1. [Morphologic evaluation and Ca2+ mobilization by glicose and acetylcholine in human pancreatic cells]
- Author
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Daniela Espinha, Cardoso, Lucimar Pereira de, França, Elisângela, Chinen, Andréa A F Souza, Moraes, Alice Teixeira, Ferreira, and Jerônimo Pereira de, França
- Subjects
Organelles ,Receptor, Muscarinic M3 ,Analysis of Variance ,Cell Nucleus Shape ,Cell Culture Techniques ,Cholinergic Agonists ,Immunohistochemistry ,Acetylcholine ,Islets of Langerhans ,Glucose ,Insulin-Secreting Cells ,Insulin Secretion ,Humans ,Insulin ,Calcium Signaling ,Cells, Cultured - Abstract
The proposal of this study was to analyze morphology of the organelles and cytoskeleton in human pancreatic cells cultured and the mobilization of the cytosolic calcium ([Ca2+]c) in response to glucose and ACh by fluorimetry method.The cells were plated on glass coverslips, fixed and stained with a combination of fluorophores: the nuclei were stained with DAPI and mitochondria with Mytotracker Red. It was used phalloidin and the secondary antibodies Alexa Fluor conjugated green and red-fluorescent (488 and 594) to identify the protein cell actin F and type M3 muscarinic receptor respectively. The cells also were loaded with fura-2/AM to study Ca2+ mobilization.The human pancreatic cells show characteristics morphologically preserved with great amount of mitochondria. In region major cell density was evidenced pseudo-islets and type M3 muscarinic receptors. Through increase of [Ca2+]c due to action of glucose and ACh were shown that the cells capacity to respond to these stimuli were conserved. The elevation of the [Ca2+]c depended on concentration by glucose-induced promoting sustained phase and ACh-induced a biphasic response.The morphologic characteristics of human pancreatic cells cultured were preserved. The Ca2+ mobilization in response to glucose and ACh confirmed its functionality. The expression of the M3 muscarinic receptors in human pancreatic cell cultured was demonstrated.
- Published
- 2006