Pub Date : 2026-03-01Epub Date: 2026-01-02DOI: 10.1016/j.acthis.2025.152315
Sanhua Fang , Li Liu, Dan Yang, Shuangshuang Liu, Wei Yin, Qiong Huang, Jingyao Chen
Total internal reflection fluorescence microscopy (TIRFM) enables live-cell imaging of clathrin-coated pits (CCPs) but is diffraction-limited (∼250 nm), hindering visualization of their dense nanostructures (∼150 nm). Super-Resolution Radial Fluctuations (SRRF) provides computational super-resolution; however, unoptimized parameters can disrupt the critical balance between resolution and structural fidelity. Here, we establish a comprehensive parameter optimization framework for SRRF-based nanoscopy of live CCPs. Using multi-modal metrics in CLTA-GFP HeLa cells, we identify a ring radius of 1.0 as optimal, balancing resolution (full width at half maximum = 180 ± 29 nm, p < 0.05 vs. TIRFM) with fidelity (resolution-scaled Pearson = 0.935 ± 0.018). The temporal radiality pairwise product mean (TRPPM) mode achieved superior resolution (154 ± 30 nm) while maintaining fidelity metrics comparable to Temporal radiality average (TRA) mode. In contrast, temporal radiality auto-correlations (TRAC) introduced artifactual structures and reduced fidelity. Parameters such as “remove positivity constraint” and gradient weighting induced severe artifacts and should be avoided. This optimized framework resolves the resolution–fidelity trade-off, enabling robust nanoscale imaging of clathrin-mediated endocytosis in live cells.
{"title":"Optimizing the resolution-fidelity trade-off in SRRF nanoscopy for live-cell clathrin imaging","authors":"Sanhua Fang , Li Liu, Dan Yang, Shuangshuang Liu, Wei Yin, Qiong Huang, Jingyao Chen","doi":"10.1016/j.acthis.2025.152315","DOIUrl":"10.1016/j.acthis.2025.152315","url":null,"abstract":"<div><div>Total internal reflection fluorescence microscopy (TIRFM) enables live-cell imaging of clathrin-coated pits (CCPs) but is diffraction-limited (∼250 nm), hindering visualization of their dense nanostructures (∼150 nm). Super-Resolution Radial Fluctuations (SRRF) provides computational super-resolution; however, unoptimized parameters can disrupt the critical balance between resolution and structural fidelity. Here, we establish a comprehensive parameter optimization framework for SRRF-based nanoscopy of live CCPs. Using multi-modal metrics in CLTA-GFP HeLa cells, we identify a ring radius of 1.0 as optimal, balancing resolution (full width at half maximum = 180 ± 29 nm, p < 0.05 vs. TIRFM) with fidelity (resolution-scaled Pearson = 0.935 ± 0.018). The temporal radiality pairwise product mean (TRPPM) mode achieved superior resolution (154 ± 30 nm) while maintaining fidelity metrics comparable to Temporal radiality average (TRA) mode. In contrast, temporal radiality auto-correlations (TRAC) introduced artifactual structures and reduced fidelity. Parameters such as “remove positivity constraint” and gradient weighting induced severe artifacts and should be avoided. This optimized framework resolves the resolution–fidelity trade-off, enabling robust nanoscale imaging of clathrin-mediated endocytosis in live cells.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"128 1","pages":"Article 152315"},"PeriodicalIF":2.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145880303","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-01Epub Date: 2025-12-16DOI: 10.1016/j.acthis.2025.152308
Victoria Julia Diederich , Sonja Scharf , Hendrik Schäfer , Sylvia Hartmann , Martin-Leo Hansmann , Patrick Wurzel
Lymph nodes function according to cellular and structural regulations. When these rules deviate from the benign equilibrium, dysregulations occur leading to the onset of diseases. In the development of malignant lymph node diseases, processes can be observed that consistently follow the same pattern. This study aims to define the structural and cellular parameters of the reactive lymph node and to demonstrate how lymph nodes change during tumorigenesis.
We analysed benign cases diagnosed as Lymphadenitis (8 patients). Malignant cases with the diagnosis of Nodular Sclerosis classical Hodgkin Lymphoma (5 patients), Mixed Cellularity classical Hodgkin Lymphoma (5 patients), Follicular Lymphoma (7 patients), and diffuse large B-cell Lymphoma (2 patients) were selected. Confocal microscopy was used to visualise immune cells and their compartments in 3D. Based on the fibroblastic reticular cell network we defined five compartments in reactive lymph nodes: Subcapsular sinus, marginal sinus, follicle, T-zone, and medulla. We analysed the cellular composition based on dendritic cells, macrophages, T cells and B cells and extended this analysis to include the presence of extracellular vesicles. During tumorigenesis, the compartmentalisation of the lymph node is progressively destroyed. Despite this ongoing destruction and loss of strict compartmental delineations, at least two distinct structural regions are still visible, defined as follicle-like and T-zone-like compartments. A comparison between reactive and neoplastic cases reveals a progressive cellular and structural homogenisation. Higher masses of CD8+ T cells were found under neoplastic conditions and higher masses of CD30+ were found in Hodgkin Lymphoma. The volume of CD20+ cells in follicles was consistently lower in malignant tissues compared to benign, but higher in T-zones in malignant cases. An increase in vesicles was detected in most neoplasms.
These findings offer new insights into cellular and structural remodelling, deepening our understanding of tumorigenesis and paving the way for more precise therapeutic interventions.
{"title":"Cellular landscape of reactive and neoplastic human lymph nodes in 3D","authors":"Victoria Julia Diederich , Sonja Scharf , Hendrik Schäfer , Sylvia Hartmann , Martin-Leo Hansmann , Patrick Wurzel","doi":"10.1016/j.acthis.2025.152308","DOIUrl":"10.1016/j.acthis.2025.152308","url":null,"abstract":"<div><div>Lymph nodes function according to cellular and structural regulations. When these rules deviate from the benign equilibrium, dysregulations occur leading to the onset of diseases. In the development of malignant lymph node diseases, processes can be observed that consistently follow the same pattern. This study aims to define the structural and cellular parameters of the reactive lymph node and to demonstrate how lymph nodes change during tumorigenesis.</div><div>We analysed benign cases diagnosed as Lymphadenitis (8 patients). Malignant cases with the diagnosis of Nodular Sclerosis classical Hodgkin Lymphoma (5 patients), Mixed Cellularity classical Hodgkin Lymphoma (5 patients), Follicular Lymphoma (7 patients), and diffuse large B-cell Lymphoma (2 patients) were selected. Confocal microscopy was used to visualise immune cells and their compartments in 3D. Based on the fibroblastic reticular cell network we defined five compartments in reactive lymph nodes: Subcapsular sinus, marginal sinus, follicle, T-zone, and medulla. We analysed the cellular composition based on dendritic cells, macrophages, T cells and B cells and extended this analysis to include the presence of extracellular vesicles. During tumorigenesis, the compartmentalisation of the lymph node is progressively destroyed. Despite this ongoing destruction and loss of strict compartmental delineations, at least two distinct structural regions are still visible, defined as follicle-like and T-zone-like compartments. A comparison between reactive and neoplastic cases reveals a progressive cellular and structural homogenisation. Higher masses of CD8<sup>+</sup> T cells were found under neoplastic conditions and higher masses of CD30<sup>+</sup> were found in Hodgkin Lymphoma. The volume of CD20<sup>+</sup> cells in follicles was consistently lower in malignant tissues compared to benign, but higher in T-zones in malignant cases. An increase in vesicles was detected in most neoplasms.</div><div>These findings offer new insights into cellular and structural remodelling, deepening our understanding of tumorigenesis and paving the way for more precise therapeutic interventions.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"128 1","pages":"Article 152308"},"PeriodicalIF":2.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145773141","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-01Epub Date: 2025-12-31DOI: 10.1016/j.acthis.2025.152313
Bryan Ôrtero Perez Gonçalves , Milla Reis Almeida , Vivian Vasconcelos Costa , Helen Lima Del Puerto , Alexander Birbrair , Luciana Maria Silva Lopes
Non-small cell lung cancer (NSCLC) exhibits substantial cellular and molecular heterogeneity, partly due to the presence of cancer stem cells (CSCs). CSCs can arise from a coordinated process known as epithelial–mesenchymal transition (EMT). EMT promotes a more aggressive phenotype, contributing significantly to tumor heterogeneity and drug resistance. Here, using state-of-the-art techniques—including confocal microscopy, flow cytometry, and transcript analysis—we investigated the cisplatin response of the LL/2 (LLC1) cell line cultured in both monolayer and tumorsphere (3D) models. Strikingly, LL/2 (LLC1) tumorspheres represent a model of cisplatin resistance, showing a remarkable increase in EMT and pluripotency mRNA regulators such as Zeb1, Zeb2, Snail, Twist, Tgfb1, Vimentin, FoxA2, Nanog, and Pou5f1 (Oct-4). Moreover, pseudotime trajectory analysis demonstrated that cisplatin treatment modulates a CSC-like phenotype differently in cells grown as monolayer versus tumorsphere. Our findings provide important insights into the role of cisplatin in NSCLC and highlight potential targets within the lung cancer microenvironment.
{"title":"Cisplatin treatment induces a shift toward a quiescent Ki-67⁻/CD44⁺/CD133⁺ cancer stem cell subpopulation in a tumorsphere model derived from a murine non-small cell lung cancer cell line","authors":"Bryan Ôrtero Perez Gonçalves , Milla Reis Almeida , Vivian Vasconcelos Costa , Helen Lima Del Puerto , Alexander Birbrair , Luciana Maria Silva Lopes","doi":"10.1016/j.acthis.2025.152313","DOIUrl":"10.1016/j.acthis.2025.152313","url":null,"abstract":"<div><div>Non-small cell lung cancer (NSCLC) exhibits substantial cellular and molecular heterogeneity, partly due to the presence of cancer stem cells (CSCs). CSCs can arise from a coordinated process known as epithelial–mesenchymal transition (EMT). EMT promotes a more aggressive phenotype, contributing significantly to tumor heterogeneity and drug resistance. Here, using state-of-the-art techniques—including confocal microscopy, flow cytometry, and transcript analysis—we investigated the cisplatin response of the LL/2 (LLC1) cell line cultured in both monolayer and tumorsphere (3D) models. Strikingly, LL/2 (LLC1) tumorspheres represent a model of cisplatin resistance, showing a remarkable increase in EMT and pluripotency mRNA regulators such as <em>Zeb1</em>, <em>Zeb2</em>, <em>Snail</em>, <em>Twist</em>, <em>Tgfb1</em>, <em>Vimentin</em>, <em>FoxA2</em>, <em>Nanog</em>, and <em>Pou5f1</em> (Oct-4). Moreover, pseudotime trajectory analysis demonstrated that cisplatin treatment modulates a CSC-like phenotype differently in cells grown as monolayer versus tumorsphere. Our findings provide important insights into the role of cisplatin in NSCLC and highlight potential targets within the lung cancer microenvironment.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"128 1","pages":"Article 152313"},"PeriodicalIF":2.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145880304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The study of oligodendrocyte precursor cells (OPCs) in both physiological and pathological contexts is challenging due to their capacity for self-renewal. This research aimed to examine the effects of OPC depletion on neurons. Tamoxifen-inducible Sox10/iCreERT2; netrin-1flox/flox (NTN-1 cKO) mice were used to inactivate NTN-1 in Sox10+ oligodendroglia at varying tamoxifen doses. The impact of Necrostatin-1s (Nec-1s) and cytarabine on neuronal degeneration was evaluated, along with a comparison of the effects of tamoxifen dissolved in different plant oils on lineage tracing in Sox10/iCreERT2; tdTomato mice, as well as on neuronal degeneration in NTN-1 cKO mice. Our findings showed that administering 3 mg of tamoxifen per NTN-1 cKO mouse triggered necroptosis and apoptosis in Sox10+ cells. Notably, a higher dose of 6 mg of tamoxifen resulted in the degeneration of cortical neurons, which was accompanied by astrogliosis, amyloidosis, and a reduction in microglia. Immunostaining and RNAscope analysis indicated that it was Cre recombinase, rather than Cre mRNA, that was transferred to neurons. Nec-1s and cytarabine successfully prevented cortical neuron degeneration, though through distinct mechanisms. Furthermore, administering tamoxifen dissolved in vitamin E-rich wheat germ oil reduced Cre transfer in both Sox10/iCreERT2; tdTomato mice and NTN-1 cKO mice, significantly preventing cortical neurons from being labeled with tdTomato and protecting them from degeneration. These results suggest that, under pathological conditions, Cre recombinase can transfer from oligodendroglia to neurons, a process triggered by neuronal stress. This highlights the need for careful consideration in using Cre-loxP lineage tracing and gene-editing methods involving oligodendrocyte lineage cells and neurons.
{"title":"Neuronal stress promotes Cre recombinase transfer from oligodendroglia to neurons","authors":"Ting Xu , Kairan Yang , Yuhu Feng , Yun-Cheng Wu , Lin-Yuan Zhang , Zuisu Yang , Falei Yuan , Haiyan Lyu","doi":"10.1016/j.acthis.2025.152300","DOIUrl":"10.1016/j.acthis.2025.152300","url":null,"abstract":"<div><div>The study of oligodendrocyte precursor cells (OPCs) in both physiological and pathological contexts is challenging due to their capacity for self-renewal. This research aimed to examine the effects of OPC depletion on neurons. Tamoxifen-inducible Sox10/iCreER<sup>T2</sup>; netrin-1<sup>flox/flox</sup> (NTN-1 cKO) mice were used to inactivate NTN-1 in Sox10<sup>+</sup> oligodendroglia at varying tamoxifen doses. The impact of Necrostatin-1s (Nec-1s) and cytarabine on neuronal degeneration was evaluated, along with a comparison of the effects of tamoxifen dissolved in different plant oils on lineage tracing in Sox10/iCreER<sup>T2</sup>; tdTomato mice, as well as on neuronal degeneration in NTN-1 cKO mice. Our findings showed that administering 3 mg of tamoxifen per NTN-1 cKO mouse triggered necroptosis and apoptosis in Sox10<sup>+</sup> cells. Notably, a higher dose of 6 mg of tamoxifen resulted in the degeneration of cortical neurons, which was accompanied by astrogliosis, amyloidosis, and a reduction in microglia. Immunostaining and RNAscope analysis indicated that it was Cre recombinase, rather than Cre mRNA, that was transferred to neurons. Nec-1s and cytarabine successfully prevented cortical neuron degeneration, though through distinct mechanisms. Furthermore, administering tamoxifen dissolved in vitamin E-rich wheat germ oil reduced Cre transfer in both Sox10/iCreER<sup>T2</sup>; tdTomato mice and NTN-1 cKO mice, significantly preventing cortical neurons from being labeled with tdTomato and protecting them from degeneration. These results suggest that, under pathological conditions, Cre recombinase can transfer from oligodendroglia to neurons, a process triggered by neuronal stress. This highlights the need for careful consideration in using Cre-loxP lineage tracing and gene-editing methods involving oligodendrocyte lineage cells and neurons.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"128 1","pages":"Article 152300"},"PeriodicalIF":2.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145572756","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-01Epub Date: 2025-12-13DOI: 10.1016/j.acthis.2025.152309
Zhiwei Wang , Shiteng Wang , Xiaoyu Zheng , Xia Peng , Xiaotong Wang , Xige Zhao , Yijia Wang , Jing Chen , Mingyue Meng , Juan Du
Mouse embryonic palatal mesenchyme (MEPM) cell culture is commonly used to study palate development and cleft palate (CP). However, there are few reports on the changes in the biological patterns of MEPM cells during palate development. In this study, we investigated the changes in the biological characteristics of MEPM cells during the critical period of mouse palate development from embryonic day (E) 13.5 to E16.5. First, we examined the proliferation and apoptotic factors, as well as the osteogenic ability, of palatal shelves from E13.5 to E16.5 in vivo using immunohistochemical staining and qRT-PCR. Then we conducted a comprehensive analysis of E13.5–E16.5 MEPM cells and compared their biological characteristics, including cell origin, proliferation, apoptosis, migration, osteogenesis, adipogenesis, and stemness. We found that MEPM cells from E13.5–E16.5 showed positive expression of the mesenchymal marker Vimentin and negative expression of the epithelial marker CK-14. The proliferation of MEPM cells was similar at E13.5 and E14.5, but it gradually declined at E15.5 and E16.5. There was no statistically significant difference in the apoptosis rate among MEPM cells at E13.5–E16.5. The migration ability of MEPM cells gradually decreased from E13.5 to E16.5, and the osteogenic ability of MEPM cells and palate shelves gradually increased. In addition, the expressions of stemness markers gradually decreased, accompanied by a decrease in adipogenic ability. These results indicate differences in the biological characteristics of MEPM cells at different palate development stages, which helps us understand the detailed process of palate development.
{"title":"Biological patterns of mouse embryonic palatal mesenchymal cells at different palatogenic stages","authors":"Zhiwei Wang , Shiteng Wang , Xiaoyu Zheng , Xia Peng , Xiaotong Wang , Xige Zhao , Yijia Wang , Jing Chen , Mingyue Meng , Juan Du","doi":"10.1016/j.acthis.2025.152309","DOIUrl":"10.1016/j.acthis.2025.152309","url":null,"abstract":"<div><div>Mouse embryonic palatal mesenchyme (MEPM) cell culture is commonly used to study palate development and cleft palate (CP). However, there are few reports on the changes in the biological patterns of MEPM cells during palate development. In this study, we investigated the changes in the biological characteristics of MEPM cells during the critical period of mouse palate development from embryonic day (E) 13.5 to E16.5. First, we examined the proliferation and apoptotic factors, as well as the osteogenic ability, of palatal shelves from E13.5 to E16.5 <em>in vivo</em> using immunohistochemical staining and qRT-PCR. Then we conducted a comprehensive analysis of E13.5–E16.5 MEPM cells and compared their biological characteristics, including cell origin, proliferation, apoptosis, migration, osteogenesis, adipogenesis, and stemness. We found that MEPM cells from E13.5–E16.5 showed positive expression of the mesenchymal marker Vimentin and negative expression of the epithelial marker CK-14. The proliferation of MEPM cells was similar at E13.5 and E14.5, but it gradually declined at E15.5 and E16.5. There was no statistically significant difference in the apoptosis rate among MEPM cells at E13.5–E16.5. The migration ability of MEPM cells gradually decreased from E13.5 to E16.5, and the osteogenic ability of MEPM cells and palate shelves gradually increased. In addition, the expressions of stemness markers gradually decreased, accompanied by a decrease in adipogenic ability. These results indicate differences in the biological characteristics of MEPM cells at different palate development stages, which helps us understand the detailed process of palate development.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"128 1","pages":"Article 152309"},"PeriodicalIF":2.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145747128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Clear cell renal cell carcinoma (ccRCC) is the most prevalent histological subtype of renal malignancy, associated with poor prognosis in advanced stages. Emerging evidence highlights the potential tumor-suppressive role of the anti-aging protein Klotho (KL) and its cofactor, fibroblast growth factor 23 (FGF23), both of which are implicated in phosphate metabolism and cellular homeostasis.
Methods
Using immunofluorescence and quantitative image analysis, we assessed KL and FGF23 protein levels in 20 ccRCC specimens stratified by tumor grade, alongside adjacent normal tissue. Publicly available RNA-seq and survival data from the TCGA-KIRC cohort were analyzed to complement our findings.
Results
Immunofluorescence analysis of 20 ccRCC samples and matched normal tissues revealed consistently low Klotho expression with no significant differences across tumor grades. However, Kaplan–Meier survival analysis revealed that high KL mRNA expression was significantly associated with improved overall survival and disease-free survival, highlighting its role as a protective prognostic biomarker. Multivariate Cox regression confirmed KL as an independent predictor of better overall survival. In contrast, FGF23 protein levels were significantly elevated in ccRCC samples, particularly in high-grade tumors, despite minimal expression in control tissue and no significant differences at the mRNA level in the TCGA cohort. Notably, patients with detectable FGF23 expression had significantly worse survival outcomes, and multivariate analysis identified elevated FGF23 as an independent risk factor for poor prognosis. Age and tumor stage also remain strong prognostic determinants in our models.
Conclusions
These findings suggest a dichotomous role for KL and FGF23 in ccRCC, with KL functioning as a favorable prognostic factor and FGF23 potentially contributing to disease progression and early relapse. Further mechanistic studies are warranted to elucidate their interplay and evaluate their utility as biomarkers or therapeutic targets in renal cancer.
{"title":"Immunofluorescent characterization of Klotho and FGF23 in clear cell renal cell carcinoma: A pilot study","authors":"Anita Racetin , Nela Kelam , Merica Glavina Durdov , Katarina Vukojević","doi":"10.1016/j.acthis.2025.152298","DOIUrl":"10.1016/j.acthis.2025.152298","url":null,"abstract":"<div><h3>Background/objectives</h3><div>Clear cell renal cell carcinoma (ccRCC) is the most prevalent histological subtype of renal malignancy, associated with poor prognosis in advanced stages. Emerging evidence highlights the potential tumor-suppressive role of the anti-aging protein Klotho (KL) and its cofactor, fibroblast growth factor 23 (FGF23), both of which are implicated in phosphate metabolism and cellular homeostasis.</div></div><div><h3>Methods</h3><div>Using immunofluorescence and quantitative image analysis, we assessed KL and FGF23 protein levels in 20 ccRCC specimens stratified by tumor grade, alongside adjacent normal tissue. Publicly available RNA-seq and survival data from the TCGA-KIRC cohort were analyzed to complement our findings.</div></div><div><h3>Results</h3><div>Immunofluorescence analysis of 20 ccRCC samples and matched normal tissues revealed consistently low Klotho expression with no significant differences across tumor grades. However, Kaplan–Meier survival analysis revealed that high <em>KL</em> mRNA expression was significantly associated with improved overall survival and disease-free survival, highlighting its role as a protective prognostic biomarker. Multivariate Cox regression confirmed <em>KL</em> as an independent predictor of better overall survival. In contrast, FGF23 protein levels were significantly elevated in ccRCC samples, particularly in high-grade tumors, despite minimal expression in control tissue and no significant differences at the mRNA level in the TCGA cohort. Notably, patients with detectable <em>FGF23</em> expression had significantly worse survival outcomes, and multivariate analysis identified elevated <em>FGF23</em> as an independent risk factor for poor prognosis. Age and tumor stage also remain strong prognostic determinants in our models.</div></div><div><h3>Conclusions</h3><div>These findings suggest a dichotomous role for KL and FGF23 in ccRCC, with KL functioning as a favorable prognostic factor and FGF23 potentially contributing to disease progression and early relapse. Further mechanistic studies are warranted to elucidate their interplay and evaluate their utility as biomarkers or therapeutic targets in renal cancer.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 4","pages":"Article 152298"},"PeriodicalIF":2.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145516944","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cholinergic neurons in the basal forebrain cholinergic nuclei (BFCN) and neostriatum (CPu) play key roles in learning, attention, and motor control. The loss of cholinergic neurons causes major neurodegenerative diseases such as Alzheimer’s disease. This study aimed to elucidate the molecular diversity of choline acetyltransferase immunoreactive (ChAT-ir) neurons in these brain regions. We performed immunohistochemistry to determine the co-expression of ChAT-ir neurons with two neuropeptides, calcitonin gene-related peptide (CGRP) and cholecystokinin (CCK), as well as three calcium-binding proteins, such as calbindin, calretinin, and parvalbumin, in the adult mouse brain. The results showed that ChAT, calbindin, CGRP and CCK were strongly expressed in the BFCN, including medial septal nucleus (MS), nucleus of vertical limb and horizontal limb of the diagonal band of Broca (VDB and HDB), substantia innominata basal part (SIB), and in the caudate putamen (CPu). CGRP and CCK showed a high immunoreactive co-expression with ChAT, especially in the HDB and CPu. Calbindin immunoreactivity was widely present and coincided with ChAT in the VDB, HDB, and CPu. However, calretinin immunoreactivity showed a selective co-expression with ChAT in the VDB, SIB, and CPu. Although parvalbumin immunoreactivity was observed throughout the BFCN and CPu, but there was no co-expression between ChAT and parvalbumin. The neurochemical diversity of ChAT-ir neurons in the BFCN and neostriatum suggests the specialized functions of cholinergic neurons across different circuits, especially by modulating CGRP, CCK, or calbindin. These results could provide new insight into cholinergic modulation throughout the BFCN and striatum.
{"title":"Neurochemical heterogeneity of ChAT-immunoreactive neurons in the basal forebrain cholinergic nuclei and striatum in reference to CGRP, CCK, and calcium-binding proteins","authors":"Mirza Mienur Meher , Marya Afrin , Mir Rubayet Jahan , Kanako Nozaki , Koh-hei Masumoto , Akie Yanai , Md Nabiul Islam","doi":"10.1016/j.acthis.2025.152291","DOIUrl":"10.1016/j.acthis.2025.152291","url":null,"abstract":"<div><div>Cholinergic neurons in the basal forebrain cholinergic nuclei (BFCN) and neostriatum (CPu) play key roles in learning, attention, and motor control. The loss of cholinergic neurons causes major neurodegenerative diseases such as Alzheimer’s disease. This study aimed to elucidate the molecular diversity of choline acetyltransferase immunoreactive (ChAT-ir) neurons in these brain regions. We performed immunohistochemistry to determine the co-expression of ChAT-ir neurons with two neuropeptides, calcitonin gene-related peptide (CGRP) and cholecystokinin (CCK), as well as three calcium-binding proteins, such as calbindin, calretinin, and parvalbumin, in the adult mouse brain. The results showed that ChAT, calbindin, CGRP and CCK were strongly expressed in the BFCN, including medial septal nucleus (MS), nucleus of vertical limb and horizontal limb of the diagonal band of Broca (VDB and HDB), substantia innominata basal part (SIB), and in the caudate putamen (CPu). CGRP and CCK showed a high immunoreactive co-expression with ChAT, especially in the HDB and CPu. Calbindin immunoreactivity was widely present and coincided with ChAT in the VDB, HDB, and CPu. However, calretinin immunoreactivity showed a selective co-expression with ChAT in the VDB, SIB, and CPu. Although parvalbumin immunoreactivity was observed throughout the BFCN and CPu, but there was no co-expression between ChAT and parvalbumin. The neurochemical diversity of ChAT-ir neurons in the BFCN and neostriatum suggests the specialized functions of cholinergic neurons across different circuits, especially by modulating CGRP, CCK, or calbindin. These results could provide new insight into cholinergic modulation throughout the BFCN and striatum.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 4","pages":"Article 152291"},"PeriodicalIF":2.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145009844","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Amyloid β (Aβ) accumulation in the brains of patients with Alzheimer's disease (AD) contributes to cognitive impairment and neuronal damage. Urolithin A (UA), a gut microbiota–derived metabolite of ellagic acid, has been reported to cross the blood-brain barrier to exert anti-inflammatory and anti-oxidation effects in the brain. However, the molecular mechanisms of UA in AD were still unclear. This study aims to explore the neuroprotective effect and mechanism of UA on APP/PS1 mice and Aβ1–42-injured N2a and PC12 cells.
Methods
In this study, Morris water maze was used to detect the cognitive function. Immunofluorescence was used to detect the deposition of Aβ and the expression of voltage-dependent anion channel 1 (VDAC1) in the brains of APP/PS1 mice. Western blotting was used to detect the expression of VDAC1, AMPK pathway, PI3K pathway and autophagy-related proteins. CCK8 was used to detect the viability of Aβ1–42-injured cells.
Results
In this research, we found that UA improved cognitive dysfunction and reduced Aβ deposition in APP/PS1 mice. Furthermore, UA activated autophagy and upregulated the levels of autophagy-related proteins in both APP/PS1 mice and Aβ1–42-injured N2a and PC12 cells. At the same time, UA down-regulated the phosphorylation level of PI3K/AKT/mTOR and up-regulated the phosphorylation level of AMPK in APP/PS1 mice and Aβ1–42-injured N2a cells and PC12 cells. In addition, UA down-regulated VDAC1, consistent with the effect of VDAC1 antagonist DIDS (4′-diisothiocyano-2,2′-disulfonic acid stilbene). Importantly, the UA-induced activation of autophagy and modulation of the PI3K and AMPK pathways were reversed by VDAC1 overexpression.
Conclusion
These findings demonstrated that UA down-regulated VDAC1 played a key neuroprotective role on AD by inhibiting the PI3K/AKT/mTOR pathway and activating the AMPK pathway to promote autophagy.
{"title":"Neuroprotective effect of Urolithin A via downregulating VDAC1-mediated autophagy in Alzheimer's disease","authors":"Bensi Zhang , Xiujun Zhang , Waleephan Treebupachatsakul , Rungusa Pantan , Natnicha Kampan , Manussabhorn Phatsara , Chun Shi , Suteera Narakornsak","doi":"10.1016/j.acthis.2025.152290","DOIUrl":"10.1016/j.acthis.2025.152290","url":null,"abstract":"<div><h3>Background</h3><div>Amyloid β (Aβ) accumulation in the brains of patients with Alzheimer's disease (AD) contributes to cognitive impairment and neuronal damage. Urolithin A (UA), a gut microbiota–derived metabolite of ellagic acid, has been reported to cross the blood-brain barrier to exert anti-inflammatory and anti-oxidation effects in the brain. However, the molecular mechanisms of UA in AD were still unclear. This study aims to explore the neuroprotective effect and mechanism of UA on APP/PS1 mice and Aβ<sub>1–42</sub>-injured N2a and PC12 cells.</div></div><div><h3>Methods</h3><div>In this study, Morris water maze was used to detect the cognitive function. Immunofluorescence was used to detect the deposition of Aβ and the expression of voltage-dependent anion channel 1 (VDAC1) in the brains of APP/PS1 mice. Western blotting was used to detect the expression of VDAC1, AMPK pathway, PI3K pathway and autophagy-related proteins. CCK8 was used to detect the viability of Aβ<sub>1–42</sub>-injured cells.</div></div><div><h3>Results</h3><div>In this research, we found that UA improved cognitive dysfunction and reduced Aβ deposition in APP/PS1 mice. Furthermore, UA activated autophagy and upregulated the levels of autophagy-related proteins in both APP/PS1 mice and Aβ<sub>1–42</sub>-injured N2a and PC12 cells. At the same time, UA down-regulated the phosphorylation level of PI3K/AKT/mTOR and up-regulated the phosphorylation level of AMPK in APP/PS1 mice and Aβ<sub>1–42</sub>-injured N2a cells and PC12 cells. In addition, UA down-regulated VDAC1, consistent with the effect of VDAC1 antagonist DIDS (4′-diisothiocyano-2,2′-disulfonic acid stilbene). Importantly, the UA-induced activation of autophagy and modulation of the PI3K and AMPK pathways were reversed by VDAC1 overexpression.</div></div><div><h3>Conclusion</h3><div>These findings demonstrated that UA down-regulated VDAC1 played a key neuroprotective role on AD by inhibiting the PI3K/AKT/mTOR pathway and activating the AMPK pathway to promote autophagy.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 4","pages":"Article 152290"},"PeriodicalIF":2.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144921397","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-11-06DOI: 10.1016/j.acthis.2025.152296
Wang Jia , Yonggui Wu
IgA nephropathy (IgAN) is a prevalent glomerular disease characterized by mesangial deposition of IgA1-containing immune complexes, yet its underlying molecular mechanisms remain incompletely understood. In this study, we integrated bioinformatics analyses of two public datasets (GSE104948 and GSE93798) to identify key differentially expressed genes (DEGs) associated with IgAN. Periostin (POSTN) emerged as a hub gene, exhibiting significant upregulation in IgAN samples and correlating with histopathological severity. Functional enrichment revealed that overlapping DEGs are involved in extracellular matrix organization, immune response, and signaling pathways relevant to renal pathology. Immunohistochemical and immunofluorescence analyses confirmed increased POSTN and decreased GPX4 expression in renal biopsies from IgAN patients, indicating enhanced ferroptosis. In vitro, IgA1 stimulation of human mesangial cells (HMCs) elevated POSTN expression and induced ferroptosis, evidenced by increased oxidative stress, mitochondrial damage, and reduced cell viability. Knockdown of POSTN ameliorated these effects by restoring glutathione levels and reducing lipid peroxidation, while POSTN overexpression exacerbated ferroptosis. Notably, treatment with the ferroptosis inhibitor ferrostatin-1 reversed POSTN-induced cellular damage. Our findings suggest that POSTN promotes IgAN progression by facilitating ferroptosis through GPX4 downregulation, highlighting a novel pathogenic mechanism. Targeting POSTN-mediated ferroptosis may provide promising therapeutic strategies for IgAN. This study advances our understanding of IgAN molecular pathology and offers potential biomarkers and intervention targets to improve patient outcomes.
{"title":"POSTN promotes ferroptosis and contributes to the pathogenesis of IgA nephropathy via GPX4 downregulation","authors":"Wang Jia , Yonggui Wu","doi":"10.1016/j.acthis.2025.152296","DOIUrl":"10.1016/j.acthis.2025.152296","url":null,"abstract":"<div><div>IgA nephropathy (IgAN) is a prevalent glomerular disease characterized by mesangial deposition of IgA1-containing immune complexes, yet its underlying molecular mechanisms remain incompletely understood. In this study, we integrated bioinformatics analyses of two public datasets (GSE104948 and GSE93798) to identify key differentially expressed genes (DEGs) associated with IgAN. Periostin (POSTN) emerged as a hub gene, exhibiting significant upregulation in IgAN samples and correlating with histopathological severity. Functional enrichment revealed that overlapping DEGs are involved in extracellular matrix organization, immune response, and signaling pathways relevant to renal pathology. Immunohistochemical and immunofluorescence analyses confirmed increased POSTN and decreased GPX4 expression in renal biopsies from IgAN patients, indicating enhanced ferroptosis. In vitro, IgA1 stimulation of human mesangial cells (HMCs) elevated POSTN expression and induced ferroptosis, evidenced by increased oxidative stress, mitochondrial damage, and reduced cell viability. Knockdown of POSTN ameliorated these effects by restoring glutathione levels and reducing lipid peroxidation, while POSTN overexpression exacerbated ferroptosis. Notably, treatment with the ferroptosis inhibitor ferrostatin-1 reversed POSTN-induced cellular damage. Our findings suggest that POSTN promotes IgAN progression by facilitating ferroptosis through GPX4 downregulation, highlighting a novel pathogenic mechanism. Targeting POSTN-mediated ferroptosis may provide promising therapeutic strategies for IgAN. This study advances our understanding of IgAN molecular pathology and offers potential biomarkers and intervention targets to improve patient outcomes.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 4","pages":"Article 152296"},"PeriodicalIF":2.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145462918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-08-19DOI: 10.1016/j.acthis.2025.152281
Yuxi Sun , Zhiyuan Gong , Yueh-Min Lin , Jeng-Wei Lu
{"title":"Corrigendum to “Inducible gankyrin overexpression drives hepatocarcinogenesis in a liver-specific zebrafish model” [Acta Histochem. 127 (2025) 152280]","authors":"Yuxi Sun , Zhiyuan Gong , Yueh-Min Lin , Jeng-Wei Lu","doi":"10.1016/j.acthis.2025.152281","DOIUrl":"10.1016/j.acthis.2025.152281","url":null,"abstract":"","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 4","pages":"Article 152281"},"PeriodicalIF":2.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144881816","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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