Reassessment of the proteomic composition and function of extracellular vesicles in the seminal plasma

Seminal plasma contains a high concentration of extracellular vesicles (EVs). The heterogeneity of small EVs or the presence of non-vesicular extracellular matter (NV) pose major obstacles in understanding the composition and function of seminal EVs. In this study, we employed high-resolution density gradient fractionation to accurately characterize the composition and function of seminal EVs and NV. We found that the seminal EVs could be divided into three different subtypes, namely high-density EV (EV-H), medium-density EV (EV-M), and low-density EV (EV-L) after purification using iodixanol,while NV was successfully isolated. EVs and NV display different features in size, shape and expression of some classic exosome markers. Both EV-H and NV could markedly promote sperm motility and capacitation compared with EV-M and EV-L, whereas only the NV fraction induced sperm acrosome reaction. Proteomic analysis results showed that EV-H, EV-M, EV-L, and NV had different protein components and were involved in different physiological functions. Further study showed that EV-M might reduce the production of sperm intrinsic reactive oxygen species (ROS) through Glutathione S-transferase Mu 2 (GSTM2).This study provides novel insights into important aspects of seminal EVs constituents and sounder footing to explore their functional properties in male fertility.

Histopathology of Fusarium wilt of staghorn sumac (Rhus typhina) caused by Fusarium oxysporum f. sp. callistephi race 3. I. Modes of tissue colonization and pathogen peculiarities

Light and transmission electron microscope studies of naturally infected or inoculated staghorn sumac plants by Fusarium oxysporum f. sp. callistephi race 3 are reported. Diverse extrinsic material (including latex in some instances) or elements occurred in vessel lumina. Some of this material labelled for pectin, often in association with tyloses, as did other opaque matter in paratracheal cells, related to alterations of their protective layer. Pronounced alterations of pit membranes of bordered pits occurred, with their outer portions disrupted into bodies of opaque matter, strongly labelled for cellulose, and their middle portions as unlabelled shreds. Similarly labelled opaque bodies occasionally occurred on vessel walls and lumina. Direct penetration of host cell secondary walls by the pathogen occurred, but these were degraded to any extent only following intramural invasion. Vessel walls, at all stages of infection, were lined with variously structured matter: in their thinnest forms, by single or paired, equidistant or widely spaced opaque bands, and in their thickest forms as alternating opaque and less opaque layers. Other thin elements, often enclosing opaque material, vesicular structures, or occasionally particles of ribosomal appearance were also delineated by similar but frequently infolded bands. These elements were sometimes observed to be confluent with fungal cells and to label for chitin. Many fungal elements were bound by only a thin or defective lucent wall layer, practically unlabelled for chitin, or by a locally thickened, labelled one; labelling for this substrate was also frequently associated with the fungal cell outer opaque wall layer or with some outer extracellular matter. Fine filamentous structures, connected to fungal cells, to the vessel lining matter, and to these other elements, extended into host walls. The lining itself generally did not label for cellulose or chitin. These observations are discussed in comparison with similar observations made regarding other wilt diseases that we have studied.

Cytochemical Labeling for Fungal and Host Components in Plant Tissues Inoculated with Fungal Wilt Pathogens

Antibodies to detect pectin in present investigations attached to distinct fibrils in vessel lumina. In carnation infected with an isolate ofFusarium oxysporumf.sp., labeling of pathogen cells also occurred; in a resistant cultivar (cv.), it was coincident with proximate pectin fibrils and linked to altered fungal walls, which was the opposite in the susceptible cv., indicating that hindrance of pathogen ability to degrade pectin may be related to resistance. Labeling of the fungus in culture was nil, except in media containing pectin, showing that pectin is not native to the pathogen. Labeling of fungal walls for cellulose in elm (inoculated withOphiostoma novo-ulmi) and carnation also occurred, linked to adsorbed host wall components. The chitin probe often attached to dispersed matter, in vessel lumina, traceable to irregularly labeled fungal cells and host wall degradation products. With an anti-horseradish peroxidase probe, host and fungal walls were equally labeled, and with a glucosidase, differences of labeling between these walls were observed, depending on pH of the test solution. Fungal extracellular matter and filamentous structures, present in fungal walls, predominantly in another elm isolate (Phaeotheca dimorphospora), did not label with any of the probes used. However, in cultures of this fungus, extracellular material labeled, even at a distance from the colony margin, with an anti-fimbriae probe.