{"title":"Expression of Glial Cell Line-derived Neurotrophic Factor and Its Receptors in Glioblastoma.","authors":"Jesper Dupont Ewald, Arnon Møldrup Knudsen, Helle Wohlleben, Lone Christiansen, Signe Regner Michaelsen, Atul Anand, Bjarne Winther Kristensen","doi":"10.1369/00221554251374108","DOIUrl":"10.1369/00221554251374108","url":null,"abstract":"<p><p>Glioblastoma is the most frequent and aggressive primary brain cancer in adults, and the prognosis is poor. The neurotrophic factor glial cell-derived neurotrophic factor (GDNF) and its receptors, which are involved in neuronal development, have in experimental studies been suggested to drive tumorigenic processes in glioblastoma, but the role and expression in glioblastoma in patients is under-investigated. The aim of this study was to investigate the expression of GDNF, GDNF family receptor 1-4 (GFRA1-4), and the downstream REarranged during Transfection (RET) receptor in human glioblastoma tissue by RNA in situ hybridization, immunohistochemistry, and immunofluorescence. Expression was quantified by software-based classifiers. The results showed that GDNF was expressed in approximately 10% of tumor cells. The GFRA1 receptor was widely expressed in tumor cells, often colocalizing with the astrocytic tumor cell marker glial fibrillary acidic protein (GFAP), and in a smaller fraction of tumor cells expressing the stem cell markers oligodendrocyte transcription factor 2 (OLIG2) and SRY-Box Transcription Factor 2 (SOX2). The GFRA2 receptor expression was very limited, whereas expression of GFRA3, GFRA4, and RET, respectively, was almost absent. In conclusion, GDNF and its primary receptor GFRA1 were expressed in patient glioblastoma tissue. Potential clinical value needs further investigation.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"221554251374108"},"PeriodicalIF":1.5,"publicationDate":"2025-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12477178/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145186069","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lorraine Perciliano de Faria, Victor E Arana-Chavez, Lexie Shannon Holliday
{"title":"Alendronate Alters the Release of EVs by Raw 264.7 Osteoclasts.","authors":"Lorraine Perciliano de Faria, Victor E Arana-Chavez, Lexie Shannon Holliday","doi":"10.1369/00221554251379540","DOIUrl":"10.1369/00221554251379540","url":null,"abstract":"<p><p>Alendronate (ALN), a nitrogen-containing bisphosphonate, is widely used to treat bone disorders. While its inhibitory effect on osteoclast activity is well-established, its impact on the release of extracellular vesicles (EVs) is less understood. This study investigated the effect of ALN on the quantity and size distribution of EVs released by osteoclasts cultured on bovine bone slices pretreated with 10-µM ALN, 100-µM ALN, or vehicle. Raw 264.7 cells were differentiated into osteoclasts using RANK-ligand, and EVs were isolated from conditioned media. Tartrate-resistant acid phosphatase (TRAP) staining, phalloidin staining for actin rings, and nanoparticle tracking analysis (NTA) were performed. TRAP staining showed a significant reduction in the number of TRAP-positive multinucleated cells in the 100-µM ALN group, confirming that high-concentration ALN also impairs osteoclast formation. Phalloidin staining showed a significant decrease in actin ring formation in the 100-µM ALN group, confirming ALN's inhibitory effect on osteoclast activity. NTA revealed a lower total EV concentration in the 100-µM ALN group, with a distinct peak of smaller EVs (<100 nm), suggestive of exosomes. These findings indicate that ALN, especially at higher concentrations, alters the release profile of osteoclast-derived EVs, potentially affecting intercellular communication and bone remodeling beyond its direct inhibition of resorption.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"221554251379540"},"PeriodicalIF":1.5,"publicationDate":"2025-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12477179/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145181908","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Von Kossa Calcium Staining Procedure Revisited.","authors":"Sheila Criswell","doi":"10.1369/00221554251379535","DOIUrl":"10.1369/00221554251379535","url":null,"abstract":"<p><p>Probably the most sensitive and frequently performed stain for calcium deposits in mammalian tissues is the von Kossa, a stain used for over a century. When originally incorporated into daily use in the histopathology laboratory on paraffin tissue sections, the procedure called for immersion of tissue sections in an aqueous 5% silver nitrate solution and placed in direct sunlight for 20-30 min to make calcium deposits appear black. Although this effective method continues to be used in select locations, the modern laboratory often does not have access to bright sunlight. This study revisited the traditional von Kossa staining method with alternatives in light sources, silver nitrate concentrations, and timing of silver solution exposure and determined that a 1% silver nitrate solution is equally effective as a 5% solution in bright sunlight for 30-60 min. In addition, a 1% silver nitrate solution can be successfully used with exposure to a 3.5W light-emitting diode (LED) daylight lamp for 1 hr, sunlight through a tinted glass window for 1 hr, or overhead room lighting for 2 hr, and the staining reaction is unaffected by temperatures ranging from 1C to 40C.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"221554251379535"},"PeriodicalIF":1.5,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12463881/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145137811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Immunohistochemical Assessment of Cardiac Macrophages in the Aged Fischer 344 Rat.","authors":"Steven A Bloomer","doi":"10.1369/00221554251374725","DOIUrl":"10.1369/00221554251374725","url":null,"abstract":"<p><p>Macrophages have multiple roles in the heart including immune surveillance and extracellular matrix remodeling. Aging increases both collagen deposition and macrophage number in the heart; however, rodent models used to study cardiac macrophages have age-related comorbidities such as atherosclerosis and hypertension. The Fischer 344 rat does not develop these conditions with aging; therefore, the purpose of this study was to evaluate macrophage number and polarization in the hearts of aged (24-month) and young (6-month) Fischer 344 rats. Paraffin-embedded hearts were assessed for collagen deposition and immunolabeled for CD68, CD163, CD206, and galectin-3. Compared with young rats, significantly greater collagen deposition was observed in the old rats. There were no significant differences in CD68<sup>+</sup> or CD163<sup>+</sup> cells between age groups, but both CD206<sup>+</sup> and galectin-3<sup>+</sup> cells were more numerous in the aged animals. Double-immunofluorescence studies demonstrated that galectin-3 colocalized with both CD68 and CD163, suggesting that galectin-3 is found in cardiac macrophages. Further colocalization studies demonstrated similar proportions of CD68<sup>+</sup>/CD163<sup>-</sup>, CD68<sup>+</sup>/CD163<sup>+</sup>, and CD68<sup>-</sup>/CD163<sup>+</sup> cells between age groups, suggesting that aging does not affect macrophage polarization. As CD206<sup>+</sup> and galectin-3<sup>+</sup> cells promote fibrosis, these results warrant future studies that delineate the specific roles of these cells in the aged heart.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"221554251374725"},"PeriodicalIF":1.5,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12460319/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145131112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nayara Fernanda Costa Castro, Vitor Grigio, Cássia Regina Suzuki Caires, Mariele Ilario Zucão, Sebastião Roberto Taboga, Patrícia Simone Leite Vilamaior
{"title":"Alterations in Inflammatory Markers and Tissue Architecture in the Gerbil Prostate Following Castration.","authors":"Nayara Fernanda Costa Castro, Vitor Grigio, Cássia Regina Suzuki Caires, Mariele Ilario Zucão, Sebastião Roberto Taboga, Patrícia Simone Leite Vilamaior","doi":"10.1369/00221554251374732","DOIUrl":"10.1369/00221554251374732","url":null,"abstract":"<p><p>SummaryOrchiectomy induces atrophy of epithelial cells in the prostate gland, stimulated by androgen deprivation. The objective of the current study was to assess the changes arising from tissue remodeling during short periods of androgen deprivation in the ventral prostate of Mongolian gerbils. Adult male gerbils were divided into groups: control and castrated, euthanized on the 3rd (Ca3d), 7th (Ca7d), and 14th (Ca14d) days post-orchiectomy (n = 7/group). The ventral lobe of the prostate was submitted for histological, stereological, morphometric, serological, and immunohistochemical analyses. Castration promoted a reduction in the weight of the ventral prostate. It altered the relative proportion of prostate tissue compartments, such as a decrease in the epithelium and non-muscular stroma and an increase in the muscular stroma. In addition, we observed that the number of Cd-68 positive cells increased in the Ca3d group, which represents a period of androgen deprivation not previously reported in the literature for Mongolian gerbils. Matrix metalloproteinase-9 quantification revealed a decrease in the Ca7d group compared to the Ca3d. In addition, the frequency of Tumor Necrosis Factor alpha increased in the Ca14d group, influenced by the duration of testosterone deficiency. The findings contribute to the understanding of prostate tissue remodeling after castration, as well as highlighting the rapid alterations in the prostate microenvironment in a short period following castration.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"221554251374732"},"PeriodicalIF":1.5,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12450208/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145091948","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Limits of Vibrating Microtomy.","authors":"Alexey Klepukov","doi":"10.1369/00221554251367488","DOIUrl":"10.1369/00221554251367488","url":null,"abstract":"<p><p>The Vibratome, or vibrating microtome allows sectioning of the mouse brain with a reliably established thickness from 50 µm, but they have a strong limitation on the size of the sample suitable for cutting. Herein is described the construction with publicly available element base (from parts for a 3D printer) a so-called 3D-vibrating microtome capable of cutting larger size brain into thin sections, and use it to investigate the limit-attainable values of the minimum slice thickness. Both of these goals have been successfully achieved. Both small mouse slices with a minimum thickness of 30 μm and large whole calf brain slices with a minimum thickness of 150 μm were obtained. Critical features of the effect of blade vibration frequency and sample feed rate on the minimum slice thickness were revealed, In the same way, a clear correlation was established between the minimum achievable thickness and its area.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"221554251367488"},"PeriodicalIF":1.5,"publicationDate":"2025-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12436324/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145064711","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Natalia Pérez-Barraza, Pedro Acuña, Sebastián San Martín, Juan Varas, Claudio Cordova, Renato Casalino, Eva Madrid
{"title":"Stromal Cells, Extracellular Matrix Components, and Adenosine A3 Receptor in Prostate Biopsies: Association With Histological Grade, PSA Levels, and Clinical Tumor Stage.","authors":"Natalia Pérez-Barraza, Pedro Acuña, Sebastián San Martín, Juan Varas, Claudio Cordova, Renato Casalino, Eva Madrid","doi":"10.1369/00221554251349873","DOIUrl":"10.1369/00221554251349873","url":null,"abstract":"<p><p>Prostate cancer (PC) is the second leading cause of cancer-related death in men worldwide. Its progression is marked by significant phenotypic changes in stromal cells and alterations in extracellular matrix (ECM) composition, including variations in collagen fibers, proteoglycans (PGs), and glycosaminoglycans. Tumor cells also modulate ECM secretion through adenosine A3 receptor (A3AR) activity. This study aimed to evaluate stromal cells, ECM components, and A3AR expression in prostate biopsies and explore their association with clinicopathological variables. We analyzed tissue samples from 96 patients diagnosed with PC or benign prostatic hyperplasia (BPH), using immunohistochemistry for alpha-smooth muscle actin and A3AR, and histochemical methods for ECM components. PC samples showed reduced stromal cell content and increased PGs, collagen fibers, and A3AR levels compared with BPH. While we found associations with histological classification, no significant correlations were observed with preoperative prostate-specific antigen levels or clinical risk categories. Our findings suggest that ECM components and A3AR may be involved in PCr progression and hold potential as biomarkers. However, due to the limited number of high-grade cases, further studies are needed to confirm these associations.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"277-288"},"PeriodicalIF":1.5,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12276202/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144667808","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Expression Diversity in Endocrine Cells and Between Species Revealed by Novel Synthetic Peptide Antibodies Recognizing the Neuroendocrine Protein 7B2.","authors":"Shota Kikuchi, Seiji Torii, Masahiro Hosaka, Tadashi Yasui, Hiroshi Gomi","doi":"10.1369/00221554251365996","DOIUrl":"https://doi.org/10.1369/00221554251365996","url":null,"abstract":"<p><p>The neuroendocrine protein 7B2 plays a crucial role in the maturation and activity regulation of prohormone convertase 2 (PC2). To elucidate the relationship between 7B2 and PC2 expression in endocrine tissues, we generated synthetic peptide antibodies in guinea pigs. The antigenic peptide sequences were selected to correspond to three different positions in the rat amino acid (aa) sequence: The <i>N</i>-terminal aa 1-14 is situated immediately following the signal sequence, the middle aa 77-90 contains a part of the proPC2 activation domain, and the <i>C</i>-terminal aa 156-168 functions to suppress PC2 activity. These antibodies demonstrated specific reactivity across a diverse array of animal species. The reactivity of these antibodies differed, suggesting that the molecular form of 7B2 differs depending on the endocrine cell, and a different expression pattern was demonstrated in rat and dog pituitary intermediate cells. The colocalization of 7B2 and PC2 in prolactin (PRL) granules in rat pituitary mammotrophs supports the interaction between these proteins. However, the expression intensities of these proteins did not correspond, and epitope-related disparities were detected. These results may be indicative of alterations in the molecular state associated with the dynamics of the interaction between 7B2 and PC2.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":"73 7-8","pages":"289-314"},"PeriodicalIF":1.5,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12378109/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144957178","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Expression of Mitochondrial Uncoupling Proteins and GABA Signaling Molecules in Unstimulated and Nerve Growth Factor-Stimulated PC12 Cells: Models for Chromaffin Cells and Sympathetic Neurons.","authors":"Keita Harada, Hidetada Matsuoka, Masumi Inoue","doi":"10.1369/00221554251332981","DOIUrl":"10.1369/00221554251332981","url":null,"abstract":"<p><p>PC12 cells are a cell line originating from rat adrenal medullary chromaffin (AMC) cells. They extend a neurite-like structure in response to nerve growth factor (NGF). Thus, unstimulated and NGF-stimulated PC12 cells are used as models for AMC cells and sympathetic ganglion cells, respectively. However, how closely unstimulated and stimulated PC12 cells resemble AMC cells and sympathetic neurons, respectively, has not been elucidated sufficiently. We explored these issues by using biochemical and immunocytochemical methods. AMC cells and PC12 cells selectively expressed uncoupling protein 3 (UCP3) and uncoupling protein 4 (UCP4), respectively, and glucocorticoid activity inhibited UCP4 expression in PC12 cells. PC12 cells expressed extremely low levels of chromaffin granule-associated proteins, whereas the amount of synaptophysin, a synaptic vesicle-associated protein, was much higher than that in the adrenal medulla. Similar to AMC cells, the muscarinic receptor type 1 was located at the cell periphery in unstimulated PC12 cells, and its expression was markedly enhanced by NGF. Furthermore, NGF stimulation abolished the expression of GABA signaling molecules in PC12 cells. The results suggest that the properties of unstimulated PC12 cells are between those of AMC cells and sympathetic ganglion cells and GABA signaling is intrinsic to AMC cells.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"251-266"},"PeriodicalIF":1.9,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12037542/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143995776","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hannes Öster, Trina Chen, Ari Mennander, Timo Paavonen, Ivana Kholová
{"title":"CMKLR-1 Expression in Human Aorta Is Dominated by Smooth Muscle Cell Positivity.","authors":"Hannes Öster, Trina Chen, Ari Mennander, Timo Paavonen, Ivana Kholová","doi":"10.1369/00221554251350743","DOIUrl":"10.1369/00221554251350743","url":null,"abstract":"<p><p>Chemokine-like receptor 1 (CMKLR-1) is a G-protein-coupled receptor that functions as the binding site of chemerin. It induces chemotaxis in various immune cells, and it has recently been linked to vascular remodeling in inflammation. In this study, we investigated CMKLR-1 expression in human aorta samples, focusing on its distribution across different cell types and its potential association with clinical and histomorphological data, particularly concerning aortic dissection and aneurysms. We analyzed 62 aorta samples taken from patients undergoing dissection surgery (n = 27) or aneurysm/dilatation (n = 35) using immunohistochemical CMKLR-1 staining. CMKLR-1 expression was primarily observed in the smooth muscle cells (SMCs) of the aortic media layer, and band-like coloring appeared in the unstained center section. CMKLR-1 positivity in the inner and outer parts of the media layer was observed in only a few cases. One-third of the vasa vasorum exhibited staining. Staining in lymphocytes, macrophages, and endothelia was rare. No significant differences in CMKLR-1 expression were found between the dissection and aneurysm cases, and the clinical or histomorphological data. Although CMKLR-1 expression did not distinguish between the studied conditions, its presence in the aortic media, especially in SMCs, is noteworthy.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":" ","pages":"237-249"},"PeriodicalIF":1.9,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12187715/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144475621","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}