Correlative Analysis of [Ca](C) and Apical Secretion during Pollen Tube Growth and Reorientation

The maintenance of a cytosolic free calcium gradient (Ca(2+)](c)) and vesicle secretion in the apex of pollen tubes is essential for growth. It has been postulated that high [Ca(2+)](c) levels promote and confine vesicle fusion with the apical plasma membrane and in this study we performed a correlative analysis of both events using specific fluorescent dyes and confocal scanning microscopy. [Ca(2+)](c) was imaged with Calcium Green-1 10 kDa dextran (CG-1) while secretory events were followed with FM1-43 or FM4-64 in pollen tubes undergoing normal growth and reorientation events.During straight growth (no modification in direction), we found that changes in apical [Ca(2+)](c) accompany changes in apical FM fluorescence indicating a tight coupling between [Ca(2+)](c) and apical secretion. This coupling seems however to be perturbed during periods of reorientation of the pollen tube growth axis. Analysis of apical and sub-apical fluorescent signals during the reorientation events and subsequent re-entry in straight growth indicate that the increase in secretory events (higher fusion rate) precede the increases in [Ca(2+)](c) that should be required for the transduction of the signal.Based on these findings, we discuss a model for membrane secretion and recycling which considers the apical and sub-apical region as a functional area containing all the elements required to promote and sustain growth.