The International Journal Of Developmental Biology[JOURNAL]
Sun
200 papers
The involvement of hormone-sensitive lipase in all-trans retinoic acid induced cleft palate.
Zheng K, Ye QN
Int J Dev Biol · 2022 · PMID 36688320 · Publisher ↗
Abnormally high concentrations of all-trans retinoic acid (atRA) induce cleft palate, which is accompanied by abnormal migration and proliferation of mouse embryonic palatal mesenchyme (MEPM) cells. Hormone-sensitive lip...
Abnormally high concentrations of all-trans retinoic acid (atRA) induce cleft palate, which is accompanied by abnormal migration and proliferation of mouse embryonic palatal mesenchyme (MEPM) cells. Hormone-sensitive lipase (HSL) is involved in many embryonic development processes. The current study was designed to elucidate the mechanism of HSL in cleft palate induced by atRA. To establish a cleft palate model in Kunming mice, pregnant mice were administered atRA (70 mg/kg) by gavage at embryonic Day 10.5 (E10.5). Embryonic palates were obtained through the dissection of pregnant mice at E15.5. Hematoxylin and eosin (H&E) staining was used to evaluate growth changes in the palatal shelves. The levels of HSL in MEPM cells were detected by immunohistochemistry, quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting. RNAi was applied to construct vectors expressing HSL small interference RNAs (siRNAs). The vectors were transfected into MEPM cells. Cell proliferation and migration were evaluated by the cell counting kit-8 (CCK-8) assay and wound healing assay, respectively. The palatal shelves in the atRA group had separated at E15.5 without fusing. In MEPM cells, the expression of HSL was reversed after atRA treatment, which caused cleft palate . In the atRA group, the proliferation of HSL siRNA-transfected cells was remarkably promoted, and the migration rate significantly increased in the HSL siRNA-transfected MEPM cells. These results suggested that HSL may be involved in cleft palate induced by atRA and that atRA enhances HSL levels to inhibit embryonic palate growth.
The stem cell transcription factor ZFP296 transforms NIH3T3 cells and promotes anchorage-independent growth of cancer cells.
Mizoue Y, Ikeda T, Ikegami T … +7 more , Riabets O, Oishi Y, Tobita M, Akutsu H, Hattori K, Heissig B, Koide H
Int J Dev Biol · 2023 · PMID 38334180 · Publisher ↗
Cancer cells and embryonic stem (ES) cells share several biological properties, suggesting that some genes expressed in ES cells may play an important role in cancer cell growth. In this study, we investigated the possib...
Cancer cells and embryonic stem (ES) cells share several biological properties, suggesting that some genes expressed in ES cells may play an important role in cancer cell growth. In this study, we investigated the possible role of zinc finger protein 296 (ZFP296), a transcription factor expressed in ES cells, in cancer development. First, we found that overexpression of in NIH3T3 mouse fibroblasts induced two phenomena indicative of cell transformation: enhanced proliferation under low-serum conditions and anchorage-independent growth. We also found that expression was upregulated in the tumor area of a mouse model of colon carcinogenesis. In addition, the expression levels of in various human cell lines were generally low in normal cells and relatively high in cancer cells. Finally, using a soft agar assay, we found that overexpression of promoted the anchorage-independent growth of cancer cells, while its knockdown had the opposite effect. Overall, these results suggest a possible role of the ES-specific transcription factor ZFP296 in cancer.
Chick chorioallantoic membrane: a valuable 3D model for screening nanoformulations for tumor antiangiogenic therapeutics.
Senrung A, Tripathi T, Janjua D … +8 more , Yadav SK, Chhokar A, Aggarwal N, Yadav J, Chaudhary A, Joshi U, Sethi P, Bharti AC
Int J Dev Biol · 2023 · PMID 38334179 · Publisher ↗
Drug discovery is an extensive process. From identifying lead compounds to approval for clinical application, it goes through a sequence of labor-intensive , preclinical screening and clinical trials. Among thousands of...
Drug discovery is an extensive process. From identifying lead compounds to approval for clinical application, it goes through a sequence of labor-intensive , preclinical screening and clinical trials. Among thousands of drugs screened only a few get approval for clinical trials. Furthermore, these approved drugs are often discontinued due to systemic toxicity and comorbidity at clinically administered dosages. To overcome these limitations, nanoformulations have emerged as the most sought-after strategy to safely and effectively deliver drugs within tumors at therapeutic concentrations. Most importantly, the employment of suitably variable preclinical models is considered highly critical for the therapeutic evaluation of candidate drugs or their formulations. A review of literature from the past 10 years on antiangiogenic nanoformulations shows the employment of limited types of preclinical models mainly the 2-dimensional (2D) monolayer cell culture and murine models as the mainstay for drug uptake, toxicity and efficiency studies. To top it all, murine models are highly expensive, time-consuming and require expertise in handling them. The current review highlights the utilization of the age-old chicken chorioallantoic membrane (CAM), a well-defined angiogenic model in the investigation of antiangiogenic compounds and nanoformulations in an economic framework. For practical applicability, we have evaluated the CAM model to demonstrate the screening of antiangiogenic compounds and that tumor cells can be implanted onto developing CAM for growing xenografts by recruiting host endothelial and other cellular components. In addition, the exploitation of CAM tumor xenograft models for the evaluation of nanoparticle distribution has also been reinforced by demonstrating that intravenously administered iron oxide nanoparticles (IONPs) passively accumulate and exhibit intracellular as well as extracellular compartment accumulation in highly vascular xenografts. Finally, the ethical considerations, benefits, and drawbacks, of using CAM as an experimental model for testing potential therapeutics are also highlighted.
Regeneration and proliferation of cardiomyocytes and its microRNA regulatory mechanisms.
Guo SN, Li JH, Li J … +2 more , Ding Y, Chai YR
Int J Dev Biol · 2022 · PMID 36571201 · Publisher ↗
Myocardial regeneration is identified as a concept at histological level. The core content is to increase the number of cardiomyocytes (CMs), so as to maintain the steady state of CMs under pathological or physiological...
Myocardial regeneration is identified as a concept at histological level. The core content is to increase the number of cardiomyocytes (CMs), so as to maintain the steady state of CMs under pathological or physiological conditions and ensure the normal cardiac function. In this review, we discussed the relevant factors involved in the regeneration of CMs, generalized in mice, large mammals and human. During different development stages of mammalian hearts, CMs showed several controlling and growth modes on the physiological or pathological state: mitosis, hypertrophy, nuclear polyploidy and multinucleation, amitosis and etc. We also discussed the mechanisms of specific microRNAs implicated in the cardiac development, as well as disease-induced apoptosis in CMs and the process of re-entering cell cycle after injury. It is hoped that this review will contribute to a deeper understanding of therapeutic approaches for myocardial regeneration after injury.
Cyclooxygenase-2 plays a crucial role during myocardial patterning of developing chick.
Parmar BK, Verma UR, Vaishnav JA … +1 more , Balakrishnan S
Int J Dev Biol · 2022 · PMID 36571200 · Publisher ↗
Cyclooxygenase-2 (COX-2), a member of the Cyclooxygenase family, initiates the biosynthesis of prostanoids that regulates various cellular functions. Our pilot attempt revealed that the administration of etoricoxib, an i...
Cyclooxygenase-2 (COX-2), a member of the Cyclooxygenase family, initiates the biosynthesis of prostanoids that regulates various cellular functions. Our pilot attempt revealed that the administration of etoricoxib, an inhibitor specific for COX-2, induces abnormal looping in the chicken heart. The present study attempts to reveal the mechanistic details of etoricoxib-induced abnormal cardiac looping. The activity of COX-2 was inhibited by administering 3.5 μg of etoricoxib into the egg's air cell on day zero of incubation. The gene and protein expression patterns of key mediators of heart development were then analyzed on day 2 (HH12) and day 3 (HH20). Reduced COX-2 activity altered the expressions of upstream regulators of organogenesis like Wnt11, BMP4, and SHH in the etoricoxib-exposed embryos. The observed expression shifts in the downstream regulators of myocardial patterning (MYOCD, HAND2, GATA4, GATA5, and GATA6) in the treated embryos corroborate the above results. In addition, the reduction in COX-2 activity hampered cardiomyocyte proliferation with a concomitant increase in the apoptosis rate. In conclusion, the collective effect of altered expression of signaling molecules of myocardial patterning and compromised cardiomyocyte turnover rate could be the reason behind the looping defects observed in the heart of etoricoxib-treated chick embryos.
Intraflagellar transport 20 cilia-dependent and cilia-independent signaling pathways in cell development and tissue homeostasis.
Jin FC, Zhou MH, Chen JJ … +5 more , Lin Y, Zhang QW, Xu QX, Zhang CC, Zhang ZG
Int J Dev Biol · 2022 · PMID 35980193 · Publisher ↗
Intraflagellar transport (IFT) is an essential condition for ciliogenesis. The primary cilia protrude like antennae and act as chemical or mechanical sensory organelles that coordinate specific receptor localization and...
Intraflagellar transport (IFT) is an essential condition for ciliogenesis. The primary cilia protrude like antennae and act as chemical or mechanical sensory organelles that coordinate specific receptor localization and signal transduction. IFT20 is the smallest molecule in IFT complex B, which is located in both the cilia and the Golgi complex. Recent studies have shown that IFT20 is a key molecule in multiple signaling pathways. Importantly, in the function of IFT20, signal transduction is not restricted to cilia, but is also involved in non-ciliary functions. Here we summarize current knowledge regarding IFT20-mediated signaling pathways and their relationship with cell development and tissue homeostasis, and analyse the cilia-dependent and cilia-independent mechanisms of IFT20 coordinated signaling pathways and potential crosstalk between the mechanisms. This review provides a comprehensive perspective on IFT20 coordinates signaling mechanisms in cell development and tissue homeostasis.
Exploring the sister cells of embryo sac: developmental and functional attributes.
Kaur I, Koul M
Int J Dev Biol · 2022 · PMID 35980192 · Publisher ↗
Synergids are metabolically dynamic cells of the egg apparatus and represent an important component of the female gametophyte. Besides directing the growth of the pollen tube towards the micropylar end of the embryo sac,...
Synergids are metabolically dynamic cells of the egg apparatus and represent an important component of the female gametophyte. Besides directing the growth of the pollen tube towards the micropylar end of the embryo sac, these ephemeral structures make room for the pollen tube cytoplasm. The nature of chemotrophic substances that direct the growth of the pollen tube, the mechanism of degeneration of one of the synergids before fertilization and the molecular aspects of synergid morphogenesis have been studied in detail. Research carried out on model systems such as and has expanded our understanding of the molecular regulation of the pollen tube journey, its guidance and navigation in the pistil. Recently, the critical role of the central cell in fertilization and prevention of polytubey has also been thoroughly investigated. Interesting aspects that lead to degeneration of synergids, and the factors governing degeneration, including molecular aspects, have produced a paradigm shift in the understanding of these intriguing units. Sophisticated confocal microscopy, live cell imaging, and molecular tools have helped in furthering our knowledge of the functioning of synergids. Recent research using high throughput techniques has deciphered the role of various genes that regulate and govern the release of chemotropic substances, cell-to-cell interaction and synergid cell degeneration. Moreover, with the diversity displayed in form and function of organs in the angiosperms, and the switching of roles of the cells of egg apparatus, new insights have been provided into the involvement of synergids both pre- and post-fertilization. The present review provides a comprehensive account of synergids, their role in fertilization and the post fertilization events that have emerged using interdisciplinary approaches in recent years. We also discuss the variations observed in degeneration of synergids and the mechanisms that have been unraveled recently. Study of the dynamism exhibited by synergids reveals newer roles of these in fertilization. How synergids in angiosperm taxa where genetic transformation/alteration is carried out will respond to pollen stimuli is still unknown. Since environmental factors such as light and temperature have a significant impact on synergids and fertilization, it would be rewarding to study the role of chemo-attractants and other factors in elucidating the functional roles of synergids. Further research into developing adequate protocols for manipulating synergid functions is certainly required. This research has enormous potential in the advancement of basic science and has potential applications in agriculture, horticulture, and bioprospecting.
The mesenchymal potential of trunk neural crest cells.
Rodrigues-Da-Silva MA, de Espindola da Silveira G, Taufer CR … +1 more , Calloni GW
Int J Dev Biol · 2022 · PMID 35833524 · Publisher ↗
It has long been held that the main difference between cranial and trunk neural crest (CNC and TNC, respectively) was the potential of CNC to originate mesenchymal cell types, especially skeletogenic. This is an age-old...
It has long been held that the main difference between cranial and trunk neural crest (CNC and TNC, respectively) was the potential of CNC to originate mesenchymal cell types, especially skeletogenic. This is an age-old question that continues to challenge researchers, even today. Unfortunately, to date, no consensus has concluded the extent of TNC mesenchymal potential, nor has a systematic review been conducted to organize current knowledge about this fascinating question. However, the number of studies related to this question have expanded and deepened considerably in the last few years thanks to several new different species of vertebrates employed, the generation of transgenic animal strains, the combination of cell markers, and also the improvement of cell culture conditions through the use of different substrates and signaling molecules. Therefore, this review summarizes the literature showing that TNCCs can generate a broad range of mesenchymal cell types, including skeletogenic. This potential can be unveiled by certain favorable conditions, but it also seems to be expressed in some animal structures , to which TNCCs contribute. We also present several works that offer a contrary view and do not detect any mesenchymal/skeletogenic contribution of TNCCs . Perhaps, it is the controversy itself that makes this subject even more exciting.
Correlation between CDK1 protein and CDK1 mRNA during oocyte maturation in mouse.
Sun YT, Zhu AZ
Int J Dev Biol · 2022 · PMID 35770734 · Publisher ↗
The aim of this study was to investigate the correlation between CDK1 protein and CDK1 mRNA during oocyte maturation in mouse. GV, GVBD, MI and MII oocytes were obtained from mice, respectively. Western blot validated t...
The aim of this study was to investigate the correlation between CDK1 protein and CDK1 mRNA during oocyte maturation in mouse. GV, GVBD, MI and MII oocytes were obtained from mice, respectively. Western blot validated that the CDK1 protein expression increased continuously and significantly with oocyte maturation (P<0.05). Real-time qRT-PCR showed that CDK1 mRNA expression was down-regulated significantly during transformation from GV to MI stages (P<0.05), and up-regulated significantly during transformation from MI to MII stages (P<0.05). The level of CDK1 mRNA peaked at MII stages. Spearman correlation analysis indicated that CDK1 protein expression was poor correlation with CDK1 mRNA expression during oocyte maturation (R=0.200). This finding suggested that the increase of CDK1 protein during oocyte maturation was not entirely caused by the change of transcription level. The results provide new food for thought for further research on the molecular mechanism of oocyte maturation .
Aristotle (384-322 BC): the beginnings of Embryology.
Zagris N
Int J Dev Biol · 2022 · PMID 35583074 · Publisher ↗
Aristotle made important contributions to many fields-biology, physics, metaphysics, logic, ethics, rhetoric, psychology, aesthetics, poetry- that are now cultivated by specialized experts, but he never lost sight of the...
Aristotle made important contributions to many fields-biology, physics, metaphysics, logic, ethics, rhetoric, psychology, aesthetics, poetry- that are now cultivated by specialized experts, but he never lost sight of the aim of unifying knowledge, of understanding the world as an organized whole. Aristotle was the first to combine wet, field biology with daring cosmological thinking. He is the father of natural history and the first embryologist known to history. Aristotle's classic treatises , and "enjoyed for more than fifteen hundred years an authority altogether without parallel". Over the last four decades, the introduction of molecular techniques has gradually overturned the entire structure of the biological sciences. Biology, initially a science of inventory and classification in the hands of the 19-century comparative naturalists, has become a science of codes and regulatory circuits. Aristotle was the first to codify laws of pure logic, and so he founded what is today known as ' ' in mathematics. Aristotle was an inveterate collector and a classifier, the master scientist of his time. His main concern was to classify "the ultimate furniture of the world", under basic headings and categories, a powerful human strategy to organize knowledge for comprehension and action. This was part of Aristotle's attempt to create a theory of reality, one strongly opposed to Plato's otherworldly doctrine of the ideal ''. To many generations of thinkers in the great era of Scholastic philosophy, Aristotle was known simply as "The Philosopher".
Developmental Biology in Greece.
Zagris N
Int J Dev Biol · 2022 · PMID 35583073 · Publisher ↗
Abstract loading — click title to view on PubMed.
Hyaluronan receptor CD44: developmentally regulated expression and role in the early chick embryo.
Konstantopoulos K, Dimiropoulos A, Zagris N
Int J Dev Biol · 2022 · PMID 35238395 · Publisher ↗
CD44 is a membrane glycoprotein and is the main receptor for hyaluronan. We studied CD44 expression and spatio-temporal distribution by RT-PCR and immunofluorescence, and used an anti-CD44 blocking antibody to perturb CD...
CD44 is a membrane glycoprotein and is the main receptor for hyaluronan. We studied CD44 expression and spatio-temporal distribution by RT-PCR and immunofluorescence, and used an anti-CD44 blocking antibody to perturb CD44-depended signalling programs in the early chick embryo. The intense CD44 levels we detected in the morula embryo (XI) were of particular interest, suggestive of a maternally stored transcript. Intriguingly, the early presence of CD44 seemed to be essential for the rapid synthesis of hyaluronan. At stage XIII (blastula), CD44 expression was intense in the epiblast and hypoblast. During gastrulation (HH3-4), the cells ingressing into the primitive groove and migrating, and the blood islands, expressed CD44 intensely. At HH8, the folding neural plate showed polarity regulation of CD44 expression, and expression was also intense in neural crest, notochord, and blood islands. During early organogenesis, CD44 was expressed intensely in the developing cranial and caudal neural tube that showed polarity regulation, in optic stalks, otic vesicles, pre-and migratory neural crest cells, ganglia, notochord, pharynx, gut, liver, aortae, heart, somites, vascular area, amnion and chorion, and was distinct in extracellular matrix of cranial neural tube and otic vesicle lumens. Antibody-mediated perturbation of CD44 function resulted in unorganized extracellular matrix, loss of tissue spaces, grossly abnormal notochord, intermingling of clumped neuroectoderm and mesenchyme, absence of somites and blood vessels and inhibition of neural crest cell emigration. CD44 has various pivotal roles in matrix integrity and tissue patterning, consistent with its known biochemical features and interactions with hyaluronan, growth factors, receptors and other signaling molecules.
Epithelial-endothelial transition and endothelial-mesenchymal transition.
Ribatti D
Int J Dev Biol · 2022 · PMID 35238394 · Publisher ↗
The movement of continuous sheets of epithelial cells occurs during embryonic development, tissue repair, and cancer. Common to cellular and molecular principles of collective cell migration, invading cancers seem to rea...
The movement of continuous sheets of epithelial cells occurs during embryonic development, tissue repair, and cancer. Common to cellular and molecular principles of collective cell migration, invading cancers seem to reactivate embryonic pathways and patterns of cell movement. Epithelial cells possess the capability to become mesenchymal cells in a process called epithelial mesenchymal transition (EMT), which has been extensively studied and described. The aim of this article is to summarize the most recent literature data concerning less known epithelial-endothelial transition and endothelial-mesenchymal transition.
Pleiotrophin, nitric oxide and glutamate AMPA receptors in chick cerebellum morphogenesis.
Kommata V, Alexopoulou E, Argyrousi EK … +1 more , Dermon CR
Int J Dev Biol · 2022 · PMID 35238393 · Publisher ↗
Avian cerebellum, a highly conserved, laminated and foliated structure, provides an excellent model for developmental studies. During the intermediate embryonic stages, granule cell progenitor proliferation and the inwar...
Avian cerebellum, a highly conserved, laminated and foliated structure, provides an excellent model for developmental studies. During the intermediate embryonic stages, granule cell progenitor proliferation and the inwards migration of post-mitotic granule cells have been implicated in the morphogenesis of cerebellar cortex cytoarchitecture and foliation. The present study questioned the spatio-temporal expression pattern of pleiotrophin, an extracellular matrix growth factor, during the morphogenesis of embryonic cerebellum, and the roles of ionotropic AMPA glutamate receptors and the diffusible neuromodulator nitric oxide (NO) in the proliferation pattern of EGL granule cell progenitors. To this end, the density of proliferating cells in the developing embryonic external granule layer (EGL) was determined following acute treatment with AMPA receptor antagonist CNQX or NO synthase inhibitor L-NAME, at embryonic stages HH38-41 (E12-E15 days), by means of BrdU immunohistochemistry and double immunofluorescence. Importantly, at earlier stages, pleiotrophin-like immunoreactivity showed high expression levels in the EGL that gradually decreased, persisting within the growing folia apices, later in development. Interestingly, blockage of AMPA receptors had no effect; while NOS inhibition resulted in transient age- and region-specific increases of EGL granule progenitor cell proliferation at earlier stages, but decreased the post-mitotic granule cells at folia apices, at a later stage HH41 (E15 day). Overall, NO had a transient anti-proliferative effect in EGL similar to mammalian cerebellum, acting as a modulator of the EGL function at different stages, suggesting its possible implication in complex processes guiding cerebellar cytoarchitecture and folia formation.
Heterogeneity of quiescent and active neural stem cells in the postnatal brain.
Dimitrakopoulos D, Kakogiannis D, Kazanis I
Int J Dev Biol · 2022 · PMID 35238392 · Publisher ↗
In the postnatal mammalian brain, neurogenic activity is retained in anatomically restricted areas, driven by pools of Neural Stem Cells (NSCs). These cells and their progeny have been studied intensively as potential ta...
In the postnatal mammalian brain, neurogenic activity is retained in anatomically restricted areas, driven by pools of Neural Stem Cells (NSCs). These cells and their progeny have been studied intensively as potential targets for regenerative treatments, aiming at either their manipulation or their use as sources of cells for transplantation-based strategies. Although their full identity, heterogeneity and differentiation potential remain elusive, due to the absence of specific cell-type markers, our knowledge of their properties is constantly expanding. Here, we focus on the NSC niche that is located at the Subependymal Zone (SEZ/ also known as Subventricular Zone) of the lateral ventricles of the brain. We review, summarize and explain the different faces of the NSC, as they have been described, using a wide range of experimental approaches, over a time-frame of three decades: the primitive, definitive, quiescent or activated NSC. We also review the growing evidence of the existence of latent NSCs outside of niches, in the brain parenchyma, that constitute promising new therapeutic targets, complemented by the novel technologies of cell reprogramming.
manipulation of Nile crocodile embryos: egg windowing and potential dental research applications.
Truter JC, Myburgh JG
Int J Dev Biol · 2021 · PMID 35112710 · Publisher ↗
Crocodilians exhibit continuous tooth replacement (i.e., polyphyodonty) and have been identified as suitable models for tooth regeneration research due to the similarity in dental cavity and tooth anatomy between these c...
Crocodilians exhibit continuous tooth replacement (i.e., polyphyodonty) and have been identified as suitable models for tooth regeneration research due to the similarity in dental cavity and tooth anatomy between these creatures and humans. Various studies reporting bird embryo manipulation exist, but such reports for reptiles are virtually non-existent. Egg windowing enables direct access to oviparous vertebrate embryos and is therefore an important component of embryo manipulation experiments. The aim of the present study was to window Nile crocodile, eggs and assess the potential of direct manipulations, targeting the pharyngeal region where the maxilla and mandible originates. eggs were successfully windowed, and a limited number of individuals survived the entire gestation period. The 1st trimester of gestation was the most sensitive, and 96.78% of the mortalities occurred within this period. Our data indicate the suitable window for embryo manipulation targeting the mandibular arch and maxillary process, without a risk of damaging the chorioallantoic membrane (CAM) (which may be fatal), was between day six and eight after laying for embryos incubated at 31°C. This data will be of use for future embryo-based experiments related to jaw and tooth development in crocodiles as well as human tooth regeneration research.
Human C2a and C6a iPSC lines and H9 ESC line have less efficient cardiomyogenesis than H1 ESC line: Beating enhances cardiac differentiation.
Sepac A, Bosnjak ZJ, Seiwerth S … +5 more , Sikiric S, Dzombeta TR, Kulic A, Karsaj JM, Sedlic F
Int J Dev Biol · 2021 · PMID 35112709 · Publisher ↗
BACKGROUND: Human induced pluripotent stem cells (hiPSCs) need to be thoroughly characterized to exploit their potential advantages in various aspects of biomedicine. The aim of this study was to compare the efficiency o...
BACKGROUND: Human induced pluripotent stem cells (hiPSCs) need to be thoroughly characterized to exploit their potential advantages in various aspects of biomedicine. The aim of this study was to compare the efficiency of cardiomyogenesis of two hiPSCs and two human embryonic stem cell (hESC) lines by genetic living cardiomyocyte labeling. We also analyzed the influence of spontaneous beating on cardiac differentiation. METHODS: H1 and H9 hESC lines and C2a and C6a hiPSC lines were induced into directed cardiac differentiation. Cardiomyogenesis was evaluated by the analysis of cell cluster beating, cardiac protein expression by immunocytochemistry, ability of cells to generate calcium transients, and cardiomyocyte quantification by the myosin light chain 2v-enhanced green fluorescent protein gene construct delivered with a lentiviral vector. RESULTS: Differentiation of all cell lines yielded spontaneously beating cell clusters, indicating the presence of functional cardiomyocytes. After the cell dissociation, H1-hESC-derived cardiomyocytes exhibited spontaneous calcium transients, corresponding to autonomous electrical activity and displayed ability to transmit them between the cells. Differentiated hESC and hiPSC cells exhibited striated sarcomeres and expressed cardiac proteins sarcomeric α-actinin and cardiac troponin T. Cardiomyocytes were the most abundant in differentiated H1 hESC line (20% more than in other tested lines). In all stem cell lines, cardiomyocyte enrichment was greater in beating than in non-beating cell clusters, irrespective of cardiomyogenesis efficiency. CONCLUSION: Although C2a and C6a hiPSC and H9 hESC lines exhibited efficient cardiomyogenesis, H1 hESC line yielded the greatest cardiomyocyte enrichment of all tested lines. Beating of cell clusters promotes cardiomyogenesis in tested hESCs and hiPSCs.
Single-cell transcriptomics defines Dot1L interacting partners and downstream target genes in the mouse molar dental pulp.
Guzzo R, Enkhmandakh B, Becker T … +6 more , Joshi P, Robson P, Vijaykumar A, Mina M, Shin DG, Bayarsaihan D
Int J Dev Biol · 2022 · PMID 36942693 · Publisher ↗
Although histone methyltransferases are implicated in many key developmental processes, the contribution of individual chromatin modifiers in dental tissues is not well understood. Using single-cell RNA sequencing, we ex...
Although histone methyltransferases are implicated in many key developmental processes, the contribution of individual chromatin modifiers in dental tissues is not well understood. Using single-cell RNA sequencing, we examined the expression profiles of the disruptor of telomeric silencing 1-like ( gene in the postnatal day 5 mouse molar dental pulp. Dot1L is the only known enzyme that methylates histone 3 on lysine 79, a modification associated with gene expression. Our research revealed 15 distinct clusters representing different populations of mesenchymal stromal cells (MSCs), immune cells, pericytes, ameloblasts and endothelial cells. We documented heterogeneity in gene expression across different subpopulations of MSCs, a good indicator that these stromal progenitors undergo different phases of osteogenic differentiation. Interestingly, although was broadly expressed across all cell clusters within the molar dental pulp, our analyses indicated specific enrichment of within two clusters of MSCs, as well as cell clusters characterized as ameloblasts and endothelial cells. Moreover, we detected co-expression with protein interactors involved in epigenetic activation such as and . In addition, was co-expressed with , and , which encode epigenetic factors associated with gene silencing and heterochromatin formation. was co-expressed with downstream targets of the insulin growth factor and WNT signaling pathways, as well as genes involved in cell cycle progression. Collectively, our results suggest that Dot1L may play key roles in orchestrating lineage-specific gene expression during MSC differentiation.
Expression of two uncharacterized protein coding genes in zebrafish lateral line system.
Fatma S, Kumar R, Dixit A … +1 more , Swain RK
Int J Dev Biol · 2021 · PMID 34881803 · Publisher ↗
The lateral line system is a mechanosensory organ of fish and amphibians that detects changes in water flow and is formed by the coordinated action of many signalling pathways. These signalling pathways can easily be tar...
The lateral line system is a mechanosensory organ of fish and amphibians that detects changes in water flow and is formed by the coordinated action of many signalling pathways. These signalling pathways can easily be targeted in zebrafish using pharmacological inhibitors to decipher their role in lateral line system development at cellular and molecular level. We have identified two uncharacterized proteins, whose mRNA are expressed in the lateral line system of zebrafish. One of these proteins, uncharacterized protein LOC564095 precursor, is conserved across vertebrates and its mRNA is expressed in posterior lateral line primordium (pLLP). The other uncharacterized protein, LOC100536887, is present only in the teleost fishes and its mRNA is expressed in neuromasts. We show that inhibition of retinoic acid (RA) signalling reduces the expression of both of these uncharacterized genes. It is reported that inhibition of RA signalling during gastrulation starting at 7 hours post fertilization (hpf) abrogates pLLP formation, and inhibition of RA signalling at 10 hpf delays the initiation of pLLP migration. Here, we show that inhibition of RA signalling before and during segmentation (9-16 hpf) results in delayed initiation and reduced speed of pLLP migration, as well as inhibition of posterior neuromasts formation.
SV40T reprograms Schwann cells into stem-like cells that can re-differentiate into terminal nerve cells.
Li RF, Nan GX, Wang D … +4 more , Gao C, Yang J, He TC, Zhang ZL
Int J Dev Biol · 2021 · PMID 34881802 · Publisher ↗
BACKGROUND: The specific effect of SV40T on neurocytes has seldom been investigated by the researchers. We transfected Schwann cells (SCs) that did not have differentiation ability with MPH 86 plasmid containing SV40T, i...
BACKGROUND: The specific effect of SV40T on neurocytes has seldom been investigated by the researchers. We transfected Schwann cells (SCs) that did not have differentiation ability with MPH 86 plasmid containing SV40T, in order to explore the effects of SV40T on Schwann cells. METHODS: SCs were transfected with MPH 86 plasmid carrying the SV40T gene and cultured in different media, and also co-cultured with neural stem cells (NSCs). In our study, SCs overexpressing SV40T were defined as SV40T-SCs. The proliferation of these cells was detected by WST-1, and the expression of different biomarkers was analyzed by qPCR and immunohistochemistry. RESULTS: SV40T induced the characteristics of NSCs, such as the ability to grow in suspension, form spheroid colonies and proliferate rapidly, in the SCs, which were reversed by knocking out SV40T by the Flip-adenovirus. In addition, SV40T up-regulated the expressions of neural crest-associated markers Nestin, Pax3 and Slug, and down-regulated S100b as well as the markers of mature SCs MBP, GFAP and Olig1/2. These cells also expressed NSC markers like Nestin, Sox2, CD133 and SSEA-1, as well as early development markers of embryonic stem cells (ESCs) like BMP4, c-Myc, OCT4 and Gbx2. Co-culturing with NSCs induced differentiation of the SV40T-SCs into neuronal and glial cells. CONCLUSIONS: SV40T reprograms Schwann cells to stem-like cells at the stage of neural crest cells (NCCs) that can differentiate to neurocytes.
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