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In vitro differentiation between oxytocin- and vasopressin-secreting magnocellular neurons requires more than one experimental criterion.
da Silva, M P; Merino, R M; Mecawi, A S; Moraes, D J; Varanda, W A.
Afiliación
  • da Silva MP; Department of Physiology, Ribeirão Preto Medical School, University of São Paulo, Av. Bandeirantes, 3900, 14049-900 Ribeirão Preto, SP, Brazil.
  • Merino RM; Department of Physiology, Ribeirão Preto Medical School, University of São Paulo, Av. Bandeirantes, 3900, 14049-900 Ribeirão Preto, SP, Brazil.
  • Mecawi AS; Department of Physiology, Faculty of Medicine, University of Malaysia, Malaysia.
  • Moraes DJ; Department of Physiology, Ribeirão Preto Medical School, University of São Paulo, Av. Bandeirantes, 3900, 14049-900 Ribeirão Preto, SP, Brazil.
  • Varanda WA; Department of Physiology, Ribeirão Preto Medical School, University of São Paulo, Av. Bandeirantes, 3900, 14049-900 Ribeirão Preto, SP, Brazil. Electronic address: wvaranda@fmrp.usp.br.
Mol Cell Endocrinol ; 400: 102-11, 2015 Jan 15.
Article en En | MEDLINE | ID: mdl-25451978
The phenotypic differentiation between oxytocin (OT)- and vasopressin (VP)-secreting magnocellular neurosecretory cells (MNCs) from the supraoptic nucleus is relevant to understanding how several physiological and pharmacological challenges affect their electrical activity. Although the firing patterns of OT and VP neurons, both in vivo and in vitro, may appear different from each other, much is assumed about their characteristics. These assumptions make it practically impossible to obtain a confident phenotypic differentiation based exclusively on the firing patterns. The presence of a sustained outward rectifying potassium current (SOR) and/or an inward rectifying hyperpolarization-activated current (IR), which are presumably present in OT neurons and absent in VP neurons, has been used to distinguish between the two types of MNCs in the past. In this study, we aimed to analyze the accuracy of the phenotypic discrimination of MNCs based on the presence of rectifying currents using comparisons with the molecular phenotype of the cells, as determined by single-cell RT-qPCR and immunohistochemistry. Our results demonstrated that the phenotypes classified according to the electrophysiological protocol in brain slices do not match their molecular counterparts because vasopressinergic and intermediate neurons also exhibit both outward and inward rectifying currents. In addition, we also show that MNCs can change the relative proportion of each cell phenotype when the system is challenged by chronic hypertonicity (70% water restriction for 7 days). We conclude that for in vitro preparations, the combination of mRNA detection and immunohistochemistry seems to be preferable when trying to characterize a single MNC phenotype.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Núcleo Supraóptico / Potenciales de Acción / ARN Mensajero / Oxitocina / Vasopresinas / Neuronas Tipo de estudio: Guideline / Prognostic_studies Límite: Animals Idioma: En Revista: Mol Cell Endocrinol Año: 2015 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Irlanda
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Núcleo Supraóptico / Potenciales de Acción / ARN Mensajero / Oxitocina / Vasopresinas / Neuronas Tipo de estudio: Guideline / Prognostic_studies Límite: Animals Idioma: En Revista: Mol Cell Endocrinol Año: 2015 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Irlanda