Extracellular vesicles (EVs) are membranous compartments produced by yeast and mycelial forms of several fungal species. One of the difficulties in perceiving the role of EVs during the fungal life, and particularly in cell wall biogenesis, is caused by the presence of a thick cell wall. One alternative to have better access to these vesicles is to use protoplasts. This approach has been investigated here with Aspergillus fumigatus, one of the most common opportunistic fungal pathogens worldwide. Analysis of regenerating protoplasts by scanning electron microscopy and fluorescence microscopy indicated the occurrence of outer membrane projections in association with surface components and the release of particles with properties resembling those of fungal EVs. EVs in culture supernatants were characterized by transmission electron microscopy and nanoparticle tracking analysis. Proteomic and glycome analysis of EVs revealed the presence of a complex array of enzymes related to lipid/sugar metabolism, pathogenic processes, and cell wall biosynthesis. Our data indicate that (i) EV production is a common feature of different morphological stages of this major fungal pathogen and (ii) protoplastic EVs are promising tools for undertaking studies of vesicle functions in fungal cells.

中文翻译：

---

烟曲霉原生质体产生的细胞外囊泡的表征。

细胞外囊泡 (EV) 是由酵母和几种真菌的菌丝体形式产生的膜状隔室。在真菌生命过程中，尤其是在细胞壁生物发生中，感知 EV 的作用的困难之一是由于厚细胞壁的存在。更好地接触这些囊泡的一种替代方法是使用原生质体。这种方法已在此处用烟曲霉进行了研究，世界上最常见的机会性真菌病原体之一。通过扫描电子显微镜和荧光显微镜对再生原生质体的分析表明，外膜突起的发生与表面成分相关，以及具有类似于真菌 EV 特性的颗粒的释放。培养上清液中的 EV 通过透射电子显微镜和纳米粒子跟踪分析进行表征。EV 的蛋白质组学和糖组分析揭示了与脂质/糖代谢、致病过程和细胞壁生物合成相关的一系列复杂酶的存在。