Polarization of the Golgi apparatus and the microtubule-organizing center in cultured fibroblasts at the edge of an experimental wound.

TLDR

Cell motility mechanisms are examined in the context of prior hypotheses and experimental evidence. Experimental wounding of confluent fibroblast monolayers induces synchronous, unidirectional migration, and the positions of the Golgi apparatus and microtubule‑organizing center were mapped by double indirect immunofluorescence microscopy with organelle‑specific antibodies. At the wound edge, the Golgi apparatus and microtubule‑organizing center align closely ahead of the nucleus, reorienting within minutes of wounding and persisting during migration, indicating they may help direct cell movement.

Abstract

We have used the technique of experimental wounding of confluent monolayers of normal fibroblasts to induce essentially unidirectional and synchronous cell movement at the edge of the wound. The intracellular location of the Golgi apparatus and the microtubule-organizing center was determined by double indirect immunofluorescence microscopy, using antibodies specific for the membranes of the Golgi apparatus and antibodies specific for tubulin, respectively. In cells at the wound edge, the immunolabeled Golgi apparatus and microtubule-organizing center were in close proximity to one another and located predominantly forward of the cell nucleus facing the wound. In the same cultures in cells removed from the wound, the two organelles were also coordinately located; however, they were randomly oriented with respect to the wound edge. This reorientation of the two organelles in cells at the wound edge was evident within minutes after wounding and persisted as cell extension subsequently occurred into the wound. These results suggest that both the Golgi apparatus and the microtubule-organizing center may participate in directing cell movement. The possible mechanisms involved are discussed in the light of previous hypotheses and experimental evidence concerning cell motility.