Pub Date : 2025-02-11DOI: 10.1016/j.ydbio.2025.02.009
Abigail E. Descoteaux , Marko Radulovic , Dona Alburi , Cynthia A. Bradham
MARVEL proteins, including those of the CMTM gene family, are multi-pass transmembrane proteins that play important roles in vesicular trafficking and cell migration; however, little is understood about their role in development, and their role in skeletal patterning is unexplored. CMTM4 is the only CMTM family member found in the developmental transcriptome of the sea urchin Lytechinus variegatus. Here, we validate that LvCMTM4 is a transmembrane protein and show that perturbation of CMTM4 expression via zygotic morpholino or mRNA injection perturbs skeletal patterning, resulting in loss of secondary skeletal elements and rotational defects. We also demonstrate that normal levels of CMTM4 are required for normal PMC migration and filopodial organization, and that these effects are not due to gross mis-specification of the ectoderm. Finally, we show that CMTM4 is sufficient to mediate mesenchymal cell-cell adhesion. Taken together, these data suggest that CMTM4 controls PMC migration and biomineralization via adhesive regulation during sea urchin skeletogenesis. This is the first discovery of a functionally required adhesive gene in this skeletal patterning system.
{"title":"CMTM4 is an adhesion modulator that regulates skeletal patterning and primary mesenchyme cell migration in sea urchin embryos","authors":"Abigail E. Descoteaux , Marko Radulovic , Dona Alburi , Cynthia A. Bradham","doi":"10.1016/j.ydbio.2025.02.009","DOIUrl":"10.1016/j.ydbio.2025.02.009","url":null,"abstract":"<div><div>MARVEL proteins, including those of the CMTM gene family, are multi-pass transmembrane proteins that play important roles in vesicular trafficking and cell migration; however, little is understood about their role in development, and their role in skeletal patterning is unexplored. CMTM4 is the only CMTM family member found in the developmental transcriptome of the sea urchin <em>Lytechinus variegatus</em>. Here, we validate that LvCMTM4 is a transmembrane protein and show that perturbation of CMTM4 expression via zygotic morpholino or mRNA injection perturbs skeletal patterning, resulting in loss of secondary skeletal elements and rotational defects. We also demonstrate that normal levels of CMTM4 are required for normal PMC migration and filopodial organization, and that these effects are not due to gross mis-specification of the ectoderm. Finally, we show that CMTM4 is sufficient to mediate mesenchymal cell-cell adhesion. Taken together, these data suggest that CMTM4 controls PMC migration and biomineralization via adhesive regulation during sea urchin skeletogenesis. This is the first discovery of a functionally required adhesive gene in this skeletal patterning system.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"521 ","pages":"Pages 85-95"},"PeriodicalIF":2.5,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143413690","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 : 2025-02-11DOI: 10.1016/j.ydbio.2025.02.008
Thaís Metzker-Pinto , Yen T.H. Tran , Igor Buzzatto-Leite , Lloyd Lok , Jórdan F. Sampar , Hernandes F. Carvalho , Gonzalo del Monte-Nieto , Lúcia E. Alvares
WFDC1 encodes an extracellular matrix protein involved in cell proliferation, migration, and epithelial-mesenchymal transition in disease conditions. Despite this, Wfdc1-null mice display no discernible malformations while cattle bearing a WFDC1 mutation present multiple ocular defects, leaving the role of WFDC1 during embryonic development unclear. To address this, we used the chicken embryo as a model to investigate WFDC1 developmental roles in amniotes. We performed a comparative expression analysis during chicken and mouse development, which revealed expression in ectodermal and mesodermal derivatives, with both conserved and species-specific domains. Conserved expression was observed in the eye, otic vesicle, central and peripheral nervous systems, and neural crest cells. Chicken-specific expression was identified in mesodermal structures, including the notochord, limbs and heart. However, even in the conserved sites like the eyes, WFDC1 localizes to different retinal layers, indicating potential divergence roles in retinal development and function across species. In contrast, WFDC1 expression in the limb buds is specific to chicken, encompassing the distal mesenchyme, interdigital membranes, and blastemas. Functional enrichment analysis links WFDC1 to limb patterning, morphogenesis, and Wnt signaling. The species-specific differences likely stem from evolutionary changes in gene regulation, supported by differences in proximal cis-regulatory elements of the WFDC1 loci between chicken and mouse. The complexity of WFDC1 expression in the chicken embryo, along with WFDC1 regulatory conservation within birds, indicates that this gene may play specific roles in avian development, possibly contributing to features specific to this lineage. Future studies using the chicken model will be valuable in further uncovering the specific roles of WFDC1.
{"title":"The chicken embryo brings new insights into the evolutionary role of WFDC1 during amniote development","authors":"Thaís Metzker-Pinto , Yen T.H. Tran , Igor Buzzatto-Leite , Lloyd Lok , Jórdan F. Sampar , Hernandes F. Carvalho , Gonzalo del Monte-Nieto , Lúcia E. Alvares","doi":"10.1016/j.ydbio.2025.02.008","DOIUrl":"10.1016/j.ydbio.2025.02.008","url":null,"abstract":"<div><div><em>WFDC1</em> encodes an extracellular matrix protein involved in cell proliferation, migration, and epithelial-mesenchymal transition in disease conditions. Despite this, <em>Wfdc1-null</em> mice display no discernible malformations while cattle bearing a <em>WFDC1</em> mutation present multiple ocular defects, leaving the role of <em>WFDC1</em> during embryonic development unclear. To address this, we used the chicken embryo as a model to investigate <em>WFDC1</em> developmental roles in amniotes. We performed a comparative expression analysis during chicken and mouse development, which revealed expression in ectodermal and mesodermal derivatives, with both conserved and species-specific domains. Conserved expression was observed in the eye, otic vesicle, central and peripheral nervous systems, and neural crest cells. Chicken-specific expression was identified in mesodermal structures, including the notochord, limbs and heart. However, even in the conserved sites like the eyes, <em>WFDC1</em> localizes to different retinal layers, indicating potential divergence roles in retinal development and function across species. In contrast, <em>WFDC1</em> expression in the limb buds is specific to chicken, encompassing the distal mesenchyme, interdigital membranes, and blastemas. Functional enrichment analysis links <em>WFDC1</em> to limb patterning, morphogenesis, and Wnt signaling. The species-specific differences likely stem from evolutionary changes in gene regulation, supported by differences in proximal cis-regulatory elements of the <em>WFDC1</em> loci between chicken and mouse. The complexity of <em>WFDC1</em> expression in the chicken embryo, along with <em>WFDC1</em> regulatory conservation within birds, indicates that this gene may play specific roles in avian development, possibly contributing to features specific to this lineage. Future studies using the chicken model will be valuable in further uncovering the specific roles of <em>WFDC1</em>.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"521 ","pages":"Pages 96-107"},"PeriodicalIF":2.5,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143412828","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 : 2025-02-10DOI: 10.1016/j.ydbio.2025.02.005
Dadakhalandar Doddamani , Daniel F. Carlson , Lynn McTeir , Lorna Taylor , Sunil Nandi , Megan G. Davey , Mike J. McGrew , James D. Glover
The zinc finger transcription factor PRDM14, part of the PR domain containing protein family, is critical for mammalian primordial germ cell (PGC) specification, epigenetic reprogramming and maintaining naïve pluripotency in stem cells. However, PRDM14's role in other species is not well understood. In chicken, PRDM14 is broadly expressed in the early embryo, before becoming restricted to the forming neural plate, migratory PGCs, and later, in the adult testes. To investigate the role of PRDM14 we generated two independent targeted chicken lines and bred homozygous knockout embryos. Strikingly, we found that gastrulation was disrupted in PRDM14−/− embryos, which lacked a definitive primitive streak. Transcriptomic and in situ hybridisation analyses revealed a broad loss of anterior primitive streak marker genes, coupled with downregulation of the multifunctional antagonists CHRD and CER1, and expansion of the NODAL expression domain. Further analysis of PRDM14−/− embryos revealed PGCs were still specified but significantly reduced in number, and PRDM14−/− PGCs could not be propagated in vitro. Knockdown studies in vitro confirmed that PRDM14 is essential for PGC survival and antagonises FGF-induced somatic differentiation, similar to PRDM14's role in mammalian stem cells. Taken together, our results show that in chicken, PRDM14 plays a multifunctional and essential role during embryonic development.
{"title":"PRDM14 is essential for vertebrate gastrulation and safeguards avian germ cell identity","authors":"Dadakhalandar Doddamani , Daniel F. Carlson , Lynn McTeir , Lorna Taylor , Sunil Nandi , Megan G. Davey , Mike J. McGrew , James D. Glover","doi":"10.1016/j.ydbio.2025.02.005","DOIUrl":"10.1016/j.ydbio.2025.02.005","url":null,"abstract":"<div><div>The zinc finger transcription factor PRDM14, part of the PR domain containing protein family, is critical for mammalian primordial germ cell (PGC) specification, epigenetic reprogramming and maintaining naïve pluripotency in stem cells. However, PRDM14's role in other species is not well understood. In chicken, <em>PRDM14</em> is broadly expressed in the early embryo, before becoming restricted to the forming neural plate, migratory PGCs, and later, in the adult testes. To investigate the role of PRDM14 we generated two independent targeted chicken lines and bred homozygous knockout embryos. Strikingly, we found that gastrulation was disrupted in <em>PRDM14</em><sup><em>−/−</em></sup> embryos, which lacked a definitive primitive streak. Transcriptomic and <em>in situ</em> hybridisation analyses revealed a broad loss of anterior primitive streak marker genes, coupled with downregulation of the multifunctional antagonists <em>CHRD</em> and <em>CER1</em><em>,</em> and expansion of the <em>NODAL</em> expression domain. Further analysis of <em>PRDM14</em><sup><em>−/−</em></sup> embryos revealed PGCs were still specified but significantly reduced in number, and <em>PRDM14</em><sup><em>−/−</em></sup> PGCs could not be propagated <em>in vitro</em>. Knockdown studies <em>in vitro</em> confirmed that PRDM14 is essential for PGC survival and antagonises FGF-induced somatic differentiation, similar to PRDM14's role in mammalian stem cells. Taken together, our results show that in chicken, PRDM14 plays a multifunctional and essential role during embryonic development.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"521 ","pages":"Pages 129-137"},"PeriodicalIF":2.5,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143405319","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}
Pub Date : 2025-02-10DOI: 10.1016/j.ydbio.2025.02.007
Abi H. Crane , Claudia J. Baldry , Kathryn E. Rankin , Claire E. Clarkin , Katherine A. Williams , Neil J. Gostling
Medullary bone is a fast-growing, ephemeral bone tissue found inside the bone cavities of female birds. Identifying this tissue in the bones of fossil avian and non-avian dinosaurs has the potential to determine which specimens represent reproductively mature females. However, difficulties in distinguishing medullary bone from superficially similar bone pathologies has led to uncertainty as to whether some specimens previously thought to contain medullary bone instead represent sick or injured individuals. The most frequently mentioned of these pathologies is avian osteopetrosis, a virally-induced condition in birds causing bony lesions which can resemble medullary bone. Lists of criteria, primarily using two-dimensional osteohistology, have yet to form a comprehensive framework through which all medullary bone can be positively identified, and all pathology excluded. Here, we use high-resolution computed tomography (μCT) to characterise the three-dimensional structure of medullary bone in modern birds for the first time and make comparisons to the endosteal lesions of avian osteopetrosis. We identify both qualitative and quantitative features which we suggest to be characteristic of medullary bone, including connectivity density and osteocyte lacunar orientation, and highlight conspicuously variable features which require further investigation. We find several three-dimensional which can be used to differentiate between medullary bone and avian osteopetrosis, including structural anisotropy and trabecular thickness. These three-dimensional characters can be added to the growing framework of criteria to identify medullary bone in the fossil record and thus help determine the sex of dinosaurs.
{"title":"The three-dimensional structure of medullary bone: Novel criteria for the identification of avian sex-specific bone tissue","authors":"Abi H. Crane , Claudia J. Baldry , Kathryn E. Rankin , Claire E. Clarkin , Katherine A. Williams , Neil J. Gostling","doi":"10.1016/j.ydbio.2025.02.007","DOIUrl":"10.1016/j.ydbio.2025.02.007","url":null,"abstract":"<div><div>Medullary bone is a fast-growing, ephemeral bone tissue found inside the bone cavities of female birds. Identifying this tissue in the bones of fossil avian and non-avian dinosaurs has the potential to determine which specimens represent reproductively mature females. However, difficulties in distinguishing medullary bone from superficially similar bone pathologies has led to uncertainty as to whether some specimens previously thought to contain medullary bone instead represent sick or injured individuals. The most frequently mentioned of these pathologies is avian osteopetrosis, a virally-induced condition in birds causing bony lesions which can resemble medullary bone. Lists of criteria, primarily using two-dimensional osteohistology, have yet to form a comprehensive framework through which all medullary bone can be positively identified, and all pathology excluded. Here, we use high-resolution computed tomography (μCT) to characterise the three-dimensional structure of medullary bone in modern birds for the first time and make comparisons to the endosteal lesions of avian osteopetrosis. We identify both qualitative and quantitative features which we suggest to be characteristic of medullary bone, including connectivity density and osteocyte lacunar orientation, and highlight conspicuously variable features which require further investigation. We find several three-dimensional which can be used to differentiate between medullary bone and avian osteopetrosis, including structural anisotropy and trabecular thickness. These three-dimensional characters can be added to the growing framework of criteria to identify medullary bone in the fossil record and thus help determine the sex of dinosaurs.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"521 ","pages":"Pages 108-121"},"PeriodicalIF":2.5,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143405623","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}
Pub Date : 2025-02-09DOI: 10.1016/j.ydbio.2025.02.006
David Sedmera , Eliska Drobna Krejci , Ondrej Nanka , Adam Eckhardt
Investigating prenatal hypoxia is difficult in mammals, as there are confounding factors stemming from maternal adaptations and compensatory mechanisms. We have thus established an avian model of hypoxic incubation (starting after 2 days of normoxia, 15% O2, normobaric, until the time of sampling at embryonic day 8) to study embryonic reactions to low oxygen concentration. Our previous studies have shown increased vascularization, oedema, and ventricular wall thinning preceding the lethality at mid-gestation. Analysis of the cardiac proteome after 6 days of hypoxic incubation showed strong upregulation of enzymes involved in anaerobic glycolysis as well as an increase in apoptosis-related proteins, cell adhesion proteins, and secretory activity.
{"title":"Proteomic analysis of chick embryonic heart in experimental hypoxia","authors":"David Sedmera , Eliska Drobna Krejci , Ondrej Nanka , Adam Eckhardt","doi":"10.1016/j.ydbio.2025.02.006","DOIUrl":"10.1016/j.ydbio.2025.02.006","url":null,"abstract":"<div><div>Investigating prenatal hypoxia is difficult in mammals, as there are confounding factors stemming from maternal adaptations and compensatory mechanisms. We have thus established an avian model of hypoxic incubation (starting after 2 days of normoxia, 15% O<sub>2</sub>, normobaric, until the time of sampling at embryonic day 8) to study embryonic reactions to low oxygen concentration. Our previous studies have shown increased vascularization, oedema, and ventricular wall thinning preceding the lethality at mid-gestation. Analysis of the cardiac proteome after 6 days of hypoxic incubation showed strong upregulation of enzymes involved in anaerobic glycolysis as well as an increase in apoptosis-related proteins, cell adhesion proteins, and secretory activity.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"521 ","pages":"Pages 28-36"},"PeriodicalIF":2.5,"publicationDate":"2025-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143394369","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 : 2025-02-07DOI: 10.1016/j.ydbio.2025.02.004
Yao Le , Kavitha Rajasekhar , Tricia Y.J. Loo , Timothy E. Saunders , Thorsten Wohland , Christoph Winkler
A midline in the developing central nervous system allows symmetric distribution of neural progenitors that later establish functional, bilaterally symmetric neural circuits. In the zebrafish hindbrain, a midline forms early during neurulation as a result of coordinated cell convergence and midline-crossing cell divisions (C-divisions). These processes are controlled by the Wnt/planar cell polarity (PCP) pathway that positions progenitors close to a presumptive midline to perform C-divisions. Other upstream cues that control the extent of neural plate convergence, however, remain unclear. Midkine (Mdk) and pleiotrophin (Ptn) are structurally related heparin-binding growth factors that are dynamically expressed in the developing hindbrain. We show that two zebrafish Mdks, Mdka and Mdkb, as well as Ptn interact with distinct affinities in vivo with the protein tyrosine phosphatase receptor Ptprz1b. Zebrafish mdka and ptprz1b mutants exhibit impaired neural plate convergence along with misplaced C-divisions, defective cell polarity and transiently duplicated midlines. These defects are absent in mdka; mdkb double mutants suggesting antagonistic roles of Mdka and Mdkb to coordinate convergence and C-divisions. Overexpression of Drosophila Prickle, a key component of the Wnt/PCP pathway, rescued the midline duplications in mdka and ptprz1b mutants that exhibited significantly reduced levels of prickle pk1b, pk2a, and pk2b expression. Ptprz1b overexpression, on the other hand, up-regulated pk2a transcription. Our findings therefore suggest roles for Mdka, Mdkb and Ptprz1b in coordinating neural plate convergence, neural progenitor positioning and midline formation by controlling the levels of prickle expression.
{"title":"Midkine-a interacts with Ptprz1b to regulate neural plate convergence and midline formation in the developing zebrafish hindbrain","authors":"Yao Le , Kavitha Rajasekhar , Tricia Y.J. Loo , Timothy E. Saunders , Thorsten Wohland , Christoph Winkler","doi":"10.1016/j.ydbio.2025.02.004","DOIUrl":"10.1016/j.ydbio.2025.02.004","url":null,"abstract":"<div><div>A midline in the developing central nervous system allows symmetric distribution of neural progenitors that later establish functional, bilaterally symmetric neural circuits. In the zebrafish hindbrain, a midline forms early during neurulation as a result of coordinated cell convergence and midline-crossing cell divisions (C-divisions). These processes are controlled by the Wnt/planar cell polarity (PCP) pathway that positions progenitors close to a presumptive midline to perform C-divisions. Other upstream cues that control the extent of neural plate convergence, however, remain unclear. Midkine (Mdk) and pleiotrophin (Ptn) are structurally related heparin-binding growth factors that are dynamically expressed in the developing hindbrain. We show that two zebrafish Mdks, Mdka and Mdkb, as well as Ptn interact with distinct affinities <em>in vivo</em> with the protein tyrosine phosphatase receptor Ptprz1b. Zebrafish <em>mdka</em> and <em>ptprz1b</em> mutants exhibit impaired neural plate convergence along with misplaced C-divisions, defective cell polarity and transiently duplicated midlines. These defects are absent in <em>mdka</em>; <em>mdkb</em> double mutants suggesting antagonistic roles of Mdka and Mdkb to coordinate convergence and C-divisions. Overexpression of <em>Drosophila</em> Prickle, a key component of the Wnt/PCP pathway, rescued the midline duplications in <em>mdka</em> and <em>ptprz1b</em> mutants that exhibited significantly reduced levels of <em>prickle pk1b, pk2a,</em> and <em>pk2b</em> expression. Ptprz1b overexpression, on the other hand, up-regulated <em>pk2a</em> transcription. Our findings therefore suggest roles for Mdka, Mdkb and Ptprz1b in coordinating neural plate convergence, neural progenitor positioning and midline formation by controlling the levels of <em>prickle</em> expression.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"521 ","pages":"Pages 52-74"},"PeriodicalIF":2.5,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143381800","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}
Pub Date : 2025-02-07DOI: 10.1016/j.ydbio.2025.02.003
Rebecca Cacciottolo , Ruben J. Cauchi
Gemin3 (Gem3) or DEAD-box RNA helicase 20 (Ddx20) has been mostly implicated in the assembly of spliceosomal small nuclear ribonucleoproteins (snRNPs) as part of the SMN-Gemins complex. Nonetheless, several studies have hinted at its participation in diverse snRNP-independent activities. Here, we utilised a narrow unbiased genetic screen to discover novel Gem3 interactors in Drosophila with the aim of gaining better insights on its function in vivo. Through this approach, we identified a novel genetic interaction between Gem3 and NAT1, which encodes the Drosophila orthologue of translational regulator eIF4G2. Despite lack of a physical association, loss of NAT1 function was found to downregulate Gem3 mRNA levels. Extensive convergence in transcriptome alterations downstream of Gem3 and NAT1 silencing further supports a functional relationship between these factors in addition to showing a requirement for both in actin cytoskeleton organisation and organism development, particularly neurodevelopment. In confirmation, flies with either Gem3 or NAT1 depletion exhibited brain growth defects and reduced muscle contraction. Severe delays in developmental progression were also observed in a newly generated Gem3 hypomorphic mutant. Our data linking Gemin3 to a key component of the translational machinery support an emerging role for Gemin3 in translation that is also critical during organism development.
{"title":"A critical genetic interaction between Gemin3/Ddx20 and translation initiation factor NAT1/eIF4G2 drives development","authors":"Rebecca Cacciottolo , Ruben J. Cauchi","doi":"10.1016/j.ydbio.2025.02.003","DOIUrl":"10.1016/j.ydbio.2025.02.003","url":null,"abstract":"<div><div>Gemin3 (Gem3) or DEAD-box RNA helicase 20 (Ddx20) has been mostly implicated in the assembly of spliceosomal small nuclear ribonucleoproteins (snRNPs) as part of the SMN-Gemins complex. Nonetheless, several studies have hinted at its participation in diverse snRNP-independent activities. Here, we utilised a narrow unbiased genetic screen to discover novel Gem3 interactors in <em>Drosophila</em> with the aim of gaining better insights on its function <em>in vivo</em>. Through this approach, we identified a novel genetic interaction between <em>Gem3</em> and <em>NAT1</em>, which encodes the <em>Drosophila</em> orthologue of translational regulator eIF4G2. Despite lack of a physical association, loss of <em>NAT1</em> function was found to downregulate <em>Gem3</em> mRNA levels. Extensive convergence in transcriptome alterations downstream of <em>Gem3</em> and <em>NAT1</em> silencing further supports a functional relationship between these factors in addition to showing a requirement for both in actin cytoskeleton organisation and organism development, particularly neurodevelopment. In confirmation, flies with either <em>Gem3</em> or <em>NAT1</em> depletion exhibited brain growth defects and reduced muscle contraction. Severe delays in developmental progression were also observed in a newly generated Gem3 hypomorphic mutant. Our data linking Gemin3 to a key component of the translational machinery support an emerging role for Gemin3 in translation that is also critical during organism development.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"521 ","pages":"Pages 37-51"},"PeriodicalIF":2.5,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143381797","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 : 2025-02-06DOI: 10.1016/j.ydbio.2025.02.002
Avery J. McGinnis, Megan E. Cull, Nichole T. Peterson, Matthew K. Tang, Bryony V. Natale, David R.C. Natale
Mouse trophoblast stem (mTS) cells can be derived from the blastocyst or extraembryonic ectoderm as late as embryonic day (E) 6.5 and when cultured in vitro, can differentiate to all trophoblast subtypes of the mature placenta. Expression of the T-box transcription factor, Eomes, is required for the maintenance of, and used to identify mTS cells. During development, Eomes is restricted to the ExE and, by E7.5, to the chorion, after which its expression declines. The placental junctional zone and labyrinth layers are thought to develop exclusively from the ectoplacental cone and chorion, respectively. While it is well established that mTS cells express Eomes in vitro, it is unknown if Eomes-positive (EomesPOS) trophoblast that reside in the chorion after E6.5 are restricted in their developmental potential to the labyrinth layer in vivo. This study utilized a lineage tracing technique to evaluate the in vivo differentiation of EomesPOS trophoblast. Using an Ai6 reporter mouse crossed with a tamoxifen-inducible Eomes-Cre-ERT2 mouse, Cre was activated from E7.5 to E9.5, permanently marking all EomesPOS trophoblast and daughter cells with the ZsGreen fluorescent protein. This approach was complemented with immunofluorescence staining to assess how the EomesPOS trophoblast had contributed to the differentiated trophoblast population within the placenta by E17.5. Importantly, the results show that daughter cells of EomesPOS trophoblast in which Cre was activated, contributed to both placental layers; specifically, spongiotrophoblast and glycogen trophoblast within the junctional zone and syncytiotrophoblast and sinusoidal trophoblast giant cells within the labyrinth. This confirms that EomesPOS trophoblast maintain the capacity to contribute to both placental layers in vivo and do so after E7.5. This study expands our understanding of trophoblast differentiation in vivo and may prove useful in assessing how EomesPOS trophoblast contribute placental development later in gestation and in the context of placental pathology, where Eomes expression has been reported.
{"title":"Exploring the differentiation potential of EomesPOS mouse trophoblast cells in mid-gestation","authors":"Avery J. McGinnis, Megan E. Cull, Nichole T. Peterson, Matthew K. Tang, Bryony V. Natale, David R.C. Natale","doi":"10.1016/j.ydbio.2025.02.002","DOIUrl":"10.1016/j.ydbio.2025.02.002","url":null,"abstract":"<div><div>Mouse trophoblast stem (mTS) cells can be derived from the blastocyst or extraembryonic ectoderm as late as embryonic day (E) 6.5 and when cultured <em>in vitro,</em> can differentiate to all trophoblast subtypes of the mature placenta. Expression of the T-box transcription factor, <em>Eomes</em>, is required for the maintenance of, and used to identify mTS cells. During development, <em>Eomes</em> is restricted to the ExE and, by E7.5, to the chorion, after which its expression declines. The placental junctional zone and labyrinth layers are thought to develop exclusively from the ectoplacental cone and chorion, respectively. While it is well established that mTS cells express <em>Eomes in vitro</em>, it is unknown if Eomes-positive (Eomes<sup>POS</sup>) trophoblast that reside in the chorion after E6.5 are restricted in their developmental potential to the labyrinth layer <em>in vivo</em>. This study utilized a lineage tracing technique to evaluate the <em>in vivo</em> differentiation of Eomes<sup>POS</sup> trophoblast. Using an Ai6 reporter mouse crossed with a tamoxifen-inducible Eomes-Cre-ERT2 mouse, Cre was activated from E7.5 to E9.5, permanently marking all Eomes<sup>POS</sup> trophoblast and daughter cells with the ZsGreen fluorescent protein. This approach was complemented with immunofluorescence staining to assess how the Eomes<sup>POS</sup> trophoblast had contributed to the differentiated trophoblast population within the placenta by E17.5. Importantly, the results show that daughter cells of Eomes<sup>POS</sup> trophoblast in which Cre was activated, contributed to both placental layers; specifically, spongiotrophoblast and glycogen trophoblast within the junctional zone and syncytiotrophoblast and sinusoidal trophoblast giant cells within the labyrinth. This confirms that Eomes<sup>POS</sup> trophoblast maintain the capacity to contribute to both placental layers <em>in vivo</em> and do so after E7.5. This study expands our understanding of trophoblast differentiation <em>in vivo</em> and may prove useful in assessing how Eomes<sup>POS</sup> trophoblast contribute placental development later in gestation and in the context of placental pathology, where Eomes expression has been reported.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"521 ","pages":"Pages 75-84"},"PeriodicalIF":2.5,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143373743","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}
Pub Date : 2025-02-03DOI: 10.1016/j.ydbio.2025.01.017
Abigail Carranza, LaFreda J. Howard, Haley E. Brown, Ayawovi Selom Ametepe, Timothy A. Evans
Drosophila Robo3 is a member of the evolutionarily conserved Roundabout (Robo) receptor family and one of three Drosophila Robo paralogs. During embryonic ventral nerve cord development, Robo3 does not participate in canonical Slit-dependent midline repulsion, but instead regulates the formation of longitudinal axon pathways at specific positions along the medial-lateral axis. Longitudinal axon guidance by Robo3 is hypothesized to be Slit dependent, but this has not been directly tested. Here we create a series of Robo3 variants in which the N-terminal Ig1 domain is deleted or modified, in order to characterize the functional importance of Ig1 and Slit binding for Robo3's axon guidance activity. We show that Robo3 requires its Ig1 domain for interaction with Slit and for proper axonal localization in embryonic neurons, but deleting Ig1 from Robo3 only partially disrupts longitudinal pathway formation. Robo3 variants with modified Ig1 domains that cannot bind Slit retain proper localization and fully rescue longitudinal axon guidance. Our results indicate that Robo3 guides longitudinal axons independently of Slit, and that sequences both within and outside of Ig1 contribute to this Slit-independent activity.
{"title":"Slit-independent guidance of longitudinal axons by Drosophila Robo3","authors":"Abigail Carranza, LaFreda J. Howard, Haley E. Brown, Ayawovi Selom Ametepe, Timothy A. Evans","doi":"10.1016/j.ydbio.2025.01.017","DOIUrl":"10.1016/j.ydbio.2025.01.017","url":null,"abstract":"<div><div><em>Drosophila</em> Robo3 is a member of the evolutionarily conserved Roundabout (Robo) receptor family and one of three <em>Drosophila</em> Robo paralogs. During embryonic ventral nerve cord development, Robo3 does not participate in canonical Slit-dependent midline repulsion, but instead regulates the formation of longitudinal axon pathways at specific positions along the medial-lateral axis. Longitudinal axon guidance by Robo3 is hypothesized to be Slit dependent, but this has not been directly tested. Here we create a series of Robo3 variants in which the N-terminal Ig1 domain is deleted or modified, in order to characterize the functional importance of Ig1 and Slit binding for Robo3's axon guidance activity. We show that Robo3 requires its Ig1 domain for interaction with Slit and for proper axonal localization in embryonic neurons, but deleting Ig1 from Robo3 only partially disrupts longitudinal pathway formation. Robo3 variants with modified Ig1 domains that cannot bind Slit retain proper localization and fully rescue longitudinal axon guidance. Our results indicate that Robo3 guides longitudinal axons independently of Slit, and that sequences both within and outside of Ig1 contribute to this Slit-independent activity.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"521 ","pages":"Pages 14-27"},"PeriodicalIF":2.5,"publicationDate":"2025-02-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143254984","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 : 2025-02-02DOI: 10.1016/j.ydbio.2025.02.001
Lindsay Henderson , Yuya Okuzaki , Christophe Marcelle , Mike J. McGrew , Ken-ichi Nishijima
Biological resources are essential for research using chickens and quails, particularly in the field of developmental biology. Various lines of chickens and quails with naturally occurring genetic mutations and diverse phenotypes have been developed. Recent advancements in genetic modification techniques, such as using DNA transposons to modify cultured primordial germ cells (PGCs) and lentivirus-mediated transduction of PGCs in vivo, have enabled the creation of several transgenic chicken and quail lines. However, the relatively large body size of chickens and the need to maintain living animals due to the previous lack of reliable frozen stock methods, until the development of cultivating methods of PGCs, has caused a steady decline in the number of available lines globally. Several research facilities maintain chicken and quail lines and provide them for research purposes. This review describes the three main avian resource sites: The National Avian Research Facility at The Roslin Institute in the United Kingdom, Lyon Transgenic Quail Facility (MeLiS) in France, and Avian Bioscience Research Center at Nagoya University in Japan.
生物资源对利用鸡和鹌鹑进行研究至关重要，尤其是在发育生物学领域。目前已培育出各种具有自然发生的基因突变和不同表型的鸡和鹌鹑品系。此外，利用 DNA 转座子对培养的原始生殖细胞（PGCs）进行基因修饰，以及利用慢病毒介导的体内 PGCs 转导 DNA 转座子，这些最新进展已被用于开发各种转基因鸡和鹌鹑品系。然而，在开发出 PGCs 培育方法之前，由于鸡的体型相对较大，而且以前缺乏可靠的冷冻储存方法，因此需要维持活体动物，这导致全球可用品系的数量持续下降。一些研究机构维持着鸡和鹌鹑品系，并将其用于研究目的。本综述介绍了三个主要的禽类资源基地：英国罗斯林研究所的国家禽类研究设施、法国里昂转基因鹌鹑设施（MeLiS）和日本名古屋大学禽类生物科学研究中心。
{"title":"Avian bioresources for developmental biology: Chicken and quail resources in the United Kingdom, France, and Japan","authors":"Lindsay Henderson , Yuya Okuzaki , Christophe Marcelle , Mike J. McGrew , Ken-ichi Nishijima","doi":"10.1016/j.ydbio.2025.02.001","DOIUrl":"10.1016/j.ydbio.2025.02.001","url":null,"abstract":"<div><div>Biological resources are essential for research using chickens and quails, particularly in the field of developmental biology. Various lines of chickens and quails with naturally occurring genetic mutations and diverse phenotypes have been developed. Recent advancements in genetic modification techniques, such as using DNA transposons to modify cultured primordial germ cells (PGCs) and lentivirus-mediated transduction of PGCs in vivo, have enabled the creation of several transgenic chicken and quail lines. However, the relatively large body size of chickens and the need to maintain living animals due to the previous lack of reliable frozen stock methods, until the development of cultivating methods of PGCs, has caused a steady decline in the number of available lines globally. Several research facilities maintain chicken and quail lines and provide them for research purposes. This review describes the three main avian resource sites: The National Avian Research Facility at The Roslin Institute in the United Kingdom, Lyon Transgenic Quail Facility (MeLiS) in France, and Avian Bioscience Research Center at Nagoya University in Japan.</div></div>","PeriodicalId":11070,"journal":{"name":"Developmental biology","volume":"521 ","pages":"Pages 1-13"},"PeriodicalIF":2.5,"publicationDate":"2025-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143188589","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}
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