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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 show normal gross histological morphology.(A) Anti-β-galactosidase immunostaining of E19.5 R26R+/+:Pax2-Cre mouse kidney. (B) Western blot analysis of Kif5b levels in kidneys from E19.5 mutant (Kif5bfl/-:Pax2-Cre), heterozygous (Kif5bfl/- and Kif5bfl/+:Pax2-Cre) and wild-type (Kif5bfl/+) embryos. Actin was used as a loading control. (C-D) Immunohistochemical analysis of Kif5b expression patterns/levels in kidneys from E19.5 control (Kif5bfl/+) and mutant (Kif5bfl/-:Pax2-Cre) embryos. (E-H) H&E staining showing that the organization of the nephrogenic zone, cortex and medulla is similar between control (Kif5bfl/+) and mutant (Kif5bfl/-:Pax2-Cre) kidneys. G, glomerulus; NZ, nephrogenic zone. Images are representative of kidney tissue sections from ten E19.5 embryos. Scale bar = 50 μm.
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pone.0126002.g006: Kidneys from Kif5b-Pax2KD (Kif5bfl/-:Pax2-Cre) embryos show normal gross histological morphology.(A) Anti-β-galactosidase immunostaining of E19.5 R26R+/+:Pax2-Cre mouse kidney. (B) Western blot analysis of Kif5b levels in kidneys from E19.5 mutant (Kif5bfl/-:Pax2-Cre), heterozygous (Kif5bfl/- and Kif5bfl/+:Pax2-Cre) and wild-type (Kif5bfl/+) embryos. Actin was used as a loading control. (C-D) Immunohistochemical analysis of Kif5b expression patterns/levels in kidneys from E19.5 control (Kif5bfl/+) and mutant (Kif5bfl/-:Pax2-Cre) embryos. (E-H) H&E staining showing that the organization of the nephrogenic zone, cortex and medulla is similar between control (Kif5bfl/+) and mutant (Kif5bfl/-:Pax2-Cre) kidneys. G, glomerulus; NZ, nephrogenic zone. Images are representative of kidney tissue sections from ten E19.5 embryos. Scale bar = 50 μm.

Mentions: In order to analyze the function of Kif5b in renal epithelial cells, conditional knock-down mice Kif5b-Pax2KD (Kif5bfl/-:Pax2-Cre) were generated by crossing Kif5b+/-:Pax2-Cre mice with Kif5bfl/fl mice. Kif5b-Pax2KD mice died with limb muscle dystrophy at the perinatal stage, probably due to hypoxia and/or impairment of milk intake [23]. The Pax2-Cre activity has been proven to efficiently delete LoxP-flanked sequences in Pax2-expressing cells and their descendants [23, 38]. Our analysis showed that positive immunostaining of β-galactosidase was observed in the ureteric bud and developing renal tubules of the kidneys of E19.5 R26R+/+:Pax2-Cre embryos (Fig 6A), which overlapped with the location of expression of Kif5b. A decrease of approximately 90% in the level of Kif5b was observed in the kidneys of E16.5 [23] and E19.5 Kif5b-Pax2KD embryos (Fig 6B–6D). The overall histological morphology of kidneys was normal in E19.5 Kif5b-Pax2KD embryos (Fig 6E–6H). The three-layer arrangement of nephrogenic zone, cortex and medulla appeared to be as well organized in the mutant kidneys as in control kidneys. Thus, in the kidneys of the mutant mice, the cortex comprised both glomeruli and developing tubules (Fig 6E and 6F); and furthermore, the overall cellularity, asymmetry, size of the glomeruli in the mutant kidneys were comparable to those in control kidneys (Fig 6G and 6H). In the medullas of both groups of kidneys, the collecting ducts and loops of Henle formed rays in parallel from the cortex to the deep medulla, with the interstitial cells aligned perpendicularly to support these ducts (Fig 6G and 6H). No dilation or cyst formation was found (Fig 6E–6H). Taken together, no obvious abnormality in the morphogenesis of the glomeruli or the renal tubules could be detected by histological examination.


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 show normal gross histological morphology.(A) Anti-β-galactosidase immunostaining of E19.5 R26R+/+:Pax2-Cre mouse kidney. (B) Western blot analysis of Kif5b levels in kidneys from E19.5 mutant (Kif5bfl/-:Pax2-Cre), heterozygous (Kif5bfl/- and Kif5bfl/+:Pax2-Cre) and wild-type (Kif5bfl/+) embryos. Actin was used as a loading control. (C-D) Immunohistochemical analysis of Kif5b expression patterns/levels in kidneys from E19.5 control (Kif5bfl/+) and mutant (Kif5bfl/-:Pax2-Cre) embryos. (E-H) H&E staining showing that the organization of the nephrogenic zone, cortex and medulla is similar between control (Kif5bfl/+) and mutant (Kif5bfl/-:Pax2-Cre) kidneys. G, glomerulus; NZ, nephrogenic zone. Images are representative of kidney tissue sections from ten E19.5 embryos. Scale bar = 50 μm.
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Related In: Results  -  Collection

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pone.0126002.g006: Kidneys from Kif5b-Pax2KD (Kif5bfl/-:Pax2-Cre) embryos show normal gross histological morphology.(A) Anti-β-galactosidase immunostaining of E19.5 R26R+/+:Pax2-Cre mouse kidney. (B) Western blot analysis of Kif5b levels in kidneys from E19.5 mutant (Kif5bfl/-:Pax2-Cre), heterozygous (Kif5bfl/- and Kif5bfl/+:Pax2-Cre) and wild-type (Kif5bfl/+) embryos. Actin was used as a loading control. (C-D) Immunohistochemical analysis of Kif5b expression patterns/levels in kidneys from E19.5 control (Kif5bfl/+) and mutant (Kif5bfl/-:Pax2-Cre) embryos. (E-H) H&E staining showing that the organization of the nephrogenic zone, cortex and medulla is similar between control (Kif5bfl/+) and mutant (Kif5bfl/-:Pax2-Cre) kidneys. G, glomerulus; NZ, nephrogenic zone. Images are representative of kidney tissue sections from ten E19.5 embryos. Scale bar = 50 μm.
Mentions: In order to analyze the function of Kif5b in renal epithelial cells, conditional knock-down mice Kif5b-Pax2KD (Kif5bfl/-:Pax2-Cre) were generated by crossing Kif5b+/-:Pax2-Cre mice with Kif5bfl/fl mice. Kif5b-Pax2KD mice died with limb muscle dystrophy at the perinatal stage, probably due to hypoxia and/or impairment of milk intake [23]. The Pax2-Cre activity has been proven to efficiently delete LoxP-flanked sequences in Pax2-expressing cells and their descendants [23, 38]. Our analysis showed that positive immunostaining of β-galactosidase was observed in the ureteric bud and developing renal tubules of the kidneys of E19.5 R26R+/+:Pax2-Cre embryos (Fig 6A), which overlapped with the location of expression of Kif5b. A decrease of approximately 90% in the level of Kif5b was observed in the kidneys of E16.5 [23] and E19.5 Kif5b-Pax2KD embryos (Fig 6B–6D). The overall histological morphology of kidneys was normal in E19.5 Kif5b-Pax2KD embryos (Fig 6E–6H). The three-layer arrangement of nephrogenic zone, cortex and medulla appeared to be as well organized in the mutant kidneys as in control kidneys. Thus, in the kidneys of the mutant mice, the cortex comprised both glomeruli and developing tubules (Fig 6E and 6F); and furthermore, the overall cellularity, asymmetry, size of the glomeruli in the mutant kidneys were comparable to those in control kidneys (Fig 6G and 6H). In the medullas of both groups of kidneys, the collecting ducts and loops of Henle formed rays in parallel from the cortex to the deep medulla, with the interstitial cells aligned perpendicularly to support these ducts (Fig 6G and 6H). No dilation or cyst formation was found (Fig 6E–6H). Taken together, no obvious abnormality in the morphogenesis of the glomeruli or the renal tubules could be detected by histological examination.

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