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First published online 16 October 2007
doi: 10.1242/jcs.012138

Journal of Cell Science 120, 3804-3819 (2007)
Published by The Company of Biologists 2007
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Research Article

Distinct endocytic pathways identified in tobacco pollen tubes using charged nanogold

Alessandra Moscatelli1,*, Fabrizio Ciampolini2, Simona Rodighiero3, Elisabetta Onelli1, Mauro Cresti2, Nadia Santo4 and Aurora Idilli1

1 Dipartimento di Biologia L. Gorini, Università degli Studi di Milano, Via Celoria 26, 20133 Milan, Italy
2 Dipartimento di Scienze Ambientali G. Sarfatti, Università degli Studi di Siena, Via P. A. Mattioli 4, 53100 Siena, Italy
3 CIMAINA, Università degli Studi di Milano, Via Celoria 16, 20133 Milan, Italy
4 CIMA, Università degli Studi di Milano, Via Celoria 16, 20133 Milan, Italy

* Author for correspondence (e-mail: alessandra.moscatelli{at}unimi.it)

Accepted 9 August 2007

In an attempt to dissect endocytosis in Nicotiana tabacum L. pollen tubes, two different probes – positively or negatively charged nanogold – were employed. The destiny of internalized plasma membrane domains, carrying negatively or positively charged residues, was followed at the ultrastructural level and revealed distinct endocytic pathways. Time-course experiments and electron microscopy showed internalization of subapical plasma-membrane domains that were mainly recycled to the secretory pathway through the Golgi apparatus and a second mainly degradative pathway involving plasma membrane retrieval at the tip. In vivo time-lapse experiments using FM4-64 combined with quantitative analysis confirmed the existence of distinct internalization regions. Ikarugamycin, an inhibitor of clathrin-dependent endocytosis, allowed us to further dissect the endocytic process: electron microscopy and time-lapse studies suggested that clathrin-dependent endocytosis occurs in the tip and subapical regions, because recycling of positively charged nanogold to the Golgi bodies and the consignment of negatively charged nanogold to vacuoles were affected. However, intact positively charged-nanogold transport to vacuoles supports the idea that an endocytic pathway that does not require clathrin is also present in pollen tubes.

Key words: Nicotiana tabacum (L.), Pollen tube, Endocytosis


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JCS 2007 120: 2104. [Full Text]  



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L. Zonia and T. Munnik
Vesicle trafficking dynamics and visualization of zones of exocytosis and endocytosis in tobacco pollen tubes
J. Exp. Bot., March 1, 2008; 59(4): 861 - 873.
[Abstract] [Full Text] [PDF]


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