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Analysis of Kif5b expression during mouse kidney development.

Cui J, Li X, Duan Z, Xue W, Wang Z, Lu S, Lin R, Liu M, Zhu G, Huang JD - PLoS ONE (2015)

Bottom Line: The distribution of Kif5b was analyzed by immunostaining.In kidneys of postnatal day 20 or of older mice, however, Kif5b was localized selectively in the basolateral domain of epithelial cells of the thick ascending loop of Henle, as well as of the distal convoluted tubule, with little expression being observed in the proximal tubule or in the collecting duct.Conditional knock-down of Kif5b in mouse kidney did not result in detectable morphological defects, but it did lead to a decrease in cell proliferation rate and also to a mislocalization of Na+/K+/-ATPase, indicating that although Kif5b is non-essential for kidney morphogenesis, it is important for nephron maturation.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Geriatrics, Beijing Hospital & Beijing Institute of Geriatrics, Ministry of Health, Beijing, China; Department of Biochemistry, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China.

ABSTRACT
Recent studies showed that kidney-specific inactivation of Kif3a produces kidney cysts and renal failure, suggesting that kinesin-mediated intracellular transportation is important for the establishement and maintenance of renal epithelial cell polarity and normal nephron functions. Kif5b, one of the most conserved kinesin heavy chain, is the mouse homologue of the human ubiquitous Kinesin Heavy Chain (uKHC). In order to elucidate the role of Kif5b in kidney development and function, it is essential to establish its expression profile within the organ. Therefore, in this study, we examined the expression pattern of Kif5b in mouse kidney. Kidneys from embryonic (E) 12.5-, 16.5-dpc (days post coitus) mouse fetuses, from postnatal (P) day 0, 10, 20 pups and from adult mice were collected. The distribution of Kif5b was analyzed by immunostaining. The possible involvement of Kif5b in kidney development was investigated in conditional mutant mice by using a Cre-LoxP strategy. This study showed that the distribution of Kif5b displayed spatiotemporal changes during postnatal kidney development. In kidneys of new born mice, Kif5b was strongly expressed in all developing tubules and in the ureteric bud, but not in the glomerulus or in other early-developing structures, such as the cap mesenchyme, the comma-shaped body, and the S-shaped body. In kidneys of postnatal day 20 or of older mice, however, Kif5b was localized selectively in the basolateral domain of epithelial cells of the thick ascending loop of Henle, as well as of the distal convoluted tubule, with little expression being observed in the proximal tubule or in the collecting duct. Conditional knock-down of Kif5b in mouse kidney did not result in detectable morphological defects, but it did lead to a decrease in cell proliferation rate and also to a mislocalization of Na+/K+/-ATPase, indicating that although Kif5b is non-essential for kidney morphogenesis, it is important for nephron maturation.

No MeSH data available.


Related in: MedlinePlus

Kidneys from Kif5b-Pax2KD (Kif5bfl/-:Pax2-Cre) embryos have a reduced cell proliferation rate.(A) In situ analysis of BrdU incorporation at E18.5. Pregnant mice were injected with BrdU and sacrificed 4 hours later. BrdU incorporation was detected by an in situ BrdU incorporation assay kit. Brown-stained nuclei are BrdU-positive. Tissues were counterstained with hematoxylin. Images are representative of kidney tissue sections from three embryos. Arrows, BrdU positive cells. Scale bar = 100 μm. (B) Quantitative analysis of BrdU incorporation at E18.5. BrdU incorporation is expressed as the percentage of the total number of cells in the ureteric bud and its branches (UB), in the metanephric mesenchyme (MM) or in the interstitial cells (IC). Kif5b deficiency significantly decreased BrdU incorporation in both ureteric bud and mesenchyme-derived epithelial structures, compared to wild type. Bars represent the means ± S.D. Six sections from three wild type and three mutant embryos were imaged and counted. P values are indicated in the figure. * P<0.05.
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pone.0126002.g007: Kidneys from Kif5b-Pax2KD (Kif5bfl/-:Pax2-Cre) embryos have a reduced cell proliferation rate.(A) In situ analysis of BrdU incorporation at E18.5. Pregnant mice were injected with BrdU and sacrificed 4 hours later. BrdU incorporation was detected by an in situ BrdU incorporation assay kit. Brown-stained nuclei are BrdU-positive. Tissues were counterstained with hematoxylin. Images are representative of kidney tissue sections from three embryos. Arrows, BrdU positive cells. Scale bar = 100 μm. (B) Quantitative analysis of BrdU incorporation at E18.5. BrdU incorporation is expressed as the percentage of the total number of cells in the ureteric bud and its branches (UB), in the metanephric mesenchyme (MM) or in the interstitial cells (IC). Kif5b deficiency significantly decreased BrdU incorporation in both ureteric bud and mesenchyme-derived epithelial structures, compared to wild type. Bars represent the means ± S.D. Six sections from three wild type and three mutant embryos were imaged and counted. P values are indicated in the figure. * P<0.05.

Mentions: Dysregulation of cell proliferation and/or apoptosis results in renal dysplasia [39]. We examined the impact on cell proliferation of abolishing Kif5b-mediated intracellular transportation using BrdU labeling. As shown in Fig 7, BrdU incorporation was observed in metanephric mesenchyme cells, mesenchyme-derived epithelial structures and ureteric buds in the outer regions of kidneys, as well as interstitial cells located in the medulla region (Fig 7A). Quantitation of the number of BrdU+ cells in ureteric bud branches (UB) and in metanephric mesenchyme (MM) revealed an approximately two- to three-fold diminution of cell proliferation in the kidneys of Kif5b-Pax2KD mice. By contrast, the cell proliferation rate was not affected in interstitial cells (IC) (Fig 7B). Based on TUNEL+ analysis, there was no detectable increase in renal cell apoptotic rate (S2 Fig).


Analysis of Kif5b expression during mouse kidney development.

Cui J, Li X, Duan Z, Xue W, Wang Z, Lu S, Lin R, Liu M, Zhu G, Huang JD - PLoS ONE (2015)

Kidneys from Kif5b-Pax2KD (Kif5bfl/-:Pax2-Cre) embryos have a reduced cell proliferation rate.(A) In situ analysis of BrdU incorporation at E18.5. Pregnant mice were injected with BrdU and sacrificed 4 hours later. BrdU incorporation was detected by an in situ BrdU incorporation assay kit. Brown-stained nuclei are BrdU-positive. Tissues were counterstained with hematoxylin. Images are representative of kidney tissue sections from three embryos. Arrows, BrdU positive cells. Scale bar = 100 μm. (B) Quantitative analysis of BrdU incorporation at E18.5. BrdU incorporation is expressed as the percentage of the total number of cells in the ureteric bud and its branches (UB), in the metanephric mesenchyme (MM) or in the interstitial cells (IC). Kif5b deficiency significantly decreased BrdU incorporation in both ureteric bud and mesenchyme-derived epithelial structures, compared to wild type. Bars represent the means ± S.D. Six sections from three wild type and three mutant embryos were imaged and counted. P values are indicated in the figure. * P<0.05.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4401754&req=5

pone.0126002.g007: Kidneys from Kif5b-Pax2KD (Kif5bfl/-:Pax2-Cre) embryos have a reduced cell proliferation rate.(A) In situ analysis of BrdU incorporation at E18.5. Pregnant mice were injected with BrdU and sacrificed 4 hours later. BrdU incorporation was detected by an in situ BrdU incorporation assay kit. Brown-stained nuclei are BrdU-positive. Tissues were counterstained with hematoxylin. Images are representative of kidney tissue sections from three embryos. Arrows, BrdU positive cells. Scale bar = 100 μm. (B) Quantitative analysis of BrdU incorporation at E18.5. BrdU incorporation is expressed as the percentage of the total number of cells in the ureteric bud and its branches (UB), in the metanephric mesenchyme (MM) or in the interstitial cells (IC). Kif5b deficiency significantly decreased BrdU incorporation in both ureteric bud and mesenchyme-derived epithelial structures, compared to wild type. Bars represent the means ± S.D. Six sections from three wild type and three mutant embryos were imaged and counted. P values are indicated in the figure. * P<0.05.
Mentions: Dysregulation of cell proliferation and/or apoptosis results in renal dysplasia [39]. We examined the impact on cell proliferation of abolishing Kif5b-mediated intracellular transportation using BrdU labeling. As shown in Fig 7, BrdU incorporation was observed in metanephric mesenchyme cells, mesenchyme-derived epithelial structures and ureteric buds in the outer regions of kidneys, as well as interstitial cells located in the medulla region (Fig 7A). Quantitation of the number of BrdU+ cells in ureteric bud branches (UB) and in metanephric mesenchyme (MM) revealed an approximately two- to three-fold diminution of cell proliferation in the kidneys of Kif5b-Pax2KD mice. By contrast, the cell proliferation rate was not affected in interstitial cells (IC) (Fig 7B). Based on TUNEL+ analysis, there was no detectable increase in renal cell apoptotic rate (S2 Fig).

Bottom Line: The distribution of Kif5b was analyzed by immunostaining.In kidneys of postnatal day 20 or of older mice, however, Kif5b was localized selectively in the basolateral domain of epithelial cells of the thick ascending loop of Henle, as well as of the distal convoluted tubule, with little expression being observed in the proximal tubule or in the collecting duct.Conditional knock-down of Kif5b in mouse kidney did not result in detectable morphological defects, but it did lead to a decrease in cell proliferation rate and also to a mislocalization of Na+/K+/-ATPase, indicating that although Kif5b is non-essential for kidney morphogenesis, it is important for nephron maturation.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Geriatrics, Beijing Hospital & Beijing Institute of Geriatrics, Ministry of Health, Beijing, China; Department of Biochemistry, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China.

ABSTRACT
Recent studies showed that kidney-specific inactivation of Kif3a produces kidney cysts and renal failure, suggesting that kinesin-mediated intracellular transportation is important for the establishement and maintenance of renal epithelial cell polarity and normal nephron functions. Kif5b, one of the most conserved kinesin heavy chain, is the mouse homologue of the human ubiquitous Kinesin Heavy Chain (uKHC). In order to elucidate the role of Kif5b in kidney development and function, it is essential to establish its expression profile within the organ. Therefore, in this study, we examined the expression pattern of Kif5b in mouse kidney. Kidneys from embryonic (E) 12.5-, 16.5-dpc (days post coitus) mouse fetuses, from postnatal (P) day 0, 10, 20 pups and from adult mice were collected. The distribution of Kif5b was analyzed by immunostaining. The possible involvement of Kif5b in kidney development was investigated in conditional mutant mice by using a Cre-LoxP strategy. This study showed that the distribution of Kif5b displayed spatiotemporal changes during postnatal kidney development. In kidneys of new born mice, Kif5b was strongly expressed in all developing tubules and in the ureteric bud, but not in the glomerulus or in other early-developing structures, such as the cap mesenchyme, the comma-shaped body, and the S-shaped body. In kidneys of postnatal day 20 or of older mice, however, Kif5b was localized selectively in the basolateral domain of epithelial cells of the thick ascending loop of Henle, as well as of the distal convoluted tubule, with little expression being observed in the proximal tubule or in the collecting duct. Conditional knock-down of Kif5b in mouse kidney did not result in detectable morphological defects, but it did lead to a decrease in cell proliferation rate and also to a mislocalization of Na+/K+/-ATPase, indicating that although Kif5b is non-essential for kidney morphogenesis, it is important for nephron maturation.

No MeSH data available.


Related in: MedlinePlus