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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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Related in: MedlinePlus

Immunolocalization of Kif5b in mouse kidney PT (A-C), TAL (D-F), CD (G-I) and DCT (J-K), 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 (AQP2-positive tubules). (L) The diagram shows the selective and asymmetric expression pattern of Kif5b in renal tubules of adult mice. Red, Kif5b; orange, LTL; yellow, THP; green, NCC; light blue, AQP2; blue, nucleus. Only cells from one side of the tubules are presented and the other side is represented by a dotted line. Arrows represent the direction of urinary flow. Cortex and medulla are separated by a dashed line. G, glomerulus; PT, proximal tubule; TDL, thick descending limbs of Henle; TS, thin segment; 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.g004: Immunolocalization of Kif5b in mouse kidney PT (A-C), TAL (D-F), CD (G-I) and DCT (J-K), 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 (AQP2-positive tubules). (L) The diagram shows the selective and asymmetric expression pattern of Kif5b in renal tubules of adult mice. Red, Kif5b; orange, LTL; yellow, THP; green, NCC; light blue, AQP2; blue, nucleus. Only cells from one side of the tubules are presented and the other side is represented by a dotted line. Arrows represent the direction of urinary flow. Cortex and medulla are separated by a dashed line. G, glomerulus; PT, proximal tubule; TDL, thick descending limbs of Henle; TS, thin segment; 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: To further confirm the nephron segments in which Kif5b was expressed in adult mice, kidney paraffin sections were double labelled for Kif5b and for markers of specific nephron segments (Fig 4). Lotus tetragonolobus agglutinin (LTL) is a lectin that specifically stains proximal tubules (green) [6]. Double immunostaining showed that Kif5b was not expressed in the proximal tubule (PT). Aquaporin-2 (AQP2) is localized mainly at the apical cell membranes of the principal cells of the CD of the kidney [25]. Lack of Kif5b expression in AQP2-positive tubules revealed that Kif5b was not expressed in the CD. Tamm-Horsfall protein, a GPI-anchored glycoprotein (THP), is found to be produced by the TAL in mammalian kidney [6]. Here, positive staining of Kif5b and THP in the same renal tubules suggested that Kif5b was expressed in the TAL. Finally, the Na-Cl cotransporter (NCC) is specifically localized at the apical surface of the DCT. Since antibodies against NCC and Kif5b were both raised in rabbit, we stained these two proteins separately in two consecutive tissue sections. Kif5b and NCC were found to be expressed in the same tubular structures, indicating that Kif5b was expressed in the DCT.


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 Kif5b in mouse kidney PT (A-C), TAL (D-F), CD (G-I) and DCT (J-K), 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 (AQP2-positive tubules). (L) The diagram shows the selective and asymmetric expression pattern of Kif5b in renal tubules of adult mice. Red, Kif5b; orange, LTL; yellow, THP; green, NCC; light blue, AQP2; blue, nucleus. Only cells from one side of the tubules are presented and the other side is represented by a dotted line. Arrows represent the direction of urinary flow. Cortex and medulla are separated by a dashed line. G, glomerulus; PT, proximal tubule; TDL, thick descending limbs of Henle; TS, thin segment; 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.g004: Immunolocalization of Kif5b in mouse kidney PT (A-C), TAL (D-F), CD (G-I) and DCT (J-K), 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 (AQP2-positive tubules). (L) The diagram shows the selective and asymmetric expression pattern of Kif5b in renal tubules of adult mice. Red, Kif5b; orange, LTL; yellow, THP; green, NCC; light blue, AQP2; blue, nucleus. Only cells from one side of the tubules are presented and the other side is represented by a dotted line. Arrows represent the direction of urinary flow. Cortex and medulla are separated by a dashed line. G, glomerulus; PT, proximal tubule; TDL, thick descending limbs of Henle; TS, thin segment; 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: To further confirm the nephron segments in which Kif5b was expressed in adult mice, kidney paraffin sections were double labelled for Kif5b and for markers of specific nephron segments (Fig 4). Lotus tetragonolobus agglutinin (LTL) is a lectin that specifically stains proximal tubules (green) [6]. Double immunostaining showed that Kif5b was not expressed in the proximal tubule (PT). Aquaporin-2 (AQP2) is localized mainly at the apical cell membranes of the principal cells of the CD of the kidney [25]. Lack of Kif5b expression in AQP2-positive tubules revealed that Kif5b was not expressed in the CD. Tamm-Horsfall protein, a GPI-anchored glycoprotein (THP), is found to be produced by the TAL in mammalian kidney [6]. Here, positive staining of Kif5b and THP in the same renal tubules suggested that Kif5b was expressed in the TAL. Finally, the Na-Cl cotransporter (NCC) is specifically localized at the apical surface of the DCT. Since antibodies against NCC and Kif5b were both raised in rabbit, we stained these two proteins separately in two consecutive tissue sections. Kif5b and NCC were found to be expressed in the same tubular structures, indicating that Kif5b was expressed in the DCT.

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