genesis最新文献

{"title":"Identification of a Novel nkx2.3 Transgenic Reporter Specifically Visualizing Craniofacial Muscle Development in Zebrafish","authors":"Yucheol Choe,&nbsp;Haewon Jeon,&nbsp;Sil Jin,&nbsp;Chong Pyo Choe","doi":"10.1002/dvg.70069","DOIUrl":"10.1002/dvg.70069","url":null,"abstract":"<div>\u0000 \u0000 <p>Transgenic reporters offer valuable opportunities to investigate the genetic and cellular basis of embryonic development in live organisms. While the specificity of transgenic reporters is essential for their effectiveness in developmental studies, many existing transgenic muscle reporters used for craniofacial muscle development in zebrafish also label trunk skeletal and cardiac muscles, as well as other head tissues. Here, we report the identification of a new transgenic reporter marking craniofacial muscles, <i>Tg(nkx2.3:Alcama-GFP)</i>, in zebrafish. This transgenic reporter exhibits Alcama-GFP expression in growing muscle fibers at embryonic stages and in grown muscle fibers at larval stages. However, endogenous <i>nkx2.3</i> transcripts are barely detected in growing craniofacial muscle fibers by in situ hybridization, and <i>nkx2.3</i> mutants display fairly normal ventral jaw muscles adjacent to the lower jaw cartilages and extraocular muscles adjacent to the eye. Although an essential role for Nkx2.3 in craniofacial muscle formation remains to be determined, the <i>Tg(nkx2.3:Alcama-GFP)</i> craniofacial muscle reporter provides better opportunities for the precise analysis of craniofacial muscle development at the cellular and genomic levels.</p>\u0000 </div>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"64 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148671110","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}

{"title":"An Efficient Method for Neuron-Like Differentiation of SH-SY5Y Neuroblastoma Cells Using Retinoic Acid and Laminin-Rich Extracellular Matrix","authors":"Marina Mantellatto Grigoli,&nbsp;Bianca Cruz Pachane,&nbsp;Angelina Maria Fuzer,&nbsp;Sabrina Dorta de Oliveira,&nbsp;Ana Beatriz Aparecida Targas,&nbsp;Vanessa Alexandre-Silva,&nbsp;Heloisa Sobreiro Selistre-de-Araujo,&nbsp;Patricia Regina Manzine,&nbsp;Marcia Regina Cominetti","doi":"10.1002/dvg.70066","DOIUrl":"10.1002/dvg.70066","url":null,"abstract":"<p>The human neuroblastoma cell line SH-SY5Y is widely used in vitro as a model due to its ability to acquire neuron-like properties. Conventional differentiation protocols often rely on retinoic acid (RA), which can lead to limited stability of neuronal-like phenotypes over time. In this study, we evaluated a differentiation approach combining RA with a laminin-rich extracellular matrix (LrECM). This approach accelerated differentiation, with neuron-like morphology and neurite outgrowth observed as early as 4 days and sustained features for up to 10 days. Immunofluorescence analysis indicated increased NeuN expression, and Western blot confirmed the sustained presence of β3-tubulin during differentiation. Furthermore, a trend toward enhanced acetylcholinesterase (AChE) activity suggested partial cholinergic features. Altogether, these findings indicate that incorporating LrECM supports the stability of RA-induced neuron-like characteristics in SH-SY5Y cells and may provide a useful alternative for studies of neuronal–phenotype differentiation.</p>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"64 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13429946/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148664690","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 Growth Factor Genes in the Body Wall of the Holothurian Eupentacta fraudatrix During Regenerative Processes","authors":"A. S. Girich","doi":"10.1002/dvg.70067","DOIUrl":"10.1002/dvg.70067","url":null,"abstract":"<div>\u0000 \u0000 <p>Research into regeneration in sea cucumbers makes a significant contribution to understanding the fundamental mechanisms of this process. However, these studies focus only on the regenerating organs—the intestine and aquapharyngeal bulb. The source of cells for regeneration remains out of sight. The cells of the body wall, under the influence of various activating molecules (growth factors), dedifferentiate, determine the direction of migration, and move toward the injury. Therefore, the aim of this study was to identify the most active growth factor genes in the body wall during the regenerative process in the sea cucumber <i>Eupentacta fraudatrix</i><i>.</i> We searched for and identified genes of the TGF-β, FGF, Neurotrophin, VEGF families. The sea cucumber <i>Eupentacta fraudatrix</i> possesses a set of growth factor genes characteristic of all echinoderms, with the exception of three orthologs of <i>gdf2</i> genes. A significant increase in the expression of <i>gdf2, bmp2/4, neurotrophin</i>, and <i>wnt5</i> genes was detected in the body wall as early as the first day of the regenerative process. A second increase in growth factor gene expression occurred on day 7, during the onset of the proliferation and growth phase. The results of this study identify the most promising genes for further investigation of their role in the signal transduction that triggers regeneration.</p>\u0000 </div>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"64 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148593931","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}

{"title":"Generation and Characterization of a Rdh1-iCre Line to Study Uterine Glandular Biology","authors":"Mana Ohtomo,&nbsp;Shunsuke Takarabe,&nbsp;Takafumi Namiki,&nbsp;Yui Kawata,&nbsp;Ryosuke Kaneko,&nbsp;Manabu Ozawa,&nbsp;Yasuhiro Yamada,&nbsp;Hisashi Mori,&nbsp;Atsuko Kageyama,&nbsp;Maki Kamoshita,&nbsp;Jumpei Terakawa,&nbsp;Junya Ito","doi":"10.1002/dvg.70068","DOIUrl":"10.1002/dvg.70068","url":null,"abstract":"<p>The uterus is an essential organ for fetal development in most mammals. Uterine glands, highly conserved structures in the mammalian uterus, play critical roles in the establishment and maintenance of pregnancy and have been implicated in the pathogenesis of uterine diseases, including endometrial cancer and endometriosis. Previous studies have shown that Retinol dehydrogenase 1 (<i>Rdh1</i>) is specifically expressed in the glandular epithelium (GE) from the onset of gland formation through adulthood. In this study, to develop a GE-specific Cre driver line, we generated <i>Rdh1-iCre</i> mice by introducing an improved Cre recombinase (<i>iCre</i>) into the <i>Rdh1</i> locus using the CRISPR/Cas9 system. To evaluate the utility of this model, <i>Rdh1-iCre</i> mice were crossed with <i>ROSA26-H2B-mCherry</i> reporter mice, and Cre-dependent reporter expression was analyzed. Robust mCherry fluorescence was observed throughout the uterine glands at 2 weeks after birth, coinciding with the active elongation and branching of the GE. These results demonstrate that the <i>Rdh1-iCre</i> mouse line is a valuable and highly efficient tool for investigating the physiological roles of uterine glands during development and pregnancy, as well as their contribution to the progression of GE-derived uterine diseases.</p>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"64 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13396816/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148580933","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":"Divergent Spatiotemporal Expression Patterns of Histone Deacetylases During Mouse Embryonic Tongue, Palate, and Mandible Development","authors":"Chubo Yang,&nbsp;Mingxing Wang,&nbsp;Jiaqi Kong,&nbsp;Jingru Wang,&nbsp;Huishu Li,&nbsp;Xinru Guo,&nbsp;Yuanbo Zhan","doi":"10.1002/dvg.70065","DOIUrl":"10.1002/dvg.70065","url":null,"abstract":"<div>\u0000 \u0000 <p>Epigenetic regulation is indispensable for embryonic development, yet the specific roles of histone deacetylases (HDACs)—a core family of epigenetic regulatory enzymes-in orchestrating craniofacial morphogenesis remain incompletely elucidated. C57BL/6 mouse embryos spanning embryonic Day 12.5 (E12.5) to E16.5 were analyzed. Histological staining (hematoxylin–eosin and Masson staining) was used to characterize morphological development of the tongue, palate, and mandible. Immunohistochemistry (IHC) was performed to map the spatiotemporal expression patterns of HDAC1–11. HDAC1 expression was scarcely detectable throughout the entire E12.5–E16.5 period. In contrast, HDAC2 showed robust expression in the mandible and tongue from E13.5 to E15.5. Notably, HDAC2 also exhibited strong positive staining in palatal epithelial cells at E14.5. By E16.5, HDAC1–10 expression became barely detectable in tongue, palate, and mandible, whereas HDAC11 retained intense expression specifically in the mandible at this late maturation stage. HDAC family members exhibit divergent spatiotemporal expression profiles during craniofacial development, with distinct subtype-specific patterns that align with key stages of tongue, palate and mandible morphogenesis. These findings establish a comprehensive spatiotemporal expression atlas of HDAC1–11 in developing craniofacial organs, providing essential foundational data to guide future investigations into the epigenetic regulation of craniofacial development.</p>\u0000 </div>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"64 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148383020","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}

{"title":"Ectopic Bulged Structures in Nkx2-1-KO and FoxE-KO Juveniles of Ciona robusta: Insights Into the Parallel Gene Regulatory System Before the Evolution of Thyroid Follicular Cells","authors":"Hiroki Kawamorita,&nbsp;Mizuki Ito,&nbsp;Yasunori Sasakura,&nbsp;Michio Ogasawara","doi":"10.1002/dvg.70064","DOIUrl":"10.1002/dvg.70064","url":null,"abstract":"<div>\u0000 \u0000 <p>Follicular thyroid cells in vertebrates are considered to have evolved from the endostyle of invertebrate chordates based on the shared features of the concentration of iodine and expression of thyroid-related genes, including <i>Nkx2-1</i>, <i>FoxE</i>, and <i>TPO</i>. We previously demonstrated using gene knockout (KO) experiments on the ascidian <i>Ciona robusta</i> that the thyroid-related transcription factors, Nkx2-1 and FoxE, were associated with endostyle development, and also that Nkx2-1 regulated the expression of <i>TPO</i> via FoxE, even though the expression of <i>Pax2/5/8</i> homologs was not affected. In the present study, which involved detailed observations of <i>Ciona Nkx2-1</i>-KO and <i>FoxE</i>-KO specimens, we found that ectopic bulged structures neighboring the hypoplastic endostyle occasionally emerged. In situ hybridization revealed that the ectopic bulged structures expressed <i>TPO</i> and <i>Pax2/5/8</i> homologs. Further KO experiments on <i>Ciona Pax2/5/8</i> homologs revealed no effects on endostyle development or <i>TPO</i> expression. These results suggest that, in addition to the established “Nkx2-1 –&gt; <i>FoxE</i>-FoxE –&gt; <i>TPO</i>” pathway, the ventral pharynx offers an insight into a potential relationship between Pax2/5/8 homologs and <i>TPO</i>. Accordingly, ascidians may show that the ventral pharyngeal endoderm possessed multiple gene regulatory networks for <i>TPO</i> expression prior to the evolutionary divergence leading to the vertebrate follicular thyroid.</p>\u0000 </div>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"64 4","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148382985","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}

{"title":"Meet Our Editorial Board—Genesis. An Interview With Anna Di Gregorio, New York University, New York, USA","authors":"Paul Trevorrow,&nbsp;Anna Di Gregorio","doi":"10.1002/dvg.70061","DOIUrl":"10.1002/dvg.70061","url":null,"abstract":"","PeriodicalId":12718,"journal":{"name":"genesis","volume":"64 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148310323","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}

{"title":"Multiplexed Retinoic Acid Signal Reporters for In Vivo and In Vitro Use","authors":"Craig Smith,&nbsp;Bianca Black,&nbsp;Ric Broadhurst,&nbsp;Stacey Parbhu,&nbsp;Peter L. Pfeffer","doi":"10.1002/dvg.70063","DOIUrl":"10.1002/dvg.70063","url":null,"abstract":"<p>Here we describe the optimisation of a modular, generally applicable and rapid method to make multimerised signaling-molecule reporters for detecting different retinoic acid concentrations. The technical plasmid design allows for the easy exchange of reporters, promoters, signal-responsive elements as well as for the multimerization of entire cassettes of these modular units. These constructs are ideal for pronuclear injection resulting in random integration into genomic DNA so avoiding the use of embryonic stem cells for generating transgenic lines or embryos. The use of insulator elements was shown to isolate reporter cassettes from integration-specific influences and from adjacent cassettes. Such insulators were found to be beneficial in transient transfections of primary cells, where DNA would not have integrated, as seen by more accurate ligand concentration responsiveness. The choice of promoter is important as weak promoters in in vitro experiments did not function in transgenic embryos. Notably, the core binding site and surrounding nucleotides could be used to modulate retinoic acid responsiveness in vitro. However, in vivo, the construct not only reflected retinoic acid concentrations but also affected tissue specificity. Increasing the number of retinoic acid receptor binding sites from 4 to 12 copies increased in vitro RA sensitivity, but had a lesser effect on in vivo sensitivity. We observed robust expression in the primitive streak of presomitic embryos and in the somites, neural tube and presomitic mesoderm of 3–10-somite embryos. Notably, use of only two copies of the binding site resulted in construct-inherent variability in cell type specificity and RA concentration sensitivity. This was conclusively demonstrated using dual receptors, which inherently control for confounding integration or copy-number dependent effects. In conclusion, the use of dual reporters with insulators is a useful approach for concurrently measuring concentration and context dependent responses to one signaling pathway and also lends itself for examining the interaction of two signaling pathways within one embryo.</p>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"64 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/dvg.70063","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148273519","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":"Generation and Characterization of Twist1 Acetyl-Mimic and Acetyl-Deficient Mouse Models","authors":"Mary Elmeniawi,&nbsp;Xiaobin Yu,&nbsp;Samuel Wen,&nbsp;Gunjan A. Bhatia,&nbsp;Lan Liao,&nbsp;Jianming Xu,&nbsp;Walid D. Fakhouri","doi":"10.1002/dvg.70062","DOIUrl":"10.1002/dvg.70062","url":null,"abstract":"<p><i>TWIST1</i> encodes a highly conserved basic helix–loop–helix transcription factor essential for embryonic development from Drosophila to humans. TWIST1 activity is regulated by post-translational modifications, including phosphorylation during development and cancer metastasis. Recent cancer studies identified acetylation of lysines K73 and K76 as a novel regulatory modification that shifts TWIST1 from a repressive to an activating state during epithelial-to-mesenchymal transition (EMT). However, the developmental and in vivo functions of TWIST1 acetylation remain unknown. To investigate the physiological role, we generated the first acetyl-deficient <i>Twist1</i>^{<i>K73,76R K73,76R</i>} and acetyl-mimic <i>Twist1</i>^{<i>K73,76Q K73,76Q</i>} mouse models using CRISPR/Cas9-mediated genome editing. Targeted sequencing confirmed substitutions, and founders were backcrossed onto a C57BL/6J genetic background. Phenotypic analysis revealed that <i>Twist1</i> acetyl-deficient mice exhibited highly penetrant craniofacial abnormalities, including severe mandibular hypoplasia, mandibular bone fusion, replacement of premaxillary, maxillary, and palatine bones with cartilage, ectopic cartilage-like structures, ocular malformations, and reduced skull mineralization. In contrast, acetyl-mimic mutant embryos displayed milder craniofacial defects characterized by reduced mandibular condylar processes and palatine bones, along with decreased skull mineralization. These findings demonstrate that TWIST1 acetylation at K73 and K76 plays a crucial role in normal craniofacial skeletogenesis, supporting prior cancer studies showing reduced oncogenic activity following loss of TWIST1 acetylation.</p>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"64 3","pages":""},"PeriodicalIF":2.8,"publicationDate":"2026-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/dvg.70062","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148273562","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":"","authors":"","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":12718,"journal":{"name":"genesis","volume":"64 3","pages":""},"PeriodicalIF":2.4,"publicationDate":"2026-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148074758","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}