Experimental measurement of endocytosis in fungal hyphae

Fungal Genetics and Biology : FG & B
Salomón Bartnicki-GarcíaRosa R Mouriño-Pérez

Abstract

The present study examines the notion that polarized exocytosis in the tips of growing hyphae creates an excess of plasma membrane and thus the need for its removal by endocytosis. To measure endocytosis experimentally, we developed a photobleaching (FRAP) procedure to count endocytic events in hyphae of Neurospora crassa carrying a fluorescent tag on the actin-binding protein fimbrin (FIM-1-GFP). Given 40 nm as the average diameter of endocytic vesicles, we calculated that about 12.5% of the plasma membrane discharged in the apex becomes endocytosed in the subapex. According to our calculations, the GFP-tagged hyphae of N. crassa, measured under the constrained conditions of confocal microscopic examination, needed about 8800 vesicles/min to extend their plasma membrane or about 9800/min, if we include predicted demands for cell wall growth and extracellular secretion. Our findings support the notion that exocytosis and endocytosis operate in tandem with the latter serving as a compensatory process to remove any excess of plasma membrane generated by the intense exocytosis in the hyphal tips. Presumably, this tandem arrangement evolved to support the hallmark features of fungi namely rapid cell extension and abundant secretion...Continue Reading

Citations

May 28, 2019·F1000Research·Robert A Arkowitz, Martine Bassilana
May 24, 2020·Fungal Genetics and Biology : FG & B·Blake CommerBrian D Shaw
Feb 26, 2021·Mycology·Blake Commer, Brian D Shaw
Nov 20, 2021·The FEBS Journal·Veneta Gerganova, Sophie G Martin

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