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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

Mislocalization of Na+/K+-ATPase in Kif5b knock-down kidneys.Na+/K+-ATPase was basolaterally localized in renal epithelial cells of control E18.5 embryos (Kif5bfl/+), but it was mislocalized to the apical surface in renal epithelial cells of mutant E18.5 embryos (Kif5bfl/-:Pax2-Cre). Images are representative of kidney tissue sections from three mice. Green, Kif5b; Red, Na+/K+-ATPase.
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pone.0126002.g009: Mislocalization of Na+/K+-ATPase in Kif5b knock-down kidneys.Na+/K+-ATPase was basolaterally localized in renal epithelial cells of control E18.5 embryos (Kif5bfl/+), but it was mislocalized to the apical surface in renal epithelial cells of mutant E18.5 embryos (Kif5bfl/-:Pax2-Cre). Images are representative of kidney tissue sections from three mice. Green, Kif5b; Red, Na+/K+-ATPase.

Mentions: The tubular localization of Kif5b in un-weaned mice, and the selective and asymmetric intracellular distribution of Kif5b both in the TAL and in the DCT in weaned mice, suggests that Kif5b is involved in physiological functions in renal tubules. Unfortunately, the early death of Kif5b-Pax2KD mice makes it impossible to undertake a functional analysis of Kif5b in the kidney. We analyzed the polarity of tubules by staining tissue sections with polarity markers (Figs 8 and 9). LTL, THP, NCC and DBA are apical markers of PT, TAL, DCT and CD, respectively. As in the kidneys of Kif5bfl/+ mice, all these markers were localized in the apical region of tubular epithelial cells in the mutant kidneys, indicating that the apical localizations of LTL, THP, NCC and DBA were not affected following the knock-down of Kif5b (Fig 8). However, the basolateral marker Na+/K+-ATPase was found to be present on the apical surface of tubular epithelial cells following the knock-down of Kif5b (Fig 9), suggesting that Kif5b is the motor protein involved in the transportation of Na+/K+-ATPase-containing vesicles in renal 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)

Mislocalization of Na+/K+-ATPase in Kif5b knock-down kidneys.Na+/K+-ATPase was basolaterally localized in renal epithelial cells of control E18.5 embryos (Kif5bfl/+), but it was mislocalized to the apical surface in renal epithelial cells of mutant E18.5 embryos (Kif5bfl/-:Pax2-Cre). Images are representative of kidney tissue sections from three mice. Green, Kif5b; Red, Na+/K+-ATPase.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0126002.g009: Mislocalization of Na+/K+-ATPase in Kif5b knock-down kidneys.Na+/K+-ATPase was basolaterally localized in renal epithelial cells of control E18.5 embryos (Kif5bfl/+), but it was mislocalized to the apical surface in renal epithelial cells of mutant E18.5 embryos (Kif5bfl/-:Pax2-Cre). Images are representative of kidney tissue sections from three mice. Green, Kif5b; Red, Na+/K+-ATPase.
Mentions: The tubular localization of Kif5b in un-weaned mice, and the selective and asymmetric intracellular distribution of Kif5b both in the TAL and in the DCT in weaned mice, suggests that Kif5b is involved in physiological functions in renal tubules. Unfortunately, the early death of Kif5b-Pax2KD mice makes it impossible to undertake a functional analysis of Kif5b in the kidney. We analyzed the polarity of tubules by staining tissue sections with polarity markers (Figs 8 and 9). LTL, THP, NCC and DBA are apical markers of PT, TAL, DCT and CD, respectively. As in the kidneys of Kif5bfl/+ mice, all these markers were localized in the apical region of tubular epithelial cells in the mutant kidneys, indicating that the apical localizations of LTL, THP, NCC and DBA were not affected following the knock-down of Kif5b (Fig 8). However, the basolateral marker Na+/K+-ATPase was found to be present on the apical surface of tubular epithelial cells following the knock-down of Kif5b (Fig 9), suggesting that Kif5b is the motor protein involved in the transportation of Na+/K+-ATPase-containing vesicles in renal 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