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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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Embryonic developmental expression of Kif5b in mouse kidney.(A-D) Immunolocalization of Kif5b protein in the fetal mouse kidney at embryonic days (E) 12.5, 16.5. (A, B) Kif5b was not detectable in the E12.5 fetal kidney. (C, D) At E16.5, Kif5b was highly expressed in the ureteric bud (UB) and tubules (T). Images are representative of kidney tissue sections from three mice. (E) Quantitative real-time RT-PCR analysis showing the expression of Kif5 members: Kif5a, Kif5b and Kif5c mRNAs during kidney development. Br E18.5, brain of E18.5 embryo; KY E12.5, kidney of E12.5 embryo. mRNAs of brain tissues from E18.5 embryos were used as positive controls in the real-time PCR analysis. Bars represent the means ± S.D. (n = 3). One-way ANOVA was performed to compare specific gene expression levels in the kidneys between different stages. * P<0.05 versus KY E12.5. Scale bar = 25 μm.
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pone.0126002.g002: Embryonic developmental expression of Kif5b in mouse kidney.(A-D) Immunolocalization of Kif5b protein in the fetal mouse kidney at embryonic days (E) 12.5, 16.5. (A, B) Kif5b was not detectable in the E12.5 fetal kidney. (C, D) At E16.5, Kif5b was highly expressed in the ureteric bud (UB) and tubules (T). Images are representative of kidney tissue sections from three mice. (E) Quantitative real-time RT-PCR analysis showing the expression of Kif5 members: Kif5a, Kif5b and Kif5c mRNAs during kidney development. Br E18.5, brain of E18.5 embryo; KY E12.5, kidney of E12.5 embryo. mRNAs of brain tissues from E18.5 embryos were used as positive controls in the real-time PCR analysis. Bars represent the means ± S.D. (n = 3). One-way ANOVA was performed to compare specific gene expression levels in the kidneys between different stages. * P<0.05 versus KY E12.5. Scale bar = 25 μm.

Mentions: To characterize the spatial dynamics of Kif5b expression during kidney development, we examined the expression pattern of Kif5b in E12.5, E16.5, P0, P10, P20 and adult kidneys. Expression of Kif5b was not detectable in the kidneys of E12.5 embryos (Fig 2A and 2B); however, in E16.5 kidneys, Kif5b was highly expressed in the ureteric bud and tubular structures (Fig 2C and 2D). The quantitative levels of Kif5b mRNA during embryonic kidney development were determined by real-time reverse transcription PCR (qRT-PCR) (Fig 2E). The Kif5b mRNA was not detected in the early embryonic kidney (E12.5), but its level increased significantly in the kidneys of E16.5 embryos and thereafter. Although Kif5a and Kif5b were reported to be neuronal specific [8], both of them could be detected in E12.5 kidneys. The expression levels of Kif5a and Kif5c declined significantly during later kidney development, and could not be detected at all in the kidneys of newborn mice (Fig 2E and S1D 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)

Embryonic developmental expression of Kif5b in mouse kidney.(A-D) Immunolocalization of Kif5b protein in the fetal mouse kidney at embryonic days (E) 12.5, 16.5. (A, B) Kif5b was not detectable in the E12.5 fetal kidney. (C, D) At E16.5, Kif5b was highly expressed in the ureteric bud (UB) and tubules (T). Images are representative of kidney tissue sections from three mice. (E) Quantitative real-time RT-PCR analysis showing the expression of Kif5 members: Kif5a, Kif5b and Kif5c mRNAs during kidney development. Br E18.5, brain of E18.5 embryo; KY E12.5, kidney of E12.5 embryo. mRNAs of brain tissues from E18.5 embryos were used as positive controls in the real-time PCR analysis. Bars represent the means ± S.D. (n = 3). One-way ANOVA was performed to compare specific gene expression levels in the kidneys between different stages. * P<0.05 versus KY E12.5. Scale bar = 25 μm.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4401754&req=5

pone.0126002.g002: Embryonic developmental expression of Kif5b in mouse kidney.(A-D) Immunolocalization of Kif5b protein in the fetal mouse kidney at embryonic days (E) 12.5, 16.5. (A, B) Kif5b was not detectable in the E12.5 fetal kidney. (C, D) At E16.5, Kif5b was highly expressed in the ureteric bud (UB) and tubules (T). Images are representative of kidney tissue sections from three mice. (E) Quantitative real-time RT-PCR analysis showing the expression of Kif5 members: Kif5a, Kif5b and Kif5c mRNAs during kidney development. Br E18.5, brain of E18.5 embryo; KY E12.5, kidney of E12.5 embryo. mRNAs of brain tissues from E18.5 embryos were used as positive controls in the real-time PCR analysis. Bars represent the means ± S.D. (n = 3). One-way ANOVA was performed to compare specific gene expression levels in the kidneys between different stages. * P<0.05 versus KY E12.5. Scale bar = 25 μm.
Mentions: To characterize the spatial dynamics of Kif5b expression during kidney development, we examined the expression pattern of Kif5b in E12.5, E16.5, P0, P10, P20 and adult kidneys. Expression of Kif5b was not detectable in the kidneys of E12.5 embryos (Fig 2A and 2B); however, in E16.5 kidneys, Kif5b was highly expressed in the ureteric bud and tubular structures (Fig 2C and 2D). The quantitative levels of Kif5b mRNA during embryonic kidney development were determined by real-time reverse transcription PCR (qRT-PCR) (Fig 2E). The Kif5b mRNA was not detected in the early embryonic kidney (E12.5), but its level increased significantly in the kidneys of E16.5 embryos and thereafter. Although Kif5a and Kif5b were reported to be neuronal specific [8], both of them could be detected in E12.5 kidneys. The expression levels of Kif5a and Kif5c declined significantly during later kidney development, and could not be detected at all in the kidneys of newborn mice (Fig 2E and S1D 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