Cytoskeletal aspects of nuclear migration during tip-growth in the fernAdiantum protonemal cell

Summary In the tip-growing protonemal cell, the nucleus migrates with the tip as it grows, keeping a constant distance between them. Cytoskeletal control of this nuclear migration was analyzed inAdiantum capillus-veneris. Using rhodamine-phalloidin (Rh-Phal), tubulin antibodies and confocal laser scanning microscopy, we found the presence of microtubule (MT) and microfilament (MF) strands connecting the cell nucleus to the cortex of the growing apex. The strands come from the apical end of the spindle-shaped nucleus and run through the endoplasm, arriving at the apical cortex, where a circular arrangement of MTs and MFs is present. Strands of MFs and MTs were also found to emanate from the proximal end of the nucleus and extend towards the cortex of the basal part of the cell. Double staining of MTs and MFs revealed a co-localization of these cytoskeletal elements. When MF strands were disrupted by cytochalasin B (CB), tip-growth ceased and nuclear movement stopped. After the application of colchicine, MT structures disappeared, tip-growth was largely inhibited, and the nucleus moved towards the basal part of the cell. When both CB and colchicine were applied to the cell, no basipetal migration of cell nucleus was observed. These results suggest that the MT strands between the apex and the nucleus may have a role in the anchorage of the cell nucleus to the tip during tip-growth, and that the MF strands may be important for basipetal movement of the nucleus. When the nucleus was dislocated basipetally by centrifugation, cytoskeletal strands between the cell apex and the nucleus were still observed, and by acropetal movement the nucleus resumed its previous position. The acropetal movement of the nucleus was inhibited by the application of both CB and colchicine but not by CB alone nor by colchicine alone, indicating that both cytoskeletal elements are involved in the forward movement of cell nucleus.Abbreviations CB cytochalasin B - DAPI4 6-diamino-2-phenylin-dole - DMSO dimethylsulfoxide - PIPES piperazine-N,N-bis(2-ethane-sulfonic acid) - EGTA ethyleneglycol-bis-(-aminoethyl-ether)-N,N,N,N-tetraacetic acid - MBS m-maleimidobenzoic acid N-hydroxysuccinimide ester - MF microfilament - MT microtubule - PMSF phenylmethylsulfonyl fluoride - PSM polyoxyethylene sorbitan monolaurate - Rh-Phal rhodamine-labeled phalloidin