Pub Date : 2024-09-12DOI: 10.1101/2024.09.11.612381
Atsuko Kageyama, Narumi Ogonuki, Takuya Wakai, Takafumi Namiki, Yui Kawata, Manabu Ozawa, Yasuhiro Yamada, Toshiyuki Fukada, Atsuo Ogura, Rafael A. Fissore, Naomi Kashiwazaki, Junya Ito
In all vertebrates studied to date, a rise(s) in intracellular calcium is indispensable for successful fertilization and further embryonic development. Recent studies demonstrated that zinc is ejected to the extracellular milieu, the 'zinc spark', and follows the first few calcium rises of fertilization. However, the role of the zinc sparks in fertilization and development, and the supporting influx mechanism(s) are unknown. In this study in mouse oocytes, we investigated the role of zinc transporters Zip6/Slc39a6 and Zip10/Slc39a10. Zip10 mRNA or ZIP10 was expressed throughout folliculogenesis in oocyte or the plasma membrane, respectively. ZIP6 was also expressed in nuclear localization in oocytes and granulosa cells throughout folliculogenesis. The number of ovulated oocytes was examined in oocyte-specific Zip6 (Zip6d/d: Zip6flox/flox Gdf9Cre/+) and Zip10 (Zip10d/d: Zip10flox/flox Gdf9Cre/+) knockout mice, and no change was observed for either strain. Zip10d/d oocytes matured and formed normal metaphase II spindles, but had lower labile zinc levels as suggested by the zinc indicator, Fluozin-3 intensity. The levels of zinc fluorescence intensity in the Zip6d/d group were not different from the Zip6f/f. Fertilization-induced calcium oscillations were present in both Zip6d/d and Zip10d/d oocytes, but zinc sparks were observed in Zip6d/d but not in Zip10d/d oocytes. Despite other events of egg activation proceeding normally in Zip10d/d oocytes, embryo development into blastocysts was compromised. We show here for the first time that the zinc transporter ZIP10 contributes to zinc homeostasis in oocytes and embryos, highlighting the pivotal role of this divalent cation in early development.
{"title":"The oocyte zinc transporter Slc39a10/Zip10 is a regulator of zinc sparks during fertilization in mice.","authors":"Atsuko Kageyama, Narumi Ogonuki, Takuya Wakai, Takafumi Namiki, Yui Kawata, Manabu Ozawa, Yasuhiro Yamada, Toshiyuki Fukada, Atsuo Ogura, Rafael A. Fissore, Naomi Kashiwazaki, Junya Ito","doi":"10.1101/2024.09.11.612381","DOIUrl":"https://doi.org/10.1101/2024.09.11.612381","url":null,"abstract":"In all vertebrates studied to date, a rise(s) in intracellular calcium is indispensable for successful fertilization and further embryonic development. Recent studies demonstrated that zinc is ejected to the extracellular milieu, the 'zinc spark', and follows the first few calcium rises of fertilization. However, the role of the zinc sparks in fertilization and development, and the supporting influx mechanism(s) are unknown. In this study in mouse oocytes, we investigated the role of zinc transporters <em>Zip6/Slc39a6</em> and <em>Zip10/Slc39a10</em>. Zip10 mRNA or ZIP10 was expressed throughout folliculogenesis in oocyte or the plasma membrane, respectively. ZIP6 was also expressed in nuclear localization in oocytes and granulosa cells throughout folliculogenesis. The number of ovulated oocytes was examined in oocyte-specific <em>Zip6</em> (<em>Zip6<sup>d/d</sup></em>: <em>Zip6<sup>flox/flox</sup> Gdf9<sup>Cre/+</sup></em>) and <em>Zip10</em> (<em>Zip10<sup>d/d</sup></em>: <em>Zip10<sup>flox/flox</sup> Gdf9<sup>Cre/+</sup></em>) knockout mice, and no change was observed for either strain. <em>Zip10<sup>d/d</sup></em> oocytes matured and formed normal metaphase II spindles, but had lower labile zinc levels as suggested by the zinc indicator, Fluozin-3 intensity. The levels of zinc fluorescence intensity in the <em>Zip6<sup>d/d</sup></em> group were not different from the <em>Zip6<sup>f/f</sup></em>. Fertilization-induced calcium oscillations were present in both <em>Zip6<sup>d/d</sup></em> and <em>Zip10<sup>d/d</sup></em> oocytes, but zinc sparks were observed in <em>Zip6<sup>d/d</sup></em> but not in <em>Zip10<sup>d/d</sup></em> oocytes. Despite other events of egg activation proceeding normally in <em>Zip10<sup>d/d</sup></em> oocytes, embryo development into blastocysts was compromised. We show here for the first time that the zinc transporter ZIP10 contributes to zinc homeostasis in oocytes and embryos, highlighting the pivotal role of this divalent cation in early development.","PeriodicalId":501269,"journal":{"name":"bioRxiv - Developmental Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142211574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1101/2024.09.11.612512
Yiting Deng, Yuanhang He, Juan Xu, Haoting He, Manling Zhang, Guang Li
The fibroblast (FB), cardiomyocyte (CM), and vascular endothelial cell (Vas_EC) are the three major cell types in the heart, yet their relationships during development are largely unexplored. To address this gap, we employed RNA staining of the FB marker gene Col1a1 together with the CM marker gene Actn2 and the Vas_EC marker gene Cdh5 at different stages. This approach enabled us to discern the anatomical pattern of cardiac FBs and identify approximately one EC and four CMs directly interacting with each FB. Molecularly, through the analysis of single-cell mRNA sequencing (scRNA-seq) data, we unveiled collagen as the top signaling molecule derived from FBs influencing CM and Vas_EC development. Subsequently, we used a Pdgfra-CreER controlled diphtheria toxin A (DTA) system to ablate the FBs at different stages. We found that the ablation of FBs disrupted myocardium and vasculature development and led to embryonic heart defects. Using scRNA-seq, we further profiled the ablated hearts and identified molecular defects in their ventricular CMs and Vas_ECs compared to control hearts. Moreover, we identified a reduction of collagen in the ablated hearts and predicted collagen as the major signaling pathway regulating the differentially expressed genes in the ablated ventricular CMs. Finally, we performed both short-term and long-term fibroblast ablation at the neonatal stage. We found that short-term ablation caused a reduction in collagen and Vas_EC density, while long-term ablation may induce compensatory collagen expression without causing heart function reduction. In summary, our study has identified the function of fibroblasts in regulating myocardium and vasculature development and implicated an important role for the collagen pathway in this process.
{"title":"Cardiac Fibroblasts regulate myocardium and coronary vasculature development via the collagen signaling pathway","authors":"Yiting Deng, Yuanhang He, Juan Xu, Haoting He, Manling Zhang, Guang Li","doi":"10.1101/2024.09.11.612512","DOIUrl":"https://doi.org/10.1101/2024.09.11.612512","url":null,"abstract":"The fibroblast (FB), cardiomyocyte (CM), and vascular endothelial cell (Vas_EC) are the three major cell types in the heart, yet their relationships during development are largely unexplored. To address this gap, we employed RNA staining of the FB marker gene <em>Col1a1</em> together with the CM marker gene <em>Actn2</em> and the Vas_EC marker gene <em>Cdh5</em> at different stages. This approach enabled us to discern the anatomical pattern of cardiac FBs and identify approximately one EC and four CMs directly interacting with each FB. Molecularly, through the analysis of single-cell mRNA sequencing (scRNA-seq) data, we unveiled collagen as the top signaling molecule derived from FBs influencing CM and Vas_EC development. Subsequently, we used a Pdgfra-CreER controlled diphtheria toxin A (DTA) system to ablate the FBs at different stages. We found that the ablation of FBs disrupted myocardium and vasculature development and led to embryonic heart defects. Using scRNA-seq, we further profiled the ablated hearts and identified molecular defects in their ventricular CMs and Vas_ECs compared to control hearts. Moreover, we identified a reduction of collagen in the ablated hearts and predicted collagen as the major signaling pathway regulating the differentially expressed genes in the ablated ventricular CMs. Finally, we performed both short-term and long-term fibroblast ablation at the neonatal stage. We found that short-term ablation caused a reduction in collagen and Vas_EC density, while long-term ablation may induce compensatory collagen expression without causing heart function reduction. In summary, our study has identified the function of fibroblasts in regulating myocardium and vasculature development and implicated an important role for the collagen pathway in this process.","PeriodicalId":501269,"journal":{"name":"bioRxiv - Developmental Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142211573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1101/2024.09.11.612541
Ramachandran Prakasam, Julianna Determan, Mishka Narasimhan, Renata Shen, Maamoon Saleh, Gareth Chapman, Komal Kaushik, Paul Gontarz, Kesavan Meganathan, Bilal Hakim, Bo Zhang, James E Huettner, Kristen L Kroll
MYT1L is a neuronal transcription factor highly expressed in the developing and adult brain. While pathogenic MYT1L mutation causes neurodevelopmental disorders, these have not been characterized in human models of neurodevelopment. Here, we defined the consequences of pathogenic MYT1L mutation in human pluripotent stem cell-derived cortical interneurons. During differentiation, mutation reduced MYT1L expression and increased progenitor cell cycle exit and neuronal differentiation and synapse-related gene expression, morphological complexity, and synaptic puncta formation. Conversely, interneuron maturation was compromised, while variant neurons exhibited altered sodium and potassium channel activity and reduced function in electrophysiological analyses. CRISPRi-based knockdown similarly impaired interneuron differentiation and maturation, supporting loss of function-based effects. We further defined MYT1L genome-wide occupancy in interneurons and related this to the transcriptomic dysregulation resulting from MYT1L mutation, to identify direct targets that could mediate these phenotypic consequences. Together, this work delineates contributors to the etiology of neurodevelopmental disorders resulting from MYT1L mutation.
{"title":"Autism and Intellectual Disability-Associated MYT1L Mutation Alters Human Cortical Interneuron Differentiation, Maturation, and Physiology","authors":"Ramachandran Prakasam, Julianna Determan, Mishka Narasimhan, Renata Shen, Maamoon Saleh, Gareth Chapman, Komal Kaushik, Paul Gontarz, Kesavan Meganathan, Bilal Hakim, Bo Zhang, James E Huettner, Kristen L Kroll","doi":"10.1101/2024.09.11.612541","DOIUrl":"https://doi.org/10.1101/2024.09.11.612541","url":null,"abstract":"MYT1L is a neuronal transcription factor highly expressed in the developing and adult brain. While pathogenic MYT1L mutation causes neurodevelopmental disorders, these have not been characterized in human models of neurodevelopment. Here, we defined the consequences of pathogenic MYT1L mutation in human pluripotent stem cell-derived cortical interneurons. During differentiation, mutation reduced MYT1L expression and increased progenitor cell cycle exit and neuronal differentiation and synapse-related gene expression, morphological complexity, and synaptic puncta formation. Conversely, interneuron maturation was compromised, while variant neurons exhibited altered sodium and potassium channel activity and reduced function in electrophysiological analyses. CRISPRi-based knockdown similarly impaired interneuron differentiation and maturation, supporting loss of function-based effects. We further defined MYT1L genome-wide occupancy in interneurons and related this to the transcriptomic dysregulation resulting from MYT1L mutation, to identify direct targets that could mediate these phenotypic consequences. Together, this work delineates contributors to the etiology of neurodevelopmental disorders resulting from MYT1L mutation.","PeriodicalId":501269,"journal":{"name":"bioRxiv - Developmental Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142211543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1101/2024.09.12.612224
Ximena Ibarra-Soria, Elizabeth Webb, John Mulley
We analyzed the International Mouse Phenotyping Consortium (IMPC) release 19 set of 8,539 phenotyped whole-gene knockouts to identify 204 genes that alter vertebral development. These genes are broadly grouped into six categories based on their phenotype: "vertebral number" (22 genes); "vertebral processes" (35 genes); "spine shape" (16 genes); "tail morphology" (73 genes); "vertebral form" (62 genes); and "somitogenesis" (24 genes), with minimal overlap between groups. Gene expression analysis of somite trios across six developmental stages show that 182 of these genes are expressed in somites, and 60% of them show variable expression during somite maturation. A further 54% show expression changes between developmental stages. Fourteen of the 204 genes affecting vertebral development have a vertebral phenotype as their only phenotype, and for 34 genes vertebral phenotypes represent ≥50% of their total phenotypes. We find no evidence for a previous association of the majority of these genes with vertebral defects, and have therefore identified an extensive set of novel candidate genes for association with vertebral malformations in humans, including vertebral fusions, numerical variation, and scoliosis.
{"title":"Large-scale mouse mutagenesis identifies novel genes affecting vertebral development.","authors":"Ximena Ibarra-Soria, Elizabeth Webb, John Mulley","doi":"10.1101/2024.09.12.612224","DOIUrl":"https://doi.org/10.1101/2024.09.12.612224","url":null,"abstract":"We analyzed the International Mouse Phenotyping Consortium (IMPC) release 19 set of 8,539 phenotyped whole-gene knockouts to identify 204 genes that alter vertebral development. These genes are broadly grouped into six categories based on their phenotype: \"vertebral number\" (22 genes); \"vertebral processes\" (35 genes); \"spine shape\" (16 genes); \"tail morphology\" (73 genes); \"vertebral form\" (62 genes); and \"somitogenesis\" (24 genes), with minimal overlap between groups. Gene expression analysis of somite trios across six developmental stages show that 182 of these genes are expressed in somites, and 60% of them show variable expression during somite maturation. A further 54% show expression changes between developmental stages. Fourteen of the 204 genes affecting vertebral development have a vertebral phenotype as their only phenotype, and for 34 genes vertebral phenotypes represent ≥50% of their total phenotypes. We find no evidence for a previous association of the majority of these genes with vertebral defects, and have therefore identified an extensive set of novel candidate genes for association with vertebral malformations in humans, including vertebral fusions, numerical variation, and scoliosis.","PeriodicalId":501269,"journal":{"name":"bioRxiv - Developmental Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142226678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1101/2024.09.11.612484
Davys Lopez, Kevin D. Rostam, Sumaira Zamurrad, Shuwa Xu, Richard S Mann
For flies to walk properly, motor neurons (MNs) from the ventral nerve cord (VNC) need to reach the correct muscle, and arborize appropriately during development. The canonical view of how this is achieved is that cell surface proteins are expressed pre- and post-synaptically that bind to each other like molecular lock-and-keys that guide neurons to their targets. The binding affinities of these molecules can vary by more than 100-fold. In the fly leg neuromuscular system, three MNs express DIP-α and their target muscles express its cognate partner, dpr10, both of which encode members of the Immunoglobulin superfamily (IgSF). Although, both of these molecules are necessary for the maintenance of MN-muscle contacts, the role that specific affinities play in this process has not been examined. Here we use knock-in mutations into DIP-α and dpr10 that either decrease or increase the affinity between these two proteins. Compared to control animals, decreasing the affinity results in phenotypes similar to DIP-α or dpr10 null animals, where MN axons fail to maintain contacts with their muscle targets and retract their filopodia, resulting in stunted and/or branchless axons. We also find that the three DIP-α-expressing motor neurons behave differently to the loss of affinity. Surprisingly, if the affinity increases past a certain threshold, a similar branchless phenotype is observed in adult legs. Live imaging during pupal development shows that MN filopodia are unable to productively engage their muscle targets and behavioral assays suggest that these defects lead to locomotor deficits. These data suggest that CAM affinities are tuned to a specific range to achieve proper neuronal morphology.
{"title":"A critical affinity window for IgSF proteins DIP-alpha and Dpr10 is required for proper motor neuron arborization","authors":"Davys Lopez, Kevin D. Rostam, Sumaira Zamurrad, Shuwa Xu, Richard S Mann","doi":"10.1101/2024.09.11.612484","DOIUrl":"https://doi.org/10.1101/2024.09.11.612484","url":null,"abstract":"For flies to walk properly, motor neurons (MNs) from the ventral nerve cord (VNC) need to reach the correct muscle, and arborize appropriately during development. The canonical view of how this is achieved is that cell surface proteins are expressed pre- and post-synaptically that bind to each other like molecular lock-and-keys that guide neurons to their targets. The binding affinities of these molecules can vary by more than 100-fold. In the fly leg neuromuscular system, three MNs express DIP-α and their target muscles express its cognate partner, dpr10, both of which encode members of the Immunoglobulin superfamily (IgSF). Although, both of these molecules are necessary for the maintenance of MN-muscle contacts, the role that specific affinities play in this process has not been examined. Here we use knock-in mutations into DIP-α and dpr10 that either decrease or increase the affinity between these two proteins. Compared to control animals, decreasing the affinity results in phenotypes similar to DIP-α or dpr10 null animals, where MN axons fail to maintain contacts with their muscle targets and retract their filopodia, resulting in stunted and/or branchless axons. We also find that the three DIP-α-expressing motor neurons behave differently to the loss of affinity. Surprisingly, if the affinity increases past a certain threshold, a similar branchless phenotype is observed in adult legs. Live imaging during pupal development shows that MN filopodia are unable to productively engage their muscle targets and behavioral assays suggest that these defects lead to locomotor deficits. These data suggest that CAM affinities are tuned to a specific range to achieve proper neuronal morphology.","PeriodicalId":501269,"journal":{"name":"bioRxiv - Developmental Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142211541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1101/2024.09.08.611877
Morgan Zych, Natalie Lo, Kate A Patton, Kewei Wang, Brian J Cox
The placenta an essential extraembryonic organ that supports the fetus throughout gestation. The interactions between the placenta and the maternal immune system during the first trimester have not been wholly characterized despite their close physical association and hemi-allogeneic relationship. The most abundant type of immune cell in the uterus in the first trimester is the decidual natural killer cell (dNK). Despite their name, dNKs play supportive roles during pregnancy by remodelling uterine spiral arteries. We present evidence suggesting that the matrix metalloproteinases (MMPs) that dNKs secrete to promote this remodelling also drive placental development. This study used a novel co-culture system of dNKs and trophoblast organoids, which are mini-organs representing two to three different cell types of the human placenta. We found that co-cultures for one week led to significant (p=0.020) increases in the organoid area. We also observed significant decreases in trophoblast stemness markers and upregulation of gene sets associated with extravillous trophoblast (EVT) development through bulk RNA sequencing and immunohistochemical examinations. These changes were accompanied by significant (p<0.001) increases in collagen subunit gene expression in the organoids, with simultaneous significant decreases (p<0.001) in the proportion of organoid area occupied by collagen as determined through Massons Trichrome. Cultures containing dNKs also contained significantly higher MMP1, 3, 9, and 10 levels in their culture media, each of which can break down collagen. These findings demonstrate that dNKs promote changes concordant with trophoblast differentiation towards EVTs and villous branching morphogenesis.
{"title":"Decidual natural killer cells promote extravillous trophoblast developmental pathways: evidence from trophoblast organoid co-cultures","authors":"Morgan Zych, Natalie Lo, Kate A Patton, Kewei Wang, Brian J Cox","doi":"10.1101/2024.09.08.611877","DOIUrl":"https://doi.org/10.1101/2024.09.08.611877","url":null,"abstract":"The placenta an essential extraembryonic organ that supports the fetus throughout gestation. The interactions between the placenta and the maternal immune system during the first trimester have not been wholly characterized despite their close physical association and hemi-allogeneic relationship. The most abundant type of immune cell in the uterus in the first trimester is the decidual natural killer cell (dNK). Despite their name, dNKs play supportive roles during pregnancy by remodelling uterine spiral arteries. We present evidence suggesting that the matrix metalloproteinases (MMPs) that dNKs secrete to promote this remodelling also drive placental development. This study used a novel co-culture system of dNKs and trophoblast organoids, which are mini-organs representing two to three different cell types of the human placenta. We found that co-cultures for one week led to significant (p=0.020) increases in the organoid area. We also observed significant decreases in trophoblast stemness markers and upregulation of gene sets associated with extravillous trophoblast (EVT) development through bulk RNA sequencing and immunohistochemical examinations. These changes were accompanied by significant (p<0.001) increases in collagen subunit gene expression in the organoids, with simultaneous significant decreases (p<0.001) in the proportion of organoid area occupied by collagen as determined through Massons Trichrome. Cultures containing dNKs also contained significantly higher MMP1, 3, 9, and 10 levels in their culture media, each of which can break down collagen. These findings demonstrate that dNKs promote changes concordant with trophoblast differentiation towards EVTs and villous branching morphogenesis.","PeriodicalId":501269,"journal":{"name":"bioRxiv - Developmental Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142211542","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1101/2024.09.11.612396
Thomas Daniele, Jeanne Cury, Marie-Charlotte Morin, Arnaud Ahier, Davide Isaia, Sophie Jarriault
Cell identity can be reprogrammed, naturally or experimentally, albeit with low frequency. Why given cells, but not their neighbours, undergo a cell identity conversion remains unclear. We find that Notch signalling plays a key role to promote natural transdifferentiation in C. elegans. Endogenous Notch signal endows a cell with the competence to transdifferentiate by promoting plasticity factors expression (hlh-16/Olig and sem-4/Sall). Strikingly, exogenous Notch can trigger ectopic transdifferentiation in vivo. However, Notch signalling can both promote and block transdifferentiation depending on its activation timing. Notch only promotes transdifferentiation during a precise window of opportunity and signal duration must be tightly controlled in time. Our findings emphasise the importance of temporality and dynamics of the underlying molecular events preceding the initiation of natural cell reprogramming. Finally, our results support a model where both an extrinsic signal and the intrinsic cellular context combine to empower a cell with the competence to transdifferentiate.
{"title":"Essential and dual effects of Notch activity on a natural transdifferentiation event","authors":"Thomas Daniele, Jeanne Cury, Marie-Charlotte Morin, Arnaud Ahier, Davide Isaia, Sophie Jarriault","doi":"10.1101/2024.09.11.612396","DOIUrl":"https://doi.org/10.1101/2024.09.11.612396","url":null,"abstract":"Cell identity can be reprogrammed, naturally or experimentally, albeit with low frequency. Why given cells, but not their neighbours, undergo a cell identity conversion remains unclear. We find that Notch signalling plays a key role to promote natural transdifferentiation in C. elegans. Endogenous Notch signal endows a cell with the competence to transdifferentiate by promoting plasticity factors expression (hlh-16/Olig and sem-4/Sall). Strikingly, exogenous Notch can trigger ectopic transdifferentiation in vivo. However, Notch signalling can both promote and block transdifferentiation depending on its activation timing. Notch only promotes transdifferentiation during a precise window of opportunity and signal duration must be tightly controlled in time. Our findings emphasise the importance of temporality and dynamics of the underlying molecular events preceding the initiation of natural cell reprogramming. Finally, our results support a model where both an extrinsic signal and the intrinsic cellular context combine to empower a cell with the competence to transdifferentiate.","PeriodicalId":501269,"journal":{"name":"bioRxiv - Developmental Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142226680","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1101/2024.09.10.612241
Leah F. Cabo, Liheng Yang, Mingze Gao, Rafaela J. da Silva, NyJaee N. Washington, Sarah M. Reilly, Christina J. Megli, Carolyn B. Coyne, Jon P. Boyle
Pregnancy is a critical point of vulnerability to infection and other insults that could compromise proper fetal development. The placenta acts as a protective and nutrient-permeable barrier to most infectious agents, but a few are capable of bypassing its defenses. Remarkably little is known about how exposure to these select pathogens might impact ongoing placental development. Here we demonstrate that Toxoplasma gondii entirely misdirects the developmental program of trophoblast stem cells. Infection of progenitor cytotrophoblasts prevents fusion and differentiation to infection-resistant syncytiotrophoblast. Rather, T. gondii elicits a unique transcriptional identity that polarizes cytotrophoblasts to the infection-permissive extravillous trophoblast fate. Strong evidence of developmental disruption is found in multiple orthogonal models, including trophoblast stem cells, trophoblast organoids, and chorionic villi. Manipulation of cell fate by the parasite is most dramatic in trophoblast organoids, where we see robust outgrowth of HLA-G(+) extravillous trophoblasts. Collectively, these data show that Toxoplasma antagonizes differentiation of an infection-resistant cell type by inducing formation of an infection-permissive cell type, therefore potentiating its own transmission to the fetus.
{"title":"Toxoplasma gondii infection misdirects placental trophoblast lineage specification","authors":"Leah F. Cabo, Liheng Yang, Mingze Gao, Rafaela J. da Silva, NyJaee N. Washington, Sarah M. Reilly, Christina J. Megli, Carolyn B. Coyne, Jon P. Boyle","doi":"10.1101/2024.09.10.612241","DOIUrl":"https://doi.org/10.1101/2024.09.10.612241","url":null,"abstract":"Pregnancy is a critical point of vulnerability to infection and other insults that could compromise proper fetal development. The placenta acts as a protective and nutrient-permeable barrier to most infectious agents, but a few are capable of bypassing its defenses. Remarkably little is known about how exposure to these select pathogens might impact ongoing placental development. Here we demonstrate that <em>Toxoplasma gondii</em> entirely misdirects the developmental program of trophoblast stem cells. Infection of progenitor cytotrophoblasts prevents fusion and differentiation to infection-resistant syncytiotrophoblast. Rather, <em>T. gondii</em> elicits a unique transcriptional identity that polarizes cytotrophoblasts to the infection-permissive extravillous trophoblast fate. Strong evidence of developmental disruption is found in multiple orthogonal models, including trophoblast stem cells, trophoblast organoids, and chorionic villi. Manipulation of cell fate by the parasite is most dramatic in trophoblast organoids, where we see robust outgrowth of HLA-G(+) extravillous trophoblasts. Collectively, these data show that <em>Toxoplasma</em> antagonizes differentiation of an infection-resistant cell type by inducing formation of an infection-permissive cell type, therefore potentiating its own transmission to the fetus.","PeriodicalId":501269,"journal":{"name":"bioRxiv - Developmental Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142211575","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1101/2024.09.12.612608
Ru Hong, Cedric Finet, Antonia Monteiro
Insect cuticle is normally deposited outside the plasma membrane of epidermal cells, making it unclear how cuticular pillars (trabeculae) are found inside butterfly wing scale cells. By co-labelling the cuticle and the plasma membrane, we found evidence that the plasma membrane invaginates towards the interior of the scale during development, and that chitin pillars form within these invaginations within the cell, but topologically outside it. Furthermore, we found that Calreticulin, a multifunctional protein, is essential for the formation of these trabeculae. This protein was found colocalized with chitin outside the cell membrane, as scales were developing, and its knockout led to loss of chitin pillars, disruption of other scale morphologies, and loss of pigmentation. Our results implicate this multifunctional protein in butterfly wing scale coloration and morphology.
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Pub Date : 2024-09-12DOI: 10.1101/2024.09.10.612284
Christian J. Bellissimo, Erica Yeo, Tatiane A. Ribeiro, Patrycja A. Jazwiec, Chethana Ellewela, Jaskiran Bains, Ali A. Ashkar, Alexander G. Beristain, Dawn M.E. Bowdish, Deborah M. Sloboda
Excess maternal adiposity (i.e., overweight and obesity) during pregnancy has been linked to impaired uteroplacental perfusion, compromised placental development, and a higher risk of adverse pregnancy outcomes. Owing to the nature of chronic inflammation and immune dysregulation accompanying excess adiposity, disruption of leukocyte-mediated tissue remodelling and immunoregulation within the decidua have emerged as likely drivers contributing to suboptimal placental function in pregnancies impacted by maternal overweight or obesity. However, the impacts of excess adiposity on major populations of innate lymphoid cells (ILCs) and macrophages which orchestrate these processes and the environment that these cells occupy remain vastly understudied. Here, we used a mouse model of chronic high-fat, high-sucrose (HFHS) diet-feeding to characterize the impacts of an obesogenic milieu on decidual immune dynamics during placental development at mid-gestation (E10.5). HFHS pregnancies exhibited marked increases in total decidual leukocyte abundance, driven by population-level increases in tissue-resident and conventional NK cells, and MHC-II+ macrophages. This was not associated with abnormalities in implantation site morphology or decidual spiral artery remodelling but was coincident with histological patterns of local inflammation. In line with this, expression of canonical proinflammatory cytokines and chemokines were moderately upregulated in bulk decidual tissue of HFHS dams. This was accompanied by more potent elevations in multiple mediators of angiogenesis, endothelial activation, and coagulation in HFHS decidual tissue. Collectively, these findings point towards pathological vascular inflammation and possibly dysregulated decidual angiogenesis in the first half of pregnancy as factors predisposing to reduced placental efficiency, malperfusion, and inflammation seen in pregnancies affected by maternal overweight and obesity.
妊娠期母体脂肪过多（即超重和肥胖）与子宫胎盘灌注受损、胎盘发育受损和不良妊娠结局的风险较高有关。由于伴随着过度肥胖的慢性炎症和免疫失调的性质，蜕膜内白细胞介导的组织重塑和免疫调节的破坏已成为导致受母体超重或肥胖影响的孕妇胎盘功能不佳的可能驱动因素。然而，过多的脂肪对先天性淋巴细胞（ILCs）和巨噬细胞等主要细胞群的影响以及这些细胞所处的环境仍未得到充分研究。在这里，我们利用长期高脂肪、高蔗糖（HFHS）饮食喂养的小鼠模型来描述肥胖环境对妊娠中期（E10.5）胎盘发育过程中蜕膜免疫动态的影响。在组织驻留细胞、常规 NK 细胞和 MHC-II+ 巨噬细胞群体水平增加的驱动下，HFHS 妊娠显示出蜕膜白细胞总丰度的显著增加。这与植入部位形态或蜕膜螺旋动脉重塑的异常无关，但与局部炎症的组织学模式相吻合。与此相一致的是，HFHS 母体的大量蜕膜组织中，典型促炎细胞因子和趋化因子的表达中度上调。与此同时，HFHS蜕膜组织中血管生成、内皮活化和凝血的多种介质也出现了更强的升高。总之，这些研究结果表明，妊娠前半期病理血管炎症和蜕膜血管生成失调可能是导致胎盘效率降低、灌注不良和炎症的因素，这在受孕妇超重和肥胖影响的妊娠中很容易见到。
{"title":"Maternal obesogenic diet disrupts mid-gestation decidual immune and vascular homeostasis without impairing spiral artery remodelling","authors":"Christian J. Bellissimo, Erica Yeo, Tatiane A. Ribeiro, Patrycja A. Jazwiec, Chethana Ellewela, Jaskiran Bains, Ali A. Ashkar, Alexander G. Beristain, Dawn M.E. Bowdish, Deborah M. Sloboda","doi":"10.1101/2024.09.10.612284","DOIUrl":"https://doi.org/10.1101/2024.09.10.612284","url":null,"abstract":"Excess maternal adiposity (i.e., overweight and obesity) during pregnancy has been linked to impaired uteroplacental perfusion, compromised placental development, and a higher risk of adverse pregnancy outcomes. Owing to the nature of chronic inflammation and immune dysregulation accompanying excess adiposity, disruption of leukocyte-mediated tissue remodelling and immunoregulation within the decidua have emerged as likely drivers contributing to suboptimal placental function in pregnancies impacted by maternal overweight or obesity. However, the impacts of excess adiposity on major populations of innate lymphoid cells (ILCs) and macrophages which orchestrate these processes and the environment that these cells occupy remain vastly understudied. Here, we used a mouse model of chronic high-fat, high-sucrose (HFHS) diet-feeding to characterize the impacts of an obesogenic milieu on decidual immune dynamics during placental development at mid-gestation (E10.5). HFHS pregnancies exhibited marked increases in total decidual leukocyte abundance, driven by population-level increases in tissue-resident and conventional NK cells, and MHC-II+ macrophages. This was not associated with abnormalities in implantation site morphology or decidual spiral artery remodelling but was coincident with histological patterns of local inflammation. In line with this, expression of canonical proinflammatory cytokines and chemokines were moderately upregulated in bulk decidual tissue of HFHS dams. This was accompanied by more potent elevations in multiple mediators of angiogenesis, endothelial activation, and coagulation in HFHS decidual tissue. Collectively, these findings point towards pathological vascular inflammation and possibly dysregulated decidual angiogenesis in the first half of pregnancy as factors predisposing to reduced placental efficiency, malperfusion, and inflammation seen in pregnancies affected by maternal overweight and obesity.","PeriodicalId":501269,"journal":{"name":"bioRxiv - Developmental Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142211602","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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