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Volume changes in single N1E-115 neuroblastoma cells measured with a fluorescent probe
W E Crowe1, J Altamirano, L Huerto
1Departamento de Neurobiología, Instituto Mexicano de Psiquiatría, México, D.F.
Neuroscience
|November 1, 1995
Summary
N1E-115 neuroblastoma cells exhibit Ca2+-sensitive volume control, shrinking in response to increased intracellular calcium. This study details a novel microspectrofluorimetric technique for measuring cell water volume changes in single cells.
Area of Science:
- Cell Biology
- Neuroscience
- Biophysics
Background:
- Cell volume regulation is crucial for cellular function.
- Understanding calcium's role in cell volume control is important for neurological research.
- N1E-115 murine neuroblastoma cells are a model for neuronal studies.
Purpose of the Study:
- To investigate experimentally induced changes in cell water volume in N1E-115 cells.
- To characterize the osmotic behavior of N1E-115 cells.
- To introduce and validate a new microspectrofluorimetric technique for measuring single-cell water volume.
Main Methods:
- Utilized a non-invasive microspectrofluorimetric technique with calcein as an intracellular water marker.
- Exposed N1E-115 cells to various osmolalities (hyperosmotic and hyposmotic solutions).
- Induced increases in intracellular calcium ([Ca2+]i) using ionomycin in isosmotic medium.
Main Results:
- N1E-115 cells showed reversible volume changes in response to osmotic challenges, behaving near-osmometrically.
- Increased intracellular calcium ([Ca2+]i) induced cell shrinkage at a rate of -1.2%/min.
- The novel technique accurately measured cell water volume changes (<5%) in single cells.
Conclusions:
- N1E-115 cells possess Ca2+-sensitive mechanisms for volume control.
- These mechanisms can induce cell shrinkage under isosmotic conditions.
- The described microspectrofluorimetric technique is a sensitive and reliable method for studying cell water volume dynamics.