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Expr14579
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We observed Pswm-1::mCherry::H2B reporter expression in six to eight of the 12 vas deferens cuboidal cells. We also observed Pswm-1::mCherry::H2B in posterior body wall muscle cells and in the male-specific diagonal muscle cells, both of which overlie the seminal vesicle and vas deferens region. Expression of the Pswm-1 reporter in both gonadal and muscle cells began at late larval stages and persisted in adult animals until at least 48 h post L4. In addition, we observed reporter expression at low levels in two cells in the head, which were likely neurons. These data indicate that swm-1 is not expressed in sperm. Instead, it is produced in somatic tissues: the vas deferens, which is involved in the production, release, and transfer of seminal fluid, and muscle cells, outside the gonad. |
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Expr14580
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In adult males, the SWM-1::mCherry protein was present, as expected, in cells in which we observed the transcriptional reporter [Expr14579]: vas deferens cuboidal cells, diagonal muscles and body wall muscles. Within cuboidal cells, SWM-1::mCherry was contained within large vesicles that were closely associated with the apical membrane, which are not well-characterized but have been shown to contain seminal fluid (Smith and Stanfield, 2011; Lints and Hall, 2009; Kimble and Hirsh, 1979). In muscle cells, we observed SWM-1::mCherry at low levels that appeared to be concentrated near the cell cortex. Interestingly, in jn62[swm-1::mCherry], SWM-1::mCherry was also visible in anterior body wall muscles, rather than being restricted to the posterior region, corresponding to slightly higher levels of rescue and protein expression as compared with the Pswm-1 MosSCI reporter strain. |
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Expr9863
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In worms carrying the Ptry-5::TRY-5::GFP reporter, the TRY-5::GFP fusion protein exhibited a localization pattern consistent with secretion from the vas deferens. Within the valve and cuboidal cells, TRY-5::GFP was localized to globular foci. In L4 larvae, most globules aligned with the apical domain that lines the developing sperm channel. In mature adults, very large globules were present that tended to cluster apically, and additional small globules were present throughout the cytoplasm. We sometimes observed TRY-5::GFP within the lumen of the seminal vesicle, likely as a result of release from the adjacent, highly-expressing valve cells. |
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Expr15983
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We also observed that ENPP-1::mScarlet localized to the secretory cuboidal cells of the vas deferens, where it accumulated at the membranes of giant vesicles filled with seminal fluid. |
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Expr14583
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We observed that SWM-1::mCherry and TRY-5::GFP were both present in the cuboidal cells' vesicles. |
