Developmental biology最新文献

{"title":"Application of the Magnet-Cre optogenetic system in the chicken model.","authors":"Michael Pfann, Enbal Ben-Tal Cohen, Dalit Sela-Donenfeld, Yuval Cinnamon","doi":"10.1016/j.ydbio.2025.04.003","DOIUrl":"https://doi.org/10.1016/j.ydbio.2025.04.003","url":null,"abstract":"<p><p>Chickens serve as an excellent model organism for developmental biology, offering unique opportunities for precise spatiotemporal access to embryos within eggs. Optogenes are light-activated proteins that regulate gene expression, offering a non-invasive method to activate genes at specific locations and developmental stages, advancing developmental biology research. This study employed the Magnet-Cre optogenetic system to control gene expression in developing chicken embryos. Magnet-Cre consists of two light-sensitive protein domains that dimerize upon light activation, each attached to an inactive half of the Cre recombinase enzyme, which becomes active upon dimerization. We developed an all-in-one plasmid containing a green fluorescent protein marker, the Magnet-Cre system, and a light-activated red fluorescent protein gene. This plasmid was electroporated into the neural tube of Hamburger and Hamilton (H&H) stage 14 chicken embryos. Embryo samples were cleared using the CUBIC protocol and imaged with a light sheet microscope to analyze optogenetic activity via red-fluorescent cells. We established a pipeline for Magnet-Cre activation in chicken embryos, demonstrating that a single 3-minute exposure to blue light following incubation at 28°C was sufficient to trigger gene activity within the neural tube, with increased activity upon additional light exposure. Finally, we showed a spatiotemporal control of gene activity using a localized laser light induction. This research lays the groundwork for further advancements in avian developmental biology and poultry research, enabling spatiotemporal control of genes in both embryos and transgenic chickens.</p>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143787906","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}

{"title":"ASCL1 protein domains with distinct functions in neuronal differentiation and subtype specification.","authors":"Yuji Nakada, Madison J Martinez, Jane E Johnson","doi":"10.1016/j.ydbio.2025.04.001","DOIUrl":"https://doi.org/10.1016/j.ydbio.2025.04.001","url":null,"abstract":"<p><p>ASCL1 is a neural basic helix-loop-helix (bHLH) transcription factor that plays essential roles during neural development, including neural differentiation and neuronal subtype specification. bHLH factors are defined by their motifs, including a basic region interacting with DNA and an HLH domain involved in protein-protein interactions. We previously defined specific regions within the bHLH domain of ASCL1 as important for its specific functions directing neuronal differentiation in the chick neural tube. Here, we build upon these findings to show how specific mutations within the basic region block DNA binding but not heterodimer formation with E-protein partners TCF3 (E12/E47) and TCF12 (HEB) yet have differential abilities to show dominant negative phenotypes. Additionally, truncating domains outside the bHLH define a nuclear localization signal, a requirement for the C-terminal acidic residues, and the non-essentiality of the N-terminal glutamine/alanine repeats. This structure/function analysis identifies functional domains for ASCL1 activity.</p>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143787907","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}

{"title":"Identification of the dysregulated let-7c-Sox2 network in the facial prominences of mouse embryos with early retinoid acid exposure","authors":"Chao Song,&nbsp;Junjie Lu,&nbsp;Ya Wang,&nbsp;Yi Zou","doi":"10.1016/j.ydbio.2025.03.018","DOIUrl":"10.1016/j.ydbio.2025.03.018","url":null,"abstract":"<div><div>RA signaling is crucial for the anteroposterior pattern formation during neural crest induction and acts as a key environmental cue for cranial neural crest cell migration as well as the subsequent mesenchymal proliferation and differentiation. Congenital malformations including cleft lip and palate have been shown associated with altered embryonic RA signaling both in human and in animal models. In this study, a dysregulated <em>let-7c-Sox2</em> network was identified in the altered transcriptomic profiles of the facial prominences of E12.5 mouse embryos induced by early RA exposure. Ubiquitously increased expression of <em>let-7c</em> was observed in the epithelium and the mesenchyme of facial prominences of the RA treated mouse and chick embryos. Direct binding and regulation between <em>let-7c</em> and <em>Sox2</em> was verified using luciferase assay and significant negative correlation between <em>let-7c</em> and <em>Sox2</em> expression was observed <em>in vitro</em>. Reduced <em>Sox2</em> expression was predominantly identified in the epithelium of maxillary and palate shelves from E10.5 to E12.5 in RA-induced mouse embryos, resulted in oral adhesion and hypoplasia of palatal shelves that could partly be explained by the reduced mesenchymal proliferation due to upregulation of <em>let-7c</em>, as shown by the results of cell proliferation assay <em>in vitro</em>.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"523 ","pages":"Pages 9-19"},"PeriodicalIF":2.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143779387","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}

{"title":"Cellular dedifferentiation. Revisiting Betty Hay's legacy","authors":"Alejandra Beltrán-Rivera,&nbsp;José E. García-Arrarás","doi":"10.1016/j.ydbio.2025.03.017","DOIUrl":"10.1016/j.ydbio.2025.03.017","url":null,"abstract":"<div><div>The concept of mature specialized cells and the stability of the differentiated state was fundamentally challenged by Elizabeth Hay's groundbreaking observations on amphibian limb regeneration, published in 1959. Building on previous work by C.S. Thornton, she discovered that muscle cells could dedifferentiate and transform into progenitor cells within the regeneration blastema reshaping our understanding of cell differentiation. This pivotal finding reshaped our understanding of cell differentiation, opening new avenues of research. Though controversial, her findings significantly advanced the fields of cell plasticity and regenerative biology.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"523 ","pages":"Pages 1-8"},"PeriodicalIF":2.5,"publicationDate":"2025-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143751708","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}

{"title":"Cereblon E3 ligase complex genes are expressed in tissues sensitive to thalidomide in chicken and zebrafish embryos but are unchanged following thalidomide exposure","authors":"Lucas Rosa Fraga ,&nbsp;Jessica Reeves,&nbsp;Chris Mahony ,&nbsp;Lynda Erskine,&nbsp;Neil Vargesson","doi":"10.1016/j.ydbio.2025.03.014","DOIUrl":"10.1016/j.ydbio.2025.03.014","url":null,"abstract":"<div><div>Thalidomide is an infamous drug used initially as a sedative until it was tragically discovered it has highly teratogenic properties. Despite this it is now being used to successfully treat a range of clinical conditions including erythema nodosum leprosum (ENL) and multiple myeloma (MM). Cereblon (CRBN), a ubiquitin ligase, is a binding target of thalidomide for both its therapeutic and teratogenic activities and forms part of an CRL4-E3 ubiquitin ligase complex with the proteins Damaged DNA Binding protein 1 (DDB1) and Cullin-4A (CUL4A). This complex mediates degradation of the zinc-finger transcription factors Ikaros (IKZF1) and Aiolos (IKZF3), to mediate thalidomide's anti-myeloma response. To better understand the importance of CRBN and its binding partners for thalidomide teratogenesis here we analysed the expression patterns of <em>CRBN</em> and some of its known E3 complex binding partners in wildtype and thalidomide-treated chicken and zebrafish embryos. <em>CRBN</em> and <em>DDB1</em> are expressed in many tissues throughout development including those that are thalidomide-sensitive while <em>CUL4A</em> and targets of the CRL4-CRBN E3 Ligase Complex <em>IKZF1</em> and <em>IKZF3</em> are expressed at different timepoints and in fewer tissues in the body than <em>CRBN</em>. Furthermore, <em>IKZF3</em> is expressed in tissues of the eye that <em>CRBN</em> is not. However, although we observed rapid changes to the chicken yolk-sac membrane vasculature following thalidomide exposure, we did not detect CRL4-CRBN E3 Ligase Complex expression in the yolk-sac membrane vessels. Furthermore, we did not detect any changes in <em>CRBN</em>, <em>DDB1</em>, <em>CUL4</em>, <em>IKZF1</em> and <em>IKZF3</em> expression following thalidomide exposure in chicken and zebrafish embryos. These findings demonstrate that the anti-angiogenic activities of thalidomide may occur independent of <em>CRBN</em> and that thalidomide does not regulate CRL4-CRBN E3 Ligase Complex gene expression at the mRNA level.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"522 ","pages":"Pages 156-170"},"PeriodicalIF":2.5,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143746595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The transcription factor Capicua maintains the oocyte polarity in the panoistic ovary of the German cockroach","authors":"Nashwa Elshaer ,&nbsp;Jorge Escudero,&nbsp;Maria-Dolors Piulachs","doi":"10.1016/j.ydbio.2025.03.016","DOIUrl":"10.1016/j.ydbio.2025.03.016","url":null,"abstract":"<div><div>The establishment of the symmetry axis is crucial for the development of all organisms. In insects, this process begins early in oogenesis with the correct distribution of the mRNAs and proteins in the oocyte. One protein that plays a role in organizing this distribution is the transcription factor Capicua (Cic). Cic has been studied in the context of oogenesis and embryonic development in <em>Drosophila melanogaster</em>. It is maternally expressed, begins essential for establishing the dorsoventral axis, and functions as a transcriptional repressor. Although the Cic sequences are conserved across species, their function in other types of insect ovaries is still little known. We wondered whether the function of Cic in insects has been maintained through evolution despite the ovary type or if it has been modified in parallel to the ovary evolution. To address this, we studied the Cic function in a phylogenetically basal insect, the cockroach <em>Blattella germanica</em>, a species with panoistic ovaries. Our findings show that <em>B. germanica</em> Cic is essential for oocyte development and the maturation of ovarian follicles. A loss of Cic function leads to disrupted cytoskeletal organization, defects in anterior-posterior polarity, and compromised follicle integrity. The conservation and functional divergence of Cic across different species suggest evolutionary adaptations in the mechanisms of insect oogenesis.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"522 ","pages":"Pages 125-135"},"PeriodicalIF":2.5,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143738795","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}

{"title":"Wnt3a is an early regulator of the Wolffian duct directionality via the regulation of apicobasal cell polarity","authors":"Shinichi Hayashi ,&nbsp;Hitomi Suzuki ,&nbsp;Shinji Takada ,&nbsp;Tatsuya Takemoto","doi":"10.1016/j.ydbio.2025.03.015","DOIUrl":"10.1016/j.ydbio.2025.03.015","url":null,"abstract":"<div><div>The Wolffian duct is a pair of epithelial ductal structures along the body axis that induces nephron development by interaction with the metanephric mesenchyme. The interaction between the mesenchyme and the ureteric bud derived from the Wolffian duct is mediated by Wnt ligands, the loss of which results in kidney agenesis. Nonetheless, the early contribution of Wnt signaling to Wolffian duct formation remains unclear. We therefore examined these dynamics in knockout and transgenic mouse embryos. The Wnt signal reporter was active in the extending Wolffian duct, and <em>Wnt3a</em>-knockout embryos exhibited a fragmented and misdirectional Wolffian duct. Apicobasal polarity was disrupted under <em>Wnt3a</em>-deficiency. These findings suggest that <em>Wnt3a</em> plays an important role in Wolffian duct development by regulating apicobasal polarity.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"522 ","pages":"Pages 136-142"},"PeriodicalIF":2.5,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143738796","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}

{"title":"Keratinization and cornification of avian skin appendages during development. Insights from immunolabeling and electron microscopic studies","authors":"Lorenzo Alibardi","doi":"10.1016/j.ydbio.2025.02.019","DOIUrl":"10.1016/j.ydbio.2025.02.019","url":null,"abstract":"<div><div>The basal cytoskeleton of avian keratinocytes consists in a number of Intermediate Filament Keratins (IFKs, also indicated as alpha-keratins), poor (soft) or rich (hard) in cysteine. In keratinocytes of developing skin appendages Corneous Beta Proteins (CBPs, formerly termed beta-keratins), build most of the corneous material of developing scutate scales, claws, beak and feathers. CBPs derive from a gene locus termed Epidermal Differentiation Complex (EDC), unrelated to genes for IFKs. CBPs and IFKs belong to two different gene families that evolved independently during the evolution of birds. The evolution of feathers derived from the initial morphogenesis of barb ridges containing specialized proteins. During feather development, the framework of IFKs that combine with CBPs in differentiating keratinocytes, barb and barbule cells, give rise to resistant but flexible corneocytes in feathers and hard corneocytes in scales, claws and beaks. Here, we mainly deal with avian IFKs that are accumulated during the development of skin derivatives of birds, especially downfeathers. The latter are corneous appendages and, when mature, are composed from a prevalent mass of feather-CBPs (fCBPs, formerly indicated as feather beta-keratins). During development fCBPs are deposited over a IFKs cytoskeleton formed in barb and barbule cells, and these small beta-proteins rapidly overcame in amount IFKs, generating the corneous barbs and barbules of downfeathers. This process likely occurs through electrostatic interactions between acidic IFKs and basic CBPs, and later by the formation of covalent bonds (-S-S- and epsilon-bonds). Proteome and molecular studies have sequenced most of IFKs and CBPs of feathers in some species of birds. Most of the proteins extracted from feathers are fCBPs, while a lower amount is constituted from IFKs and other minor proteins.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"522 ","pages":"Pages 196-219"},"PeriodicalIF":2.5,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143742534","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}

{"title":"Drosophila grainyhead gene and its neural stem cell specific enhancers show epigenetic synchrony in the cells of the central nervous system","authors":"Rashmi Sipani ,&nbsp;Yamini Rawal ,&nbsp;Jiban Barman ,&nbsp;Prakeerthi Abburi ,&nbsp;Vishakha Kurlawala ,&nbsp;Rohit Joshi","doi":"10.1016/j.ydbio.2025.03.012","DOIUrl":"10.1016/j.ydbio.2025.03.012","url":null,"abstract":"<div><div>Enhancers are the epicentres of tissue-specific gene regulation. In this study, we have used the central nervous system (CNS) specific expression of the <em>Drosophila grainyhead</em> (<em>grh</em>) gene to make a case for deleting the enhancers in a sensitised background of other enhancer deletion, to functionally validate their role in tissue-specific gene regulation. We identified novel enhancers for <em>grh</em> and subsequently deleted two of them, to establish their collective importance in regulating <em>grh</em> expression in CNS. This showed that <em>grh</em> relies on multiple enhancers for its robust expression in neural stem cells (NSCs), with different combinations of enhancers playing a critical role in regulating its expression in various subset of these cells. We also found that these enhancers and the <em>grh</em> gene show epigenetic synchrony across the three cell types (NSCs, intermediate progenitors and neurons) of the developing CNS; and <em>grh</em> is not transcribed in intermediate progenitor cells, which inherits the Grh protein from the NSCs. We propose that this could be a general mechanism for regulating the expression of cell fate determinant protein in intermediate progenitor cells. Lastly, our results underline that enhancer redundancy results in phenotypic robustness in <em>grh</em> gene expression, which seems to be a consequence of the cumulative activity of multiple enhancers.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"522 ","pages":"Pages 227-239"},"PeriodicalIF":2.5,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143742464","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}

{"title":"Retinoic acid promotes second heart field addition and regulates ventral aorta patterning in zebrafish","authors":"Austin H.C. Griffin ,&nbsp;Allison M. Small ,&nbsp;Riley D. Johnson ,&nbsp;Anna M. Medina ,&nbsp;Kiki T. Kollar ,&nbsp;Ridha A. Nazir ,&nbsp;Acasia M. McGuire ,&nbsp;Jennifer A. Schumacher","doi":"10.1016/j.ydbio.2025.03.013","DOIUrl":"10.1016/j.ydbio.2025.03.013","url":null,"abstract":"<div><div>Retinoic acid (RA) signaling is used reiteratively during vertebrate heart development. Its earliest known role is to restrict formation of the earlier-differentiating first heart field (FHF) progenitors, while promoting the differentiation of second heart field (SHF) progenitors that give rise to the arterial pole of the ventricle and outflow tract (OFT). However, requirements for RA signaling at later stages of cardiogenesis remain poorly understood. Here, we investigated the role of RA signaling after the later differentiating SHF cells have begun to add to the OFT. We found that inhibiting RA production in zebrafish beginning at 26 hours post fertilization (hpf) produced embryos that have smaller ventricles with fewer ventricular cardiomyocytes, and reduced number of smooth muscle cells in the bulbus arteriosus (BA) of the OFT. Our results suggest that the deficiency of the ventricular cardiomyocytes is due to reduced SHF addition to the arterial pole. In contrast to smaller ventricles and BA, later RA deficiency also results in a dramatically elongated posterior branch of the adjacent ventral aorta, which is surrounded by an increased number of smooth muscle cells. Altogether, our results reveal that RA signaling is required during the period of SHF addition to promote addition of ventricular cardiomyocytes, partition smooth muscle cells onto the BA and posterior ventral aorta, and to establish proper ventral aorta anterior-posterior patterning.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"522 ","pages":"Pages 143-155"},"PeriodicalIF":2.5,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143729178","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}