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Related Experiment Videos

Microtubules and the endoplasmic reticulum are highly interdependent structures.

M Terasaki, L B Chen, K Fujiwara

    The Journal of Cell Biology
    |October 1, 1986
    PubMed
    Summary

    This study explores how microtubules and the endoplasmic reticulum (ER) interact in the outer regions of cultured cells. Using fluorescent and immunofluorescent staining, the researchers observed that both structures extend into lamellipodia before intermediate filaments are present. They found that ER and microtubules polymerize together in these regions. When microtubules were disrupted for short periods, the ER network remained stable. However, prolonged microtubule absence caused the ER network to retract toward the cell center. The findings suggest that microtubules and ER tubules are highly interdependent in both their extension and long-term maintenance.

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    Area of Science:

    • Cell biology
    • Membrane and cytoskeleton dynamics
    • Microtubule and ER interactions

    Background:

    The endoplasmic reticulum (ER) and microtubules are essential components of the cell's internal architecture. While their roles in transport and signaling are well known, their spatial and functional interdependence remains less clear. Previous studies have shown that microtubules guide organelle positioning, but the ER's dependency on microtubules is less understood. Research has established that the ER extends into lamellipodia, but the timing of this extension relative to microtubules is unclear. No prior work has directly compared ER and microtubule dynamics in the same cell using dual fluorescent techniques. This gap motivated the use of combined staining methods to study ER and microtubule interactions in cultured cells. The uncertainty around whether ER extension requires microtubules led to experiments with microtubule disruption. The question of whether ER retraction occurs after microtubule loss remained unresolved. This study aimed to clarify the interdependence of ER and microtubules in cell periphery.

    Purpose Of The Study:

    Keywords:
    Endoplasmic reticulumMicrotubule dynamicsER network retractionCell motility

    Frequently Asked Questions

    The study suggests that microtubules and ER tubules polymerize together in lamellipodia regions of cultured cells.

    They used fluorescent dyes to label the ER and immunofluorescence to label microtubules in the same cell.

    The ER network remained stable for up to 15 minutes after microtubule depolymerization.

    Prolonged microtubule absence leads to ER network retraction toward the cell center over time.

    Related Experiment Videos

    This study aimed to investigate the spatial and functional relationship between microtubules and the endoplasmic reticulum in the peripheral regions of cultured cells. The researchers sought to determine whether ER and microtubule extension occur simultaneously or sequentially in lamellipodia. They also wanted to test if microtubules are necessary for maintaining the ER network's extended state. The study focused on fibroblasts, epithelial cells, and vascular endothelial cells, which are known for dynamic cytoskeletal changes. The team used fluorescent and immunofluorescent staining to visualize both structures in the same cell. They aimed to observe how ER distribution changes when microtubules are disrupted. The goal was to clarify whether ER and microtubules polymerize together or if ER extension depends on microtubule stability. The study aimed to address unresolved questions about ER and microtubule interdependence in cell motility.

    Main Methods:

    The researchers used cultured fibroblasts, epithelial cells, and vascular endothelial cells as model systems. They applied fluorescent dyes to visualize the endoplasmic reticulum in live cells. Immunofluorescence techniques were used to label microtubules and intermediate filaments in the same cells. The cells were observed in peripheral regions, particularly lamellipodia, where cytoskeletal changes are prominent. Microtubule disruption was induced using nocodazole, cold temperature, or hypotonic shock. ER and microtubule distributions were compared in both untreated and treated cells. The team analyzed the spatial overlap and temporal sequence of ER and microtubule extension in lamellipodia. They measured the effects of microtubule depolymerization on ER network integrity over time.

    Main Results:

    Microtubules and ER tubules showed nearly identical distributions in lamellipodia regions of cultured cells. Both structures extended into lamellipodia before intermediate filaments were present. ER and microtubules polymerized together at the resolution of fluorescence microscopy. Short-term microtubule disruption (15 minutes) did not cause ER retraction. Prolonged microtubule absence (2 hours) led to ER network retraction toward the cell center. ER extension into lamellipodia occurred independently of intermediate filaments. ER and microtubules were sparse in lamellipodia, but ER tubules were more prevalent than microtubules. The ER network remained stable for up to 15 minutes after microtubule depolymerization.

    Conclusions:

    The study suggests that microtubules and ER tubules polymerize together in lamellipodia regions of cultured cells. ER extension into lamellipodia occurs independently of intermediate filaments. Microtubules are not immediately required for ER network stability. Prolonged microtubule absence leads to ER network retraction over time. ER and microtubules advance into lamellipodia before intermediate filaments. ER and microtubule distributions are nearly identical in lamellipodia. ER extension and microtubule polymerization occur at the same spatial resolution. The ER network requires microtubules for long-term maintenance.

    Yes, ER extension into lamellipodia occurred independently of intermediate filaments.

    The study suggests that microtubules and ER tubules have nearly identical distributions in lamellipodia.