Osteosarcoma (OS) cells that deviate from the normal osteogenic differentiation pathway from mesenchymal stem cells (MSCs) to less migratory osteocytes are in an undifferentiated and highly malignant state. β-glycerophosphate (β-gp) is commonly used to induce the osteogenic differentiation of MSCs, inorganic phosphate (Pi) is also widely used to promote MSC differentiation into osteocytes. Recently, OS cells were found to differentiate into an osteocyte-like state by culturing in osteocyte differentiation medium containing β-gp, dexamethasone, and ascorbate. However, whether Pi can induce the osteogenic differentiation of OS cells and its underlying mechanisms remain unclear. In this study, we evaluated the ability of two types of Pi (i.e., disodium phosphate [Na2HPO4] and monosodium phosphate [NaH2PO4]) to promote the osteogenic differentiation of OS cells. Culturing OS cells in Pi-supplemented medium resulted in increased osteogenic marker gene expression and calcium deposition and reduced cell motility. Notably, Na2HPO4 exhibited particularly strong differentiation-inducing effects. Furthermore, our data suggest that WNT5b, a key factor of the noncanonical Wnt signaling pathway, is involved in the Na2HPO4-induced osteogenic differentiation of OS cells. These findings suggest that above 3 mM of Na2HPO4 function as an inducer of osteocyte-like differentiation in OS cells and that targeting this pathway may offer new therapeutic strategies to suppress OS metastasis.
{"title":"Osteocyte-like differentiation of osteosarcoma by inorganic phosphate","authors":"Yuya Suzuki , Makoto Takeuchi , Sumie Koike , Satoshi Takagi , Ryohei Katayama","doi":"10.1016/j.diff.2025.100912","DOIUrl":"10.1016/j.diff.2025.100912","url":null,"abstract":"<div><div>Osteosarcoma (OS) cells that deviate from the normal osteogenic differentiation pathway from mesenchymal stem cells (MSCs) to less migratory osteocytes are in an undifferentiated and highly malignant state. β-glycerophosphate (β-gp) is commonly used to induce the osteogenic differentiation of MSCs, inorganic phosphate (Pi) is also widely used to promote MSC differentiation into osteocytes. Recently, OS cells were found to differentiate into an osteocyte-like state by culturing in osteocyte differentiation medium containing β-gp, dexamethasone, and ascorbate. However, whether Pi can induce the osteogenic differentiation of OS cells and its underlying mechanisms remain unclear. In this study, we evaluated the ability of two types of Pi (i.e., disodium phosphate [Na<sub>2</sub>HPO<sub>4</sub>] and monosodium phosphate [NaH<sub>2</sub>PO<sub>4</sub>]) to promote the osteogenic differentiation of OS cells. Culturing OS cells in Pi-supplemented medium resulted in increased osteogenic marker gene expression and calcium deposition and reduced cell motility. Notably, Na<sub>2</sub>HPO<sub>4</sub> exhibited particularly strong differentiation-inducing effects. Furthermore, our data suggest that WNT5b, a key factor of the noncanonical Wnt signaling pathway, is involved in the Na<sub>2</sub>HPO<sub>4</sub>-induced osteogenic differentiation of OS cells. These findings suggest that above 3 mM of Na<sub>2</sub>HPO<sub>4</sub> function as an inducer of osteocyte-like differentiation in OS cells and that targeting this pathway may offer new therapeutic strategies to suppress OS metastasis.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"146 ","pages":"Article 100912"},"PeriodicalIF":2.6,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145057165","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-01Epub Date: 2025-08-13DOI: 10.1016/j.diff.2025.100900
Yinyu Fu, Bo Yang, Ling Lai, Jun He, Xinzhu Li , Jin Hou
The excision of introns and subsequent ligation of exons in precursor messenger RNA (pre-mRNA) is a fundamental mechanism requisite for the expression of eukaryotic genes. Alternative splicing (AS) serves as a potent amplifying factor, augmenting the spectrum of protein isoforms that can emanate from a singular genetic locus, thereby bolstering proteomic diversity. Perturbations in the regulatory framework of pre-mRNA splicing have been associated with an extensive array of pathological conditions. Oral and maxillofacial morphogenesis, a multifaceted process governed by intricate molecular interactions during embryonic development, is also susceptible to the modulating influence of alternative splicing. Aberrations or dysfunctions in the components responsible for alternative splicing during this critical developmental window can culminate in abnormal craniofacial architectures. In this scholarly review, our emphasis is placed on the exploration of RNA splicing as an emergent feature in oral and maxillofacial development. Importantly, we spotlight instances of splicing dysregulation that contribute to either non-syndromic or syndromic manifestations of cleft palate and enamel developmental anomalies. Deepening our understanding of the role of RNA splicing components and potential downstream effectors in oral and maxillofacial development may provide invaluable insights for prenatal diagnostic modalities.
{"title":"Emerging role of alternative splicing in oral and maxillofacial development","authors":"Yinyu Fu, Bo Yang, Ling Lai, Jun He, Xinzhu Li , Jin Hou","doi":"10.1016/j.diff.2025.100900","DOIUrl":"10.1016/j.diff.2025.100900","url":null,"abstract":"<div><div>The excision of introns and subsequent ligation of exons in precursor messenger RNA (pre-mRNA) is a fundamental mechanism requisite for the expression of eukaryotic genes. Alternative splicing (AS) serves as a potent amplifying factor, augmenting the spectrum of protein isoforms that can emanate from a singular genetic locus, thereby bolstering proteomic diversity. Perturbations in the regulatory framework of pre-mRNA splicing have been associated with an extensive array of pathological conditions. Oral and maxillofacial morphogenesis, a multifaceted process governed by intricate molecular interactions during embryonic development, is also susceptible to the modulating influence of alternative splicing. Aberrations or dysfunctions in the components responsible for alternative splicing during this critical developmental window can culminate in abnormal craniofacial architectures. In this scholarly review, our emphasis is placed on the exploration of RNA splicing as an emergent feature in oral and maxillofacial development. Importantly, we spotlight instances of splicing dysregulation that contribute to either non-syndromic or syndromic manifestations of cleft palate and enamel developmental anomalies. Deepening our understanding of the role of RNA splicing components and potential downstream effectors in oral and maxillofacial development may provide invaluable insights for prenatal diagnostic modalities.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100900"},"PeriodicalIF":2.6,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144864608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-01Epub Date: 2025-07-25DOI: 10.1016/j.diff.2025.100897
Jianchun Zhang , Xiexin Wang , Xuanhe Feng , Yan Chen , Ke Yu , Zhiwei Jiang
piRNAs are considered to be plentiful and heterogeneous non-coding RNAs, playing a significant role in embryonic patterning and stem cells. In recent research, scientists have explored how piRNAs contribute to both biological processes and roles in stem cells. Their regulatory action is essential for enabling stem cells to both renew themselves and differentiate into specialized types. piRNAs mediate transposable element (TE) silencing to ensure gene integrity via epigenetic repression mechanisms, ‘ping-pong’ mechanism, and splicing. Recent findings reveal that piRNAs are involved in silencing TE and regulating protein-coding mRNAs. This regulation suppresses the target mRNAs either via cleavage mediated by PIWI proteins or through recruiting the CCR4-NOT complex. Thereby the processes of stem cell self-renewal and differentiation have been lightened. piRNAs together with PIWI proteins contribute to controlling the translation of factors. As biological translation regulators, piRNAs in germline stem cells (GSCs) can trigger cell fate by specific regulation of mRNA targets. This role of piRNAs in regulating translation appears to be conserved across stem cells and is likely essential for maintaining stem cell balance. Furthermore, piRNAs along with PIWI proteins can be the biomarkers of stem cells. This review concentrates on recent findings that explore the developmental and biological functions of piRNAs, with a particular emphasis on their contributions to early embryonic organization and the regulation of stem cell behavior.
{"title":"Roles and mechanisms of piRNAs in self-renewal and differentiation of germline stem cells","authors":"Jianchun Zhang , Xiexin Wang , Xuanhe Feng , Yan Chen , Ke Yu , Zhiwei Jiang","doi":"10.1016/j.diff.2025.100897","DOIUrl":"10.1016/j.diff.2025.100897","url":null,"abstract":"<div><div>piRNAs are considered to be plentiful and heterogeneous non-coding RNAs, playing a significant role in embryonic patterning and stem cells. In recent research, scientists have explored how piRNAs contribute to both biological processes and roles in stem cells. Their regulatory action is essential for enabling stem cells to both renew themselves and differentiate into specialized types. piRNAs mediate transposable element (TE) silencing to ensure gene integrity via epigenetic repression mechanisms, ‘ping-pong’ mechanism, and splicing. Recent findings reveal that piRNAs are involved in silencing TE and regulating protein-coding mRNAs. This regulation suppresses the target mRNAs either via cleavage mediated by PIWI proteins or through recruiting the CCR4-NOT complex. Thereby the processes of stem cell self-renewal and differentiation have been lightened. piRNAs together with PIWI proteins contribute to controlling the translation of factors. As biological translation regulators, piRNAs in germline stem cells (GSCs) can trigger cell fate by specific regulation of mRNA targets. This role of piRNAs in regulating translation appears to be conserved across stem cells and is likely essential for maintaining stem cell balance. Furthermore, piRNAs along with PIWI proteins can be the biomarkers of stem cells. This review concentrates on recent findings that explore the developmental and biological functions of piRNAs, with a particular emphasis on their contributions to early embryonic organization and the regulation of stem cell behavior.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100897"},"PeriodicalIF":2.6,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144763779","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-01Epub Date: 2025-08-07DOI: 10.1016/j.diff.2025.100899
Naomi M. Calhoun, Richard R. Behringer
WNT7A regulates numerous developmental processes. It can activate canonical and non-canonical signaling depending on context. It is expressed in the developing central nervous system, limb buds, reproductive organs, and other tissues. Spontaneous and targeted Wnt7a mutations in mouse models resulted in abnormal limbs, defects in male and female reproductive tract organs, infertility, and defects in cerebellar axon remodeling. In zebrafish, wnt7aa mutants exhibited neurogenesis and angiogenesis defects in the central nervous system. In humans, recessive WNT7A missense and nonsense mutations resulted in severe limb and pelvic bone defects. Alterations in WNT7A expression correlated with multiple types of cancer.
{"title":"WNT7A","authors":"Naomi M. Calhoun, Richard R. Behringer","doi":"10.1016/j.diff.2025.100899","DOIUrl":"10.1016/j.diff.2025.100899","url":null,"abstract":"<div><div>WNT7A regulates numerous developmental processes. It can activate canonical and non-canonical signaling depending on context. It is expressed in the developing central nervous system, limb buds, reproductive organs, and other tissues. Spontaneous and targeted <em>Wnt7a</em> mutations in mouse models resulted in abnormal limbs, defects in male and female reproductive tract organs, infertility, and defects in cerebellar axon remodeling. In zebrafish, <em>wnt7aa</em> mutants exhibited neurogenesis and angiogenesis defects in the central nervous system. In humans, recessive <em>WNT7A</em> missense and nonsense mutations resulted in severe limb and pelvic bone defects. Alterations in <em>WNT7A</em> expression correlated with multiple types of cancer.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100899"},"PeriodicalIF":2.6,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144841502","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-01Epub Date: 2025-07-12DOI: 10.1016/j.diff.2025.100896
Kristen Russell , Christina A. Kearns , Macie B. Walker , Christopher S. Knoeckel , Angeles B. Ribera , Caleb A. Doll , Bruce Appel
Myelin, a specialized membrane produced by oligodendroglial cells in the central nervous system, wraps axons to enhance conduction velocity and maintain axon health. Not all axons are myelinated, and not all myelinated axons are uniformly wrapped along their lengths. Several lines of evidence indicate that neuronal activity can influence myelination, however, the cellular and molecular mechanisms that mediate communication between axons and oligodendrocytes remain poorly understood. Prior research showed that the neurotrophic growth factor Bdnf and its receptor Ntrk2 promote myelination in rodents, raising the possibility that Bdnf and Ntrk2 convey myelin-promoting signals from neurons to oligodendrocytes. We explored this possibility using a combination of gene expression analyses, gene function tests, and myelin sheath formation assays in zebrafish larvae. Altogether, our data indicate that, although not essential for myelination, Bdnf-Ntrk2 signaling contributes to the timely formation of myelin in the developing zebrafish spinal cord.
{"title":"Bdnf and Ntrk2 functions promote but are not essential for spinal cord myelination in larval zebrafish","authors":"Kristen Russell , Christina A. Kearns , Macie B. Walker , Christopher S. Knoeckel , Angeles B. Ribera , Caleb A. Doll , Bruce Appel","doi":"10.1016/j.diff.2025.100896","DOIUrl":"10.1016/j.diff.2025.100896","url":null,"abstract":"<div><div>Myelin, a specialized membrane produced by oligodendroglial cells in the central nervous system, wraps axons to enhance conduction velocity and maintain axon health. Not all axons are myelinated, and not all myelinated axons are uniformly wrapped along their lengths. Several lines of evidence indicate that neuronal activity can influence myelination, however, the cellular and molecular mechanisms that mediate communication between axons and oligodendrocytes remain poorly understood. Prior research showed that the neurotrophic growth factor Bdnf and its receptor Ntrk2 promote myelination in rodents, raising the possibility that Bdnf and Ntrk2 convey myelin-promoting signals from neurons to oligodendrocytes. We explored this possibility using a combination of gene expression analyses, gene function tests, and myelin sheath formation assays in zebrafish larvae. Altogether, our data indicate that, although not essential for myelination, Bdnf-Ntrk2 signaling contributes to the timely formation of myelin in the developing zebrafish spinal cord.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100896"},"PeriodicalIF":2.2,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144633198","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-01Epub Date: 2025-07-13DOI: 10.1016/j.diff.2025.100895
Alannah Morse, Hailey Kaba, Corinne Leighty, Emily MacLean, Rosemarie Mirabella, Mary Reinaker, Rohan Harris, Jazmyn Moodie, Antonio Rockwell
The m6A modification is responsible for regulating several aspects of RNA metabolism. The enzyme that catalyzes this modification Mettl3, is highly conserved and required for numerous biological processes such as spermatogenesis. Here we examine the role of Mettl3 in germline function during Drosophila spermatogenesis. We find that depletion of Mettl3 in the germline results in errors in late-stage spermatogenesis, the process known as spermiogenesis. In germline knockdowns, actin cones bound to spermatids fail to remain tightly packed during the individualization process. Issues with actin cone assembly appear to disrupt progression through spermiogenesis resulting in waste bag deficiency, an indicator of abnormal spermatid individualization. These errors result in little to no sperm in seminal vesicles culminating in reduced fertility in germline knockdowns. Furthermore, our findings suggest Hsp60B is misregulated in knockdowns, which potentially explains at least some observed phenotypes. Collectively, the data presented in this investigation suggests Mettl3 has a prominent role in regulating spermatid differentiation during Drosophila spermatogenesis.
{"title":"Mettl3 is required for germline function during Drosophila spermatogenesis","authors":"Alannah Morse, Hailey Kaba, Corinne Leighty, Emily MacLean, Rosemarie Mirabella, Mary Reinaker, Rohan Harris, Jazmyn Moodie, Antonio Rockwell","doi":"10.1016/j.diff.2025.100895","DOIUrl":"10.1016/j.diff.2025.100895","url":null,"abstract":"<div><div>The m<sup>6</sup>A modification is responsible for regulating several aspects of RNA metabolism. The enzyme that catalyzes this modification Mettl3, is highly conserved and required for numerous biological processes such as spermatogenesis. Here we examine the role of Mettl3 in germline function during <em>Drosophila</em> spermatogenesis. We find that depletion of Mettl3 in the germline results in errors in late-stage spermatogenesis, the process known as spermiogenesis. In germline knockdowns, actin cones bound to spermatids fail to remain tightly packed during the individualization process. Issues with actin cone assembly appear to disrupt progression through spermiogenesis resulting in waste bag deficiency, an indicator of abnormal spermatid individualization. These errors result in little to no sperm in seminal vesicles culminating in reduced fertility in germline knockdowns. Furthermore, our findings suggest <em>Hsp60B</em> is misregulated in knockdowns, which potentially explains at least some observed phenotypes. Collectively, the data presented in this investigation suggests Mettl3 has a prominent role in regulating spermatid differentiation during <em>Drosophila</em> spermatogenesis.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100895"},"PeriodicalIF":2.2,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144665708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Fibronectin adhesion assay progenitors (FAA-CPs) and migratory assay progenitors (MCPs), subsets of mesenchymal-like stromal cells (MSCs), exhibit superior in-vitro chondrogenic potential compared to bone marrow (BM)-MSCs and chondrocytes. To assess this potential, differentiation studies followed by confirmatory staining for collagen deposition are utilized. Polarized light microscopy (PLM), based on birefringence principles, is a valuable tool for visualizing organized collagen fibers. Its use as a predictive tool for assessing chondrogenesis and osteogenesis has not been reported.
Methods
This study involved FAA-CPs, MCPs, chondrocytes, and BM-MSCs derived from osteoarthritic knee joints (n = 3). After phenotypic characterization, the cells underwent chondrogenic and osteogenic differentiation, followed by Picrosirius red staining and PLM analysis, including immunohistochemical analysis for collagen types I, II, and X.
Results
Birefringence assessment revealed greater collagen fibril alignment and significant remodeling in the BM-MSC group, which exhibited an arcade-like pattern. The MCP group displayed well-organized collagen fibrils in pericellular zones and as a peripheral band, while chondrocytes and FAA-CPs exhibited lower intensity birefringence, indicating random alignment. Areas with higher collagen type II deposition corresponded to reduced collagen type I and the absence of collagen type X, highlighting the unique fibrillar network seen with PLM was indicative of collagen type II.
Conclusion
While its application for osteogenesis was limited, probably due to the non-fibrillar nature of collagen type X, its value for chondrogenesis is notable. Although not directly reflecting chondrogenesis, PLM can serve as a valuable tool for gaining insights into collagen remodeling, particularly concerning collagen type II during chondrogenic differentiation.
{"title":"Evaluation of chondrogenesis and osteogenesis in human mesenchymal stem cells, chondrocytes, and chondroprogenitors using molecular markers, cellular markers and polarized microscopy","authors":"Archa Suresh , Ayshath Ruksana C , Ganesh Parasuraman , Mohana Priya , Grace Rebekah , Christo Jeyaraj , Elizabeth Vinod","doi":"10.1016/j.diff.2025.100898","DOIUrl":"10.1016/j.diff.2025.100898","url":null,"abstract":"<div><h3>Purpose</h3><div>Fibronectin adhesion assay progenitors (FAA-CPs) and migratory assay progenitors (MCPs), subsets of mesenchymal-like stromal cells (MSCs), exhibit superior in-vitro chondrogenic potential compared to bone marrow (BM)-MSCs and chondrocytes. To assess this potential, differentiation studies followed by confirmatory staining for collagen deposition are utilized. Polarized light microscopy (PLM), based on birefringence principles, is a valuable tool for visualizing organized collagen fibers. Its use as a predictive tool for assessing chondrogenesis and osteogenesis has not been reported.</div></div><div><h3>Methods</h3><div>This study involved FAA-CPs, MCPs, chondrocytes, and BM-MSCs derived from osteoarthritic knee joints (n = 3). After phenotypic characterization, the cells underwent chondrogenic and osteogenic differentiation, followed by Picrosirius red staining and PLM analysis, including immunohistochemical analysis for collagen types I, II, and X.</div></div><div><h3>Results</h3><div>Birefringence assessment revealed greater collagen fibril alignment and significant remodeling in the BM-MSC group, which exhibited an arcade-like pattern. The MCP group displayed well-organized collagen fibrils in pericellular zones and as a peripheral band, while chondrocytes and FAA-CPs exhibited lower intensity birefringence, indicating random alignment. Areas with higher collagen type II deposition corresponded to reduced collagen type I and the absence of collagen type X, highlighting the unique fibrillar network seen with PLM was indicative of collagen type II.</div></div><div><h3>Conclusion</h3><div>While its application for osteogenesis was limited, probably due to the non-fibrillar nature of collagen type X, its value for chondrogenesis is notable. Although not directly reflecting chondrogenesis, PLM can serve as a valuable tool for gaining insights into collagen remodeling, particularly concerning collagen type II during chondrogenic differentiation.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100898"},"PeriodicalIF":2.6,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144722381","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-01Epub Date: 2025-08-20DOI: 10.1016/j.diff.2025.100901
Xiu-Rong Wei , Dan Hu , Zi-Jiang Yang , Lv-Bin Yan , Guang-Yu Xu , Rui-Gang Zhang , Xiu-Juan Zhang
Non-POU domain containing octamer binding protein (NONO) is a multifunctional nuclear protein which plays important roles in regulating nuclear processes such as transcription and splicing. We aimed to delineate the effects and the underlying mechanisms of NONO on monocyte-macrophage lineage differentiation. By depolying a phorbol 12-myristate 13-acetate (PMA)-induced THP-1 cell differentiation model and a macrophage colony-stimulating factor (M-CSF)-induced mouse bone marrow cell differentiation model, we examined the expression pattern and the effects of NONO during monocyte-macrophage lineage differentiation. The research revealed that the expression of NONO protein progressively decreased during the M-CSF-induced differentiation of mouse bone marrow cells into macrophages and the PMA-induced differentiation of THP-1 human monocytic leukemia cells into macrophages. The monocyte-macrophage lineage differentiation process was enhanced in Nono gene knockout (Nono K.O.) mouse bone marrow cells as well as NONO knockdown (NONO K.D.) THP-1 cells. The study also found that reduced NONO expression enhanced the AKT phosphorylation during macrophage lineage differentiation. At the same time, the PI3K inhibitor suppressed THP-1 cell differentiation into macrophages and attenuated the AKT phosphorylation activation by PMA and NONO knockdown during PMA-induced differentiation of THP-1 cells into macrophages. These results suggested an important role of NONO in regulating monocyte-macrophage lineage differentiation and this process was mediated, at least partially, through PI3K/AKT signaling pathway.
{"title":"NONO regulates monocyte-macrophage lineage differentiation through a potential PI3K/AKT-dependent mechanism","authors":"Xiu-Rong Wei , Dan Hu , Zi-Jiang Yang , Lv-Bin Yan , Guang-Yu Xu , Rui-Gang Zhang , Xiu-Juan Zhang","doi":"10.1016/j.diff.2025.100901","DOIUrl":"10.1016/j.diff.2025.100901","url":null,"abstract":"<div><div>Non-POU domain containing octamer binding protein (NONO) is a multifunctional nuclear protein which plays important roles in regulating nuclear processes such as transcription and splicing. We aimed to delineate the effects and the underlying mechanisms of NONO on monocyte-macrophage lineage differentiation. By depolying a phorbol 12-myristate 13-acetate (PMA)-induced THP-1 cell differentiation model and a macrophage colony-stimulating factor (M-CSF)-induced mouse bone marrow cell differentiation model, we examined the expression pattern and the effects of NONO during monocyte-macrophage lineage differentiation. The research revealed that the expression of NONO protein progressively decreased during the M-CSF-induced differentiation of mouse bone marrow cells into macrophages and the PMA-induced differentiation of THP-1 human monocytic leukemia cells into macrophages. The monocyte-macrophage lineage differentiation process was enhanced in <em>Nono</em> gene knockout (Nono K.O.) mouse bone marrow cells as well as <em>NONO</em> knockdown (NONO K.D.) THP-1 cells. The study also found that reduced NONO expression enhanced the AKT phosphorylation during macrophage lineage differentiation. At the same time, the PI3K inhibitor suppressed THP-1 cell differentiation into macrophages and attenuated the AKT phosphorylation activation by PMA and NONO knockdown during PMA-induced differentiation of THP-1 cells into macrophages. These results suggested an important role of NONO in regulating monocyte-macrophage lineage differentiation and this process was mediated, at least partially, through PI3K/AKT signaling pathway.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100901"},"PeriodicalIF":2.6,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144895308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-01Epub Date: 2025-08-23DOI: 10.1016/j.diff.2025.100902
Tingwei Gao , Jiankun Wang , Jiajia Wang , Xi Li , Zhanhao Xiao
Sirolimus can inhibit osteoclastogenesis. But sirolimus-activated autophagy is a favorable factor for osteoclastogenesis. This study aimed to explore the significance of autophagy in sirolimus-regulated osteoclastogenesis. Our results confirmed that sirolimus inhibited osteoclastic differentiation (including the number and size of osteoclasts as well as the expression of osteoclastic genes) and promotes osteoclast precursor (OCP) autophagy (including LC3 conversion and autophagosome/autolysosome formation). As expected, OCP autophagy (including LC3 conversion and LC3-puncta formation) promoted by sirolimus was reversed by autophagy inactivation with 3-MA or Atg13 silencing. Importantly, compared with single intervention of sirolimus, the combination of sirolimus and 3-MA or Atg13 silencing more effectively inhibited osteoclastic differentiation and OCP proliferation. In vivo experiments also demonstrated that the combination of sirolimus and Atg13-silencing adeno-associated viruses (AAVs) was more effective than sirolimus alone in improving decreased bone density and damaged bone microstructure (including Micro-CT imaging results, bone tissue parameters and trabecular area), and attenuating osteoclastic activity (including the abundance of osteoclasts in trabecular bones and the production of osteoclastic markers in serum) in ovariectomized (OVX) mice. In conclusion, repressing Atg13-related autophagy can effectively enhance the function of sirolimus in inhibiting osteoclastogenesis by counteracting its pro-autophagic activity. Therefore, the combination of sirolimus and Atg13-targeting therapy is expected to enhance the efficacy of sirolimus in ameliorating osteoclastic osteoporosis.
{"title":"Inhibition of Atg13-mediated autophagy enhances the anti-osteoclastogenic effect of sirolimus by counteracting its pro-autophagic activity","authors":"Tingwei Gao , Jiankun Wang , Jiajia Wang , Xi Li , Zhanhao Xiao","doi":"10.1016/j.diff.2025.100902","DOIUrl":"10.1016/j.diff.2025.100902","url":null,"abstract":"<div><div>Sirolimus can inhibit osteoclastogenesis. But sirolimus-activated autophagy is a favorable factor for osteoclastogenesis. This study aimed to explore the significance of autophagy in sirolimus-regulated osteoclastogenesis. Our results confirmed that sirolimus inhibited osteoclastic differentiation (including the number and size of osteoclasts as well as the expression of osteoclastic genes) and promotes osteoclast precursor (OCP) autophagy (including LC3 conversion and autophagosome/autolysosome formation). As expected, OCP autophagy (including LC3 conversion and LC3-puncta formation) promoted by sirolimus was reversed by autophagy inactivation with 3-MA or Atg13 silencing. Importantly, compared with single intervention of sirolimus, the combination of sirolimus and 3-MA or Atg13 silencing more effectively inhibited osteoclastic differentiation and OCP proliferation. <em>In vivo</em> experiments also demonstrated that the combination of sirolimus and Atg13-silencing adeno-associated viruses (AAVs) was more effective than sirolimus alone in improving decreased bone density and damaged bone microstructure (including Micro-CT imaging results, bone tissue parameters and trabecular area), and attenuating osteoclastic activity (including the abundance of osteoclasts in trabecular bones and the production of osteoclastic markers in serum) in ovariectomized (OVX) mice. In conclusion, repressing Atg13-related autophagy can effectively enhance the function of sirolimus in inhibiting osteoclastogenesis by counteracting its pro-autophagic activity. Therefore, the combination of sirolimus and Atg13-targeting therapy is expected to enhance the efficacy of sirolimus in ameliorating osteoclastic osteoporosis.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100902"},"PeriodicalIF":2.6,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144903192","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-07-01Epub Date: 2025-06-13DOI: 10.1016/j.diff.2025.100881
Pavani Ponnimbaduge Perera , Kaitlyn Webster , Misty R. Riddle
The enteric nervous system (ENS) regulates gastrointestinal (GI) functions such as secretion, blood flow, and motility, yet how its structure and function evolve with dietary adaptations remains unclear. Astyanax mexicanus, a teleost fish with surface and cave morphotypes, offers a model to explore these changes; cavefish exhibit altered GI motility and transit that may help them adapt to their unique diet. We compared early ENS development in surface fish and cavefish, tracking enteric neural crest cell (ENCC) migration and differentiation using phox2bb and HuC/D markers. We found that ENCCs reach the gut by 36 hours post-fertilization (hpf) in both morphotypes but migrate and differentiate more rapidly along the gut in cavefish. To explore the genetic basis of this difference, we used available genomic datasets to compare the predicted peptide sequences of genes important for ENS development in other species and identified mutations that could impact protein function, for example in the endothelin signaling genes important for ENCC migration and differentiation. We specifically examined the expression of endothelin-3 (edn3) and endothelin receptor-b a (ednrba) during ENCC migration and found that the localization of edn3, but not ednrba, is consistent with a potential role in ENS development. Overall, our findings establish A. mexicanus as a model for studying evolution of ENS development.
肠神经系统（ENS）调节胃肠道（GI）功能，如分泌、血流和运动，但其结构和功能如何随着饮食适应而进化尚不清楚。Astyanax mexicanus，一种具有表面和洞穴形态的硬骨鱼，为探索这些变化提供了一个模型；洞穴鱼表现出改变的胃肠道运动和运输，这可能有助于它们适应独特的饮食。我们比较了表层鱼和洞穴鱼的早期ENS发育，利用phox2bb和HuC/D标记跟踪了肠神经嵴细胞（enteric neural crest cell， ENCC）的迁移和分化。我们发现，两种形态的ENCCs在受精后36小时到达肠道，但在洞穴鱼中，ENCCs沿着肠道迁移和分化的速度更快。为了探索这种差异的遗传基础，我们使用现有的基因组数据集来比较其他物种ENS发育重要基因的预测肽序列，并确定可能影响蛋白质功能的突变，例如对ENCC迁移和分化重要的内皮素信号基因。我们专门检查了内皮素-3 （edn3）和内皮素受体-b - a （ednrba）在ENCC迁移过程中的表达，发现edn3的定位，而不是ednrba，与ENS发展的潜在作用一致。总的来说，我们的研究结果建立了墨西哥拟南猿作为研究ENS发展进化的模型。
{"title":"Enteric neural crest development in Astyanax mexicanus surface fish and cavefish","authors":"Pavani Ponnimbaduge Perera , Kaitlyn Webster , Misty R. Riddle","doi":"10.1016/j.diff.2025.100881","DOIUrl":"10.1016/j.diff.2025.100881","url":null,"abstract":"<div><div>The enteric nervous system (ENS) regulates gastrointestinal (GI) functions such as secretion, blood flow, and motility, yet how its structure and function evolve with dietary adaptations remains unclear. <em>Astyanax mexicanus</em>, a teleost fish with surface and cave morphotypes, offers a model to explore these changes; cavefish exhibit altered GI motility and transit that may help them adapt to their unique diet. We compared early ENS development in surface fish and cavefish, tracking enteric neural crest cell (ENCC) migration and differentiation using <em>phox2bb</em> and HuC/D markers. We found that ENCCs reach the gut by 36 hours post-fertilization (hpf) in both morphotypes but migrate and differentiate more rapidly along the gut in cavefish. To explore the genetic basis of this difference, we used available genomic datasets to compare the predicted peptide sequences of genes important for ENS development in other species and identified mutations that could impact protein function, for example in the endothelin signaling genes important for ENCC migration and differentiation. We specifically examined the expression of <em>endothelin-3 (edn3)</em> and <em>endothelin receptor-b a (ednrba)</em> during ENCC migration and found that the localization of <em>edn3</em>, but not <em>ednrba</em>, is consistent with a potential role in ENS development. Overall, our findings establish <em>A. mexicanus</em> as a model for studying evolution of ENS development.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"144 ","pages":"Article 100881"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144365759","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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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