Chao Li, Jingwen Wang, Jun Feng, Gaoliang Zhou, Yongjin Jiang, Chunmiao Luo, Ziping Cheng, Jiehua Li
{"title":"Circ-JA760602 promotes the apoptosis of hypoxia-induced cardiomyocytes by transcriptionally suppressing BCL2.","authors":"Chao Li, Jingwen Wang, Jun Feng, Gaoliang Zhou, Yongjin Jiang, Chunmiao Luo, Ziping Cheng, Jiehua Li","doi":"10.1387/ijdb.220150jl","DOIUrl":"https://doi.org/10.1387/ijdb.220150jl","url":null,"abstract":"<p><p>Acute myocardial infarction (AMI) is myocardial necrosis caused by the complete or partial obstruction of a coronary artery. Circular RNAs (circRNAs) have been proven as regulators in the progression of various human diseases, including AMI. However, the role of novel circ-JA760602 in AMI remains unknown. Here, we investigated the role of circ-JA760602 in modulating the apoptosis of hypoxia-induced AMI cells using the AC16 cardiomyocyte in vitro cell model. The expression of circ-JA760602 in AC16 cardiomyocytes subjected to hypoxia was measured by quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability was measured by cell counting kit-8 (CCK-8) assay. Apoptosis of cardiomyocytes was evaluated by TUNEL assay and flow cytometry analysis. The cellular location of circ-JA760602 was identified through fluorescence in situ hybridization (FISH) assay and subcellular fractionation assay. The downstream molecular mechanisms of circ-JA760602 were demonstrated by luciferase reporter assay, RNA binding protein immunoprecipitation (RIP) assay and chromatin immunoprecipitation (ChIP) assay. Rescue assays were performed to demonstrate the effects of BCL2 knockdown on circ-JA760602 silencing-mediated cardiomyocyte apoptosis. Circ-JA760602 expression was elevated after hypoxia treatment. Knockdown of circ-JA760602 enhanced viability and curbed apoptosis of hypoxia-treated cardiomyocytes. EGR1 and E2F1 could activate BCL2 transcription. Cytoplasmic circ-JA760602 bound with EGR1 and E2F1 to thus inhibit their nuclear translocation. BCL2 knockdown reversed the effects of circ-JA760602 silencing on the apoptosis of hypoxia-treated AC16 cells. Circ-JA760602 promotes hypoxia-induced apoptosis of cardiomyocytes by binding with EGR1 and E2F1 to inhibit the transcriptional activation of BCL2.</p>","PeriodicalId":50329,"journal":{"name":"International Journal of Developmental Biology","volume":"67 1","pages":"9-17"},"PeriodicalIF":0.7,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10002337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Importance of the filamin A-Sav1 interaction in organ size control: evidence from transgenic mice.","authors":"Huaguan Zhang, Ziwei Yang, Fumihiko Nakamura","doi":"10.1387/ijdb.230054fn","DOIUrl":"https://doi.org/10.1387/ijdb.230054fn","url":null,"abstract":"<p><p>The nucleocytoplasmic translocation of yes-associated protein 1 (YAP1) controls the growth of animal tissues and organs. YAP1 binds to transcription factors in the nucleus to activate the transcription of proliferation and anti-apoptotic genes. The Hippo pathway prevents the nuclear translocation of YAP1 by phosphorylating YAP1, while mechanical forces promote it by opening the nuclear pore complex and stimulating other signaling pathways. Recently we found that Protein salvador homolog 1 (SAV1), a component of the Hippo pathway, interacts with filamin A (FLNA) in a force-dependent manner, raising a possibility that the Hippo pathway is regulated by mechanical force through the FLNA-SAV1 axis. To test this hypothesis, we generated conditional knock-in (KI) mice expressing non-Flna-binding mutant Sav1 in hepatocytes by crossing with mice carrying Cre recombinase driven by the serum albumin (alb) gene promoter. Unexpectedly, the insertion of the flox cassette skipped exon 2, resulting in a shorter Sav1 in all the transgenic mice. Since exon 2 encodes a fragment containing a Flna-binding domain, we analyzed both point mutant KI and exon 2-deleted mutant mice. Here we show that disruption of the Flna-Sav1 interaction in the mouse liver promotes apoptosis and suppresses tissue and organ growth without affecting the phosphorylation level of Yap1. These results provide evidence that the growth of animal tissues and organs is regulated by apoptosis downstream of the force-dependent FLNA-SAV1 interaction, at least in part.</p>","PeriodicalId":50329,"journal":{"name":"International Journal of Developmental Biology","volume":"67 2","pages":"27-37"},"PeriodicalIF":0.7,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10318904","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jason Charish, Hidekiyo Harada, Xiaoyan Chen, Thomas Wälchli, Cathy L Barr, Philippe P Monnier
{"title":"The Dyslexia-associated gene <i>KIAA0319L</i> is involved in neuronal migration in the developing chick visual system.","authors":"Jason Charish, Hidekiyo Harada, Xiaoyan Chen, Thomas Wälchli, Cathy L Barr, Philippe P Monnier","doi":"10.1387/ijdb.230052pm","DOIUrl":"https://doi.org/10.1387/ijdb.230052pm","url":null,"abstract":"<p><p>The gene KIAA0319-Like (KIAA0319L) is thought to confer susceptibility for developmental dyslexia. Dyslexia may be caused by alterations in neuronal migration, and in utero knockdown of KIAA0319L in rats indicated migration errors. However, studies carried out with KIAA0319L knockout mice did not reveal an altered neuronal migration phenotype. Gene knockout may activate compensatory mechanisms to buffer against genetic mutations during development. Here we assessed the role of KIAA0319L on migrating neurons in the chick developing tectum. Whole mount in situ hybridization was performed for KIAA0319L on embryonic day (E)3 - E5 chick embryos and in situ hybridization on sections was performed at later stages. The specificity and efficiency of engineered microRNA (miRNA) constructs targeting KIAA0319L for knocking down KIAA0319L were verified. miRNAs were electroporated into E5 chick optic tecta. Our studies demonstrate that KIAA0319L is expressed in the developing chick visual system, as well as in the otic vesicles. Knockdown of KIAA0319L in the optic tectum results in abnormal neuronal migration, strengthening the argument that KIAA0319L is involved in this developmental process.</p>","PeriodicalId":50329,"journal":{"name":"International Journal of Developmental Biology","volume":"67 2","pages":"49-56"},"PeriodicalIF":0.7,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9966566","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"DNA methyltransferase (Dnmt) silencing causes increased Cdx2 and Nanog levels in surviving embryos.","authors":"Fatma Uysal, Gozde Sukur, Nazlican Bozdemir, Ozgur Cinar","doi":"10.1387/ijdb.230040oc","DOIUrl":"https://doi.org/10.1387/ijdb.230040oc","url":null,"abstract":"<p><p>Epigenetic mechanisms are one of the essential regulators of gene expression which do not involve altering the primary nucleotide sequence. DNA methylation is considered among the most prominent epigenetic mechanisms in controlling the functions of genes related to cell differentiation, cell cycle, cell survival, autophagy, and embryo development. DNA methyl transferases (Dnmts) control DNA methylation, the levels of which are differentially altered during embryonic development, and may determine cell differentiation fate as in the case of pluripotent inner cell mass (ICM) or trophectoderm (TE). In this study, we aimed to analyze the role of Dnmt1 and Dnmt3a enzymes in ICM (using the Nanog marker) and TE (using the Cdx2 marker) differentiation, autophagy (using p62 marker), reactive oxygen species (ROS) production, and apoptosis (using TUNEL) during mouse preimplantation embryo development. Following knockdown of <i>Dnmt1</i> and <i>Dnmt3a</i> in zygotes, expression levels of Cdx2 in the trophectoderm and Nanog in the inner cell mass were measured, as well as p62 levels, reactive oxygen species (ROS) production, and apoptosis levels after 96 hours in embryo culture. We found that knockdown of Dnmt1 or Dnmt3a significantly induced Cdx2 and Nanog expression. Similarly, p62 expression, ROS levels and apoptosis significantly increased after silencing. This study shows that Dnmt genes are highly crucial for embryonic fate determination and survival. Further studies are required to reveal the specific targets of these methylation processes related to cell differentiation, survival, autophagy, and ROS production in mouse and human preimplantation embryos.</p>","PeriodicalId":50329,"journal":{"name":"International Journal of Developmental Biology","volume":"67 1","pages":"1-8"},"PeriodicalIF":0.7,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9831253","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Single-cell transcriptome profiling reveals distinct expression patterns among genes in the mouse incisor dental pulp.","authors":"Badam Enkhmandakh, Dashzeveg Bayarsaihan","doi":"10.1387/ijdb.220173db","DOIUrl":"https://doi.org/10.1387/ijdb.220173db","url":null,"abstract":"<p><p>SOX transcription factors play key roles in cell differentiation and cell fate determination during development. Using single-cell RNA-sequencing data, we examined the expression profiles of <i>Sox</i> genes in the mouse incisor dental pulp. Our analysis showed that <i>Sox4</i>, <i>Sox5</i>, <i>Sox9</i>, <i>Sox11</i>, and <i>Sox12</i> are mainly expressed in mesenchymal stem/stromal cells (MSCs) representing osteogenic cells at different stages of differentiation. We found that in several MSCs, <i>Sox</i> genes co-expressed with regulatory genes such as <i>Sp7</i>, <i>Satb2</i>, <i>Msx1</i>, <i>Snai2</i>, <i>Dlx1</i>, <i>Twist2</i>, and <i>Tfap2a</i>. In addition, <i>Sox</i> family genes colocalized with <i>Runx2</i> and <i>Lef1</i>, which are highly enriched in MSCs undergoing osteoblast differentiation. A protein interaction network analysis uncovered that CREBBP, CEBPB, TLE1, TWIST1, and members of the HDAC and SMAD families are interacting partners of RUNX2 and LEF1 during skeletal development. Collectively, the distinct expression patterns of the SOX transcription factors suggest that they play essential regulatory roles in directing lineage-specific gene expression during differentiation of MSCs.</p>","PeriodicalId":50329,"journal":{"name":"International Journal of Developmental Biology","volume":"67 1","pages":"19-25"},"PeriodicalIF":0.7,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9585281","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Batoul Chami, Bella Bates, Luke Shaheen, John Abramyan
{"title":"Characterization of the developing axolotl nasal cavity supports multiple evolution of the vertebrate choana.","authors":"Batoul Chami, Bella Bates, Luke Shaheen, John Abramyan","doi":"10.1387/ijdb.230098ja","DOIUrl":"https://doi.org/10.1387/ijdb.230098ja","url":null,"abstract":"<p><p>All tetrapods (mammals, birds, reptiles, and amphibians) share the ability to breathe with their mouths closed due to the formation of choanae, which are openings that allow communication between the nasal and oral cavities. In most fishes, the nasal cavities serve a strictly olfactory function, possessing incurrent and excurrent nares that lie outside of the mouth and therefore, never communicate with the respiratory system. It is not until the evolution of tetrapods, in which the nasal cavities consistently open into the mouth, that they are used both for olfaction and for respiration. However, this developmental transition is poorly understood, with no consensus on the evolutionary origin of the choana in various groups despite decades of debate. Here, we use high-contrast 3D imaging in conjunction with histology and apoptotic cell analysis in non-mineralized embryonic tissues to study the formation of the choana in the axolotl (<i>Ambystoma mexicanum</i>), an aquatic salamander species. We show that the axolotl choana forms from an extension of the embryonic nasal sac, which pushes through intervening mesenchyme and connects with the palate epithelium of the oral cavity, eventually breaking through. This mechanism differs from caecilians, mammals and reptiles, where fusion across a bucconasal groove plays an active role in choana formation. Nevertheless, caecilians, mammals and axolotls converge on the development of a transient epithelial tissue that has to break down in order to develop a patent choana, adding another twist to the intriguing arguments on the evolutionary history of the choana.</p>","PeriodicalId":50329,"journal":{"name":"International Journal of Developmental Biology","volume":"67 2","pages":"57-63"},"PeriodicalIF":0.7,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9964652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Geovanna Carla Zacheo Coelho, Gabriella Braga Carvalho, Paulo Sergio Monzani, Lucia Suarez Lopez, Nivaldo Ferreira do Nascimento, José Augusto Senhorini, George Shigueki Yasui
{"title":"Conditions for transplantation of primordial germ cells in the yellowtail tetra, <i>Astyanax altiparanae</i>.","authors":"Geovanna Carla Zacheo Coelho, Gabriella Braga Carvalho, Paulo Sergio Monzani, Lucia Suarez Lopez, Nivaldo Ferreira do Nascimento, José Augusto Senhorini, George Shigueki Yasui","doi":"10.1387/ijdb.230009gc","DOIUrl":"https://doi.org/10.1387/ijdb.230009gc","url":null,"abstract":"<p><p>Biotechniques, including surrogate propagation derived from primordial germ cell (PGC) transplantation, are valuable tools for the reconstitution of endangered fish species. Although promising, there are no previous studies reporting such approaches using neotropical fish species. The aim of this study was to establish germline chimeras in neotropical fish by using the yellowtail tetra <i>Astyanax altiparanae</i> as a model species of the order Characiformes. Germline chimeras were obtained after transplantation of PGCs cultivated under different conditions: saline medium and supplemented with DMEM, amino acids, vitamins, glutamine, pyruvate, and fetal bovine serum, and subsequently transplanted into <i>A. altiparanae</i> triploids and triploid hybrids from the cross between <i>A. altiparanae</i> (♀) and <i>A. fasciatus</i> (♂). The results indicate ectopic migration in host embryos after transplantation of PGCs cultivated in saline medium. However, PGCs cultivated in supplemented medium migrated to the region of the gonadal ridge in 4.5% of triploid and 19.3% in triploid hybrid. In addition, the higher expression of <i>dnd1, ddx4</i> and <i>dazl</i> genes was found in PGCs cultivated in supplemented culture medium. This indicates that the culture medium influences the maintenance and development of the cultivated cells. The expression levels of <i>nanos</i> and <i>cxcr4b</i> (related to the differentiation and migration of PGCs) were decreased in PGCs from the supplemented culture medium, supporting the results of ectopic migration. This is the first study to report the transplantation of PGCs to obtain germline chimera in neotropical species. The establishment of micromanipulation procedures in a model neotropical species will open new insights for the conservation of endangered species.</p>","PeriodicalId":50329,"journal":{"name":"International Journal of Developmental Biology","volume":"67 2","pages":"39-48"},"PeriodicalIF":0.7,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10336536","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"<i>Cend1</i> and <i>Neurog2</i> efficiently reprogram human cortical astrocytes to neural precursor cells and induced-neurons.","authors":"Katerina Aravantinou-Fatorou, Seyyedeh Vejdani, Dimitra Thomaidou","doi":"10.1387/ijdb.210148dt","DOIUrl":"https://doi.org/10.1387/ijdb.210148dt","url":null,"abstract":"<p><p>Direct reprogramming of glial cells into induced-neurons is a promising strategy for CNS repair after acute injury or neurodegenerative diseases. Grey matter astrocytes, which exhibit features of neural stem cells when activated, are an ideal cell source for direct neuronal conversion. The aim of the study is the investigation of the neuronal reprogramming capacity of <i>CEND1</i> and/or <i>Neurogenin-2</i> (<i>NEUROG2</i>) upon their overexpression on primary human adult cortical astrocytes. Our data indicate that adult human cortical astrocytes can be directly reprogrammed by either <i>CEND1</i> or <i>NEUROG2</i> to cells with differentiated neuronal morphology, exhibiting long neurites and branched processes. Exploration of gene expression dynamics along the conversion process revealed that neuronal genes are significantly up-regulated while astrocytic genes are down-regulated. Differentiated induced-neurons (iNs) exhibit either GABAergic or glutamatergic/dopaminergic identity upon <i>CEND1</i> and <i>NEUROG2</i> overexpression respectively. Co-expression of <i>CEND1</i> and <i>NEUROG2</i> in double-transduced cultures induced elevated expression levels of neural progenitor/stem genes and appearance of highly proliferative spheres with neural progenitor cell (NPC) properties in culture.</p>","PeriodicalId":50329,"journal":{"name":"International Journal of Developmental Biology","volume":"66 1-2-3","pages":"199-209"},"PeriodicalIF":0.7,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39438258","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Despoina Beris, Varvara Podia, Irene Dervisi, Georgios Kapolas, Ioannis Isaioglou, Vasiliki Tsamadou, Lambrini Pikoula, Mantalena Rovoli, Alexandra Vallianou, Andreas Roussis, Dimitra Milioni, Helen Giannoutsou, Kosmas Haralampidis
{"title":"RNAi silencing of the <i>Arabidopsis thaliana ULCS1</i> gene results in pleiotropic phenotypes during plant growth and development.","authors":"Despoina Beris, Varvara Podia, Irene Dervisi, Georgios Kapolas, Ioannis Isaioglou, Vasiliki Tsamadou, Lambrini Pikoula, Mantalena Rovoli, Alexandra Vallianou, Andreas Roussis, Dimitra Milioni, Helen Giannoutsou, Kosmas Haralampidis","doi":"10.1387/ijdb.210114kh","DOIUrl":"https://doi.org/10.1387/ijdb.210114kh","url":null,"abstract":"<p><p>WD40-repeat-containing proteins (WDRs) are highly abundant in all eukaryotes. Several have been implicated as subunits of multi-protein CRL E3 ligase complexes that regulate ubiquitination mediated protein degradation and thus various cellular and developmental processes. Impairment of the WDR protein ULCS1 from Arabidopsis causes pleiotropic phenotypes during plant development, including reduced lignification, anther indehiscence, and sterility. Here we show that RNAi-mediated downregulation of <i>ULCS1</i> results in a fast-growing phenotype during vegetative development. Due to accelerated growth, <i>ulcs1i</i> mutants reach their vegetative to reproductive transition point earlier than WT plants. However, their comparable germination rate and their similar number of secondary branches and rosette leaves at bolting indicate that <i>ulcs1i</i> is not an early flowering time mutant. GUS staining of progeny, obtained from crosses between <i>ulcs1i</i> and <i>CYCB1::GUS</i> plants, revealed an increased number of mitotic cell divisions in the root meristems of <i>ulcs1i</i> compared to WT. Immunolabeling of homogalacturonans (HGAs) epitopes showed significant fluorescent signal differences at the cell walls and the mucilage of the seeds between <i>ulcs1i</i> and WT. Furthermore, we demonstrate that ULCS1 interacts with the UBA-like protein in a yeast two-hybrid assay, suggesting a direct or indirect physical coupling of these proteins in Arabidopsis.</p>","PeriodicalId":50329,"journal":{"name":"International Journal of Developmental Biology","volume":"66 1-2-3","pages":"177-186"},"PeriodicalIF":0.7,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39438847","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sophia Karouzaki, Charoula Peta, Emmanouella Tsirimonaki, George Leondaritis, Kostas Vougas, George T Tsangaris, Dimitra Mangoura
{"title":"Lipid rafts integrity is essential for prolactin-induced mitogenesis in mouse embryonic stem cells.","authors":"Sophia Karouzaki, Charoula Peta, Emmanouella Tsirimonaki, George Leondaritis, Kostas Vougas, George T Tsangaris, Dimitra Mangoura","doi":"10.1387/ijdb.210194dm","DOIUrl":"https://doi.org/10.1387/ijdb.210194dm","url":null,"abstract":"<p><p>Embryonic stem cells, ESCs, retain the capacity to self-renew, yet, the protein machinery essential in maintaining this undifferentiated status remains largely undefined. Signalling interactions are initiated and enhanced at the plasma membrane lipid rafts, within constraints and regulations applied by the actin and tubulin cytoskeleton systems. First, we undertook a comprehensive approach using two-dimensional gel electrophoresis and mass spectrometry analysis combined with Western blotting and immunofluorescence analyses at the single cell level to compile the proteome profile of detergent-free preparations of lipid rafts of E14 mouse embryonic stem cells. In comparison with the proteomic profiles of other membrane fractions, recovery of actin and tubulin network proteins, including folding chaperones, was impressively high. At equally high frequency, we detected annexins, pleiotropic proteins that may bind membrane lipids and actin filaments to regulate important membrane processes, and we validated their expression in lipid rafts. Next, we tested whether lipid raft integrity is required for completion of mitogenic signalling pathways. Disruption of the rafts with the cholesterol sequestering methyl-β-cyclodextrin (MCD) greatly downregulated the mitotic index of ESCs, in a dose- and time of exposure-dependent manner. Moreover, MCD greatly reduced the mitogenic actions of prolactin, a hormone known to stimulate proliferation in a great variety of stem and progenitor cells. Taken together, our data postulate that lipid rafts in ESCs act in close association with the actin and tubulin cytoskeletons to support signal compartmentalization, especially for signalling pathways pertinent to symmetric divisions for self-renewal.</p>","PeriodicalId":50329,"journal":{"name":"International Journal of Developmental Biology","volume":"66 1-2-3","pages":"187-197"},"PeriodicalIF":0.7,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39794483","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}