Detection of extracellular vesicles in the mouse vaginal fluid: Their delivery of sperm proteins that stimulate capacitation and modulate fertility

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 a...

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Bibliographic Details
Published in:Journal of cellular physiology 2019-08, Vol.234 (8), p.12745-12756
Main Authors: Fereshteh, Zeinab, Bathala, Pradeepthi, Galileo, Deni S., Martin‐DeLeon, Patricia A.
Format: Article
Language:English
Subjects:
Acrosome - metabolism
Acrosome - physiology
Acrosome reaction
acrosome reaction (AR) uterosomes (UTS)
Adenosine triphosphatase
Animals
Biochemical markers
Ca2+ efflux pumps
Calcium
Calcium ions
Capacitation
CD9 antigen
Cell Membrane - metabolism
Cell Membrane - physiology
Exosomes - metabolism
Exosomes - physiology
Extracellular vesicles
Extracellular Vesicles - metabolism
Extracellular Vesicles - physiology
Female
Fertility
Fertility - physiology
Flow cytometry
Hsc70 protein
Infertility
Male
Medical treatment
Mice
Mice, Inbred C57BL
Progesterone
Progesterone - metabolism
Proteins
Sperm
sperm capacitation
Sperm Capacitation - physiology
Spermatozoa - metabolism
Spermatozoa - physiology
Transmission electron microscopy
Tyrosine
tyrosine‐phosphorylated proteins
Ultracentrifugation
Vagina
Vagina - metabolism
Vagina - physiology
vaginal exosomes
Very Low Frequencies
Vesicles
Western blotting
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Summary: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 
ISSN:0021-9541
1097-4652
DOI:10.1002/jcp.27894