Altered Cytoskeletal Structure of Smooth Muscle Cells in Ureteropelvic Junction Obstruction

Abstract Background: Ptch1 and Ptch2 are expressed in tubular epithelium and stromal cells adjacent to the UPJ. They mediate inhibition of Smoothened, a transmembrane protein expressed on the cell surface. If the pathway is disturbed, UPJOcan occur. This aim study aimed to determine the expression of Ptch1 (P1) and Ptch2 (P2) in stenotic segments in children with congenital ureteropelvic junction obstruction (UPJO) compared with normal control subjects. Methods: Stenotic segments of ureter tissues were obtained from 20 UPJO patients.UPJO caused by other pathogenies, such as vessel and ureteral polyps, were excluded. The control ureter specimens were obtained from 10 patients with Wilm’s tumor, and the tissues were confirmed histologically to be unaffected. Immunofluorescence, western blot and real-time PCR were used to investigate the expression of P1 and P2. Statistical methods were used to find the differences between the two groupsResults: P1 and P2 were identified in the cytoplasm of smooth muscle in two groups through immunohistochemistry. However, there were no statistical differences between the two groups in P1 and P2 with immunohistochemistry (P=0.31 and P=0.3, respectively). There were also no statistical differences with western blot (P=0.75 and P=0.9, respectively) and real-time PCR (P=0.52 and P=0.45, respectively). However, with the immunofluorescence it was found that red-stained P1 were diffused in the controls group, but were mainly located in the intracellular perinuclear compartment of smooth muscle cells in UPJO. Conclusions: The expression of P1 and P2 between the two groups had no statistical significant. P1 were mainly located in the intracellular perinuclear compartment of smooth muscle cells in UPJO. The P1 pathway might be disturbed by the abnormal distribution rather than the quantity, which might be one probable pathogenesis of UPJO.

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Changes in structure, interstitial Cajal-like cells and apoptosis of smooth muscle cells in congenital ureteropelvic junction obstruction

Pathology ◽

10.1097/01.pat.0000454556.77877.27 ◽

2014 ◽

Vol 46 ◽

pp. S134 ◽

Cited By ~ 1

Author(s):

Mitra Mehrazma ◽

Parin Tanzifi ◽

Nasser Rakhshani

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Ureteropelvic Junction Obstruction ◽

Muscle Cells ◽

Ureteropelvic Junction

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The Expression of Ptch1 and Ptch2 in the Stenotic Tissue of Congenital Ureteropelvic Junction Obstruction in Children

10.21203/rs.2.21792/v1 ◽

2020 ◽

Author(s):

Yanfang Yang ◽

wenwen han ◽

Weiping Zhang ◽

Ning Sun

Keyword(s):

Smooth Muscle ◽

Western Blot ◽

Smooth Muscle Cells ◽

Real Time ◽

Real Time Pcr ◽

Ureteropelvic Junction Obstruction ◽

Transmembrane Protein ◽

Muscle Cells ◽

Ureteropelvic Junction ◽

Abnormal Distribution

Abstract Objective Ptch1 and Ptch2 are expressed in tubular epithelium and stromal cells adjacent to the UPJ. They mediated inhibition of smoothened, a transmembrane protein expressed on the cell surface. If the pathway was disturbed, UPJO might be onset. The aim of this study was to determine the expression of Ptch1 (P1) and Ptch2 (P2) in stenotic segments in children with congenital ureteropelvic junction obstruction (UPJO) versus in normal control subjects. Materials and methods Stenotic segments of ureter tissues were obtained from 20 UPJO patients. Extrinsic stenosis, such as vessel and ureteral polyp, were excluded. The control ureter specimens were obtained from 10 patients with Wilm’s tumor, and the tissues were confirmed histologically to be unaffected. Immunofluorescence, Western blot and real-time PCR were used to investigate the expression of P1 and P2. Statistical methods were used to find the differences between the two groups Results P1 and P2 were identified that located in the cytoplasm of smooth muscle in two groups through Immunohistochemistry. However, there were no statistical differences between the two groups in P1 and P2 with immunohistochemistry (P=0.31 and P=0.3, respectively). Meanwhile there were no statistical differences with Western blot (P=0.75 and P=0.9, respectively) and real-time PCR (P=0.52 and P=0.45, respectively). But we found that in the immunofluorescence P1 were diffused in controls and mainly surrounding the nucleuses of smooth muscle cells in UPJO. Conclusions The expression of P1 and P2 between the two group had no statistically significant. P1 mainly surrounding the nucleuses of smooth muscle cells in UPJO. The P1 pathway might be disturbed by the abnormal distribution of P1 rather than the quantity.

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Changes in the cytoskeletal structure of cultured smooth muscle cells induced by calyculin-A

Journal of Cell Science ◽

10.1242/jcs.105.4.883 ◽

1993 ◽

Vol 105 (4) ◽

pp. 883-890 ◽

Cited By ~ 1

Author(s):

N. Hosoya ◽

M. Mitsui ◽

F. Yazama ◽

H. Ishihara ◽

H. Ozaki ◽

...

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Kinase Inhibitor ◽

Muscle Cells ◽

Stress Fibers ◽

Calyculin A ◽

Microtubule Severing ◽

Cytoskeletal Structure ◽

Focal Contacts ◽

Cultured Smooth Muscle Cells

Changes in the cytoskeletal structure of cultured A10 smooth muscle cells induced by calyculin-A (CL-A), a potent inhibitor of types 1 and 2A protein phosphatases, were analyzed using indirect fluorescence techniques. In the presence of 1 × 10(−7) M CL-A the cells became round and subsequently detached from the substratum. The effect of CL-A was inhibited by a non-selective kinase inhibitor, K-252a, but not by EGTA. In rounded cells stress fibers were absent and staining for F-actin appeared in patches. Vinculin, one of the components of focal contacts, was localized at the periphery of control cells. CL-A treatment moved the focal contacts towards the inside of the cell along the stress fibers, and this was followed by the rounding up of the cell. In addition, rapid and marked changes in microtubule structure were observed in CL-A-treated cells. Many ‘nicks’ or ‘gaps’ were observed along the microtubules in the attached, spread cells. A filamentous network of microtubules was not observed in the detached cells, i.e. after longer exposure to CL-A. These results suggest that CL-A may change the structure of focal contacts, resulting in the rounding up of the cell, and inducing a microtubule-severing activity. These effects were independent of the external Ca2+ concentration. The changes in cytoskeletal structure may be caused by disturbing the balance of phosphorylation and dephosphorylation in the cell.

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Electrical Characterization of Interstitial Cells of Cajal-Like Cells and Smooth Muscle Cells Isolated From the Mouse Ureteropelvic Junction

The Journal of Urology ◽

10.1016/j.juro.2006.11.073 ◽

2007 ◽

Vol 177 (4) ◽

pp. 1573-1580 ◽

Cited By ~ 29

Author(s):

Richard J. Lang ◽

Beata Z. Zoltkowski ◽

Jocelyn M. Hammer ◽

William F. Meeker ◽

Igor Wendt

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Interstitial Cells Of Cajal ◽

Electrical Characterization ◽

Muscle Cells ◽

Interstitial Cells ◽

Ureteropelvic Junction ◽

Cells Of Cajal

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Immunogold localization of HMB-45 antigen in pulmonary lymphangiomyomatosis

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100141366 ◽

1995 ◽

Vol 53 ◽

pp. 998-999

Author(s):

J.M. Minda ◽

E. Dessy ◽

G. G. Pietra

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Room Temperature ◽

Osmium Tetroxide ◽

Muscle Cells ◽

Ultrastructural Localization ◽

Reproductive Age ◽

Thin Sections ◽

Smooth Muscle Proliferation ◽

Hmb 45

Pulmonary lymphangiomyomatosis (PLAM) is a rare disease occurring exclusively in women of reproductive age. It involves the lungs, lymph nodes and lymphatic ducts. In the lungs, it is characterized by the proliferation of smooth muscle cells around lymphatics in the bronchovascular bundles, lobular septa and pleura The nature of smooth muscle proliferation in PLAM is still unclear. Recently, reactivity of the smooth muscle cells for HMB-45, a melanoma-related antigen has been reported by immunohistochemistry. The purpose of this study was the ultrastructural localization of HMB-45 immunoreactivity in these cells using gold-labeled antibodies.Lung tissue from three cases of PLAM, referred to our Institution for lung transplantation, was embedded in either Poly/Bed 812 post-fixed in 1% osmium tetroxide, or in LR White, without osmication. For the immunogold technique, thin sections were placed on Nickel grids and incubated with affinity purified, monoclonal anti-melanoma antibody HMB-45 (1:1) (Enzo Diag. Co) overnight at 4°C. After extensive washing with PBS, grids were treated with Goat-anti-mouse-IgG-Gold (5nm) (1:10) (Amersham Life Sci) for 1 hour, at room temperature.

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