scholarly journals Expression of Cathepsin L and Its Intrinsic Inhibitors in Glomeruli of Rats With Puromycin Aminonucleoside Nephrosis

2018 ◽  
Vol 66 (12) ◽  
pp. 863-877 ◽  
Author(s):  
Ayano Kubo ◽  
Isao Shirato ◽  
Teruo Hidaka ◽  
Miyuki Takagi ◽  
Yu Sasaki ◽  
...  
Keyword(s):  
Cystatin C ◽  
Cysteine Proteinase ◽  
Cathepsin L ◽  
Puromycin Aminonucleoside ◽  
Podocyte Injury ◽  
Almost All ◽  
Critical Process ◽  
Aminonucleoside Nephrosis ◽  
Puromycin Aminonucleoside Nephrosis

Cathepsin L, a lysosomal cysteine proteinase, may have a key role in various biological and disease processes by intracellular and extracellular degradation of proteins. We examined the levels of cathepsin L and its intrinsic inhibitors in glomeruli of rats with puromycin aminonucleoside (PAN) nephrosis. In contrast to the weak levels of cathepsin L in normal glomeruli, on days 4 and 8, strong immunostaining was detected in almost all podocytes when proteinuria and pathological changes of the podocytes developed. Cathepsin L was reduced after day 28, but remained in a focal and segmental manner. Cystatin β, an intracellular inhibitor, was not detected in podocytes. However, cystatin C, an extracellular inhibitor, was detected in podocytes after day 4, coincident with cathepsin L. Cystatin C levels were gradually reduced but sustained in many podocytes on day 28, while cystatin C was not detected in podocytes sustained cathepsin L. These results demonstrated that cathepsin L levels are not always accompanied by the levels of its inhibitors in podocytes of PAN nephrosis, suggesting a potential role of cathepsin L in podocyte injury, which is a critical process for the development and progression of tuft adhesion and sclerosis.

scholarly journals APOL1 risk variants enhance podocyte necrosis through compromising lysosomal membrane permeability

2014 ◽  
Vol 307 (3) ◽  
pp. F326-F336 ◽  
Author(s):  
Xiqian Lan ◽  
Aakash Jhaveri ◽  
Kang Cheng ◽  
Hongxiu Wen ◽  
Moin A. Saleem ◽  
...  
Keyword(s):  
Membrane Permeability ◽  
Disease Risk ◽  
Lucifer Yellow ◽  
Cathepsin L ◽  
Expression System ◽  
Lysosomal Membrane ◽  
Puromycin Aminonucleoside ◽  
Podocyte Injury ◽  
Risk Variants

Development of higher rates of nondiabetic glomerulosclerosis (GS) in African Americans has been attributed to two coding sequence variants (G1 and G2) in the APOL1 gene. To date, the cellular function and the role of APOL1 variants (Vs) in GS are still unknown. In this study, we examined the effects of overexpressing wild-type (G0) and kidney disease risk variants (G1 and G2) of APOL1 in human podocytes using a lentivirus expression system. Interestingly, G0 inflicted podocyte injury only at a higher concentration; however, G1 and G2 promoted moderate podocyte injury at lower and higher concentrations. APOL1Vs expressing podocytes displayed diffuse distribution of both Lucifer yellow dye and cathepsin L as manifestations of enhanced lysosomal membrane permeability (LMP). Chloroquine attenuated the APOL1Vs-induced increase in podocyte injury, consistent with targeting lysosomes. The chloride channel blocker DIDS prevented APOL1Vs- induced injury, indicating a role for chloride influx in osmotic swelling of lysosomes. Direct exposure of noninfected podocytes with conditioned media from G1- and G2-expressing podocytes also induced injury, suggesting a contributory role of the secreted component of G1 and G2 as well. Adverse host factors (AHFs) such as hydrogen peroxide, hypoxia, TNF-α, and puromycin aminonucleoside augmented APOL1- and APOL1Vs-induced podocyte injury, while the effect of human immunodeficiency virus (HIV) on podocyte injury was overwhelming under conditions of APOLVs expression. We conclude that G0 and G1 and G2 APOL1 variants have the potential to induce podocyte injury in a manner which is further augmented by AHFs, with HIV infection being especially prominent.

scholarly journals Role of Fat1 in cell-cell contact formation of podocytes in puromycin aminonucleoside nephrosis and neonatal kidney

2005 ◽  
Vol 68 (2) ◽  
pp. 542-551 ◽  
Author(s):  
Eishin Yaoita ◽  
Hidetake Kurihara ◽  
Yutaka Yoshida ◽  
Tsutomu Inoue ◽  
Asako Matsuki ◽  
...  
Keyword(s):  
Cell Contact ◽  
Puromycin Aminonucleoside ◽  
Contact Formation ◽  
Neonatal Kidney ◽  
Aminonucleoside Nephrosis ◽  
Puromycin Aminonucleoside Nephrosis ◽  
Cell Cell

p21-Activated kinases regulate actin remodeling in glomerular podocytes

2010 ◽  
Vol 298 (4) ◽  
pp. F951-F961 ◽  
Author(s):  
Jianxin Zhu ◽  
Ortal Attias ◽  
Lamine Aoudjit ◽  
Ruihua Jiang ◽  
Hiroshi Kawachi ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Tyrosine Phosphorylation ◽  
Adaptor Proteins ◽  
Dominant Negative ◽  
Puromycin Aminonucleoside ◽  
Actin Remodeling ◽  
Cellular Processes ◽  
Aminonucleoside Nephrosis ◽  
Puromycin Aminonucleoside Nephrosis

The tyrosine phosphorylation of nephrin is reported to regulate podocyte morphology via the Nck adaptor proteins. The Pak family of kinases are regulators of the actin cytoskeleton and are recruited to the plasma membrane via Nck. Here, we investigated the role of Pak in podocyte morphology. Pak1/2 were expressed in cultured podocytes. In mouse podocytes, Pak2 was predominantly phosphorylated, concentrated at the tips of the cellular processes, and its expression and/or phosphorylation were further increased when differentiated. Overexpression of rat nephrin in podocytes increased Pak1/2 phosphorylation, which was abolished when the Nck binding sites were mutated. Furthermore, dominant-negative Nck constructs blocked the Pak1 phosphorylation induced by antibody-mediated cross linking of nephrin. Transient transfection of constitutively kinase-active Pak1 into differentiated mouse podocytes decreased stress fibers, increased cortical F-actin, and extended the cellular processes, whereas kinase-dead mutant, kinase inhibitory construct, and Pak2 knockdown by shRNA had the opposite effect. In a rat model of puromycin aminonucleoside nephrosis, Pak1/2 phosphorylation was decreased in glomeruli, concomitantly with a decrease of nephrin tyrosine phosphorylation. These results suggest that Pak contributes to remodeling of the actin cytoskeleton in podocytes. Disturbed nephrin-Nck-Pak interaction may contribute to abnormal morphology of podocytes and proteinuria.

The Immunosuppressive Drug Mizoribine Directly Prevents Podocyte Injury in Puromycin Aminonucleoside Nephrosis

2010 ◽  
Vol 116 (1) ◽  
pp. e3-e10 ◽  
Author(s):  
Shigeru Takeuchi ◽  
Keiju Hiromura ◽  
Mai Tomioka ◽  
Satoshi Takahashi ◽  
Toru Sakairi ◽  
...  
Keyword(s):  
Immunosuppressive Drug ◽  
Puromycin Aminonucleoside ◽  
Podocyte Injury ◽  
Aminonucleoside Nephrosis ◽  
Puromycin Aminonucleoside Nephrosis

scholarly journals Novel expression of sodium/myo-inositol co-transporter in podocytes in puromycin aminonucleoside nephrosis

2004 ◽  
Vol 19 (4) ◽  
pp. 817-822 ◽  
Author(s):  
Y. Watanabe ◽  
T. Kobayashi ◽  
E. Yaoita ◽  
H. Kawachi ◽  
A. Yamauchi ◽  
...  
Keyword(s):  
Puromycin Aminonucleoside ◽  
Aminonucleoside Nephrosis ◽  
Puromycin Aminonucleoside Nephrosis

Glomerular Expression of Smooth-Muscle Myosin Heavy-Chain Isoforms in Aminonucleoside Nephrosis in Rats

1995 ◽  
Vol 89 (1) ◽  
pp. 45-52 ◽  
Author(s):  
Tsukasa Nakamura ◽  
Kenjiro Kimura ◽  
Isao Ebihara ◽  
Toshimasa Takahashi ◽  
Yasuhiko Tomino ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Myosin Heavy Chain ◽  
Heavy Chain ◽  
Proliferating Cell Nuclear Antigen ◽  
Smooth Muscle Actin ◽  
Nuclear Antigen ◽  
Puromycin Aminonucleoside ◽  
Proliferating Cell ◽  
Aminonucleoside Nephrosis ◽  
Puromycin Aminonucleoside Nephrosis

1. We investigated the glomerular expression of three types of myosin heavy-chain isoforms, including S-myosin heavy-chain 40 (SM1), S-myosin heavy-chain 29 (SM2) and FS-myosin heavy-chain 34 (SMemb) in puromycin aminonucleoside nephrosis. 2. There was little change in SM1 and SM2 mRNA levels throughout the experiment. In contrast, glomerular SMemb mRNA increased on days 2 and 4 (before and soon after the onset of proteinuria, respectively), but declined on day 8 (the peak of proteinuria). 3. Histological myosin heavy-chain expression was examined using three antibodies against SM1, SM2 and SMemb. Immunohistochemically, SM1 and SM2 were absent in the glomeruli associated with puromycin aminonucleoside nephrosis until day 20. The SMemb isoform was barely detectable in normal glomeruli, but substantial amounts of SMemb were demonstrated in the glomeruli of rats with puromycin aminonucleoside nephrosis. In the puromycin aminonucleoside-treated rats, the number of SMemb-positive glomerular cells increased on days 2 and 4. 4. We examined whether levels of α-smooth-muscle actin or proliferating cell nuclear antigen correlated with myosin heavy-chain levels in the glomeruli of rats with puromycin aminonucleoside nephrosis. None of the cellular components in the glomeruli was positive for either α-smooth-muscle actin or proliferating cell nuclear antigen in puromycin aminonucleoside nephrosis. 5. Administration of methylprednisolone to puromycin aminonucleoside-treated rats resulted in the rapid disappearance of proteinuria. However, methylprednisolone did not affect SMemb mRNA or immunostaining in the glomeruli of rats with puromycin aminonucleoside nephrosis. 6. These data suggest that SMemb may be a molecular marker for phenotypic change in early glomerular injury, and demonstrate that SMemb regulation differs from that of SM1, SM2, α-smooth-muscle actin and proliferating cell nuclear antigen in the glomeruli of rats with puromycin aminonucleoside nephrosis.

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