Nature Protocols, Journal Year: 2020, Volume and Issue: 16(1), P. 239 - 262
Published: Nov. 27, 2020
Language: Английский
iScience, Journal Year: 2021, Volume and Issue: 24(4), P. 102258 - 102258
Published: March 2, 2021
The fundamental morphology of the endometrial glands is not sufficiently understood by 2D observation because these have complicated winding and branching patterns. To construct a large picture gland structure, we performed tissue-clearing-based 3D imaging human uterine tissue. Our immunohistochemistry layer analyses revealed that form plexus network in stratum basalis expand horizontally along muscular layer, similar to rhizome grass. We then extended our method assess tissue affected adenomyosis, representative "endometrium-related disease," observed its morphological features, including direct invasion into myometrium an ant colony-like ectopic within myometrium. Thus, further understanding endometrium based on analysis will lead identification pathogenesis endometrium-related diseases.
Language: Английский
Citations
92
iScience, Journal Year: 2021, Volume and Issue: 24(3), P. 102178 - 102178
Published: Feb. 16, 2021
Advanced optical methods combined with various probes pave the way toward molecular imaging within living cells. However, major challenges are associated need to enhance resolution even further subcellular level for of larger tissues, as well in vivo studies. High scattering and absorption opaque tissues limit penetration light into deep thus depth. Tissue clearing technique provides an innovative perform deep-tissue imaging. Recently, have been developed, which provide tissue based on similar physical principles via different chemical approaches. Here, we introduce mechanisms current from fundamental perspectives, including main principle, refractive index matching approaches, such dissociation collagen, delipidation, decalcification, dehydration, hyperhydration, reduce scattering, decolorization absorption.
Language: Английский
Citations
89
Kidney International, Journal Year: 2019, Volume and Issue: 97(5), P. 934 - 950
Published: Dec. 25, 2019
Hypoxia-inducible factor (HIF) prolyl hydroxylase inhibitors, also known as HIF stabilizers, increase endogenous erythropoietin production and serve novel therapeutic agents against anemia in chronic kidney disease. induces the expression of various genes related to energy metabolism an adaptive response hypoxia. However, it remains obscure how metabolic reprogramming renal tissue by stabilization affects pathophysiology diseases. Previous studies suggest that systemic disorders such hyperglycemia dyslipidemia cause alterations metabolism, leading dysfunction including diabetic Here, we analyze effects enarodustat (JTZ-951), oral stabilizer, on early stages disease, using streptozotocin-induced rats alloxan-induced mice. Transcriptome analysis revealed counteracts metabolism. showed fatty acid amino metabolisms were upregulated downregulated enarodustat, whereas glucose was upregulated. These symmetric changes confirmed metabolome analysis. Whereas glycolysis tricarboxylic cycle metabolites accumulated acids reduced animals, these disturbances mitigated enarodustat. Furthermore, increased glutathione disulfide ratio relieved oxidative stress animals. Thus, occurring incipient see commentary page 855 Translational StatementHypoxia-inducible inhibitors (also stabilizers) Our transcriptome analyses rat mouse models have The results provide important data for extrapolating stabilizers clinical settings, although further are needed clarify this progression disease.1Hasegawa S. Tanaka T. Nangaku M. treating disease.Curr Opin Nephrol Hypertens. 2018; 27: 331-338Crossref PubMed Scopus (30) Google Scholar Cells endowed with a defensive mechanism hypoxia, is master regulator defense.2Maxwell P. HIF-1: oxygen system special relevance kidney.J Am Soc Nephrol. 2003; 14: 2712-2722Crossref (118) Scholar,3Marx J. Cell biology: cells endure low oxygen.Science. 2004; 303: 1454-1456Crossref (69) Kidneys physiologically exposed hypoxia recognized final common pathway end-stage disease.4Nangaku Chronic tubulointerstitial injury: failure.J 2006; 17: 17-25Crossref (877) Scholar,5Tanaka Miyata Inagi R. et al.Hypoxia disease proteinuria and/or glomerular hypertension.Am J Pathol. 165: 1979-1992Abstract Full Text PDF (89) Considering responses, might pleiotropic diseases well improvement Interestingly, glycolytic pyruvate dehydrogenase kinase 1, which inhibits from fuel mitochondrial (TCA) cycle.6Kim J.W. Tchernyshyov I. Semenza G.L. al.HIF-1-mediated kinase: switch required cellular adaptation hypoxia.Cell Metab. 3: 177-185Abstract (2593) Scholar,7Papandreou Cairns R.A. Fontana L. al.HIF-1 mediates actively downregulating consumption.Cell 187-197Abstract (1614) This TCA represses consumption critical hypoxic environments. Diabetic (DKD) major disease.8Thomas M.C. Cooper M.E. Zimmet Changing epidemiology type 2 diabetes mellitus associated disease.Nat Rev 2016; 12: 73-81Crossref (314) Systemic alterations, DKD.9Hasegawa Jao T.M. Dietary disease.Nutrients. 2017; 9: 358Crossref (28) shown flux accumulation cortical tissue,10Sas K.M. Kayampilly Byun al.Tissue-specific drives nutrient complications.JCI Insight. 1e86976Crossref (142) Scholar,11Tanaka Sugiura Y. Saito H. al.Sodium-glucose cotransporter inhibition normalizes suppresses kidneys mice.Kidney Int. 94: 912-925Abstract be DKD progression.12Sharma K. Mitochondrial kidney.Adv Exp Med Biol. 982: 553-562Crossref We hypothesized reverse tissue, considering HIF, reduces repress consumption. utilizing analyses, conducted proof-of-concept study comprehensively understand (JTZ-951),13Ogoshi Matsui Mitani al.Discovery JTZ-951: inhibitor treatment anemia.ACS Chem Lett. 8: 1320-1325Crossref (38) Scholar,14Akizawa Yamaguchi al.A placebo-controlled, randomized trial patients followed long-term trial.Am 2019; 49: 165-174Crossref (51) DKD. As cortex mainly composed proximal tubules, first examined tubule vitro. First, Mito Stress Test (Agilent Technologies, Inc., Santa Clara, CA) Glycolytic Rate Assay (Agilent) cultured HK-2 cells,15Ryan M.J. Johnson G. Kirk al.HK-2: immortalized epithelial cell line normal adult human kidney.Kidney 1994; 45: 48-57Abstract (721) (Figure 1a). Enarodustat significantly respiration (TCA cycle) basal glycolysis, indicating 1; Supplementary Figure S1). experiment small interfering RNA (siRNA) HIF-1 2; S2). knockdown siRNA reversed (basal respiration, maximal spare respiratory capacity, adenosine triphosphate [ATP] production) induced through Pyruvate activity, use cycle, 2f), compatible previously published observations.6Kim ScholarFigure 2Metabolic occurs hypoxia-inducible factor-1 (HIF-1) stabilization. (a) Study protocols shown. (b) Western blotting protein at time successfully types small, (siHIF-1; #1 #2). (c) O2 rates (OCR) measured real under conditions indicated (oligomycin, carbonyl cyanide-p-trifluoromethoxyphenylhydrazone [FCCP], rotenone + antimycin A [Rot/AA]). (d,e) OCR decrease (n = 18 each group, ∗∗∗∗P < 0.0001). experiments another siHIF-1 (#2) same result (see (f) (PDH) activities PDH activity 8, ∗∗P 0.01, All expressed mean ± SD. μM, μmol/l; siNC, negative control. To optimize viewing image, please online version article www.kidney-international.org.View Large Image ViewerDownload Hi-res image Download (PPT) From vitro experiments, alleviate glycolysis. chose streptozotocin (STZ)-induced model test above-mentioned hypothesis. In model, rapidly induced, allowing us observe net within short period time. protocol basic STZ-induced 3a. divided into 3 groups: group (sham), B (DKD), C (DKD enarodustat). Blood plasma glucose, glycosylated hemoglobin HbA1c, triglyceride, total cholesterol levels day 14 groups compared A, there no significant differences between 3d–g). Although creatinine not different groups, urinary albumin excretion glomerulomegaly noticeable tended 4). urea nitrogen higher reflecting dehydration due diabetes. summary, STZ-treated our represent rats.Figure 4Basic parameters pathologies rats. (BUN), (Cre), (d) area pathological images markedly B, periodic acid–Schiff staining images). (e) Representative (Upper) Periodic images. Bar 100 μm. (Lower) Electron microscopy Glomerular basement membrane thickening B. 1 For multiplex comparisons, 1-way variance applied, Tukey multiple comparisons test, if appropriate (∗P 0.05, Figures 5 6. Principal component hierarchical clustering separated clusters, respectively 5a b). Differentially selected |log2 fold-change (FC)| ≥ 0.5 Q value 0.05. Gene ontology canonical fatty-acid A. contrast, network 5c d).Figure 6Gene set enrichment (GSEA) data. GSEA (diabetes vs. enarodustat): amino-acid (diabetic [DKD]) (sham). they downregulated, enarodustat) Detailed Tables 2.View gene transcriptomics 6, 2). sets 6). showing Moreover, (Table 2), notion enarodustat-induced tubules observed study. counteracted transcriptomic perspectives.Table 1Results (DKD)/A (sham)KEGG (FDR 0.25)Gene setsNES-Log10 (FDR)KEGG_PPAR signaling pathway2.26>10KEGG_Complement coagulation cascades2.22>10KEGG_Drug cytochrome P4502.002.3KEGG_Metabolism xenobiotics P4501.982.3KEGG_P53 pathway1.821.4KEGG_Porphyrin chlorophyll metabolism1.791.3KEGG_Bladder cancer1.660.9KEGG_Tyrosine metabolismaGene metabolism.1.620.8KEGG_ABC transporters1.610.8KEGG_Adipocytokine pathway1.590.8KEGG_Biosynthesis unsaturated acidsaGene metabolism.1.580.8KEGG_Drug other enzymes1.560.8KEGG_Fatty-acid metabolism.1.560.8KEGG_Prion diseases1.550.8KEGG_Alanine, aspartate, glutamate metabolism.1.520.7KEGG_Cytokine–cytokine receptor interaction1.510.7KEGG_Valine, leucine, isoleucine degradationaGene metabolism.1.510.7KEGG_Chronic myeloid leukemia1.480.7KEGG_Cell cycle1.470.7KEGG_ErbB pathway1.440.6KEGG_Starch sucrose metabolism.1.440.7KEGG_Peroxisome1.430.7KEGG_Toll-like pathway1.430.7KEGG_Primary immunodeficiency1.420.7KEGG_Glutathione metabolism.1.400.6KEGG_O-glycan biosynthesis−1.660.7KEGG_Selenoamino metabolism.−1.740.7ABC, binding cassette; DKD, disease; FDR, false discovery rate; GSEA, analysis; KEGG, Kyoto Encyclopdia Genes Genomes; NES, normalized score; PPAR, peroxisome proliferator activated receptor.Gene FDR 0.25 raised table. NES > 0 those (sham).a Open table new tab Table 2Results enarodustat)/B (DKD)KEGG setsNES−Log10 (FDR)KEGG_Pentose phosphate pathwayaGene metabolism.2.22>10KEGG_RIG-I–like pathway1.881.4KEGG_Glycolysis, gluconeogenesisaGene metabolism.1.881.6KEGG_Fructose mannose metabolism.1.791.3KEGG_Primary immunodeficiency1.761.3KEGG_DNA replication1.731.3KEGG_Systemic lupus erythematosus1.701.2KEGG_Complement cascades1.661.1KEGG_Cytosolic DNA-sensing pathway1.641.1KEGG_Prion diseases1.601.0KEGG_P53-signaling pathway1.581.0KEGG_Galactose metabolism.1.581.0KEGG_Cytokine–cytokine interaction1.541.0KEGG_Hematopoietic lineage1.541.0KEGG_Toll-like pathway1.521.0KEGG_Cell cycle1.490.9KEGG_Cell adhesion molecules (CAMs)1.410.7KEGG_Progesterone-mediated oocyte maturation1.360.6KEGG_Proximal bicarbonate reclamation–1.470.8KEGG_Retinol metabolism–1.470.8KEGG_Butanoate metabolism–1.490.8KEGG_Spliceosome–1.490.8KEGG_Histidine metabolism.–1.490.8KEGG_Fatty-acid metabolism.–1.500.8KEGG_Tyrosine metabolism.–1.581.1KEGG_One carbon pool folateaGene metabolism.–1.591.1KEGG_Alanine, metabolism.–1.611.1KEGG_Peroxisome–1.641.2KEGG_RNA polymerase–1.641.2KEGG_Propanoate metabolism–1.661.2KEGG_Oxidative phosphorylationaGene metabolism.–1.661.2KEGG_Steroid hormone biosynthesis–1.711.4KEGG_Metabolism P450–1.751.6KEGG_Pyruvate metabolism.–1.781.7KEGG_Glycine, serine, threonine metabolism.–1.801.8KEGG_Valine, metabolism.–1.821.8KEGG_Citrate cycle)aGene metabolism.–1.892.2KEGG_Aminoacyl-tRNA biosynthesis–1.922.4KEGG_Drug P450–1.932.2KEGG_Tryptophan metabolism.–2.152.9DKD, RIG-I, retinoic inducible I; TCA, acid; tRNA, transfer RNA.Gene (DKD).a ABC, receptor. RNA. (DKD). absolute concentrations 116 energy-related 4, group) randomly (Supplementary S1; S3). Partial least squares discriminant (PLS-DA) performed assess significance class discrimination 7). metabolite (MSEA) PLS-DA variable importance projection (VIP) score 1. Differences acids, glycine, methionine, glutamate, arginine, proline, β-alanine noted. C. processes, gluconeogenesis, trends visualized comprehensive understanding 8). Glycolysis found excessive inflow upregulation Amino concentration 8a). facilitated flow diabetes-induced 8b). alleviated (GSSG) thus glutathione/GSSG ratio, suggested 8a reduction lipid peroxidation marker malondialdehyde tissue: S4). integration has demonstrated mice, animal diabetes, confirm findings model. background 9. Urinary 9d). addition, applied 3-dimensional (Clear, Unobstructed Brain/Body Imaging Cocktails, Computational [CUBIC]–kidney)16Hasegawa Susaki E.A. al.Comprehensive three-dimensional (CUBIC-kidney) visualizes abnormal sympathetic nerves after ischemia/reperfusion injury.Kidney 96: 129-138Abstract visualize glomeruli kidney. Glomerulomegaly 9e). mice way 10): model.Figure 10Transcriptome Symmetric (GO) differentially (DEGs) [DKD])/A DEGs 0.05 (1085 probes). Fatty-acid Canonical enarodustat)/B. described (65 Glucose upregulated, HIF-1, NAD, nicotinamide adenine dinucleotide; NADP, NAD phosphate.View study, pathways additionally 5, 7, 9, 10). It been fully elucidated whether protective or not. hypoxia17Laustsen Lycke Palm F. al.High altitude may alter availability diabetics hyperpolarized [1-(13)C]pyruvate magnetic resonance imaging.Kidney 2014; 86: 67-74Abstract Te
Language: Английский
Citations
87
Advanced Science, Journal Year: 2020, Volume and Issue: 7(8)
Published: Feb. 25, 2020
Abstract Tissue optical clearing techniques have provided important tools for large‐volume imaging. Aqueous‐based methods are known good fluorescence preservation and scalable size maintenance, but limited by long incubation time, insufficient performance, or requirements specialized devices. Additionally, few compatible with widely used lipophilic dyes while maintaining high performance. Here, to address these issues, m‐xylylenediamine (MXDA) is firstly introduced into tissue develop a rapid, highly efficient aqueous method robust compatibility, termed MXDA‐based Aqueous Clearing System (MACS). MACS can render whole adult brains transparent within 2.5 days also applicable other intact organs. Meanwhile, possesses ideal compatibility multiple probes, especially dyes. achieves 3D imaging of the neural structures labeled various techniques. Combining DiI labeling, allows reconstruction detailed vascular organs generates pathology glomeruli tufts in healthy diabetic kidneys. Therefore, provides useful mapping tissues expected facilitate morphological, physiological, pathological studies
Language: Английский
Citations
87
Nature Protocols, Journal Year: 2020, Volume and Issue: 16(1), P. 239 - 262
Published: Nov. 27, 2020
Language: Английский
Citations
86