ABSTRACT
The low effectiveness of symptomatic pharmacotherapy for Parkinson's disease (PD), which compensates for dopamine (DA) deficiency under degeneration of nigrostriatal dopaminergic (DAergic) neurons, could apparently be improved with neuroprotective therapy, which slows down neurodegeneration and PD progression. For this, it is necessary to have a DAergic cell line for the development of a PD model to screen neuroprotectors. We used immortalized human embryonic mesencephalon LUHMES cells (LCs) differentiated into DAergic neurons. The aim of this study was to characterize the phenotype of differentiated LCs and develop an 1-methyl-4-phenylpyridinium iodide (MPP+)-based test system for screening neuroprotectors. Using polymerase chain reaction (PCR) and immunocytochemistry, it has been shown that all differentiated LCs express genes and synthesize proteins characteristic of all neurons (microtubule-associated protein 2, bIII-tubulin, synaptotagmin 1) and specifically of DAergic neurons (tyrosine hydroxylase, aromatic L-amino acid decarboxylase, DA transporter, vesicular monoamine transporter 2). Furthermore, LCs are able to produce a small amount of DA, but under special conditions. To assess the mechanisms of neurodegeneration and neuroplasticity under the influence of toxins and antiparkinsonian drugs, including neuroprotectors, we have developed an LCs-based MPP+ PD model and proposed an original panel of markers for testing functional and structural cell disorders.
Subject(s)
1-Methyl-4-phenylpyridinium , Parkinson Disease , Humans , 1-Methyl-4-phenylpyridinium/pharmacology , Dopamine/metabolism , Dopaminergic Neurons/metabolism , Parkinson Disease/drug therapy , Parkinson Disease/metabolism , Antiparkinson Agents/metabolism , PhenotypeABSTRACT
This is the first study aiming to develop a method for the long-term visualization of living nigrostriatal dopaminergic neurons using 1-(2-(bis(4-fluorophenyl)methoxy)ethyl)-4-(3-phenylpropyl)piperazine-BODIPY (GBR-BP), the original fluorescent substance, which is a derivative of GBR-12909, a dopamine uptake inhibitor. This method is based on the authors' hypothesis about the possibility of specifically internalizing into dopaminergic neurons substances with a high affinity for the dopamine transporter (DAT). Using a culture of mouse embryonic mesencephalic and LUHMES cells (human embryonic mesencephalic cells), as well as slices of the substantia nigra of adult mice, we have obtained evidence that GBR-BP is internalized specifically into dopaminergic neurons in association with DAT via a clathrin-dependent mechanism. Moreover, GBR-BP has been proven to be nontoxic. As we have shown in a primary culture of mouse metencephalon, GBR-BP is also specifically internalized into some noradrenergic and serotonergic neurons, but is not delivered to nonmonoaminergic neurons. Our data hold great promise for visualization of dopaminergic neurons in a mixed cell population to study their functioning, and can also be considered a new approach for the development of targeted drug delivery to dopaminergic neurons in pathology, including Parkinson's disease.
Subject(s)
Dopaminergic Neurons , Membrane Glycoproteins , Animals , Dopamine Uptake Inhibitors/pharmacology , Dopaminergic Neurons/metabolism , Membrane Glycoproteins/metabolism , Mesencephalon/metabolism , Mice , Nerve Tissue ProteinsABSTRACT
BACKGROUND: The Multidimensional Alcohol Craving Scale (MACS) and Single Photon Emission Computerized Tomography (SPECT) with (123)I-iodobenzamide ((123)I-IBZM) can be useful tools for assessing relapse risk in early recovery from alcohol-dependency. The aim of this study was to assess possible relationships between MACS score, (123)I-IBZM binding and time to first heavy drinking day (TFHD) after detoxification treatment. METHODS: Nineteen alcohol-dependent in-patients were evaluated by MACS scale and an 123I-IBZM-SPECT, performed following alcohol detoxification treatment. At discharge, participants were advised to take naltrexone 50 mg/day for relapse prevention. TFHD was assessed over a 12-week follow up. RESULTS: The MACS score at the beginning of the detoxification process and naltrexone treatment after detoxification were independent predictive factors for TFHD. CONCLUSIONS: The MACS scale is a better predictor of TFHD than IBZM binding. It is simple, non-invasive and inexpensive and appears to be a useful instrument both for clinical practice and for research.
Subject(s)
Alcoholism/diagnostic imaging , Alcoholism/psychology , Iodobenzenes , Tomography, Emission-Computed, Single-Photon , Adult , Humans , Male , Middle Aged , Predictive Value of Tests , Recurrence , Severity of Illness Index , Time FactorsABSTRACT
Objetivos: La Escala Multidimensional de Craving de Alcohol (EMCA) yla Tomografía Computarizada por emisión de Fotón Simple (SPECT) con123I-yodobenzamida (IBZM) pueden ser instrumentos válidos para evaluar el riesgo de recaída, durante la etapa inicial de la recuperación del trastorno por dependencia del alcohol. El objetivo de este estudio es evaluarlas posibles relaciones entre la escala EMCA y la captación de IBZM y el tiempo hasta el primer consumo excesivo de alcohol (TPCEA) una vez finalizado el tratamiento de desintoxicación. Metodología: Diecinueve pacientes hospitalizados han sido evaluados mediante la escala EMCA y la SPECT con IBZM al finalizar el tratamiento de desintoxicación del alcohol. En el momento del alta se les aconsejó seguir un tratamiento con naltrexona 50 mg/día para la prevención de recaídas. El TPCEA ha sido evaluado durante 12 semanas de seguimiento. Resultados: La puntuación de la escala EMCA, al inicio del proceso de desintoxicación, y el seguimiento de un tratamiento con naltrexona, posteriormente a dicho proceso, fueron factores predictivos independientes del TPCEA. Conclusiones: La escala EMCA se ha mostrado como un buen predictor del TPCEA mientras que la captación de IBZM parece no serlo. La escala EMCA parece presentar una mayor utilidad, tanto clínica como para la investigación, frente a evaluaciones más complejas, invasivas y costosas (AU)
Background: The Multidimensional Alcohol Craving Scale (MACS) and Single Photon Emission Computerized Tomography (SPECT) with 123I-iodobenzamide (123I-IBZM) can be useful tools for assessing relapse risk in early recovery from alcohol-dependency. The aim of this study was to assess possible relationships between MACS score, 123I-IBZM binding and time to first heavy drinking day (TFHD) after detoxification treatment. Methods: Nineteen alcohol-dependent in-patients were evaluated by MACS scale and an 123I-IBZM-SPECT, performed following alcohol detoxification treatment. At discharge, participants were advised to take naltrexone 50 mg/day for relapse prevention. TFHD was assessed over a 12-week follow up. Results: The MACS score at the beginning of the detoxification process and naltrexone treatment after detoxification were independent predictive factors for TFHD. Conclusions: The MACS scale is a better predictor of TFHD than IBZM binding. It is simple, non-invasive and inexpensive and appears to be a useful instrument both for clinical practice and for research (AU)
