Mechanical stress reduces podocyte proliferation in vitro

Arndt T. Petermann, Keiju Hiromura, Mary Blonski, Jeffrey Pippin, Toshiaki Monkawa, Raghu Durvasula, William G. Couser, Stuart J. Shankland

Research output: Contribution to journalArticle

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Abstract

Background. Mechanical stretch, a consequence of capillary glomerular hypertension, is thought to be the common final pathway for glomerulosclerosis in systemic hypertension, diabetes, reduced nephron number and focal segmental glomerulosclerosis. However, the effects of stretch on podocyte growth and the mechanisms that underlie this have not been elucidated. Methods. Mouse podocyte growth (3H-thymidine, MTT-assay, FACS) was measured following the application of mechanical stretch created by vacuum. The expression of specific cell cycle regulatory proteins was examined by RNAse protection assay and Western blot analysis. Control cells were grown under similar conditions, but were not exposed to stretch. Results. Mechanical stretch decreased DNA-synthesis (3H-thymidine incorporation) and cell number (MTT-assay) in podocytes at 24, 48 and 72 hours (P lt; 0.001 vs. control non-stretched cells), which was not due to apoptosis (Hoechst staining) nor cell detachment. Stretch decreased the mRNA and protein levels of cyclins D1, A and B1 within 24 hours. Stretching cells decreased the activity of Cdk2 (measured by histone H1 kinase assay) at 48 and 72 hours and Cdc2 at 72 hours. In contrast, stretch increased the protein levels of the cyclin dependent kinase inhibitors (CKI) p21cip/Kip/waf (p21) and p27Kip1 (p27) within the first 24 hours, and increased the mRNA levels of p57Kip2 (p57) at 72 hours. To examine the role of p21 in inhibiting proliferation induced by stretch, we studied p21 -/- podocytes in culture. Stretch did not reduce proliferation in p21-/- podocytes (P > 0.05 vs. non-stretched podocytes; P < 0.001 vs. stretched p21 +/+ podocytes). Conclusions. In contrast to mesangial cells, mechanical stretch decreases the growth of podocytes. This effect is mediated through the regulation of specific cell cycle regulatory proteins. These events may explain the apparent lack of podocyte proliferation in diseases correlated with capillary glomerular hypertension.

Original languageEnglish
Pages (from-to)40-50
Number of pages11
JournalKidney International
Volume61
Issue number1
DOIs
Publication statusPublished - 2002
Externally publishedYes

Fingerprint

Podocytes
Mechanical Stress
Cell Cycle Proteins
Hypertension
Thymidine
Cyclin-Dependent Kinase Inhibitor Proteins
Growth
Cyclin B1
In Vitro Techniques
Focal Segmental Glomerulosclerosis
Messenger RNA
Mesangial Cells
Cyclin D1
Nephrons
Vacuum
Cell Count
Western Blotting
Apoptosis
Staining and Labeling

Keywords

  • Capillary glomerular hypertension
  • Cell cycle
  • Cyclin dependent kinase
  • Kidney
  • p21
  • Stretch
  • Visceral glomerular epithelial cell podocyre

ASJC Scopus subject areas

  • Nephrology

Cite this

Petermann, A. T., Hiromura, K., Blonski, M., Pippin, J., Monkawa, T., Durvasula, R., ... Shankland, S. J. (2002). Mechanical stress reduces podocyte proliferation in vitro. Kidney International, 61(1), 40-50. https://doi.org/10.1046/j.1523-1755.2002.00102.x

Mechanical stress reduces podocyte proliferation in vitro. / Petermann, Arndt T.; Hiromura, Keiju; Blonski, Mary; Pippin, Jeffrey; Monkawa, Toshiaki; Durvasula, Raghu; Couser, William G.; Shankland, Stuart J.

In: Kidney International, Vol. 61, No. 1, 2002, p. 40-50.

Research output: Contribution to journalArticle

Petermann, AT, Hiromura, K, Blonski, M, Pippin, J, Monkawa, T, Durvasula, R, Couser, WG & Shankland, SJ 2002, 'Mechanical stress reduces podocyte proliferation in vitro', Kidney International, vol. 61, no. 1, pp. 40-50. https://doi.org/10.1046/j.1523-1755.2002.00102.x
Petermann, Arndt T. ; Hiromura, Keiju ; Blonski, Mary ; Pippin, Jeffrey ; Monkawa, Toshiaki ; Durvasula, Raghu ; Couser, William G. ; Shankland, Stuart J. / Mechanical stress reduces podocyte proliferation in vitro. In: Kidney International. 2002 ; Vol. 61, No. 1. pp. 40-50.
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AU - Petermann, Arndt T.

AU - Hiromura, Keiju

AU - Blonski, Mary

AU - Pippin, Jeffrey

AU - Monkawa, Toshiaki

AU - Durvasula, Raghu

AU - Couser, William G.

AU - Shankland, Stuart J.

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N2 - Background. Mechanical stretch, a consequence of capillary glomerular hypertension, is thought to be the common final pathway for glomerulosclerosis in systemic hypertension, diabetes, reduced nephron number and focal segmental glomerulosclerosis. However, the effects of stretch on podocyte growth and the mechanisms that underlie this have not been elucidated. Methods. Mouse podocyte growth (3H-thymidine, MTT-assay, FACS) was measured following the application of mechanical stretch created by vacuum. The expression of specific cell cycle regulatory proteins was examined by RNAse protection assay and Western blot analysis. Control cells were grown under similar conditions, but were not exposed to stretch. Results. Mechanical stretch decreased DNA-synthesis (3H-thymidine incorporation) and cell number (MTT-assay) in podocytes at 24, 48 and 72 hours (P lt; 0.001 vs. control non-stretched cells), which was not due to apoptosis (Hoechst staining) nor cell detachment. Stretch decreased the mRNA and protein levels of cyclins D1, A and B1 within 24 hours. Stretching cells decreased the activity of Cdk2 (measured by histone H1 kinase assay) at 48 and 72 hours and Cdc2 at 72 hours. In contrast, stretch increased the protein levels of the cyclin dependent kinase inhibitors (CKI) p21cip/Kip/waf (p21) and p27Kip1 (p27) within the first 24 hours, and increased the mRNA levels of p57Kip2 (p57) at 72 hours. To examine the role of p21 in inhibiting proliferation induced by stretch, we studied p21 -/- podocytes in culture. Stretch did not reduce proliferation in p21-/- podocytes (P > 0.05 vs. non-stretched podocytes; P < 0.001 vs. stretched p21 +/+ podocytes). Conclusions. In contrast to mesangial cells, mechanical stretch decreases the growth of podocytes. This effect is mediated through the regulation of specific cell cycle regulatory proteins. These events may explain the apparent lack of podocyte proliferation in diseases correlated with capillary glomerular hypertension.

AB - Background. Mechanical stretch, a consequence of capillary glomerular hypertension, is thought to be the common final pathway for glomerulosclerosis in systemic hypertension, diabetes, reduced nephron number and focal segmental glomerulosclerosis. However, the effects of stretch on podocyte growth and the mechanisms that underlie this have not been elucidated. Methods. Mouse podocyte growth (3H-thymidine, MTT-assay, FACS) was measured following the application of mechanical stretch created by vacuum. The expression of specific cell cycle regulatory proteins was examined by RNAse protection assay and Western blot analysis. Control cells were grown under similar conditions, but were not exposed to stretch. Results. Mechanical stretch decreased DNA-synthesis (3H-thymidine incorporation) and cell number (MTT-assay) in podocytes at 24, 48 and 72 hours (P lt; 0.001 vs. control non-stretched cells), which was not due to apoptosis (Hoechst staining) nor cell detachment. Stretch decreased the mRNA and protein levels of cyclins D1, A and B1 within 24 hours. Stretching cells decreased the activity of Cdk2 (measured by histone H1 kinase assay) at 48 and 72 hours and Cdc2 at 72 hours. In contrast, stretch increased the protein levels of the cyclin dependent kinase inhibitors (CKI) p21cip/Kip/waf (p21) and p27Kip1 (p27) within the first 24 hours, and increased the mRNA levels of p57Kip2 (p57) at 72 hours. To examine the role of p21 in inhibiting proliferation induced by stretch, we studied p21 -/- podocytes in culture. Stretch did not reduce proliferation in p21-/- podocytes (P > 0.05 vs. non-stretched podocytes; P < 0.001 vs. stretched p21 +/+ podocytes). Conclusions. In contrast to mesangial cells, mechanical stretch decreases the growth of podocytes. This effect is mediated through the regulation of specific cell cycle regulatory proteins. These events may explain the apparent lack of podocyte proliferation in diseases correlated with capillary glomerular hypertension.

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KW - Cell cycle

KW - Cyclin dependent kinase

KW - Kidney

KW - p21

KW - Stretch

KW - Visceral glomerular epithelial cell podocyre

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