دورية أكاديمية
Visualizing Shifts on Neuron-Glia Circuit with the Calcium Imaging Technique.
| العنوان: | Visualizing Shifts on Neuron-Glia Circuit with the Calcium Imaging Technique. |
|---|---|
| المؤلفون: | Tempone MH; Laboratory of Neurochemistry, Institute of Biophysics Carlos Chagas Filho, Universidade Federal do Rio de Janeiro., Freitas HR; Laboratory of Neuroenergetics and Inborn Errors of Metabolism, Institute of Medical Biochemistry Leopoldo de Meis, Universidade Federal do Rio de Janeiro., Schitine CS; Laboratory of Neurochemistry and Cell Biology, Institute of Life Sciences, Universidade Federal da Bahia., de Melo Reis RA; Laboratory of Neurochemistry, Institute of Biophysics Carlos Chagas Filho, Universidade Federal do Rio de Janeiro; ramreis@biof.ufrj.br. |
| المصدر: | Journal of visualized experiments : JoVE [J Vis Exp] 2022 Apr 08 (182). Date of Electronic Publication: 2022 Apr 08. |
| نوع المنشور: | Journal Article; Video-Audio Media; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: MYJoVE Corporation Country of Publication: United States NLM ID: 101313252 Publication Model: Electronic Cited Medium: Internet ISSN: 1940-087X (Electronic) Linking ISSN: 1940087X NLM ISO Abbreviation: J Vis Exp Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: [Boston, Mass. : MYJoVE Corporation, 2006]- |
| مواضيع طبية MeSH: | Calcium* , Neuroglia*, Astrocytes ; Cell Communication/physiology ; Neurons |
| مستخلص: | Here, we report on selective in vitro models of circuits based on glia (astrocytes, oligodendrocytes, and microglia) and/or neurons from peripheral (dorsal root ganglia) and central tissues (cortex, subventricular zone, organoid) that are dynamically studied in terms of calcium shifts. The model chosen to illustrate the results is the retina, a simple tissue with complex cellular interactions. Calcium is a universal messenger involved in most of the important cellular roles. We explain in a step-by-step protocol how retinal neuron-glial cells in culture can be prepared and evaluated, envisioning calcium shifts. In this model, we differentiate neurons from glia based on their selective response to KCl and ATP. Calcium permeable receptors and channels are selectively expressed in different compartments. To analyze calcium responses, we use ratiometric fluorescent dies such as Fura-2. This probe quantifies free Ca 2+ concentration based on Ca 2+ -free and Ca 2+ -bound forms, presenting two different peaks, founded on the fluorescence intensity perceived on two wavelengths. |
| المشرفين على المادة: | SY7Q814VUP (Calcium) |
| تواريخ الأحداث: | Date Created: 20220425 Date Completed: 20220427 Latest Revision: 20220523 |
| رمز التحديث: | 20231215 |
| DOI: | 10.3791/63338 |
| PMID: | 35467648 |
| قاعدة البيانات: | MEDLINE |
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