understanding of their respective connections and function. We additionally outline parallels with the entopeduncular nucleus and the globus pallidus that may help develop a more comprehensive understanding of basal ganglia organization.

NADPH oxidases constitute a major source of superoxide anion (⋅O2 -) in hypertension. Several studies suggest an important role of NADPH oxidases in different effects mediated by TGF-β 1. In this study we show that chronic administration of P144, a peptide synthesized from type III TGF-β 1 receptor, significantly reduced the cardiac NADPH oxidase expression and activity as well as in the nitrotyrosine levels observed in control spontaneously hypertensive rats (V-SHR) to levels similar to control normotensive Wistar Kyoto rats. In addition, P144 was also able to reduce the significant increases in the expression of collagen type I protein and mRNA observed in hearts from V-SHR. In addition, positive correlations between collagen expression, NADPH oxidase activity, and nitrotyrosine levels were found in all animals. Finally, TGF-β 1-stimulated Rat-2 exhibited significant increases in NADPH oxidase activity that was inhibited in the presence of P144. It could be concluded that the blockade of TGF-β 1 with P144 inhibited cardiac NADPH oxidase in SHR, thus adding new data to elucidate the involvement of this enzyme in the profibrotic actions of TGF-β 1.

Resume Objetivo Analizar la aceptacion de la sustitucion por medicamentos genericos en la farmacia comunitaria y estudiar la variabilidad de la respuesta en funcion de las caracteristicas del usuario, la prescripcion y la oficina de farmacia (OF). Diseno Observacional, transversal. Emplazamiento Oficinas de farmacia de Albacete y provincia. Participantes Seleccion consecutiva de 769 usuarios en 21 farmacias durante febrero de 2005. Metodo Usuarios a los que, en caso de que el medicamento prescrito no se encontrara disponible en la OF, se les proponia la sustitucion por un generico. Las propuestas fueron reflejadas en una hoja-registro. Resultados Un 90,8% de la poblacion acepta la sustitucion (n = 698; intervalo de confianza [IC] del 95%, 88,5-92,7%). La principal causa de rechazo fue que el paciente no quiere cambios en la prescripcion medica (50,7%). La aceptacion del cambio fue inversamente proporcional a la edad del paciente (p = 0,005) y directamente proporcional al numero de habitantes del nucleo de poblacion donde se encuentra ubicada la OF (p Conclusiones Se constata una elevada aceptacion de la sustitucion por medicamentos genericos en las OF (90,8%). La principal causa que genera un rechazo a la sustitucion es que el paciente no quiere cambios en la prescripcion.

We have examined the morphology of the somatodendritic domain of projection neurons located in different sectors of rat substantia nigra pars reticulata (SNr) or having distinct axonal arborizations. Forty-three neurons - 23 located in the dorsal half and 20 in the ventral half of SNr - were injected with biotinylated dextran amine and their somatodendritic domain was reconstructed from serial sagittal sections with a camera lucida. The axonal arborization of 14 neurons was also reconstructed. Dorsally located SNr neurons had a larger perikaryon, a higher number of primary dendrites and a more extensive dendritic arbor than the ventrally located ones. However, irrespective of their location in the SNr, the somatodendritic domain was always longer along the rostrocaudal axis than along the dorsoventral and mediolateral axes. Specific correlations between somatodendritic morphology and axonal arborization could be established for some SNr neurons, but among SNr neurons with similar efferent projections, those lying dorsally always exhibited a larger perikaryon and a more widespread dendritic arbor than those located ventrally. These results indicate that the morphology of the somatodendritic domain of SNr projection neurons is related to the location of their perikaryon within the structure rather than to the pattern of their axonal projections.

Neuromelanins (NM) are a family of compounds occurring in all brain regions of human brain. These pigments are contained in special lysosomes together with lipid bodies, protein matrix, and accumulate in aging (Zucca et al. Neurotox Res 2014). The synthesis of NM is a protective process because the melanic component is generated through the removal of reactive/toxic quinones that would otherwise cause neurotoxicity (Sulzer et al. PNAS 2000). NM serves an additional protective role through its ability to chelate and accumulate metals, including environmentally toxic metals such as mercury and lead. Other metals like Fe, Zn, Al, Cr and Mo are also accumulated by NM (Zecca et al. PNAS 2008). However NM can play also a toxic role in Parkinson’s disease (PD), when it is released by dying neurons of substantia nigra. Extracellular NM particles induce microglial activation with production of superoxide, nitric oxide, hydrogen peroxide, pro-inflammatory factors and causes neurodegeneration (Zhang et al. Neurotox Res 2011). A high content of major histocompatibility class I complex (MHC-I) was found in NM-containing organelles of the neurons in substantia nigra (SN) and locus coeruleus (LC) which degenerate in PD, while other neurons not targeted by PD has low MHC-I. The latter can bind antigens derived from foreign proteins, presenting them on neuronal membrane. Then CD8+ cytotoxic T-cells, which were observed in proximity of MHC-I presenting neurons of SN and LC in PD subjects, can target these neurons inducing neuronal death. Infiltration of T-cells occurs in SN and LC of PD subjects. The presence of MHC-I in catecholamine neurons containing NM could explain their selective vulnerability in PD, revealing a novel inflammatory T-cell mediated neurodegenerative process of PD (Cebrian et al. Nat Commun 2014). In conclusion NM can play a protective or toxic role depending on the molecular/cellular context.

The motor symptoms of Parkinson's disease are due to the progressive degeneration of dopaminergic neurons in the substantia nigra. Multiple neuroinflammatory processes are exacerbated in Parkinson's disease, including glial-mediated reactions, increased expression of proinflammatory substances, and lymphocytic infiltration, particularly in the substantia nigra. Neuroinflammation is also implicated in the neurodegeneration and consequent behavioral symptoms of many Parkinson's disease animal models, although it is not clear whether these features emulate pathogenic steps in the genuine disorder or if some inflammatory features provide protective stress responses. Here, we compare and summarize findings on neuroinflammatory responses and effects on behavior in a wide range of toxin-based, inflammatory and genetic Parkinson's disease animal models.

Neuronal expression of major histocompatibility complex I (MHC-I) has been implicated in developmental synaptic plasticity and axonal regeneration in the central nervous system (CNS), but recent findings demonstrate that constitutive neuronal MHC-I can also be involved in neurodegenerative diseases by playing a neuroinflammtory role. Recent reports demonstrate its expression in vitro and in human postmortem samples and support a role in neurodegeneration involving proinflammatory cytokines, activated microglia and increased cytosolic oxidative stress. Major histocompatibility complex I may be important for both normal development and pathogenesis of some CNS diseases including Parkinson’s.

Background: An accumulating body of evidence points to the significance of neuroinflammation and immunogenetics in schizophrenia, and an imbalance of cytokines in the central nervous system (CNS) has been suggested to be associated with the disorder. Munc18-overexpressing mice (Munc18-OE) have provided a model for the study of the alterations that may underlie the symptoms of subjects with schizophrenia. The aim of the present study was to elucidate the involvement of neuroinflammation and cytokine imbalance in this model. Methods: Cytokines were evaluated in the cortex and the striatum of Munc18-OE and wild-type (WT) mice by enzyme-linked immunosorbent assay (ELISA). Protein levels of specific microglia and macrophage, astrocytic and neuroinflammation markers were quantified by western blot in the cortex and the striatum of Munc18-OE and WT mice. Results: Each cytokine evaluated (Interferon-gamma (IFN-gamma), Tumor Necrosis Factor-alpha (TNF-alpha), Interleukin-2 (IL-2) and CCL2 chemokine) was present at higher levels in the striatum of Munc18-OE mice than WT. Cortical TNF-alpha and IL-2 levels were significantly lower in Munc18-OE mice than WT mice. The microglia and macrophage marker CD11b was lower in the cortexes of Munc18-OE mice than WT, but no differences were observed in the striatum. Glial Fibrillary Acidic Protein (GFAP) and Nuclear Factor-kappaB (NF-kappa B)p65 levels were not different between the groups. Interleukin-1beta (IL-1 beta) and IL-6 levels were beneath detection limits. Conclusions: The disrupted levels of cytokines detected in the brain of Munc18-OE mice was found to be similar to clinical reports and endorses study of this type for analysis of this aspect of the disorder. The lower CD11b expression in the cortex but not in the striatum of the Munc18-OE mice may reflect differences in physiological activity. The cytokine expression pattern observed in Munc18-OE mice is similar to a previously published model of schizophrenia caused by maternal immune activation. Together, these data suggest a possible role for an immune imbalance in this disorder. The study was supported by Spanish MINECO (SAF2009-08460) and the Basque Government (IT616-13) to JJM. Experiments were performed in DS's laboratory. CC was supported by the Caja Madrid Foundation and the Parkinson's Disease Foundation. DS's laboratory was supported by NIDA07418 and DA10154 and the Parkinson's, Simons and JPB Foundations.

Subsets of rodent neurons are reported to express major histocompatibility complex class I (MHC-I), but such expression has not been reported in normal adult human neurons. Here we provide evidence from immunolabel, RNA expression and mass spectrometry analysis of postmortem samples that human catecholaminergic substantia nigra and locus coeruleus neurons express MHC-I, and that this molecule is inducible in human stem cell-derived dopamine (DA) neurons. Catecholamine murine cultured neurons are more responsive to induction of MHC-I by gamma-interferon than other neuronal populations. Neuronal MHC-I is also induced by factors released from microglia activated by neuromelanin or alpha-synuclein, or high cytosolic DA and/or oxidative stress. DA neurons internalize foreign ovalbumin and display antigen derived from this protein by MHC-I, which triggers DA neuronal death in the presence of appropriate cytotoxic T cells. Thus, neuronal MHC-I can trigger antigenic response, and catecholamine neurons may be particularly susceptible to T-cell-mediated cytotoxic attack.

The motor symptoms of Parkinson’s disease are due to the progressive degeneration of dopaminergic neurons in the substantia nigra. Multiple neuroinflammatory processes are exacerbated in Parkinson’s disease, including glial-mediated reactions, increased expression of proinflammatory substances, and lymphocytic infiltration, particularly in the substantia nigra. Neuroinflammation is also implicated in the neurodegeneration and consequent behavioral symptoms of many Parkinson’s disease animal models, although it is not clear whether these features emulate pathogenic steps in the genuine disorder or if some inflammatory features provide protective stress responses. Here, we compare and summarize findings on neuroinflammatory responses and effects on behavior in a wide range of toxin-based, inflammatory and genetic Parkinson’s disease animal models.

The patterns of axonal collateralization of nucleus accumbens (Acb) projection neurons were investigated in the rat by means of single-axon tracing techniques using the anterograde tracer biotinylated dextran amine. Seventy-three axons were fully traced, originating from either the core (AcbC) or shell (AcbSh) compartment, as assessed by differential calbindin D28kimmunoreactivity. Axons from AcbC and AcbSh showed a substantial segregation in their targets; target areas were either exclusively or preferentially innervated from AcbC or AcbSh. Axon collaterals in the subthalamic nucleus were found at higher than expected frequencies; moreover, these originated exclusively in the dorsal AcbC. Intercompartmental collaterals were observed from ventral AcbC axons into AcbSh, and likewise, interconnections at pallidal and mesencephalic levels were also observed, although mostly from AcbC axons toward AcbSh targets, possibly supporting crosstalk between the two subcircuits at several levels. Cell somata giving rise to short-range accumbal axons, projecting to the ventral pallidum (VP), were spatially intermingled with others, giving rise to long-range axons that innervated VP and more caudal targets. This anatomical organization parallels that of the dorsal striatum and provides the basis for possible dual direct and indirect actions from a single axon on either individual or small sets of neurons. J. Comp. Neurol. 518:4649– 4673, 2010.

The most caudally located dopaminergic (DA) ventral tier neurons of the substantia nigra pars compacta (SNc) form typical cell clusters that are deeply embedded in the substantia nigra pars reticulata (SNr). Here we examine the efferent projections of 35 neurons located in the SNr region where these SNc cell clusters reside. The neuronal cell body was injected with biotinylated dextran amine so as to trace each complete axon in the sagittal or the coronal plane. Electrophysiological guidance guaranteed that the tracer was ejected among neurons displaying a typical SNc discharge pattern. Furthermore, double immunofluorescence and immunohistochemical labeling ensured that the tracer deposits were placed within the DA cell clusters. Three types of projection neurons occurred in the SNc ventral tier cell cluster region: type I neurons, projecting to basal ganglia; type II neurons, targeting both the basal ganglia and thalamus; and type III neurons, projecting only to the thalamus. The striatum was targeted by most of the type I and II neurons and the innervation reached both the striosome/subcallosal streak and matrix compartments. Many nigrostriatal fibers provided collaterals to the globus pallidus and, less frequently, to the subthalamic nucleus. At a thalamic level, type II and III neurons preferentially targeted the reticular, ventral posterolateral, and ventral medial nuclei. Our results reveal that the SNr region where DA ventral tier cell clusters reside harbors neurons projecting to the basal ganglia and/or the thalamus, thus suggesting that neurodegeneration of nigral neurons in Parkinson′s disease might affect various extrastriatal basal ganglia structures and multiple thalamic nuclei. J. Comp. Neurol. 518:1283–1300, 2010. © 2009 Wiley-Liss, Inc.

This paper reviews two of the major features of the nigrostriatal pathway, its axonal collateralization, and compartmental specificity, as revealed by single-axon labeling experiments in rodents and immunocytological analysis of human postmortem tissue. The dorsal and ventral tiers of the substantia nigra pars compacta harbor various types of neurons the axons of which branch not only within the striatum but also in other major components of the basal ganglia. Furthermore, some nigrostriatal axons send collaterals both to thalamus and to brainstem pedunculopontine tegmental nucleus. In humans, the compartmental specificity of the nigrostriatal pathway is revealed by the fact that the matrix compartment is densely innervated by dopaminergic fibers, whereas the striosomes display different densities of dopaminergic terminals depending on their location within the striatum. The nigral neurons most severely affected in Parkinson's disease are the ventral tier cells that project to the matrix and form deep clusters in the substantia nigra pars reticulata.

OBJECTIVES: . To analyse the acceptance of the replacement of patent medicines by generic medicines at community pharmacies and to study the variability of the answers in terms of the characteristics of the user, prescription and community pharmacy. DESIGN: Observational, cross-sectional study. SETTING: Community pharmacies in Albacete and its province, Spain. PARTICIPANTS: Consecutive selection of 769 users in 21 pharmacies during February 2005. METHOD: Users of community pharmacies who were asked to replace a prescribed drug by a generic one when the prescribed one was unavailable. The proposals were recorded in a register and the data were processed with the SPSS statistical package. RESULTS: 90.8% of the population accepted the replacement (n=698; 95% CI, 88.5%-92.7%). The main cause of non-acceptance was that the patient did not want changes in his/her medical prescription (50.7%). The acceptance of the change was inversely proportional to the age of the patient (P=.005) and directly proportional to the number of inhabitants of the population centre where the pharmacy was located (P

Axons from neurons of the rat substantia nigra pars reticulata (SNr) and pars lateralis (SNl) were traced after injecting their cell body with biotinylated dextran amine. Thirty-two single axons were reconstructed from serial sagittal sections with a camera lucida, whereas four other SNr axons were reconstructed in the coronal plane to determine whether they innervate the contralateral hemisphere. Four distinct types of SNr projection neurons were identified based on their main axonal targets: type I neurons that project to the thalamus; type II neurons that target the thalamus, the superior colliculus (SC), and the pedunculopontine tegmental nucleus (PPTg); type III neurons that project to the periaqueductal gray matter and the thalamus; and type IV neurons that target the deep mesencephalic nucleus (DpMe) and the SC. The axons of the SNl showed the same branching patterns as SNr axons of types I, II, and IV. The coronal reconstructions demonstrated that SNr neurons innervate the thalamus, the SC, and the DpMe bilaterally. At the thalamic level, SNr and SNl axons targeted preferentially the ventral medial, ventral lateral, paracentral, parafascicular, and mediodorsal nuclei. Axons reaching the SC arborized selectively within the deep layers of this structure. Our results reveal that the SNr and SNl harbor several subtypes of projection neurons endowed with a highly patterned set of axon collaterals. This organization allows single neurons of these output structures of the basal ganglia to exert a multifaceted influence on a wide variety of diencephalic and midbrain structures. J. Comp. Neurol. 492:349 –369, 2005. © 2005 Wiley-Liss, Inc.