Journal of Histochemistry & Cytochemistry最新文献

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Expression of Protein Markers in Spermatogenic and Supporting Sertoli Cells Affected by High Abdominal Temperature in Cryptorchidism Model Mice. 隐睾症模型小鼠的生精细胞和支持细胞受高腹温影响的蛋白标记表达
IF 1.9 4区 生物学
Journal of Histochemistry & Cytochemistry Pub Date : 2023-07-01 Epub Date: 2023-07-10 DOI: 10.1369/00221554231185626
Arunothai Wanta, Kazuhiro Noguchi, Taichi Sugawara, Kayoko Sonoda, Suthat Duangchit, Tomohiko Wakayama
{"title":"Expression of Protein Markers in Spermatogenic and Supporting Sertoli Cells Affected by High Abdominal Temperature in Cryptorchidism Model Mice.","authors":"Arunothai Wanta, Kazuhiro Noguchi, Taichi Sugawara, Kayoko Sonoda, Suthat Duangchit, Tomohiko Wakayama","doi":"10.1369/00221554231185626","DOIUrl":"10.1369/00221554231185626","url":null,"abstract":"<p><p>Cryptorchidism is a congenital abnormality resulting in increased rates of infertility and testicular cancer. We used cryptorchidism model mice that presented with the translocation of the left testis from the scrotum to the abdominal cavity. Mice underwent the surgical procedure of the left testis at day 0 and were sacrificed at days 3, 5, 7, 14, 21, and 28 post-operatively. The weight of the left cryptorchid testis decreased significantly at days 21 and 28. The morphological changes were observed after 5 days and showed detached spermatogenic cells and abnormal formation of acrosome at day 5, multinucleated giant cells at day 7, and atrophy of seminiferous tubules at days 21 and 28. The high abdominal temperature disrupted the normal expression of cell adhesion molecule-1, Nectin-2, and Nectin-3 which are essential for spermatogenesis. In addition, the pattern and alignment of acetylated tubulin in cryptorchid testes were also changed at days 5, 7, 14, 21, and 28. Ultrastructure of cryptorchid testes revealed giant cells that had been formed by spermatogonia, spermatocytes, and round and elongating spermatids. The study's findings reveal that cryptorchidism's duration is linked to abnormal changes in the testis, impacting protein marker expression in spermatogenic and Sertoli cells. These changes stem from the induction of high abdominal temperature.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10363907/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9869843","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}
引用次数: 0
Absence of E-Cadherin and β-Catenin in the Basal Plasma Membrane of Collecting Duct Cells During NDI Development and Recovery. 在 NDI 发育和恢复过程中,集导管细胞基底浆膜中缺少 E-Cadherin 和 β-Catenin。
IF 1.9 4区 生物学
Journal of Histochemistry & Cytochemistry Pub Date : 2023-07-01 Epub Date: 2023-07-13 DOI: 10.1369/00221554231185809
Xabier Sørtvedt, Rikke Nielsen, Jeppe Praetorius, Birgitte M Christensen
{"title":"Absence of E-Cadherin and β-Catenin in the Basal Plasma Membrane of Collecting Duct Cells During NDI Development and Recovery.","authors":"Xabier Sørtvedt, Rikke Nielsen, Jeppe Praetorius, Birgitte M Christensen","doi":"10.1369/00221554231185809","DOIUrl":"10.1369/00221554231185809","url":null,"abstract":"<p><p>Lithium (Li) induces severe polyuria and polydipsia in up to 40% of patients undergoing Li treatment. In rats, Li treatment induces a reversible cellular remodeling of the collecting duct (CD), decreasing the fraction of principal-to-intercalated cells. To investigate the potential role of adherens junction proteins, we performed immunohistochemistry on kidney cross-sections from rats treated with Li as well as rats undergoing recovery on a normal diet following 4 weeks of Li-treatment. We performed immunoelectron microscopy on cryosections to determine the ultrastructural localizations. Immunohistochemistry showed that E-cadherin and β-catenin were present in both the lateral and basal plasma membrane domains of CD cells. Immunoelectron microscopy confirmed that β-catenin was localized both to the lateral and the basal plasma membrane. The basal localization of both proteins was absent from a fraction of mainly principal cells after 10 and 15 days of Li-treatment. After 4 weeks of Li-treatment few to no cells were absent of E-cadherin and β-catenin at the basal plasma membrane. After 12 and 19 days of recovery some cells exhibited an absence of basal localization of both proteins. Thus, the observed localizational changes of E-cadherin and β-catenin appear before the cellular remodeling during both development and recovery from Li-NDI.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10363910/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9871810","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}
引用次数: 0
Commentary on a Classic JHC Article on the Histochemical Measurement of DNA Content in Cells. 评论 JHC 有关细胞中 DNA 含量的组织化学测量的经典文章。
IF 3.2 4区 生物学
Journal of Histochemistry & Cytochemistry Pub Date : 2023-06-01 Epub Date: 2023-06-13 DOI: 10.1369/00221554231182467
Cornelis J F van Noorden
{"title":"Commentary on a Classic JHC Article on the Histochemical Measurement of DNA Content in Cells.","authors":"Cornelis J F van Noorden","doi":"10.1369/00221554231182467","DOIUrl":"10.1369/00221554231182467","url":null,"abstract":"<p><p>This article comments on the significance of a highly cited review article on DNA cytochemical quantitation that was published in the <i>Journal of Histochemistry and Cytochemistry</i> in 2002 (David C. Hardie, T. Ryan Gregory, and Paul D.N. Hebert. From pixels to picograms: A beginners' guide to genome quantification by Feulgen image analysis densitometry.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10315993/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9752612","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}
引用次数: 0
Developmental Expression of the Cell Cycle Regulator p16INK4a in Retinal Glial Cells: A Novel Marker for Immature Ocular Astrocytes? 视网膜胶质细胞中细胞周期调节因子 p16INK4a 的发育表达:未成熟眼星形胶质细胞的新标记?
IF 3.2 4区 生物学
Journal of Histochemistry & Cytochemistry Pub Date : 2023-06-01 Epub Date: 2023-06-23 DOI: 10.1369/00221554231184286
Cristina Martinez-Fernandez de la Camara, Tina Storm, Ahmed Salman, Thomas Burgoyne, Martin Qvist Rasmussen, Harry O Orlans, Angela J Russell, Stephen G Davies, Alun R Barnard, Robert E MacLaren
{"title":"Developmental Expression of the Cell Cycle Regulator p16<sup>INK4a</sup> in Retinal Glial Cells: A Novel Marker for Immature Ocular Astrocytes?","authors":"Cristina Martinez-Fernandez de la Camara, Tina Storm, Ahmed Salman, Thomas Burgoyne, Martin Qvist Rasmussen, Harry O Orlans, Angela J Russell, Stephen G Davies, Alun R Barnard, Robert E MacLaren","doi":"10.1369/00221554231184286","DOIUrl":"10.1369/00221554231184286","url":null,"abstract":"<p><p>Retinal astrocytes are vital for neuronal homeostasis in the retina. Together with Müller glia, they provide retinal cells with neurotrophic factors, antioxidative support, and defense mechanisms such as the formation of the blood-retinal barrier. Substantial heterogeneity of astrocyte morphology and function represents a challenge for identification of distinct subtypes which may be potential targets for therapeutic purposes. Hence, identification of novel markers of astrocyte subpopulations is highly relevant to better understand the molecular mechanisms involved in retinal development, homeostasis, and pathology. In this study, we observed that the cell cycle regulator, p16<sup>INK4a</sup>, is expressed in immature astrocytes in the mouse retina. Immunohistochemical analysis showed p16<sup>INK4a</sup> expression in the optic nerve of wild-type mice from 3 days to 3 months of age and in the nerve fiber layer of the adult mouse retina. Colocalization of p16<sup>INK4a</sup> expression and glial fibrillary acidic protein (immature/mature astrocyte marker) tends to decrease with age. However, colocalization of p16<sup>INK4a</sup> expression and vimentin (immature astrocyte marker) remains high in the optic nerve from the early postnatal period to adulthood. The observations from this study provide a valuable tool for further investigations of ocular astrocytes in the developing retina as well as in degenerative retinopathies.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/0a/be/10.1369_00221554231184286.PMC10315990.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10127383","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}
引用次数: 0
Expression of JCAD and EGFR in Perineurial Cell-Cell Junctions of Human Inferior Alveolar Nerve. 人下肺泡神经周围细胞-细胞连接中 JCAD 和表皮生长因子受体的表达
IF 3.2 4区 生物学
Journal of Histochemistry & Cytochemistry Pub Date : 2023-06-01 Epub Date: 2023-06-13 DOI: 10.1369/00221554231182193
Yujiro Hiraoka, Megumi Matsumura, Yasumasa Kakei, Daisuke Takeda, Manabu Shigeoka, Akira Kimoto, Takumi Hasegawa, Masaya Akashi
{"title":"Expression of JCAD and EGFR in Perineurial Cell-Cell Junctions of Human Inferior Alveolar Nerve.","authors":"Yujiro Hiraoka, Megumi Matsumura, Yasumasa Kakei, Daisuke Takeda, Manabu Shigeoka, Akira Kimoto, Takumi Hasegawa, Masaya Akashi","doi":"10.1369/00221554231182193","DOIUrl":"10.1369/00221554231182193","url":null,"abstract":"<p><p>Although perineurium has an important role in maintenance of the blood-nerve barrier, understanding of perineurial cell-cell junctions is insufficient. The aim of this study was to analyze the expression of junctional cadherin 5 associated (JCAD) and epidermal growth factor receptor (EGFR) in the perineurium of the human inferior alveolar nerve (IAN) and investigate their roles in perineurial cell-cell junctions using cultured human perineurial cells (HPNCs). In human IAN, JCAD was strongly expressed in endoneurial microvessels. JCAD and EGFR were expressed at various intensities in the perineurium. In HPNCs, JCAD was clearly expressed at cell-cell junctions. EGFR inhibitor AG1478 treatment changed cell morphology and the ratio of JCAD-positive cell-cell contacts of HPNCs. Therefore, JCAD and EGFR may have a role in the regulation of perineurial cell-cell junctions.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10315992/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9752611","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}
引用次数: 0
Colonic Myenteric Plexus Neurodegeneration and Minor Colon Inflammation in Trimethyltin-induced Rat Model of Neurodegeneration. 三甲基锡诱导的大鼠神经变性模型中的结肠肠系膜神经丛神经变性和轻微结肠炎症
IF 3.2 4区 生物学
Journal of Histochemistry & Cytochemistry Pub Date : 2023-06-01 Epub Date: 2023-06-15 DOI: 10.1369/00221554231182195
Dian Eurike Septyaningtrias, Hilizza Awalina Zulfa, Mahayu Firsty Ramadhani, Sumaryati, Dewi Sulistyawati, Dewi Kartikawati Paramita, Yustina Andwi Ari Sumiwi, Rina Susilowati
{"title":"Colonic Myenteric Plexus Neurodegeneration and Minor Colon Inflammation in Trimethyltin-induced Rat Model of Neurodegeneration.","authors":"Dian Eurike Septyaningtrias, Hilizza Awalina Zulfa, Mahayu Firsty Ramadhani, Sumaryati, Dewi Sulistyawati, Dewi Kartikawati Paramita, Yustina Andwi Ari Sumiwi, Rina Susilowati","doi":"10.1369/00221554231182195","DOIUrl":"10.1369/00221554231182195","url":null,"abstract":"<p><p>Gastrointestinal symptoms are common health problems found during aging and neurodegenerative diseases. Trimethyltin-induced rat is known as an animal model of hippocampal degeneration with no data on enteric neurodegeneration. This study aimed to investigate the effect of trimethyltin (TMT) induction on the gastrointestinal tract. A 28-day animal study with male Sprague-Dawley rats (3 months old, 150-200 g) given a single TMT injection (8 mg/kg body weight, intraperitoneal) was conducted. The number of neurons in the colonic myenteric plexus was measured using stereological estimation. Histological scoring of colon inflammation, immunohistochemistry of tumor necrosis factor-α (TNF-α), and quantitative PCR were conducted. This study showed neuronal loss in the colonic myenteric plexus of TMT-induced rat model of neurodegeneration. Minor colon inflammation characterized by inflammatory cell infiltration and slightly higher expression of TNF-α in the colon mucosa were observed in the TMT-induced rat. However, the gut microbiota composition of the TMT-induced rat was not different from that of the control rats. This study demonstrates that TMT induces colonic myenteric plexus neurodegeneration and minor colon inflammation, which suggests the potential of this animal model to elucidate the communication between the gastrointestinal tract and central nervous system in neurodegenerative diseases.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10315991/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9750668","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}
引用次数: 0
Improving Yields in Multi-analyte Extractions by Utilizing Post-homogenized Tissue Debris. 利用均质化后的组织残渣提高多分析物萃取的产量
IF 3.2 4区 生物学
Journal of Histochemistry & Cytochemistry Pub Date : 2023-05-01 Epub Date: 2023-04-29 DOI: 10.1369/00221554231172823
Ala Petersons, Joseph Carlson, William Mathieson
{"title":"Improving Yields in Multi-analyte Extractions by Utilizing Post-homogenized Tissue Debris.","authors":"Ala Petersons, Joseph Carlson, William Mathieson","doi":"10.1369/00221554231172823","DOIUrl":"10.1369/00221554231172823","url":null,"abstract":"<p><p>In multi-analyte extractions, tissue is typically homogenized in a lysis buffer, and then DNA, RNA, and protein are purified from the supernatant. However, yields are typically lower than in dedicated, single-analyte extractions. In a two-part experiment, we assessed whether yields could be improved by revisiting the normally discarded, post-homogenized tissue debris. We initially performed additional homogenizations, each followed by a simultaneous extraction. These yielded no additional RNA, 13% additional DNA (which became progressively more degraded), and 161.7% additional protein (which changed in proteome when analyzed using SDS-PAGE). We then digested post-homogenized tissue debris from a simultaneous extraction using proteinase K and extracted DNA using silica spin columns or alcohol precipitation. An average additional DNA yield of 27.1% (silica spin columns) or 203.9% (alcohol precipitation) was obtained with/without compromising DNA integrity (assessment by long-range PCR, DNA Integrity Numbers, and size at peak fluorescence of electropherogram). Validation using a cohort of 65 tissue blocks returned an average additional DNA yield of 31.6% (silica columns) and 54.8% (alcohol precipitation). Users can therefore refreeze the homogenized remnants of tissue blocks rather than disposing of them and then perform additional DNA extractions if yields in the initial multi-analyte extractions were low.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10227881/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9659320","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}
引用次数: 0
Prematurity and Hyperoxia Have Different Effects on Alveolar and Microvascular Lung Development in the Rabbit. 早产和高氧对兔肺泡和微血管肺发育的影响不同。
IF 3.2 4区 生物学
Journal of Histochemistry & Cytochemistry Pub Date : 2023-05-01 DOI: 10.1369/00221554231177757
Giacomo Rößler, Jonas Labode, Yannick Regin, Thomas Salaets, André Gie, Jaan Toelen, Christian Mühlfeld
{"title":"Prematurity and Hyperoxia Have Different Effects on Alveolar and Microvascular Lung Development in the Rabbit.","authors":"Giacomo Rößler,&nbsp;Jonas Labode,&nbsp;Yannick Regin,&nbsp;Thomas Salaets,&nbsp;André Gie,&nbsp;Jaan Toelen,&nbsp;Christian Mühlfeld","doi":"10.1369/00221554231177757","DOIUrl":"https://doi.org/10.1369/00221554231177757","url":null,"abstract":"<p><p>Bronchopulmonary dysplasia (BPD) is a developmental disorder of infants born prematurely, characterized by disrupted alveolarization and microvascular maturation. However, the sequence of alveolar and vascular alterations is currently not fully understood. Therefore, we used a rabbit model to evaluate alveolar and vascular development under preterm birth and hyperoxia, respectively. Pups were born by cesarean section 3 days before term and exposed for 7 days to hyperoxia (95% O<sub>2</sub>) or normoxia (21% O<sub>2</sub>). In addition, term-born rabbits were exposed to normoxia for 4 days. Rabbit lungs were fixed by vascular perfusion and prepared for stereological analysis. Normoxic preterm rabbits had a significantly lower number of alveoli than term rabbits. The number of septal capillaries was lower in preterm rabbits but less pronounced than the alveolar reduction. In hyperoxic preterm rabbits, the number of alveoli was similar to that in normoxic preterm animals; however, hyperoxia had a severe additional negative effect on the capillary number. In conclusion, preterm birth had a strong effect on alveolar development, and hyperoxia had a more pronounced effect on capillary development. The data provide a complex picture of the vascular hypothesis of BPD which rather seems to reflect the ambient oxygen concentration than the effect of premature birth.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/bc/b1/10.1369_00221554231177757.PMC10227883.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10016331","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}
引用次数: 0
Commentary on a Classic JHC Article on the Development of Highly Sensitive Fluorochrome-labeled Tyramides for Immunocytochemistry. 评论 JHC 有关开发用于免疫细胞化学的高灵敏度荧光标记酪胺的经典文章。
IF 3.2 4区 生物学
Journal of Histochemistry & Cytochemistry Pub Date : 2023-05-01 Epub Date: 2023-05-18 DOI: 10.1369/00221554231178068
Kevin A Roth
{"title":"Commentary on a Classic JHC Article on the Development of Highly Sensitive Fluorochrome-labeled Tyramides for Immunocytochemistry.","authors":"Kevin A Roth","doi":"10.1369/00221554231178068","DOIUrl":"10.1369/00221554231178068","url":null,"abstract":"<p><p>This commentary reflects on the significance and impact of the highly cited companion article that was published in the <i>Journal of Histochemistry and Cytochemistry</i> in 1997 (Gijlswijk RPM et al. Fluorochrome-labeled Tyramides: Use in Immunocytochemistry and Fluorescence In Situ Hybridization. <i>Journal of Histochemistry & Cytochemistry</i>. 1997;45(3):375-382).</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10227882/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10033807","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}
引用次数: 0
Epitope Lability of Phosphorylated Biomarkers of the DNA Damage Response Pathway Results in Increased Vulnerability to Effects of Delayed or Incomplete Formalin Fixation. DNA 损伤应答途径磷酸化生物标记物的表位易变性导致更易受延迟或不完全福尔马林固定的影响。
IF 3.2 4区 生物学
Journal of Histochemistry & Cytochemistry Pub Date : 2023-05-01 Epub Date: 2023-04-29 DOI: 10.1369/00221554231174069
Elizabeth J Wiseman, Jennifer I Moss, James Atkinson, Hana Baakza, Emily Hayes, Sophie E Willis, Paul M Waring, Jaime Rodriguez Canales, Gemma N Jones
{"title":"Epitope Lability of Phosphorylated Biomarkers of the DNA Damage Response Pathway Results in Increased Vulnerability to Effects of Delayed or Incomplete Formalin Fixation.","authors":"Elizabeth J Wiseman, Jennifer I Moss, James Atkinson, Hana Baakza, Emily Hayes, Sophie E Willis, Paul M Waring, Jaime Rodriguez Canales, Gemma N Jones","doi":"10.1369/00221554231174069","DOIUrl":"10.1369/00221554231174069","url":null,"abstract":"<p><p>Phosphorylated biomarkers are crucial for our understanding of drug mechanism of action and dose selection during clinical trials, particularly for drugs that target protein kinases, such as DNA-damage-response (DDR) inhibitors. However, tissue fixation conditions needed to preserve DDR-specific phospho-biomarkers have not been previously investigated. Using xenograft tissues and tightly controlled formalin fixation conditions, we assessed how preanalytical factors affect phosphorylated DDR biomarkers pRAD50(Ser635), ɣH2AX(Ser139), pKAP1(Ser824), and non-phosphorylated biomarkers cMYC and ATM. Cold ischemia times ranged from 15 min to 6 hr, and the fixation duration ranged from 24 hr to 4 weeks. Epitopes pRAD50 and pKAP1 appeared the most labile assessed with staining loss after just 15 min of cold ischemia time, while ATM was more robust showing consistent expression up to 1 hr of cold ischemia. Notably, ɣH2AX expression was lost with formalin fixation over 48 hr. The use of core needle biopsies where possible and novel fixation methods such as the 2-step temperature-controlled formalin approach may improve phosphorylated biomarker preservation; however, practical challenges may affect wider clinical application. The most essential tissue-processing step when downstream analysis includes DDR phosphorylated biomarkers is immediate tissue submersion in formalin, without delay, upon excision from the patient, followed by room temperature fixation for 24 hr.</p>","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10227880/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9659316","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}
引用次数: 0
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