Display Settings:

Items per page
We are sorry, but NCBI web applications do not support your browser and may not function properly. More information

Results: 7

1.
FIGURE 5:

FIGURE 5:. From: The nucleoporin Nup205/NPP-3 is lost near centrosomes at mitotic onset and can modulate the timing of this process in Caenorhabditis elegans embryos.

NPP-3 localization at mitotic onset. (A–D) Wild-type, one-cell-stage embryos in prophase (A and B), prometaphase (C), or metaphase (D) stained for NPP-3 (shown alone in the insets on the right and in red in the merged images), α-tubulin (green), and DNA (blue). (E–G) Wild-type, one-cell-stage embryos stained for NPP-3 (shown alone in the insets in the middle and in red in the merged images), lamin, NPP-13, or NPP-5, as indicated (shown alone in the insets on the right and in green in the merged images). DNA is shown in blue.

Virginie Hachet, et al. Mol Biol Cell. 2012 August 15;23(16):3111-3121.
2.
FIGURE 7:

FIGURE 7:. From: The nucleoporin Nup205/NPP-3 is lost near centrosomes at mitotic onset and can modulate the timing of this process in Caenorhabditis elegans embryos.

Model for the role of centrosomes and AIR-1 in promoting NPP-3 local loss and timely mitotic entry in C. elegans embryos. Green circle: AIR-1 at centrosomes. Note that cytoplasmic AIR-1 is not depicted in this figure but likely contributes to overall AIR-1 activity. Red: NPP-3; light blue: lamina; gray lines: microtubules; filled line: intact NPP-3 signal; dashed line: disrupted NPP-3 signal. Centrosomes and AIR-1 promote NPP-3 local loss from the NE, potentially facilitating NEBD and timely mitotic entry.

Virginie Hachet, et al. Mol Biol Cell. 2012 August 15;23(16):3111-3121.
3.
FIGURE 3:

FIGURE 3:. From: The nucleoporin Nup205/NPP-3 is lost near centrosomes at mitotic onset and can modulate the timing of this process in Caenorhabditis elegans embryos.

Defective nuclear permeability does not drastically alter timing of mitotic entry. (A–C) Nuclear exclusion of fluorescently labeled dextrans of 70 kDa (green) and 155 kDa (red). Confocal images of wild-type (A), npp-3(RNAi) (B), and npp-4(RNAi) (C) live embryos are shown. Vertical arrowheads indicate nuclear exclusion; horizontal arrowheads indicate lack of exclusion. (D) Nuclear-to-cytoplasmic (N/C) ratio of fluorescently labeled dextrans (70 kDa, green; 155 kDa, red) plotted against the average time difference between NEBD in the male and female pronuclei (Δt (NEBD)). See also Figure S2.

Virginie Hachet, et al. Mol Biol Cell. 2012 August 15;23(16):3111-3121.
4.
FIGURE 4:

FIGURE 4:. From: The nucleoporin Nup205/NPP-3 is lost near centrosomes at mitotic onset and can modulate the timing of this process in Caenorhabditis elegans embryos.

Decreased asynchrony correlates with loss of NPP-3 at the NE. (A) Western blot of extracts from wild-type or npp-3(RNAi) embryos probed with NPP-3 antibodies. (B–I) NPP-3 distribution at the NE in embryos of the indicated genotypes stained for NPP-3 (shown alone in the insets on the right and in red in the merged images), α-tubulin (green), and DNA (blue). Note that depletion of some nucleoporins affects nuclear size as previously observed (Galy et al., 2003). (J) NE-to-cytoplasmic (NE/C) ratio of NPP-3 signal plotted against the average time difference between NEBD in the male and female pronuclei in seconds. See also Table S4.

Virginie Hachet, et al. Mol Biol Cell. 2012 August 15;23(16):3111-3121.
5.
FIGURE 6:

FIGURE 6:. From: The nucleoporin Nup205/NPP-3 is lost near centrosomes at mitotic onset and can modulate the timing of this process in Caenorhabditis elegans embryos.

NPP-3 local loss is centrosome- and AIR-1–dependent. (A–J) One-cell-stage embryos of the indicated genotypes stained for NPP-3 (shown alone in the insets on the right and in red in the merged images), α-tubulin (green), and DNA (blue). Note in (E) and (F) that the female pronucleus is much larger and located next to the male pronucleus, as reported previously for tba-2(RNAi) embryos (Sonneville and Gönczy, 2004). The numbers of embryos analyzed, which all exhibited the phenotype illustrated in the panels, except when mentioned otherwise, are as follows: spd-5(RNAi): 17; zyg-12(RNAi): 8; tbg-1(RNAi): 15, 1 of which did not exhibit NPP-3 local loss; tba-2(RNAi): 16; tba-2 (RNAi) zyg-12(ct350): 7; ncc-1(RNAi): 12; air-1(RNAi): 11; GFP-AIR-1R: 9; GFP-AIR-1R T201A: 19.

Virginie Hachet, et al. Mol Biol Cell. 2012 August 15;23(16):3111-3121.
6.
FIGURE 1:

FIGURE 1:. From: The nucleoporin Nup205/NPP-3 is lost near centrosomes at mitotic onset and can modulate the timing of this process in Caenorhabditis elegans embryos.

RNAi-based modifier screen for modulators of mitotic entry. (A and B) Images from time-lapse DIC microscopy of wild-type (A) and zyg-9(b244) (B) one-cell-stage embryos. In all figures, anterior is to the left, posterior to the right; F and M designate the female and male pronuclei, respectively. Arrowheads with thick lines: intact pronucleus; arrowheads with dashed lines: pronucleus undergoing NEBD. Scale bars, 10 μm. The onset of NEBD, as defined by the loss of the smooth line corresponding to the NE is defined as time 0. (C and D) Average time difference between NEBD in the male and female pronuclei in zyg-9(b244) + candidate gene(RNAi) in the initial screen (C; see also Table S2) or selected after the confirmation round using a threshold of ± 35 s as compared with the zyg-9(b244) timing (D). The time corresponding to zyg-9(b244) alone is shown in red. The following candidate genes that were selected in the initial screen did not give a reproducible phenotype in the confirmation round: T01C3.1, C03C10.3, ZC308.1a, ZK1037.5, nsh-1, T2308.9, T20B12.8, F53G12.5, ZK1127.7, and T13F2.7. In addition, glucosamine (F07A11.2) and sqv-4 (F29F11.1) were not retained because RNAi-treated embryos exhibited severe pleiotropic phenotypes, precluding a proper analysis of NEBD. Finally, the following genes could not be analyzed for their impact on asynchronous NEBD as their depletion rescued pronuclear migration to some extent: cyb-1, cyb-3, cye-1, ncc-1, cdc25.1, F09G2.4, gpr-2, rga-3, C03C10.3, F59B2.6, and hmg-3.

Virginie Hachet, et al. Mol Biol Cell. 2012 August 15;23(16):3111-3121.
7.
FIGURE 2:

FIGURE 2:. From: The nucleoporin Nup205/NPP-3 is lost near centrosomes at mitotic onset and can modulate the timing of this process in Caenorhabditis elegans embryos.

NPP-3 depletion abolishes asynchronous NEBD of separated male and female pronuclei. (A and C) Images from time-lapse DIC microscopy of zyg-9(b244) (A) and zyg-9(b244) npp-3(RNAi) (C) one-cell-stage embryos. See also corresponding Movies M1 and M2. (E and G) Time-lapse fluorescence microscopy of zyg-9(RNAi) (E) and zyg-9(RNAi) npp-3(RNAi) (G) one-cell-stage embryos expressing YFP-lamin. See also corresponding Movies M3 and M4. Lamina disassembly is apparent by loss of the smooth appearance characteristic of earlier stages. Note that the 6% neutral density filter was removed in panel G (see also legends of Movies M3 and M4). Similar results were obtained with zyg-9(b244) npp-3(RNAi). (B, D, F, and H) Average time difference between NEBD onset in the male and female pronuclei in seconds ± SEM in zyg-9(b244) (B) or zyg-9(RNAi) (F), as well as zyg-9(b244) npp-3(RNAi) (D) or zyg-9(RNAi) npp-3(RNAi) (H) one-cell-stage embryos, monitored by DIC (B and D) or YFP-lamin (F and H). Numbers of embryos and statistical analysis are given in Table S3 for B and D. (I and J) Monitoring NCC-1 activation in the male and female pronuclei of zyg-9(b244) (I) and zyg-9(b244) npp-3(RNAi) (J) one-cell-stage embryos by staining for Cdk1P-Tyr15 (shown alone on the right and in red in the merged images), α-tubulin (green), and DNA (blue). Histograms of pixel intensities of the male (red) and female (blue) pronucleus from the two genotypes are depicted on the right. (K) Time separating the exit of meiosis II from NEBD of the male and female pronuclei in seconds ± SEM, determined from DIC time-lapse recordings. Extrusion of the second polar body was used as a reference for the exit from meiosis (time 0 s). Embryos were timed until NEBD in one-cell-stage embryos. The number of embryos analyzed (n) and p values (Student's t test, time from polar body extrusion until female NEBD, compared with the wild type) are as follows: wild type (n = 13); p zyg-9(b244) = 0.0025 (n = 14); p zyg-9(b244) npp-3(RNAi) = 0.74 (n = 14).

Virginie Hachet, et al. Mol Biol Cell. 2012 August 15;23(16):3111-3121.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk