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Identification of multiple microtubule initiating sites in mouse neuroblastoma cells
Abstract:
Mouse neuroblastoma N-18 cells can be induced by serum deprivation to sprout multiple neurite-like processes which contain many microtubules. Mitotic drugs such as colcemid and colchicine depolymerize these microtubules and the cells lose their processes. Reappearance of microtubules after removal of the drugs was followed by immunofluorescence microscopy using tubulin specific antibodies. At early recovery times multiple star-like structures which contained tubulin were detected in the perinuclear are and in the cytoplasm of individual cells. The mean number seen per cell as approximately 5. Their formation preceeded the organization of the complex microtubular networks typical of N-18 cells. The probable action of these structures as microtubular organization centers (MTOCs) is discussed. Multiple structures were detected during recovery from the influence of mitotic drugs both in previously induced and non-induced N-18 cells, suggesting that N-18 cells harbour the potential of formation of multiple organization centers even without previous induction. We discuss the possibility that differentiation of neuroblastoma N-18 cells may require microtubular organization centers.
Insights
Mouse neuroblastoma cells form multiple microtubule organizing centers (MTOCs) during recovery from mitotic drugs. These structures precede complex microtubule networks and may be crucial for cell differentiation.
Area of Science:
- Cell Biology
- Neuroscience
- Biochemistry
Background:
- Mouse neuroblastoma N-18 cells extend neurite-like processes rich in microtubules.
- Mitotic drugs like colcemid and colchicine depolymerize microtubules, causing process retraction.
Purpose of the Study:
- To investigate the formation and potential role of microtubule organizing centers (MTOCs) during microtubule regrowth in N-18 cells.
- To explore whether N-18 cells possess the inherent capacity to form multiple MTOCs.
Main Methods:
- Serum deprivation to induce neurite-like processes in N-18 cells.
- Treatment with colcemid or colchicine to depolymerize microtubules.
- Immunofluorescence microscopy with tubulin-specific antibodies to visualize microtubule regrowth.
- Quantitative analysis of star-like structures (potential MTOCs).
Main Results:
- Following drug removal, multiple (approx. 5 per cell) star-like tubulin-containing structures appeared in the perinuclear and cytoplasmic regions.
- These structures formed before the organization of complex microtubule networks.
- Multiple MTOCs formed even in non-induced cells, indicating an intrinsic potential.
Conclusions:
- The observed star-like structures likely function as microtubule organizing centers (MTOCs).
- N-18 cells can form multiple MTOCs, suggesting this is a general capability, not solely dependent on prior induction.
- The formation of MTOCs may be a prerequisite for the differentiation of neuroblastoma N-18 cells.