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. 2020 Jul 6;30(13):R755-R756.
doi: 10.1016/j.cub.2020.05.079.

Release and targeting of polycystin-2-carrying ciliary extracellular vesicles

Juan Wang  1 , Inna A Nikonorova  1 , Amanda Gu  1 , Paul W Sternberg  2 , Maureen M Barr  3
Affiliations

Release and targeting of polycystin-2-carrying ciliary extracellular vesicles

Juan Wang et al. Curr Biol. .
No abstract available

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Conflict of interest statement

DECLARATION OF INTERESTS: The authors declare no conflicts of interests.

Figures

Figure 1.
Figure 1.. Polycystin-2 extracellular vesicles (EVs) are released upon mechanical stimulation of ciliary tip of male-specific sensory neurons and are directionally transferred to hermaphrodite’s vulva during C. elegans mating.
(A) Experimental design for tracking the PKD-2::GFP EVs and MitoTracker labeled spermatozoa during mating. Unlabeled hermaphrodites were mated with labeled males and examined under the microscope post-copulation to score for male-derived PKD-2::GFP-labeled EVs and MitoTracker labeled sperm. Insert diagram shows essential anatomical structures of hermaphrodite to aid in interpretation of images on panel B. (B) Projected Z-stack of confocal sections through the inseminated hermaphrodite. The vulva is outlined with solid white; PKD-2 carrying EVs, sperm, and intestine are outlined with green, magenta, and yellow dashed lines, respectively. Autofluorescence from intestinal granules is marked with asterisks; autofluorescence is visible through both channels. (C) Surface rendering of the confocal optical sections from (B) performed by Imaris software, showing that EVs are located on the cuticle around vulva. No PKD-2::GFP EVs were found in the uterus. (D) Diagram of male tail shows identities of sensory ray cilia protruding into environment: 1/5/7/9 protrude from a dorsal side, 2/4/8 protrude from a ventral side of the male tail. Ray 3 is lateral; its cilium protrudes from the edge of the male tail. Ray 6 is a closed ray; its neurons do not express the pkd-2 gene. Confocal optical sections show EV release (indicated with arrows) upon contact of either dorsal or ventral side of a tail with a bare coverslip. Contact with a padded coverslip rarely triggers EV release. The posterior-most edge of the male fan is autofluorescent (yellow asterisks). (E) Quantitative analysis of ciliary EV release upon contact with either padded or bare coverslip. Each male was first imaged being covered with an agarose-padded coverslip, then the padded coverslip was replaced with a bare coverslip and imaging was repeated for that male. The box plots show median values, top and bottom hinges correspond to the first and third quartiles (the 25th and 75th percentiles), and the whiskers extend to the smallest and the largest value within 1.5 distance of the interquartile range. (F) Quantitative analysis of the propensity of dorsal and ventral cilia to release the PKD-2 EVs when positioned on a side that is either adjacent or opposite to the coverslip. * p-value <0.01, n=39 in the Kruskal-Wallis test for (E); n=56 for dorsal rays, n=42 for ventral rays in the two proportion Z-test for (F).
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