Vacuolation in Renal Tubular Epithelium of Mice: an Incidental Lesion

1997 ◽  
Vol 3 (S2) ◽  
pp. 261-262
Author(s):  
B.J. Dovey-Hartman ◽  
R.C. Johnson ◽  
M.W. Leach ◽  
D.W. Frank ◽  
D. E. Sinha ◽  
...  
Keyword(s):  
Osmium Tetroxide ◽  
Tubular Epithelial Cells ◽  
Outer Medulla ◽  
Drug Induced ◽  
Test Article ◽  
Renal Tubular Epithelium ◽  
Cytoplasmic Vacuoles ◽  
Transmission Electron ◽  
Vacuolar Change ◽  
Renal Tubular

Prominent cytoplasmic vacuoles in renal tubular epithelial cells of the outer medulla were observed in several kidneys from test article-dosed mice during routine light microscopic (LM) examination. Because the vacuolar change was seen infrequently by LM examination, and was not found in any control mice from that study, it was not clear whether the vacuolation represented a drug-induced change. To address this question, kidney sections from multiple unrelated mouse studies (214 control mice and 541 test article-dosed mice representing six different compounds) were examined by LM for similar vacuolar changes. Vacuolation was seen by LM in 2.3% of the control and 2.8% of the test article-dosed mice. Transmission electron microscopy (TEM) was performed on kidneys with prominent vacuoles seen by LM in 5 control mice and 2 test article-dosed mice to further characterize the vacuoles. Additionally, kidneys from 4 randomly selected control mice lacking vacuolation by LM were examined by TEM. Samples from the outer medulla of kidney that had been fixed by immersion in 10% formalin were post-fixed in 2.5% glutaraldehyde and 2% osmium tetroxide and processed routinely for TEM.

scholarly journals Mesenchymal stem cell-derived exosome miR-21-3p prevents renal fibrosis

2020 ◽  
Author(s):  
Zongying Li ◽  
Shuyi Cao
Keyword(s):  
Renal Fibrosis ◽  
Rt Pcr ◽  
Paracrine Effects ◽  
Downstream Target ◽  
Renal Tubular Epithelium ◽  
Pathological Result ◽  
Transmission Electron ◽  
Renal Tubular ◽  
Tgf Β1

Abstract Background: Renal fibrosis is the pathological result of excessive deposition of extracellular matrix (ECM) in the process of chronic kidney disease, but its mechanism is not clear. Mesenchymal stem cells (MSCs) exerts its therapeutic effect mainly through paracrine effects, such as exosome, to change the cellular microenvironment. Here, we explore the function of exosome derived MSCs in renal fibrosis.Methods: UUO model was constructed to simulate renal fibrosis in mice. Heat map and RT-PCR were used to explore the differential expression of miRNAs. RT-PCR and western blot were performed to detect the expression levels of fibrosis-associated genes and proteins in vivo and vitro. Transmission electron microscope and particle size detection were used to confirm the exosome construct. Then we forced expression of miR-21-3p in MSCs and isolated the exosomes. Then the fibrosis-associated genes and proteins were explored after exosomes injection.Results: In this study, we observed that exogenous miR-21-3p, interacted with smad2, the downstream target of miR-21-3p, which prevented renal fibrosis in UUO mice, and alleviate fibrosis in TGF-β1-induced renal tubular epithelium cells (HK-2). The extractive exosome-miR-21-3p treatment blocked renal fibrosis in UUO mice and alleviated fibrosis in TGF-β1-induced HK-2 cells and renal fibrosis mice. Conclusion: Taken together, Overexpression of miR-21-3p prevented CKD-induced renal fibrosis via exosome-mediated miR-21-3p transfer. These results suggest possible therapeutic strategies for using exosome delivery of miR-21-3p to treat complications of CKD.

Integrin receptors in renal tubular epithelium: new insights into pathophysiology of acute renal failure

1993 ◽  
Vol 264 (1) ◽  
pp. F1-F8 ◽  
Author(s):  
M. S. Goligorsky ◽  
W. Lieberthal ◽  
L. Racusen ◽  
E. E. Simon
Keyword(s):  
Renal Failure ◽  
Acute Renal Failure ◽  
Epithelial Cells ◽  
Apical Surface ◽  
Tubular Epithelium ◽  
Tubular Epithelial Cells ◽  
Renal Tubular Epithelial Cells ◽  
Integrin Receptors ◽  
Renal Tubular Epithelium ◽  
Renal Tubular

This review summarizes the existing evidence implicating disordered adhesion of renal tubular epithelial cells to the basement membrane in the pathophysiology of acute renal failure. The following three major lines of investigation are discussed: 1) exfoliation of renal tubular epithelial cells as a potential mechanism of tubular obstruction, 2) normal distribution of integrin receptors along the tubular apparatus, and 3) redistribution of integrin receptors and remodeling of the cytoskeleton following acute injury to renal tubular epithelium. We advance the hypothesis that the loss of the basolateral expression of integrin receptors is responsible for the exfoliation of viable proximal epithelial cells and that the redistribution of integrin receptors from the basolateral to the apical surface of epithelial cells facilitates adhesion of detached cells to the in situ cells. These two processes culminate in tubular obstruction.

scholarly journals Caspase 3/GSDME-dependent pyroptosis contributes to chemotherapy drug-induced nephrotoxicity

2021 ◽  
Vol 12 (2) ◽  
Author(s):  
Xiujin Shen ◽  
Haibing Wang ◽  
Chunhua Weng ◽  
Hong Jiang ◽  
Jianghua Chen
Keyword(s):  
Epithelial Cells ◽  
Caspase 3 ◽  
Tubular Epithelial Cell ◽  
Tubular Epithelial Cells ◽  
Chemotherapeutic Drug ◽  
Renal Tubular Epithelial Cells ◽  
Renal Tubular Epithelial Cell ◽  
Drug Induced ◽  
Jnk Signaling ◽  
Renal Tubular

AbstractChemotherapy drug-induced nephrotoxicity limits clinical applications for treating cancers. Pyroptosis, a newly discovered programmed cell death, was recently reported to be associated with kidney diseases. However, the role of pyroptosis in chemotherapeutic drug-induced nephrotoxicity has not been fully clarified. Herein, we demonstrate that the chemotherapeutic drug cisplatin or doxorubicin, induces the cleavage of gasdermin E (GSDME) in cultured human renal tubular epithelial cells, in a time- and concentration-dependent manner. Morphologically, cisplatin- or doxorubicin-treated renal tubular epithelial cells exhibit large bubbles emerging from the cell membrane. Furthermore, activation of caspase 3, not caspase 9, is associated with GSDME cleavage in cisplatin- or doxorubicin-treated renal tubular epithelial cells. Meanwhile, silencing GSDME alleviates cisplatin- or doxorubicin-induced HK-2 cell pyroptosis by increasing cell viability and decreasing LDH release. In addition, treatment with Ac-DMLD-CMK, a polypeptide targeting mouse caspase 3-Gsdme signaling, inhibits caspase 3 and Gsdme activation, alleviates the deterioration of kidney function, attenuates renal tubular epithelial cell injury, and reduces inflammatory cytokine secretion in vivo. Specifically, GSDME cleavage depends on ERK and JNK signaling. NAC, a reactive oxygen species (ROS) inhibitor, reduces GSDME cleavage through JNK signaling in human renal tubular epithelial cells. Thus, we speculate that renal tubular epithelial cell pyroptosis induced by chemotherapy drugs is mediated by ROS-JNK-caspase 3-GSDME signaling, implying that therapies targeting GSDME may prove efficacious in overcoming chemotherapeutic drug-induced nephrotoxicity.

Induction of apoptosis in ischemia-reperfusion model of mouse kidney: possible involvement of Fas.

1998 ◽  
Vol 9 (4) ◽  
pp. 620-631 ◽  
Author(s):  
S Nogae ◽  
M Miyazaki ◽  
N Kobayashi ◽  
T Saito ◽  
K Abe ◽  
...  
Keyword(s):  
Cell Death ◽  
Epithelial Cells ◽  
Ischemia Reperfusion ◽  
Mouse Kidney ◽  
Renal Tubules ◽  
Tubular Epithelial Cells ◽  
Renal Tubular Epithelial Cells ◽  
Outer Medulla ◽  
Fas Mrna ◽  
Renal Tubular

Although ischemia-reperfusion of mouse kidney is known to cause severe renal failure due to tubular cell death, the exact cellular mechanism responsible for this phenomenon is not clear. To investigate the spatial and temporal development of renal cell death and the role of Fas/APO-1/CD95 (Fas) in this process, the left renal vessels were occluded in a group of mice for 30, 60, or 120 min followed by reperfusion for 24 h (n = 4 for each group). Analysis of the isolated DNA in agarose-gel electrophoresis revealed a typical ladder pattern of bands consisting of multiples of 180 to 200 bp, considered the hallmark of apoptosis. The intensity of the bands increased proportionately with the duration of ischemia. Histochemical analysis using terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling showed the presence of nuclei with DNA double-strand breaks specifically in distal renal tubules of the outer medulla. The presence of apoptosis was also confirmed by electron microscopy. Analysis of total RNA by Northern blotting revealed one appropriate-sized band for Fas mRNA in the normal kidney, which intensified in the ischemia-reperfused kidney. Moreover, nonradioactive in situ hybridization revealed that distal renal tubular epithelial cells were positive for Fas mRNA in the outer medulla. Fas antigen was also localized to the renal tubular epithelial cells of the outer medulla by immunohistochemistry. The number of apoptotic cells in the ischemia-reperfusion kidney of the lpr/lpr mouse was low. These findings strongly indicate that ischemia-reperfusion of the kidney induces apoptosis of a specific area of tubular epithelial cells in the outer medulla through the Fas system.

scholarly journals Use of 3D Human Liver Organoids to Predict Drug-Induced Phospholipidosis

2020 ◽  
Vol 21 (8) ◽  
pp. 2982
Author(s):  
Ji-Young Lee ◽  
Hyo-Jeong Han ◽  
Sang-Joon Lee ◽  
Eun-Ho Cho ◽  
Han-Byul Lee ◽  
...  
Keyword(s):  
Hepg2 Cells ◽  
Human Liver ◽  
Three Dimensional ◽  
Lamellar Bodies ◽  
Albumin Secretion ◽  
Drug Induced ◽  
Cytoplasmic Vacuoles ◽  
Transmission Electron ◽  
Liver Organoids

Drug-induced phospholipidosis (PL) is a storage disorder caused by the formation of phospholipid-drug complexes in lysosomes. Because of the diversity of PL between species, human cell-based assays have been used to predict drug-induced PL in humans. We established three-dimensional (3D) human liver organoids as described previously and investigated their liver characteristics through multiple analyses. Drug-induced PL was initiated in these organoids and in monolayer HepG2 cultures, and cellular changes were systemically examined. Organoids that underwent differentiation showed characteristics of hepatocytes rather than HepG2 cells. The organoids also survived under PL-inducing drug conditions for 48 h and maintained a more stable albumin secretion level than the HepG2 cells. More cytoplasmic vacuoles were observed in organoids and HepG2 cells treated with more potent PL-induced drugs, but to a greater extent in organoids than in HepG2 cells. Lysosome-associated membrane protein 2, a marker of lysosome membranes, showed a stronger immunohistochemical signal in the organoids. PL-distinctive lamellar bodies were observed only in amiodarone-treated organoids by transmission electron microscopy. Human liver organoids are thus more sensitive to drug-induced PL and less affected by cytotoxicity than HepG2 cells. Since PL is a chronic condition, these results indicate that organoids better reflect metabolite-mediated hepatotoxicity in vivo and could be a valuable system for evaluating the phospholipidogenic effects of different compounds during drug development.

Destruction of Actin Filaments by Osmium Tetroxide

1977 ◽  
Vol 35 ◽  
pp. 466-467
Author(s):  
P. Maupin-Szamier ◽  
T. D. Pollard
Keyword(s):  
Actin Filaments ◽  
Time Course ◽  
Osmium Tetroxide ◽  
Rabbit Muscle ◽  
Muscle Actin ◽  
Sodium Phosphate Buffer ◽  
Transmission Electron ◽  
The Difference ◽  
Rates Of Decay ◽  
Short Time

We have studied the destruction of rabbit muscle actin filaments by osmium tetroxide (OSO4) to develop methods which will preserve the structure of actin filaments during preparation for transmission electron microscopy.Negatively stained F-actin, which appears as smooth, gently curved filaments in control samples (Fig. 1a), acquire an angular, distorted profile and break into progressively shorter pieces after exposure to OSO4 (Fig. 1b,c). We followed the time course of the reaction with viscometry since it is a simple, quantitative method to assess filament integrity. The difference in rates of decay in viscosity of polymerized actin solutions after the addition of four concentrations of OSO4 is illustrated in Fig. 2. Viscometry indicated that the rate of actin filament destruction is also dependent upon temperature, buffer type, buffer concentration, and pH, and requires the continued presence of OSO4. The conditions most favorable to filament preservation are fixation in a low concentration of OSO4 for a short time at 0°C in 100mM sodium phosphate buffer, pH 6.0.

Scanning and Transmission Microscopy of Nematocyst Batteries in Epitheliomuscular Cells of Hydra

1971 ◽  
Vol 29 ◽  
pp. 410-411 ◽  
Author(s):  
Jane A. Westfall ◽  
S. Yamataka ◽  
Paul D. Enos
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Osmium Tetroxide ◽  
External Surface ◽  
Three Dimensional ◽  
Surface Structures ◽  
Transmission Microscopy ◽  
Transmission Electron ◽  
Freeze Dried ◽  
Scanning Electron

Scanning electron microscopy (SEM) provides three dimensional details of external surface structures and supplements ultrastructural information provided by transmission electron microscopy (TEM). Animals composed of watery jellylike tissues such as hydras and other coelenterates have not been considered suitable for SEM studies because of the difficulty in preserving such organisms in a normal state. This study demonstrates 1) the successful use of SEM on such tissue, and 2) the unique arrangement of batteries of nematocysts within large epitheliomuscular cells on tentacles of Hydra littoralis.Whole specimens of Hydra were prepared for SEM (Figs. 1 and 2) by the fix, freeze-dry, coat technique of Small and Màrszalek. The specimens were fixed in osmium tetroxide and mercuric chloride, freeze-dried in vacuo on a prechilled 1 Kg brass block, and coated with gold-palladium. Tissues for TEM (Figs. 3 and 4) were fixed in glutaraldehyde followed by osmium tetroxide. Scanning micrographs were taken on a Cambridge Stereoscan Mark II A microscope at 10 KV and transmission micrographs were taken on an RCA EMU 3G microscope (Fig. 3) or on a Hitachi HU 11B microscope (Fig. 4).

Cell Reactivity Pattern of the Guinea Pig Cecal Mucosa Evidenced by the ZIO Technique

1982 ◽  
Vol 40 ◽  
pp. 50-51
Author(s):  
Juan Mora-Galindo ◽  
Jorge Arauz-Contreras
Keyword(s):  
Guinea Pig ◽  
Phase Contrast ◽  
Osmium Tetroxide ◽  
Cell Types ◽  
Cell Reactivity ◽  
Transmission Electron ◽  
Neural Tissues ◽  
Maturation Stages ◽  
Zinc Iodide ◽  
Cecal Mucosa

The zinc iodide-osmium tetroxide (ZIO) technique is presently employed to study both, neural and non neural tissues. Precipitates depends on cell types and possibly cell metabol ism as well.Guinea pig cecal mucosa, already known to be composed of epithelium with cells at different maturation stages and lamina propria which i s formed by morphologically and functionally heterogeneous cell population, was studied to determine the pat tern of ZIO impregnation. For this, adult Guinea pg cecal mucosa was fixed with buffered 1.2 5% g 1 utara 1 dehyde before incubation with ZIO for 16 hours, a t 4°C in the dark. Further steps involved a quick sample dehydration in graded ethanols, embedding in Epon 812 and sectioning to observe the unstained material under a phase contrast light microscope (LM) and a transmission electron microscope (TEM).

Ultrastructural comparison of prolactin adenomas of pituitary in men and women

1988 ◽  
Vol 46 ◽  
pp. 346-347
Author(s):  
R.C. Caughey ◽  
U.P. Kalyan-Raman
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Phosphate Buffer ◽  
Pituitary Adenomas ◽  
Osmium Tetroxide ◽  
Secretory Granules ◽  
Uranyl Acetate ◽  
En Bloc ◽  
Men And Women ◽  
Transmission Electron

Prolactin producing pituitary adenomas are ultrastructurally characterized by secretory granules varying in size (150-300nm), abundance of endoplasmic reticulum, and misplaced exocytosis. They are also subclassified as sparsely or densely granulated according to the amount of granules present. The hormone levels in men and women vary, being higher in men; so also the symptoms vary between both sexes. In order to understand this variation, we studied 21 prolactin producing pituitary adenomas by transmission electron microscope. This was out of a total of 80 pituitary adenomas. There were 6 men and 15 women in this group of 21 prolactinomas.All of the pituitary adenomas were fixed in 2.5% glutaraldehyde, rinsed in Millonig's phosphate buffer, and post fixed with 1% osmium tetroxide. They were then en bloc stained with 0.5% uranyl acetate, rinsed with Walpole's non-phosphate buffer, dehydrated with graded series of ethanols and embedded with Epon 812 epoxy resin.

TEM comparison of osmium vs osmium with potassium ferricyanide secondary fixatives and the impact of secondary fixative temperature on tissue preservation/contrast quality

1996 ◽  
Vol 54 ◽  
pp. 910-911
Author(s):  
J. W. Horn ◽  
B. J. Dovey-Hartman ◽  
V. P. Meador
Keyword(s):  
Osmium Tetroxide ◽  
Potassium Ferricyanide ◽  
Electron Microscopic ◽  
Transmission Electron ◽  
Microscopic Evaluation ◽  
Cacodylate Buffer ◽  
Transmission Electron Microscopic ◽  
Glutaraldehyde Solution ◽  
Temperature Impact ◽  
The Impact

Osmium tetroxide (OsO4) is a universally used secondary fixative for routine transmission electron microscopic evaluation of biological specimens. Use of OsO4 results in good ultrastructural preservation and electron density but several factors, such as concentration, length of exposure, and temperature, impact overall results. Potassium ferricyanide, an additive used primarily in combination with OsO4, has mainly been used to enhance the contrast of lipids, glycogen, cell membranes, and membranous organelles. The purpose of this project was to compare the secondary fixative solutions, OsO4 vs. OsO4 with potassium ferricyanide, and secondary fixative temperature for determining which combination gives optimal ultrastructural fixation and enhanced organelle staining/contrast.Fresh rat liver samples were diced to ∼1 mm3 blocks, placed into porous processing capsules/baskets, preserved in buffered 2% formaldehyde/2.5% glutaraldehyde solution, and rinsed with 0.12 M cacodylate buffer (pH 7.2). Tissue processing capsules were separated (3 capsules/secondary fixative.solution) and secondarily fixed (table) for 90 minutes. Tissues were buffer rinsed, dehydrated with ascending concentrations of ethanol solutions, infiltrated, and embedded in epoxy resin.

Sign in / Sign up

Export Citation Format

Share Document