Human Podocytes Develop “Names” to Practice Self-Avoidance
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ABSTRACT:
PROVIDER: PRJNA939188 | ENA |
REPOSITORIES: ENA
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Project description:Podocyte effacement and loss characterizes glomerulopathies such as diabetic nephropathy, lupus, and glomerular toxicity. Human primary podocyte-like epithelial cells cultured from urine (ECU) were characterized as a window to understand podocyte regeneration in these glomerulopathies. Cytokines TNF, and VEGF stimulated regrowth, whereas FGF-1 and IL1 inhibited growth, consistent with other podocyte or parietal epithelial cell models. Review of published micrographs indicated that podocyte foot processes in rodent kidneys interdigitated exclusively with foot processes from other cells, suggesting podocytes practice self-avoidance. In culture, ECU avoided close interactions with other cells arising from the same clone, until enough cells grew for the culture to become confluent. In contrast, ECU derived from multiple clones associated actively to form contiguous monolayers. Gene expression profiling of eight clonal ECU colonies using RNA sequencing revealed that each colony expressed a highly variable profile of the 53 Protocadherin genes. Variable combinations of the 53 Protocadherin PCDHA-, PCDHB-, and PCDHG- gene products are known to act homophilically to direct self-avoidance interactions in human neurons, so PCDH- genes are postulated to govern self-avoidance similarly in podocytes. Such protocadherin profiles could be considered cell names. Transcriptional profiles of ECUs suggest they may represent intermediate stages of Parietal Epithelial Cells (PECs) differentiating into podocytes. They further suggest that each podocyte practices self-avoidance, interdigitating selectively with other cells in the glomerulus. To enable further investigation of multi-clonal podocyte interactions, five podocyte clones were immortalized using temperature-sensitive SV40 Large T antigen. These results suggest that non-self interdigitation of new podocytes with remnant podocytes may facilitate replacement of dying podocytes and re-establishment of a functional filtration barrier.
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Project description:Allergen exposure was thought to play a critical role in the etiology of AR. And allergen avoidance, the practice of avoiding exposure to allergens, has been generally advised as the management of AR. However, the effect is uncertain and the underlying mechanism is far from known. We used gene expression microarrays to identify genes differentially regulated by allergen avoidance in allergic rhinitis mouse model. Affymetrix Mouse Gene 1.0 ST arrays were used to identify the expression profiling of nasal mucosa in three groups of mice: (1) mice sensitized and challenged with saline (control group); (2) mice sensitized and challenged with ovalbumin (OVA) and sacrificed 2 hours after the last challenge (OVA group); (3) mice sensitized and challenged with OVA and sacrificed 4 weeks after the last challenge (4w-after group).
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Project description:Purpose: Next-generation sequencing (NGS) was used to define the transcriptome of native mouse podocytes and non-podocytes glomerular cells as part of a project aiming to define the molecular fingerprint of mouse podocytes. Method: Glomeruli from 29 Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J x hNPHS2Cre mice at the age of 10 weeks were purified and a single cell solution was prepared to seperate GFP-expressing (podocytes) and GFP-negative (non-podocytes glomerular cells) cells by FACS sorting. RNA was extracted and prepared for further analysis using directional, polyA+ library preparation. An Illumina HiSeq2500 was used for a paired-end sequencing of 100 cycles . Salmon and Sleuth were used for downstream analysis. Results: A total of 100 Million reads each from podocytes and non-podocytes glomerular cells could be used for further analysis.
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Project description:Comparisons of hippocampi of Syracuse High Avoidance (SHA) and Syracuse Low Avoidance (SLA) learning rats Keywords: other
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