In the rodent brain neural progenitors cells are born in the subventricular zone (SVZ) migrate along the rostral migratory stream (RMS) t the olfactory bulb, and differentiate into several classes of interneurons. Tangential migration in the RMS takes place in ‘chains’ of cells as compared to individual cells in cortical radial migration. To examine the biophysics of migration in this pathway we labeled SVZ progenitors with Cell Tracker Green (CTG) in P2 and P17 mice. At 3 days post injection acute saggital slices were time-lapse imaged on a confocal microscope. The centroid of the cell soma was tracked for at least 60min. CTG is only retained by cells with intact cell membranes ensuring that imaged cells were viable. Individual cells in the RMS migrate in a salutatory manner with bursts of high speed followed by periods of slower speed (mean 26-31µm/hr). The frequency of these ‘bursts’ were similar within a cell, but varied substantially between different cells. Neurotransmitters, particularly GABA, have been implicated as modulators of neuroblasts migration in the RMS. To test the role of GABA and glutamate in this model slices were incubated with specific agonists/antagonists. Incubation of the slices with the GABAA receptor antagonist gabazine increases the migratory speed by 45% while the agonist muscimol decreases speed of 34%. NMDA/AMPA and mGluR group I receptors are also expressed along the RMS; however, inhibition of these receptors does not significantly affect migration. Migratory cells can interact and modify the extracellular matrix through expression of a family of proteins, the matrix metalloproteinases (MMPs). We identified several gelatinase and membrane type MMPs in the RMS. To probe their function the inhibitors of these MMPs (Dec-RVKR-CMK and GM1489), were used. In the presence of inhibitor neuroblasts migration in the RMS was reduced by ~35%, suggesting a role for these proteases in CNS neuroblasts migration.
Time lapse confocal microscopy on migrating neuroblasts in the mouse rostral migratory stream.
BOVETTI, Serena;BOVOLIN, Patrizia;PERROTEAU, Isabelle;
2005-01-01
Abstract
In the rodent brain neural progenitors cells are born in the subventricular zone (SVZ) migrate along the rostral migratory stream (RMS) t the olfactory bulb, and differentiate into several classes of interneurons. Tangential migration in the RMS takes place in ‘chains’ of cells as compared to individual cells in cortical radial migration. To examine the biophysics of migration in this pathway we labeled SVZ progenitors with Cell Tracker Green (CTG) in P2 and P17 mice. At 3 days post injection acute saggital slices were time-lapse imaged on a confocal microscope. The centroid of the cell soma was tracked for at least 60min. CTG is only retained by cells with intact cell membranes ensuring that imaged cells were viable. Individual cells in the RMS migrate in a salutatory manner with bursts of high speed followed by periods of slower speed (mean 26-31µm/hr). The frequency of these ‘bursts’ were similar within a cell, but varied substantially between different cells. Neurotransmitters, particularly GABA, have been implicated as modulators of neuroblasts migration in the RMS. To test the role of GABA and glutamate in this model slices were incubated with specific agonists/antagonists. Incubation of the slices with the GABAA receptor antagonist gabazine increases the migratory speed by 45% while the agonist muscimol decreases speed of 34%. NMDA/AMPA and mGluR group I receptors are also expressed along the RMS; however, inhibition of these receptors does not significantly affect migration. Migratory cells can interact and modify the extracellular matrix through expression of a family of proteins, the matrix metalloproteinases (MMPs). We identified several gelatinase and membrane type MMPs in the RMS. To probe their function the inhibitors of these MMPs (Dec-RVKR-CMK and GM1489), were used. In the presence of inhibitor neuroblasts migration in the RMS was reduced by ~35%, suggesting a role for these proteases in CNS neuroblasts migration.
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