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RHAMM deficiency disrupts folliculogenesis resulting in female hypofertility.

Li H, Moll J, Winkler A, Frappart L, Brunet S, Hamann J, Kroll T, Verlhac MH, Heuer H, Herrlich P, Ploubidou A - Biol Open (2015)

Bottom Line: This resulted in folliculogenesis defects and female hypofertility, although HA-induced signalling was not affected.Deletion of the RHAMM C-terminus in vivo abolishes its spindle association, resulting in impaired spindle orientation in the dividing granulosa cells, folliculogenesis defects and subsequent female hypofertility.These data reveal the first identified physiological function for RHAMM, during oogenesis, and the importance of this spindle-associated function for female fertility.

View Article: PubMed Central - PubMed

Affiliation: Leibniz Institute for Age Research - Fritz Lipmann Institute, Beutenbergstrasse 11, D-07745 Jena, Germany.

No MeSH data available.


Related in: MedlinePlus

The RHAMM centrosome-targeting domain is required for RHAMM association with the mitotic spindle of the granulosa cells, where it regulates the orientation of the spindle axis.(A–D) RHAMM is localised at the mitotic spindle of granulosa cells, in follicles of hmmr+/+ mice (C). This localization is abolished by the deletion of the RHAMM centrosome targeting domain in the hmmrm/m mutants (D), as shown in sections of ovaries labelled with anti-RHAMM (green) (C,D) and anti-pH3 (red) antibodies plus DAPI (blue) (A–D). No obvious spindle defects were observed in the granulosa cells of hmmrm/m mutants (B); microtubules were labelled with anti-α-tubulin antibody (green) (A,B). In A–D, the framed mitotic cells are magnified in the panels below each main image, showing the indicated single- and the merged triple-immunofluorescence labeling. (E–H) Ovarian follicle sections from 10-week-old mice stained with hematoxylin and eosin to visualise the mitotic granulosa cells (arrowheads) (E,F). Follicles with 1–2 layers of granulosa cells were used in quantification of the angle θ between oocyte-basal membrane (orange line) and spindle axes (black line), as schematically illustrated in G (see also Materials and Methods). Granulosa cells in hmmr+/+ ovaries orient their spindle axis parallel to the oocyte-granulosa cell axis, as indicated by the small θ angle (H) (median 11.1°). The orientation of division is impaired in hmmrm/m granulosa cells (H) (median 24.2°) which assemble bipolar spindles without RHAMM (B,D). The difference of the two populations is 11.7° with a 95% confidence interval. The follicles of 4 hmmr+/+ (n = 41 mitotic cells) and 8 hmmrm/m (n = 31 mitotic cells) mouse ovaries were used in the quantification. Scale bar: 20 µm; **p<0.01; red lines in H indicate the median angle.
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f05: The RHAMM centrosome-targeting domain is required for RHAMM association with the mitotic spindle of the granulosa cells, where it regulates the orientation of the spindle axis.(A–D) RHAMM is localised at the mitotic spindle of granulosa cells, in follicles of hmmr+/+ mice (C). This localization is abolished by the deletion of the RHAMM centrosome targeting domain in the hmmrm/m mutants (D), as shown in sections of ovaries labelled with anti-RHAMM (green) (C,D) and anti-pH3 (red) antibodies plus DAPI (blue) (A–D). No obvious spindle defects were observed in the granulosa cells of hmmrm/m mutants (B); microtubules were labelled with anti-α-tubulin antibody (green) (A,B). In A–D, the framed mitotic cells are magnified in the panels below each main image, showing the indicated single- and the merged triple-immunofluorescence labeling. (E–H) Ovarian follicle sections from 10-week-old mice stained with hematoxylin and eosin to visualise the mitotic granulosa cells (arrowheads) (E,F). Follicles with 1–2 layers of granulosa cells were used in quantification of the angle θ between oocyte-basal membrane (orange line) and spindle axes (black line), as schematically illustrated in G (see also Materials and Methods). Granulosa cells in hmmr+/+ ovaries orient their spindle axis parallel to the oocyte-granulosa cell axis, as indicated by the small θ angle (H) (median 11.1°). The orientation of division is impaired in hmmrm/m granulosa cells (H) (median 24.2°) which assemble bipolar spindles without RHAMM (B,D). The difference of the two populations is 11.7° with a 95% confidence interval. The follicles of 4 hmmr+/+ (n = 41 mitotic cells) and 8 hmmrm/m (n = 31 mitotic cells) mouse ovaries were used in the quantification. Scale bar: 20 µm; **p<0.01; red lines in H indicate the median angle.

Mentions: In cultured cells, RHAMM associates with the mitotic spindle (Assmann et al., 1999; Maxwell et al., 2003; Groen et al., 2004; Maxwell et al., 2011) via its C-terminal centrosome-targeting domain (Maxwell et al., 2003). Consistent with these data, RHAMM localised at the mitotic spindle of granulosa cells in wild type ovaries (Fig. 5C), while this localization was completely abolished in hmmrm/m granulosa cells (Fig. 5D). Our in vivo data, confirmed, therefore, the previous observations in cell culture experiments.


RHAMM deficiency disrupts folliculogenesis resulting in female hypofertility.

Li H, Moll J, Winkler A, Frappart L, Brunet S, Hamann J, Kroll T, Verlhac MH, Heuer H, Herrlich P, Ploubidou A - Biol Open (2015)

The RHAMM centrosome-targeting domain is required for RHAMM association with the mitotic spindle of the granulosa cells, where it regulates the orientation of the spindle axis.(A–D) RHAMM is localised at the mitotic spindle of granulosa cells, in follicles of hmmr+/+ mice (C). This localization is abolished by the deletion of the RHAMM centrosome targeting domain in the hmmrm/m mutants (D), as shown in sections of ovaries labelled with anti-RHAMM (green) (C,D) and anti-pH3 (red) antibodies plus DAPI (blue) (A–D). No obvious spindle defects were observed in the granulosa cells of hmmrm/m mutants (B); microtubules were labelled with anti-α-tubulin antibody (green) (A,B). In A–D, the framed mitotic cells are magnified in the panels below each main image, showing the indicated single- and the merged triple-immunofluorescence labeling. (E–H) Ovarian follicle sections from 10-week-old mice stained with hematoxylin and eosin to visualise the mitotic granulosa cells (arrowheads) (E,F). Follicles with 1–2 layers of granulosa cells were used in quantification of the angle θ between oocyte-basal membrane (orange line) and spindle axes (black line), as schematically illustrated in G (see also Materials and Methods). Granulosa cells in hmmr+/+ ovaries orient their spindle axis parallel to the oocyte-granulosa cell axis, as indicated by the small θ angle (H) (median 11.1°). The orientation of division is impaired in hmmrm/m granulosa cells (H) (median 24.2°) which assemble bipolar spindles without RHAMM (B,D). The difference of the two populations is 11.7° with a 95% confidence interval. The follicles of 4 hmmr+/+ (n = 41 mitotic cells) and 8 hmmrm/m (n = 31 mitotic cells) mouse ovaries were used in the quantification. Scale bar: 20 µm; **p<0.01; red lines in H indicate the median angle.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400598&req=5

f05: The RHAMM centrosome-targeting domain is required for RHAMM association with the mitotic spindle of the granulosa cells, where it regulates the orientation of the spindle axis.(A–D) RHAMM is localised at the mitotic spindle of granulosa cells, in follicles of hmmr+/+ mice (C). This localization is abolished by the deletion of the RHAMM centrosome targeting domain in the hmmrm/m mutants (D), as shown in sections of ovaries labelled with anti-RHAMM (green) (C,D) and anti-pH3 (red) antibodies plus DAPI (blue) (A–D). No obvious spindle defects were observed in the granulosa cells of hmmrm/m mutants (B); microtubules were labelled with anti-α-tubulin antibody (green) (A,B). In A–D, the framed mitotic cells are magnified in the panels below each main image, showing the indicated single- and the merged triple-immunofluorescence labeling. (E–H) Ovarian follicle sections from 10-week-old mice stained with hematoxylin and eosin to visualise the mitotic granulosa cells (arrowheads) (E,F). Follicles with 1–2 layers of granulosa cells were used in quantification of the angle θ between oocyte-basal membrane (orange line) and spindle axes (black line), as schematically illustrated in G (see also Materials and Methods). Granulosa cells in hmmr+/+ ovaries orient their spindle axis parallel to the oocyte-granulosa cell axis, as indicated by the small θ angle (H) (median 11.1°). The orientation of division is impaired in hmmrm/m granulosa cells (H) (median 24.2°) which assemble bipolar spindles without RHAMM (B,D). The difference of the two populations is 11.7° with a 95% confidence interval. The follicles of 4 hmmr+/+ (n = 41 mitotic cells) and 8 hmmrm/m (n = 31 mitotic cells) mouse ovaries were used in the quantification. Scale bar: 20 µm; **p<0.01; red lines in H indicate the median angle.
Mentions: In cultured cells, RHAMM associates with the mitotic spindle (Assmann et al., 1999; Maxwell et al., 2003; Groen et al., 2004; Maxwell et al., 2011) via its C-terminal centrosome-targeting domain (Maxwell et al., 2003). Consistent with these data, RHAMM localised at the mitotic spindle of granulosa cells in wild type ovaries (Fig. 5C), while this localization was completely abolished in hmmrm/m granulosa cells (Fig. 5D). Our in vivo data, confirmed, therefore, the previous observations in cell culture experiments.

Bottom Line: This resulted in folliculogenesis defects and female hypofertility, although HA-induced signalling was not affected.Deletion of the RHAMM C-terminus in vivo abolishes its spindle association, resulting in impaired spindle orientation in the dividing granulosa cells, folliculogenesis defects and subsequent female hypofertility.These data reveal the first identified physiological function for RHAMM, during oogenesis, and the importance of this spindle-associated function for female fertility.

View Article: PubMed Central - PubMed

Affiliation: Leibniz Institute for Age Research - Fritz Lipmann Institute, Beutenbergstrasse 11, D-07745 Jena, Germany.

No MeSH data available.


Related in: MedlinePlus