Detection of extracellular vesicles in the mouse vaginal fluid: Their delivery of sperm proteins that stimulate capacitation and modulate fertility
| Autor: | Patricia A. Martin-DeLeon, Pradeepthi Bathala, Zeinab Fereshteh, Deni S. Galileo |
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| Rok vydání: | 2018 |
| Předmět: |
Male
0301 basic medicine Physiology ATPase Clinical Biochemistry Acrosome reaction Exosomes Exosome Article Flow cytometry Andrology Extracellular Vesicles Mice 03 medical and health sciences 0302 clinical medicine Capacitation In vivo medicine Animals Progesterone medicine.diagnostic_test biology Chemistry Cell Membrane Cell Biology Spermatozoa Sperm In vitro Mice Inbred C57BL Fertility 030104 developmental biology 030220 oncology & carcinogenesis Vagina biology.protein Female Acrosome Sperm Capacitation |
| Zdroj: | J Cell Physiol |
| ISSN: | 1097-4652 0021-9541 |
| Popis: | Extracellular vesicles (EVs) were isolated by ultracentrifugation of vaginal luminal fluid (VLF) from superovulated mice and identified for the first time using transmission electron microscopy. Characterized by size and biochemical markers (CD9 and HSC70), EVs were shown to be both microvesicular and exosomal and were dubbed as "Vaginosomes" (VGS). Vaginal cross-sections were analyzed to visualize EVs in situ: EVs were present in the lumen and also embedded between squamous epithelial and keratinized cells, consistent with their endogenous origin. Western blots detected Plasma membrane Ca2+ -ATPase 1 (PMCA1) and tyrosine-phosphorylated proteins in the VGS cargo and also in uterosomes. Flow cytometry revealed that following coincubation of caudal sperm and VLF for 30 min, the frequencies of cells with the highest Sperm adhesion molecule 1 (SPAM1), PMCA1/4, and PMCA1 levels increased 16.4-, 8.2-, and 27-fold, respectively; compared with control coincubated in phosphate buffered saline (PBS). Under identical conditions, sperm tyrosine-phosphorylated proteins were elevated ~3.3-fold, after VLF coincubation. Progesterone-induced acrosome reaction (AR) rates were significantly (p < 0.001) elevated in sperm coincubated with VGS for 10-30 min, compared with PBS. Sperm artificially deposited in the vaginas of superovulated females for these periods also showed significant (p < 0.01) increases in AR rates, compared with PBS. Thus in vitro and in vivo, sperm acquire from the vaginal environment factors that induce capacitation, explaining recent findings for their acrosomal status in the isthmus. Overall, VGS appear to deliver higher levels of proteins involved in preventing premature capacitation and AR than those promoting them. Our findings which have implications for humans open the possibility of new approaches to infertility treatment with exosome therapeutics. |
| Databáze: | OpenAIRE |
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