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Rat parathyroid phosphoproteins LC-MS/MS

ABSTRACT: The mechanisms of chronic kidney disease-associated secondary hyperparathyroidism are partially understood. In this project we aimed to gain new information and propose research hypotheses by proteome and phosphoproteome profiling of normal and hyperplastic rat parathyroid glands. Glands were microdissected from rats on normal control diet or CKD-inducing high adenine diet enriched with phosphorus to produce secondary hyperparathyroidism. Protein extracts were pooled from 12 glands (6 rats) for phosphoproteome profiling (3 normal and 3 CKD pools).

INSTRUMENT(S):

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Parathyroid Gland, Endocrine Gland

DISEASE(S): Chronic Kidney Disease

SUBMITTER: Iddo Ben-Dov

LAB HEAD: Iddo Zeev Ben-Dov

PROVIDER: PXD029401 | Pride | 2022-06-28

REPOSITORIES: Pride

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			Action	DRS
	Seq58185-90_Phos_QE2.raw	Raw
	Seq58191-6_Phos_QE2.raw	Raw
	Seq60296_QE2.raw	Raw
	Seq60297_QE2.raw	Raw
	Seq60298_QE2.raw	Raw

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Rat parathyroid glands LC-MS/MS

Project description:The mechanisms of chronic kidney disease-associated secondary hyperparathyroidism are partially understood. In this project we aimed to gain new information and propose research hypotheses by proteome and phosphoproteome profiling of normal and hyperplastic rat parathyroid glands. Glands were microdissected from rats on normal control diet or CKD-inducing high adenine diet enriched with phosphorus to produce secondary hyperparathyroidism. Protein extracts were pooled from 4 glands (2 rats) for proteome profiling (3 normal and 3 CKD pools), and from 12 glands (6 rats) for phosphoproteome profiling (2 normal and 2 CKD pools).

2022-06-28 | PXD029368 | Pride

microRNA profiling in normal and hyperplastic human and murine parathyroid glands

Project description:Secondary hyperparathyroidism (SHP) is a common complication of chronic kidney disease (CKD) and correlates with morbidity and mortality. In this study we profiled microRNAs (miRNA) in parathyroids from different experimental SHP models and uremic patients and studied the function of specific miRNA using antagonizing oligonucleotides (anti-miRs). miRNA profiles established by small RNA deep sequencing showed that human, rat and mouse parathyroids share the same most abundant miRNAs. Principal component analyses clearly segregated parathyroids from SHP rats from normal rats, based on their miRNA expression profiles. Similar findings were observed in hyperplastic parathyroids from CKD patients compared to normal parathyroids from patients without kidney disease. We identifed specific parathyroid miRNAs that were dysregulated in all experimental SHP models studied. let-7i was decreased and miR-141 and miR-148a were increased in the parathyroids of rats with prolonged CKD induced by an 8 w adenine high phosphorus diet. Down-regulation of let-7 by anti-miRs increased PTH secretion in normal and in CKD rats, as well as in parathyroid organ cultures. Anti-miR-148 prevented the increase in serum PTH in CKD rats and decreased secreted PTH in parathyroid organ cultures. Our findings characterized parathyroid miRNA profiles and demonstrated conservation of the abundant miRNAs in different species. The evolutionary conservation of abundant miRNAs and their regulation in SHP suggest that miRNAs are important for parathyroid function and the development of SHP. Down-regulation of let-7 and miR-148 affects PTH secretion in vivo and in vitro, suggesting a role for these miRNAs in SHP. We propose that let-7 restrains while miR-148 promotes PTH secretion. In CKD, the decrease in parathyroid let-7 and the increase in miR-148 miRNAs may contribute to the development of SHP.

2017-03-21 | GSE79727 | GEO

Parathyroid tissue transcriptome of control, untreated uremic and Cinacalcet-treated uremic rats at two timepoints

Project description:We investigate whether Cinacalcet treatment timed to the expression of its drug target; the Calcium-Sensing Receptor, could improve efficacy of treatment in secondary hyperparathyroidism. We found that proliferation rate of parathyroid glands of uremic rats was markedly reduced when Cinacalcet was administered in the early inactive phase compared to conventional timing early in the active phase. We use RNAseq of parathyroid tissue to investigate the possible mechanism behind this enhanced anti-proliferative effect.

2024-12-19 | GSE227079 | GEO

Human Parathyroid Gland: Normal vs. Adenoma

Project description:Transcriptional profiling of genes expressed in normal parathyroid glands (Normal) and adenomatous parathyroid glands from patients with primary hyperparathyroidism (PHPT). Stratagen Human Refernece RNA, pooled from 10 human cell lines, was used to identify genes, which are specifically expressed or suppressed in parathyroid tissue. Comparison between normal and tumor parathyroids shows genes that might be involved in hyperproliferation and PTH hypersecretion in tumors.

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Investigation of Resistant Starch effect on gut microbiome in 5/6 nephrectomy mouse model of Chronic Kidney Disease

Project description:Background. Resistant Starch (RS) improves CKD outcomes. In this report we study how RS modulates host-microbiome interactions in CKD by measuring changes in abundance of proteins and bacteria in the gut. In addition, we demonstrate RS-mediated reduction in CKD-induced kidney damage. Methods. Eight mice underwent 5/6 nephrectomy to induce CKD and 8 served as healthy controls. CKD and Healthy (H) groups were further split into those receiving RS (CKDRS, n=4; HRS, n=4) and those on normal diet (CKD, n=4, H, n=4). Kidney injury was evaluated by measuring BUN/creatinine and by most-mortem histopathological evaluation. Cecal contents were analyzed mass spectrometry-based metaproteomics. PEAKS Studio was used to identify peptides via de novo sequencing. A set of R/Bioconductor packages and in house written scripts was further used to infer bacteria present, to evaluate changes in proteins and bacterial abundances, and to perform statistical analysis and hierarchical clustering. Results. The 5/6 nephrectomy compromised kidney function as seen by an increase in creatinine and BUN. Representative photomicrographs of trichrome-stained kidney sections showed reduced tubulointerstitial injury in CKD mice fed HAM-RS2 diet comparing to CKD mice fed normal diet. Identified organisms and proteins point toward a higher population of butyrate-producing bacteria, and reduced abundance of mucin-degrading bacteria. Conclusion. Resistant starch slows the progression of chronic kidney disease. Gut barrier function through maintenance of the mucin barrier plays a role in RS-associated improvements in CKD phenotype. Resistant starch supplementation leads to the active bacterial proliferation and the reduction of harmful bacterial metabolites.

2020-10-19 | PXD019623 | Pride

Proteomics and phosphoproteomics of the diurnal cycle in chow and high fat high sucrose feed rats

Project description:There is growing evidence that energy metabolism and insulin action are regulated by mechanisms that follow a diurnal rhythm and it has been proposed that defects in Akt signalling are associated with the pathophysiology of metabolic disease. It is therefore important to investigate these parameters under physiology of the free-living state. We therefore examined the insulin action in muscle of chow or high fat, high sucrose diet-fed (HFHS) rats during the normal diurnal cycle. HFHS animals displayed hyperinsulinemia, however had reduced systemic glucose disposal and impaired muscle glucose uptake during the feeding period. Proteomics and phosphoproteomics was performed over the diurnal cycle in chow and HFHS rats.

2019-05-20 | PXD010242 | Pride

LOESS-Corrected UMI Counts Enable Absolute RNA Quantification in Single-Cell Transcriptomics

Project description:The parathyroid gland is one of the endocrine glands in the body and plays a key role in regulating calcium and phosphate homeostasis through the production and release of parathyroid hormone (PTH). Despite its physiological importance, research on the parathyroid gland remains limited due to its small size and concealed anatomical location. Previous studies have suggested that the parathyroid gland consists of principal cells, acidophilic cells, and other cell types. While recent studies have applied single-cell sequencing to investigate the rat parathyroid gland, a comprehensive analysis of cell subpopulations, gene expression differences, and rare cell types is still needed. Therefore, this study aims to further utilize single-cell sequencing technology to analyze the transcriptional profiles of different cell types in the normal rat parathyroid gland, refine the characterization of cellular diversity, and provide additional insights into the molecular landscape of the rat parathyroid gland.

2025-07-01 | GSE291906 | GEO

CKD Rats

Project description:CKD Rats

| PRJNA1086545 | ENA

microRNA profiling in normal and hyperplastic human and murine parathyroid glands

Project description:microRNA profiling in normal and hyperplastic human and murine parathyroid glands

| PRJNA316852 | ENA

Metaproteomics of the Resistant Starch-fed Chronic Kidney Disease Rats

Project description:Background- Resistant starch is a prebiotic metabolized by the gut bacteria. It has been shown to attenuate chronic kidney disease (CKD) progression in rats. Previous studies employed taxonomic analysis using 16S rRNA sequencing and untargeted metabolomics profiling. Here we expand these studies by metaproteomics, gaining new insight into the host-microbiome interaction. Methods- Differences between cecum contents in CKD rats fed a diet containing resistant starch with those fed a diet containing digestible starch were examined by comparative metaproteomics analysis. Taxonomic information was obtained using unique protein sequences. Our methodology results in quantitative data covering both host and bacterial proteins. Results - 5,834 proteins were quantified, with 947 proteins originating from the host organism. Taxonomic information derived from metaproteomics data surpassed previous 16S RNA analysis, and reached species resolutions for moderately abundant taxonomic groups. In particular, the Ruminococcaceae family becomes well resolved – with butyrate producers and amylolytic species such as R. bromii clearly visible and significantly higher while fibrolytic species such as R. flavefaciens are significantly lower with resistant starch feeding. The observed changes in protein patterns are consistent with fiber-associated improvement in CKD phenotype. Several known host CKD-associated proteins and biomarkers of impaired kidney function were significantly reduced with resistant starch supplementation. Conclusions- Metaproteomics analysis of cecum contents of CKD rats with and without resistant starch supplementation reveals changes within gut microbiota at unprecedented resolution, providing both functional and taxonomic information. Proteins and organisms differentially abundant with RS supplementation point toward a shift from mucin degraders to butyrate producers.

2018-12-18 | PXD008845 | Pride

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