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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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Immunolocalization of Na+/K+-ATPase in mouse kidney PT (A-C), TAL (D-F), DCT (G-I) and CD (J-L) using antibodies against specific markers for each segment.Kif5b was expressed in TAL (THP-positive tubules) and DCT (NCC-positive tubules), but not in PT (LTL-positive tubules) or CD (DBA-positive tubules). (M-O) Co-localization of Kif5b and Na+/K+-ATPase in renal tubules of adult mice. PT, proximal tubule; TAL, thick ascending limbs of Henle; DCT: distal convoluted tubule; CD, collecting duct. Images are representative of kidney tissue sections from three mice. Scale bar = 50 μm.
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pone.0126002.g005: Immunolocalization of Na+/K+-ATPase in mouse kidney PT (A-C), TAL (D-F), DCT (G-I) and CD (J-L) using antibodies against specific markers for each segment.Kif5b was expressed in TAL (THP-positive tubules) and DCT (NCC-positive tubules), but not in PT (LTL-positive tubules) or CD (DBA-positive tubules). (M-O) Co-localization of Kif5b and Na+/K+-ATPase in renal tubules of adult mice. PT, proximal tubule; TAL, thick ascending limbs of Henle; DCT: distal convoluted tubule; CD, collecting duct. Images are representative of kidney tissue sections from three mice. Scale bar = 50 μm.

Mentions: Tubular cells in the TAL and the DCT are rich in mitochondria [28] and have a high density of Na+/K+-ATPase [29], which is essential for energy-demanding active intracellular transportation [30, 31]. It was reported that Kif5b forms functional Kinesin-1 with Kinesin light chain-2 to regulate the transportation of the Na+/K+-ATPase-containing vesicle in alveolar epithelial cells [20]. We further analyzed the expression patterns of Na+/K+-ATPase in the kidneys of adult mice by double labelling of Na+/K+-ATPase and of markers of specific nephron segments (Fig 5). It was found that Na+/K+-ATPase was basolaterally localized in the epithelial cells of THP-positive TAL and NCC-positive DCT (Fig 5D–5I), but not expressed in the LTL-positive PT or DAB (Dolichos biflorus agglutinin) positive CD (Fig 5A–5C and 5J–5L). Moreover, Kif5b and Na+/K+-ATPase were found to be co-localized in the same tubular structures (Fig 5M–5O), suggesting that Kif5b performs regulatory functions in Na+/K+-ATPase-containing vesicle transportation in renal epithelial cells of TAL and DCT that are similar to the functions that it performs in alveolar epithelial cells.


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)

Immunolocalization of Na+/K+-ATPase in mouse kidney PT (A-C), TAL (D-F), DCT (G-I) and CD (J-L) using antibodies against specific markers for each segment.Kif5b was expressed in TAL (THP-positive tubules) and DCT (NCC-positive tubules), but not in PT (LTL-positive tubules) or CD (DBA-positive tubules). (M-O) Co-localization of Kif5b and Na+/K+-ATPase in renal tubules of adult mice. PT, proximal tubule; TAL, thick ascending limbs of Henle; DCT: distal convoluted tubule; CD, collecting duct. Images are representative of kidney tissue sections from three mice. Scale bar = 50 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401754&req=5

pone.0126002.g005: Immunolocalization of Na+/K+-ATPase in mouse kidney PT (A-C), TAL (D-F), DCT (G-I) and CD (J-L) using antibodies against specific markers for each segment.Kif5b was expressed in TAL (THP-positive tubules) and DCT (NCC-positive tubules), but not in PT (LTL-positive tubules) or CD (DBA-positive tubules). (M-O) Co-localization of Kif5b and Na+/K+-ATPase in renal tubules of adult mice. PT, proximal tubule; TAL, thick ascending limbs of Henle; DCT: distal convoluted tubule; CD, collecting duct. Images are representative of kidney tissue sections from three mice. Scale bar = 50 μm.
Mentions: Tubular cells in the TAL and the DCT are rich in mitochondria [28] and have a high density of Na+/K+-ATPase [29], which is essential for energy-demanding active intracellular transportation [30, 31]. It was reported that Kif5b forms functional Kinesin-1 with Kinesin light chain-2 to regulate the transportation of the Na+/K+-ATPase-containing vesicle in alveolar epithelial cells [20]. We further analyzed the expression patterns of Na+/K+-ATPase in the kidneys of adult mice by double labelling of Na+/K+-ATPase and of markers of specific nephron segments (Fig 5). It was found that Na+/K+-ATPase was basolaterally localized in the epithelial cells of THP-positive TAL and NCC-positive DCT (Fig 5D–5I), but not expressed in the LTL-positive PT or DAB (Dolichos biflorus agglutinin) positive CD (Fig 5A–5C and 5J–5L). Moreover, Kif5b and Na+/K+-ATPase were found to be co-localized in the same tubular structures (Fig 5M–5O), suggesting that Kif5b performs regulatory functions in Na+/K+-ATPase-containing vesicle transportation in renal epithelial cells of TAL and DCT that are similar to the functions that it performs in alveolar epithelial cells.

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