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Podocytes

Podocytes are specialized epithelial cells located in the glomeruli of the kidney.
They play a critical role in the filtration barrier, responsible for preventing the loss of essential proteins and molecules from the bloodstream.
Podocytes have a complex and highly dynamic structure, with interdigitating foot processes that form the slit diaphragm.
Dysfunction or injury to podocytes can lead to proteinuria and various glomerular diseases, making them a key target for research and therapeutic interventions.
Understading the biology and pathology of podocytes is crucial for advancing our knowledge of kidney physiology and developing effective treatments for kidney disorders.

Most cited protocols related to «Podocytes»

Generating Chimeric Mouse Models

Chimeric mice were generated by aggregation of targeted ES-cells with eight-cell stage embryos as described previously (16 ). Germline transmitting chimeric males were crossed with outbred ICR females and the offspring were genotyped by Southern blotting and PCR for the transmission of the transgene.
The Cre-recombinase transgenic founder lines were generated as described previously (17 (link)). Regarding the Podocin-Cre transgene four individual founder lines were crossed with the Z/EG reporter strain (15 (link)) to determine the degree and timing of Cre-mediated DNA excision in podocytes.

Belteki G., Haigh J., Kabacs N., Haigh K., Sison K., Costantini F., Whitsett J., Quaggin S.E, & Nagy A. (2005). Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction. Nucleic Acids Research, 33(5), e51.

Publication 2005

Animals, Transgenic Cell Aggregation Cells Chimera Cre recombinase Embryo Females Germ Line Males Mice, Laboratory NPHS2 protein Podocytes Strains Transgenes Transmission, Communicable Disease

Subcellular Fractionation of Podocytes

SEC was performed in podocytes or isolated mouse glomeruli as described previously ^{15 (link)}. For a detailed method, please see the online Data supplement.
All other methods are described in the online Data Supplement at http://hyper.ahajournals.org.

Zhang C., Boini K.M., Xia M., Abais J.M., Li X., Liu Q, & Li P.L. (2012). ACTIVATION OF NALP3 INFLAMMASOMES TURNS ON PODOCYTE INJURY AND GLOMERULAR SCLEROSIS IN HYPERHOMOCYSTEINEMIA. Hypertension, 60(1), 154-162.

Publication 2012

Dietary Supplements Kidney Glomerulus Mice, House Podocytes

Isolation and Characterization of Rat Kidney Glomeruli

For the successful isolation of rat kidneys, male rats of approximately 8 weeks old are used. The kidneys are fl ushed with PBS to clean out blood and urine. In addition to the electrophysiological measurements described below, this procedure may be useful for several other applications, e.g., immunohistochemical, biochemical, or molecular analysis of the tissue, etc. Figures 1 and 2 demonstrate steps required to isolate the glomeruli. Figure 3 shows the electron microscopy of the freshly isolated glomerulus fixed with 2% glutaraldehyde buffered in 0.1 M cacodylate (pH 7.4). As seen on these images acquired by the transmission electron microscopy, a normal configuration of podocytes with coordinated pattern of foot processes was observed (Fig. 3). With the help of the patch clamp technique, we have examined the biophysical properties of native TRPC channels in the podocytes of the freshly isolated decapsulated glomeruli. Our results demonstrate that it is possible to study functional channels in these intact glomeruli. The use of specific inhibitors within the patch pipette together with the composition of the pipette and bath solutions and the parameters of the current–voltage dependencies allowed distinguishing two channel types with distinct TRPC properties. However the development of this approach and identi fi cation of the precise composition of TRPC channels in podocytes require additional studies.

Ilatovskaya D.V, & Staruschenko A. (2013). Single-Channel Analysis of TRPC Channels in the Podocytes of Freshly Isolated Glomeruli. Methods in molecular biology (Clifton, N.J.), 998, 355-369.

Publication 2013

Bath BLOOD Cacodylate Electron Microscopy Foot Glutaral inhibitors Kidney Kidney Glomerulus Males Podocytes Rattus Tissues Transmission Electron Microscopy Urine Vision

Podocyte-Specific Gene Expression in Mice

The genome-wide expression data of murine podocytes, mesangial, and endothelial cells were obtained from http://www.gudmap.org/. The identifications of data sets that have been utilized in our study are listed in Supplemental Table 5. The detailed protocol of data generation is described in a recently published paper by Brunskill et al. (2011) (link) and can also be found in our Supplemental Methods. We preprocessed and normalized data as described in Microdissected Human Kidney Data. By comparing the expression level in podocytes versus the other two major cell types in glomeruli, we define a gene to be podocyte-specific if its expression in podocytes is 4.76-fold over mesangial cells and 4.65-fold over glomerular capillary endothelial cells. The cut-off values represent three standard deviations of the average difference between podocytes and mesangial/endothelial cell transcripts, respectively. By use of HomoloGene from NCBI Entrez (Maglott et al. 2011 (link)), 102 murine genes could be mapped to their Homo sapiens ortholog (Supplemental Table 6).

Ju W., Greene C.S., Eichinger F., Nair V., Hodgin J.B., Bitzer M., Lee Y.S., Zhu Q., Kehata M., Li M., Jiang S., Rastaldi M.P., Cohen C.D., Troyanskaya O.G, & Kretzler M. (2013). Defining cell-type specificity at the transcriptional level in human disease. Genome Research, 23(11), 1862-1873.

Publication 2013

Capillary Endothelial Cells Cells Endothelial Cells Endothelium Genes Genome Homo sapiens Kidney Kidney Glomerulus Mesangial Cells, Kidney Mesangiums, Glomerular Mus Podocytes

Immortalized Podocyte Cell Lines

Adult podocyte cell lines were cloned by limiting dilution from glomerular cultures of the “immortomouse” (Jat et al., 1991 (link)) harboring a transgene for a thermosensible variant of the SV-40-T–antigen, which is under the control of the H-2K^b-promotor, whose activity can be increased by γ interferon. The cellular origin of the cloned cell lines from the podocyte lineage was proven by the detection of the Wilm's tumor gene product (WT-1), which in the adult kidney is an exclusive marker of podocytes (Mundlos et al., 1993 (link)). Cells were propagated at 33°C in RPMI 1640 containing 10% FCS (Boehringer Mannheim, Mannheim, Germany), 100 U/ml penicillin (GIBCO BRL, Karlsruhe, Germany), 100 μg/ml streptomycin (GIBCO BRL), and 10 U/ml recombinant mouse γ interferon (Sigma Chemical Co., Munich, Germany) to induce synthesis of the immortalizing T antigen. Subcultivation was done with trypsin at 37°C after cells had reached confluence. Cells were passaged after 1:5 dilution. To initiate differentiation, cells were thermoshifted to 37°C and maintained in medium without γ interferon (Mundel et al., 1997 ).

Mundel P., Heid H.W., Mundel T.M., Krüger M., Reiser J, & Kriz W. (1997). Synaptopodin: An Actin-associated Protein in Telencephalic Dendrites and Renal Podocytes. The Journal of Cell Biology, 139(1), 193-204.

Publication 1997

Adult Antigens, Synthetic Cell Lines Cells Interferon Type II Kidney Kidney Glomerulus Mus Penicillins Podocytes Simian virus 40 Streptomycin Technique, Dilution Transgenes Trypsin Viral Tumor Antigens WT1 Proteins

Most recents protocols related to «Podocytes»

Immunoperoxidase Staining of ET-1 in Kidney Tissue

Not available on PMC !

Example 3

As shown in FIG. 2, immunoperoxidase staining for ET-1 is negative in kidney tissue from a donor nephrectomy (panels a and b). A kidney biopsy from a patient with FSGS (panels c and d) exhibits increased content of ET-1 in parietal epithelium lining Bowman's space (large arrow in panel d), as well as visceral epithelial cells overlying a segmental scar (smaller arrow in panel d). Surrounding tubules also show increased cytoplasmic expression of ET-1. A kidney biopsy from a patient infected with HIV (panels e and f) shows increased expression of ET-1 in tubules, similar to that in the FSGS sample. In contrast to FSGS, however, glomerular epithelial cells evidence only trace amounts of the protein.

US11874283B2. Method and compositions for the treatment and detection of endothelin-1 related kidney diseases (2024-01-16). MOREHOUSE SCHOOL OF MEDICINE [US]. Inventors: Gale W. Newman [US], James W. Lillard, Jr. [US], Chamberlain Obialo [US].

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Patent 2024

Biopsy Cicatrix Cytoplasm Epithelial Cells Epithelium Kidney Kidney Glomerulus Nephrectomy Patients Podocytes Proteins Tissue Donors Tissues

Adriamycin-Induced Podocyte Injury Model

Mouse kidney podocytes MPC-5 (CTCC-400-0327) purchased from Meisen Chinese Tissue Culture Collections (China) was maintained in RPMI-1640 medium (PM150110, Procell, China) containing 10% fetal bovine serum (164210-50, Procell, China) and 1% penicillin-streptomycin solution (PB180120, Procell, China) at 37 °C with 5% CO₂.
To mimic FRNS in vitro, MPC-5 cells were stimulated with 10 µM adriamycin (ADR, T1020, TopScience, China) for 6 h (h).

Wang H., Shi J., Tang B., Liu Y, & Wang Q. (2023). Forecast and verification of the active compounds and latent targets of Guyuan decoction in treating frequently relapsing nephrotic syndrome based on network pharmacology. Renal Failure, 45(1), 2184654.

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Publication 2023

Adriamycin Cells Chinese Fetal Bovine Serum Fryns syndrome Kidney Mus Penicillins Podocytes Streptomycin

Comprehensive Renal Biopsy Evaluation

All of the patients underwent a renal biopsy before treatment. Each biopsy specimen was evaluated by light microscopy, direct immunofluorescence, and electron microscopy. Paraffin-embedded sections (3 μm in thickness) stained with Periodic acid–Schiff (PAS), Periodic acid–silver metheramine (PASM), and Masson were observed by light microscopy. The number of sclerotic glomeruli and crescents was presented as the percentage of the total number of glomeruli in the biopsy specimen. The chronic tubulointerstitial injury was graded as follows: grade 0 (none), grade 1 (below 25% of tubulointerstitial tissue affected), grade 2 (25% to 50% of tubulointerstitial tissue affected), grade 3 (above 50% of tubulointerstitial tissue affected). In addition, the acute tubulointerstitial injury was graded 0 (absent) or 1 (present). Glomerular lesions were assigned one of four stages, from I to IV, in accordance with Ehrenreich and Churg’s classification criteria [26 ]. Frozen sections (4 μm in thickness) were used for direct immunofluorescence to detect IgG, IgA, IgM, C3, C1q, and PLA2R. The fluorescence intensity was described by a semiquantitative method as follows: 0 (none), 1 (+), 2 (2+), 3 (3+ to 4+). Transmission electron microscopy was performed using ultrathin sections embedded in resin and stained with lead citrate and uranium acetate in order to observe the ultrastructure. In each measurement, GBM thickness was defined by the length between the endothelial cell and podocyte membrane. This included intramembranous immune deposits. The highest GBM thickness was routinely measured on at least five glomerular capillary loops and reviewed, and the mean value was obtained. Pathologists who measured GBM thickness were blinded to the patient’s clinical data and outcome. Iteratively, a review of any scoring differences between the two pathologists was performed.

Duan S., Sun L., Zhang C., Zeng M., Sun B., Yuan Y., Mao H., Xing C, & Zhang B. (2023). The thickness of glomerular basement membrane predicts complete remission in primary membranous nephropathy. Renal Failure, 45(1), 2179335.

Publication 2023

Biopsy Capillaries Citrate Electron Microscopy Endothelial Cells Fluorescence Frozen Sections Immunofluorescence, Direct Injuries Kidney Kidney Glomerulus Light Microscopy Paraffin Pathologists Patients Periodic Acid PLA2R1 protein, human Podocytes Resins, Plant Sclerosis Silver Tissue, Membrane Tissues Transmission Electron Microscopy uranyl acetate

Generating Podocyte-Specific GSK-3β-KO Mice

Mice were purchased from the Jackson Laboratory (GSK-3βFL, Stock No 029592; NPHS-Cre, Stock No 008205), and the podocyte-specific GSK-3β-KO mice were generated as described previously [35 (link)]. The grouping and treatment protocols used in the aforementioned lncRNA Dlx6-os1 overexpression study with the db/m mice were adapted for this experiment.

Guo J., Zheng W., Liu Y., Zhou M., Shi Y., Lei M., Zhang C, & Liu Z. (2023). Long non-coding RNA DLX6-AS1 is the key mediator of glomerular podocyte injury and albuminuria in diabetic nephropathy by targeting the miR-346/GSK-3β signaling pathway. Cell Death & Disease, 14(2), 172.

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Publication 2023

Hydrocephalus, Normal Pressure Mice, Laboratory Podocytes RNA, Long Untranslated Treatment Protocols

Dlx6-os1 Overexpression and Knockdown in Podocytes

The lncRNA Dlx6-os1 (full length)-overexpressing, specific knockdown shRNA, and control adenovirus (Supplementary Methods and Materials) used in this study were designed and synthesized by Hanbio Co., Ltd. (Shanghai, China). Podocytes were seeded in six-well plates at 60–70% confluence and infected with adenovirus following the manufacturer’s instructions (4 × 10⁶ podocytes; MOI = 100). The media was replaced with fresh ones after 6–8 h of viral infection. Mouse miR-346 mimics and mutants were provided by Bo Rui Biotech. Co., Ltd. (Guangzhou, China), and transfected into cells in the presence of Lipofectamine 3000 (Thermo Fisher Scientific, CA, USA). The lentivirus carrying a mouse Dlx6-os 1 fragment (exon 3 of lncRNA dlx6-os1 containing binding sites for miR-346) and the lentivirus with specific shRNA to induce knockdown were also constructed by Hanbio Co., Ltd. (Shanghai, China). The information on the vectors is listed in the Supplementary Methods and Materials section. The lentivirus was used to treat db/m and db/db mice via tail vein injections (one single injection: virus titer 10⁸ TU/ml; 80–100 µl per mouse). The sequences of the adenovirus and lentivirus packaging fragments are listed in the Supplementary Methods and Materials. Both the overexpression and shRNA lentiviruses carried the promoter for podocin (nphs-2), a podocyte marker protein. The control lentivirus was HA-tagged and also carried the promoter for podocin (nphs-2). The transfection efficiency of lentivirus with a podocyte-specific promoter in the podocytes of glomeruli was analyzed by immunofluorescence staining showing the expression of HA-tag and podocyte marker protein podocin in the OCT-embedded frozen kidney tissues.

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Publication 2023

Adenovirus Vaccine Binding Sites Cells Cloning Vectors Exons Freezing Hydrocephalus, Normal Pressure Immunofluorescence Kidney Kidney Glomerulus Lentivirus Lipofectamine MIRN346 microRNA, mouse Mus NPHS2 protein Podocytes Proteins RNA, Long Untranslated Short Hairpin RNA Tail Tissues Transfection Veins Virus Diseases

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More about "Podocytes"

Podocytes are specialized epithelial cells found in the glomeruli of the kidney.
They play a crucial role in the filtration barrier, preventing the loss of essential proteins and molecules from the bloodstream.
These highly dynamic cells have a complex structure, with interdigitating foot processes that form the slit diaphragm.
Podocyte dysfunction or injury can lead to proteinuria and various glomerular diseases, making them a key target for research and therapeutic interventions.
Understanding the biology and pathology of podocytes is essential for advancing our knowledge of kidney physiology and developing effective treatments for kidney disorders.
Researchers often utilize cell culture techniques, such as RPMI 1640 medium, Lipofectamine 2000, TRIzol reagent, and Penicillin/streptomycin, to study podocytes in vitro.
These tools help uncover the mechanisms underlying podocyte function and dysfunction, as well as explore potential therapeutic strategies.
Podocyte-related research is crucial for understanding conditions like glomerulonephritis, nephrotic syndrome, and diabetic nephropathy.
By leveraging AI-driven platforms like PubCompare.ai, scientists can optimize their research by locating the most effective protocols from literature, preprints, and patents, ultimately accelerating scientific discoveries and unlocking new insights into this vital cell type.
Key areas of focus include podocyte structure, signaling pathways, injury response, and novel therapeutic targets.