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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.

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Spatiotemporal expression pattern of Kif5b in mouse kidney.(A, B) Kif5b was widely distributed inside renal tubular epithelial cells of newborn mice. (C-H) Kif5b was gradually restricted to epithelial cells in DCT and TAL during postnatal kidney development. (I) Quantitative analysis of the relative optical density values for cells in renal tubules. Kidney sections were analyzed using ImageJ software to determine the mean optical density values per cell of positive-labeled tubules in the kidney. The relative mean optical densities of cells in the tubular structures compared with that in the DCT were calculated. Bars represent the means ± S.D. (n = 10 for each tubular structure from three mice were measured and analyzed). One-way ANOVA was performed to compare relative intensity levels of different tubular structures at indicated stages. * P<0.05 versus DCT. (J) Kif5b was evenly distributed in the cytoplasm of renal epithelial cells in newborn mice. (K) Kif5b was asymmetrically distributed in the basolateral domain in renal tubular cells of adult mice. Lu, lumen; PCT, proximal convoluted tubules; TAL, thick ascending limbs of loops of Henle; DCT, distal convoluted tubule; CD, collecting duct. Images are representative of kidney tissue sections from three mice. Scale bar: (A-H) 50 μm, (I, J) 10 μm.
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pone.0126002.g003: Spatiotemporal expression pattern of Kif5b in mouse kidney.(A, B) Kif5b was widely distributed inside renal tubular epithelial cells of newborn mice. (C-H) Kif5b was gradually restricted to epithelial cells in DCT and TAL during postnatal kidney development. (I) Quantitative analysis of the relative optical density values for cells in renal tubules. Kidney sections were analyzed using ImageJ software to determine the mean optical density values per cell of positive-labeled tubules in the kidney. The relative mean optical densities of cells in the tubular structures compared with that in the DCT were calculated. Bars represent the means ± S.D. (n = 10 for each tubular structure from three mice were measured and analyzed). One-way ANOVA was performed to compare relative intensity levels of different tubular structures at indicated stages. * P<0.05 versus DCT. (J) Kif5b was evenly distributed in the cytoplasm of renal epithelial cells in newborn mice. (K) Kif5b was asymmetrically distributed in the basolateral domain in renal tubular cells of adult mice. Lu, lumen; PCT, proximal convoluted tubules; TAL, thick ascending limbs of loops of Henle; DCT, distal convoluted tubule; CD, collecting duct. Images are representative of kidney tissue sections from three mice. Scale bar: (A-H) 50 μm, (I, J) 10 μm.

Mentions: In both the cortex and the medulla, the spatial expression of Kif5b changed with development in the postnatal kidney in two aspects. First, Kif5b expression gradually became restricted to DCT and TAL (Fig 3A–3I). This alteration became obvious at P20 (Fig 3E, 3F and 3I), at which stage the renal tubules probably became functionally mature. Secondly, the subcellular localization of Kif5b changed during development. In the newborn mouse, Kif5b was evenly distributed in the cytoplasm of renal epithelial cells (Fig 3A, 3B and 3J), whereas it was asymmetrically distributed in the basolateral domain in renal tubular cells of adult mice (Fig 3G, 3H and 3K). Although Kif5b was preferentially expressed in some tubules during postnatal kidney development, its total expression level in the whole kidney remained unchanged during this process (S1E 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)

Spatiotemporal expression pattern of Kif5b in mouse kidney.(A, B) Kif5b was widely distributed inside renal tubular epithelial cells of newborn mice. (C-H) Kif5b was gradually restricted to epithelial cells in DCT and TAL during postnatal kidney development. (I) Quantitative analysis of the relative optical density values for cells in renal tubules. Kidney sections were analyzed using ImageJ software to determine the mean optical density values per cell of positive-labeled tubules in the kidney. The relative mean optical densities of cells in the tubular structures compared with that in the DCT were calculated. Bars represent the means ± S.D. (n = 10 for each tubular structure from three mice were measured and analyzed). One-way ANOVA was performed to compare relative intensity levels of different tubular structures at indicated stages. * P<0.05 versus DCT. (J) Kif5b was evenly distributed in the cytoplasm of renal epithelial cells in newborn mice. (K) Kif5b was asymmetrically distributed in the basolateral domain in renal tubular cells of adult mice. Lu, lumen; PCT, proximal convoluted tubules; TAL, thick ascending limbs of loops of Henle; DCT, distal convoluted tubule; CD, collecting duct. Images are representative of kidney tissue sections from three mice. Scale bar: (A-H) 50 μm, (I, J) 10 μm.
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Related In: Results  -  Collection

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pone.0126002.g003: Spatiotemporal expression pattern of Kif5b in mouse kidney.(A, B) Kif5b was widely distributed inside renal tubular epithelial cells of newborn mice. (C-H) Kif5b was gradually restricted to epithelial cells in DCT and TAL during postnatal kidney development. (I) Quantitative analysis of the relative optical density values for cells in renal tubules. Kidney sections were analyzed using ImageJ software to determine the mean optical density values per cell of positive-labeled tubules in the kidney. The relative mean optical densities of cells in the tubular structures compared with that in the DCT were calculated. Bars represent the means ± S.D. (n = 10 for each tubular structure from three mice were measured and analyzed). One-way ANOVA was performed to compare relative intensity levels of different tubular structures at indicated stages. * P<0.05 versus DCT. (J) Kif5b was evenly distributed in the cytoplasm of renal epithelial cells in newborn mice. (K) Kif5b was asymmetrically distributed in the basolateral domain in renal tubular cells of adult mice. Lu, lumen; PCT, proximal convoluted tubules; TAL, thick ascending limbs of loops of Henle; DCT, distal convoluted tubule; CD, collecting duct. Images are representative of kidney tissue sections from three mice. Scale bar: (A-H) 50 μm, (I, J) 10 μm.
Mentions: In both the cortex and the medulla, the spatial expression of Kif5b changed with development in the postnatal kidney in two aspects. First, Kif5b expression gradually became restricted to DCT and TAL (Fig 3A–3I). This alteration became obvious at P20 (Fig 3E, 3F and 3I), at which stage the renal tubules probably became functionally mature. Secondly, the subcellular localization of Kif5b changed during development. In the newborn mouse, Kif5b was evenly distributed in the cytoplasm of renal epithelial cells (Fig 3A, 3B and 3J), whereas it was asymmetrically distributed in the basolateral domain in renal tubular cells of adult mice (Fig 3G, 3H and 3K). Although Kif5b was preferentially expressed in some tubules during postnatal kidney development, its total expression level in the whole kidney remained unchanged during this process (S1E 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