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The histone H3K9 demethylase Kdm3b is required for somatic growth and female reproductive function.

Liu Z, Chen X, Zhou S, Liao L, Jiang R, Xu J - Int. J. Biol. Sci. (2015)

Bottom Line: We found that Kdm3b ablation decreased IGFBP-3 expressed in the kidney by 53% and significantly reduced IGFBP-3 in the blood, which caused an accelerated degradation of IGF-1 and a 36% decrease in circulating IGF-1 concentration.We also found Kdm3b was highly expressed in the female reproductive organs including ovary, oviduct and uterus.Importantly, these female reproductive phenotypes were associated with significantly increased levels of H3K9me1/2/3 in the ovary and uterus.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas, USA. ; 3. Institute of Cancer Prevention and Treatment, Harbin Medical University, Harbin, China.

ABSTRACT
Kdm3b is a Jumonji C domain-containing protein that demethylates mono- and di-methylated lysine 9 of histone H3 (H3K9me1 and H3K9me2). Although the enzyme activity of Kdm3b is well characterized in vitro, its genetic and physiological function remains unknown. Herein, we generated Kdm3b knockout (Kdm3bKO) mice and observed restricted postnatal growth and female infertility in these mice. We found that Kdm3b ablation decreased IGFBP-3 expressed in the kidney by 53% and significantly reduced IGFBP-3 in the blood, which caused an accelerated degradation of IGF-1 and a 36% decrease in circulating IGF-1 concentration. We also found Kdm3b was highly expressed in the female reproductive organs including ovary, oviduct and uterus. Knockout of Kdm3b in female mice caused irregular estrous cycles, decreased 45% of the ovulation capability and 47% of the fertilization rate, and reduced 44% of the uterine decidual response, which were accompanied with a more than 50% decrease in the circulating levels of the 17beta-estradiol. Importantly, these female reproductive phenotypes were associated with significantly increased levels of H3K9me1/2/3 in the ovary and uterus. These results demonstrate that Kdm3b-mediated H3K9 demethylation plays essential roles in maintenance of the circulating IGF-1, postnatal somatic growth, circulating 17beta-estradiol, and female reproductive function.

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Knockout of Kdm3b increases the levels of H3K9 methylation in the ovary and uterus. (A) & (B). IHC analyses of H3K9me1/2/3 in the ovaries and uteri of WT and Kdm3bKO (KO) mice. F, follicular cells; I, interstitial cells; E, endometrial epithelium; S, endometrial stroma; M, myometrium. Two mice and multiple sections from each mouse in each genotype group were examined. (C). Analysis of H3K9me1/2/3 in the uteri of WT and KO mice by Western blotting. Histone H3 served as a loading control.
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Figure 5: Knockout of Kdm3b increases the levels of H3K9 methylation in the ovary and uterus. (A) & (B). IHC analyses of H3K9me1/2/3 in the ovaries and uteri of WT and Kdm3bKO (KO) mice. F, follicular cells; I, interstitial cells; E, endometrial epithelium; S, endometrial stroma; M, myometrium. Two mice and multiple sections from each mouse in each genotype group were examined. (C). Analysis of H3K9me1/2/3 in the uteri of WT and KO mice by Western blotting. Histone H3 served as a loading control.

Mentions: In the ovaries of WT mice with synchronized estrous cycle induced by PMSG and hCG treatments, relatively high levels of H3K9me1 were detected in the follicular granulosa, interstitial stromal cells and corpus luteum. Knockout of Kdm3b further increased the signals of H3K9me1 immunostaining in these cells. In WT ovaries, low levels of H3K9me2 were observed in the follicular granulosa and interstitial stromal cells. However, in the ovaries isolated from Kdm3bKO mice, H3K9me2 levels were increased in both of granulosa and interstitial stromal cells. H3K9me3 levels were detected at medium levels in WT granulosa cells, but at high levels in Kdm3bKO granulosa cells. On the other hand, H3K9me3 levels were similar in both WT and Kdm3bKO interstitial stromal cells (Fig. 5A and data not shown). In WT uteri, H3K9me1 immunoreactivity was present in the nuclei of luminal and glandular epithelial cells, some endometrial stromal and myometrial smooth muscle cells. H3K9me2 was mainly detected in the luminal epithelial cells, some endometrial stromal cells and most myometrial smooth muscle cells, but only a few glandular epithelial cells were H3K9me2 positive. H3K9me3 immunoreactivity was weak and only presented in small numbers of luminal epithelial, endometrial stromal and myometrial smooth muscle cells (Fig. 5B and data not shown). Importantly, in Kdm3bKO uteri, H3K9me1 was elevated in the endometrial stromal and myometrial smooth muscle cells, H3K9me2 was induced in all glandular epithelial and most endometrial stromal cells, and H3K9me3 was increased in nearly all uterine cells, especially in the glandular epithelial cells (Fig. 5B and data not shown). In agreement with these IHC results, Western blot analysis also demonstrated that the levels of H3K9me1, H3K9me2 and H3K9me3 were significantly increased in Kdm3bKO uteri versus WT uteri (Fig. 5C). Taken together, these results indicate that Kdm3b is required for maintaining normal patterns of H3K9 methylation in multiple types of cells in the female reproductive organs. Kdm3b deficiency altered these epigenetic codes and disrupted the female reproductive function.


The histone H3K9 demethylase Kdm3b is required for somatic growth and female reproductive function.

Liu Z, Chen X, Zhou S, Liao L, Jiang R, Xu J - Int. J. Biol. Sci. (2015)

Knockout of Kdm3b increases the levels of H3K9 methylation in the ovary and uterus. (A) & (B). IHC analyses of H3K9me1/2/3 in the ovaries and uteri of WT and Kdm3bKO (KO) mice. F, follicular cells; I, interstitial cells; E, endometrial epithelium; S, endometrial stroma; M, myometrium. Two mice and multiple sections from each mouse in each genotype group were examined. (C). Analysis of H3K9me1/2/3 in the uteri of WT and KO mice by Western blotting. Histone H3 served as a loading control.
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Related In: Results  -  Collection

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Figure 5: Knockout of Kdm3b increases the levels of H3K9 methylation in the ovary and uterus. (A) & (B). IHC analyses of H3K9me1/2/3 in the ovaries and uteri of WT and Kdm3bKO (KO) mice. F, follicular cells; I, interstitial cells; E, endometrial epithelium; S, endometrial stroma; M, myometrium. Two mice and multiple sections from each mouse in each genotype group were examined. (C). Analysis of H3K9me1/2/3 in the uteri of WT and KO mice by Western blotting. Histone H3 served as a loading control.
Mentions: In the ovaries of WT mice with synchronized estrous cycle induced by PMSG and hCG treatments, relatively high levels of H3K9me1 were detected in the follicular granulosa, interstitial stromal cells and corpus luteum. Knockout of Kdm3b further increased the signals of H3K9me1 immunostaining in these cells. In WT ovaries, low levels of H3K9me2 were observed in the follicular granulosa and interstitial stromal cells. However, in the ovaries isolated from Kdm3bKO mice, H3K9me2 levels were increased in both of granulosa and interstitial stromal cells. H3K9me3 levels were detected at medium levels in WT granulosa cells, but at high levels in Kdm3bKO granulosa cells. On the other hand, H3K9me3 levels were similar in both WT and Kdm3bKO interstitial stromal cells (Fig. 5A and data not shown). In WT uteri, H3K9me1 immunoreactivity was present in the nuclei of luminal and glandular epithelial cells, some endometrial stromal and myometrial smooth muscle cells. H3K9me2 was mainly detected in the luminal epithelial cells, some endometrial stromal cells and most myometrial smooth muscle cells, but only a few glandular epithelial cells were H3K9me2 positive. H3K9me3 immunoreactivity was weak and only presented in small numbers of luminal epithelial, endometrial stromal and myometrial smooth muscle cells (Fig. 5B and data not shown). Importantly, in Kdm3bKO uteri, H3K9me1 was elevated in the endometrial stromal and myometrial smooth muscle cells, H3K9me2 was induced in all glandular epithelial and most endometrial stromal cells, and H3K9me3 was increased in nearly all uterine cells, especially in the glandular epithelial cells (Fig. 5B and data not shown). In agreement with these IHC results, Western blot analysis also demonstrated that the levels of H3K9me1, H3K9me2 and H3K9me3 were significantly increased in Kdm3bKO uteri versus WT uteri (Fig. 5C). Taken together, these results indicate that Kdm3b is required for maintaining normal patterns of H3K9 methylation in multiple types of cells in the female reproductive organs. Kdm3b deficiency altered these epigenetic codes and disrupted the female reproductive function.

Bottom Line: We found that Kdm3b ablation decreased IGFBP-3 expressed in the kidney by 53% and significantly reduced IGFBP-3 in the blood, which caused an accelerated degradation of IGF-1 and a 36% decrease in circulating IGF-1 concentration.We also found Kdm3b was highly expressed in the female reproductive organs including ovary, oviduct and uterus.Importantly, these female reproductive phenotypes were associated with significantly increased levels of H3K9me1/2/3 in the ovary and uterus.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas, USA. ; 3. Institute of Cancer Prevention and Treatment, Harbin Medical University, Harbin, China.

ABSTRACT
Kdm3b is a Jumonji C domain-containing protein that demethylates mono- and di-methylated lysine 9 of histone H3 (H3K9me1 and H3K9me2). Although the enzyme activity of Kdm3b is well characterized in vitro, its genetic and physiological function remains unknown. Herein, we generated Kdm3b knockout (Kdm3bKO) mice and observed restricted postnatal growth and female infertility in these mice. We found that Kdm3b ablation decreased IGFBP-3 expressed in the kidney by 53% and significantly reduced IGFBP-3 in the blood, which caused an accelerated degradation of IGF-1 and a 36% decrease in circulating IGF-1 concentration. We also found Kdm3b was highly expressed in the female reproductive organs including ovary, oviduct and uterus. Knockout of Kdm3b in female mice caused irregular estrous cycles, decreased 45% of the ovulation capability and 47% of the fertilization rate, and reduced 44% of the uterine decidual response, which were accompanied with a more than 50% decrease in the circulating levels of the 17beta-estradiol. Importantly, these female reproductive phenotypes were associated with significantly increased levels of H3K9me1/2/3 in the ovary and uterus. These results demonstrate that Kdm3b-mediated H3K9 demethylation plays essential roles in maintenance of the circulating IGF-1, postnatal somatic growth, circulating 17beta-estradiol, and female reproductive function.

Show MeSH
Related in: MedlinePlus