Journal of Developmental Biology最新文献

{"title":"CRISPR/Cas9-Targeted <i>Myostatin</i> Deletion Improves the Myogenic Differentiation Parameters for Muscle-Derived Stem Cells in Mice.","authors":"Mohamed I Elashry, Victoria C Schneider, Manuela Heimann, Sabine Wenisch, Stefan Arnhold","doi":"10.3390/jdb13010005","DOIUrl":"10.3390/jdb13010005","url":null,"abstract":"<p><p>Skeletal muscle plays a pivotal role in physical activity, protein storage and energy utilization. Skeletal muscle wasting due to immobilization, aging, muscular dystrophy and cancer cachexia has negative impacts on the quality of life. The deletion of myostatin, a growth and differentiation factor-8 (GDF-8) augments muscle mass through hyperplasia and hypertrophy of muscle fibers. The present study examines the impact of myostatin deletion using CRISPR/Cas9 editing on the myogenic differentiation (MD) of C2C12 muscle stem cells. A total of five myostatin loci were targeted using guided RNAs that had been previously cloned into a vector. The clones were transfected in C2C12 cells via electroporation. The cell viability and MD of myostatin-edited clones (Mstn^-/-) were compared with C2C12 (Mstn^+/+) using a series of assays, including MTT, sulforhodamine B, immunocytochemistry, morphometric analysis and RT-qPCR. The clones sequenced showed evidence of nucleotides deletion in Mstn^-/- cells. Mstn^-/- cells demonstrated a normal physiological performance and lack of cytotoxicity. Myostatin depletion promoted the myogenic commitment as evidenced by upregulated MyoD and myogenin expression. The number of MyoD-positive cells was increased in the differentiated Mstn^-/- clones. The Mstn^-/- editing upregulates both mTOR and MyH expression, as well as increasing the size of myotubes. The differentiation of Mstn^-/- cells upregulates ActRIIb; in contrast, it downregulates decorin expression. The data provide evidence of successful CRISPR/Cas9-mediated myostatin deletion. In addition, targeting myostatin could be a beneficial therapeutic strategy to promote MD and to restore muscle loss. In conclusion, the data suggest that myostatin editing using CRISPR/Cas9 could be a potential therapeutic manipulation to improve the regenerative capacity of muscle stem cells before in vivo application.</p>","PeriodicalId":15563,"journal":{"name":"Journal of Developmental Biology","volume":"13 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11843916/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143468228","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Utilizing <i>C. elegans</i> Spermatogenesis and Fertilization Mutants as a Model for Human Disease.","authors":"Sofia M Perez, Helena S Augustineli, Matthew R Marcello","doi":"10.3390/jdb13010004","DOIUrl":"10.3390/jdb13010004","url":null,"abstract":"<p><p>The nematode <i>C. elegans</i> is a proven model for identifying genes involved in human disease, and the study of <i>C. elegans</i> reproduction, specifically spermatogenesis and fertilization, has led to significant contributions to our understanding of cellular function. Approximately 70 genes have been identified in <i>C. elegans</i> that control spermatogenesis and fertilization (<i>spe</i> and <i>fer</i> mutants). This review focuses on eight genes that have human orthologs with known pathogenic phenotypes. Using <i>C. elegans</i> to study these genes has led to critical developments in our understanding of protein domain function and human disease, including understanding the role of <i>OTOF</i> (the ortholog of <i>C. elegans fer-1</i>) in hearing loss, the contribution of the <i>spe-39</i> ortholog <i>VIPAS39</i> in vacuolar protein sorting, and the overlapping functions of <i>spe-26</i> and <i>KLHL10</i> in spermatogenesis. We discuss the cellular function of both the <i>C. elegans</i> genes and their human orthologs and the impact that <i>C. elegans</i> mutants and human variants have on cellular function and physiology. Utilizing <i>C. elegans</i> to understand the function of the genes reviewed here, and additional understudied and undiscovered genes, represents a unique opportunity to understand the function of variants that could lead to better disease diagnosis and clinical decision making.</p>","PeriodicalId":15563,"journal":{"name":"Journal of Developmental Biology","volume":"13 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11843878/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143468271","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Neuronal Populations Involved in Motor Function Show Prominent Expression of Sbno1 During Postnatal Brain Development.","authors":"Sunjidmaa Zolzaya, Dai Ihara, Munkhsoyol Erkhembaatar, Shinsuke Ochiai, Ayaka Isa, Mariko Nishibe, Jean-Pierre Bellier, Takahiro Shimizu, Satoshi Kikkawa, Ryo Nitta, Yu Katsuyama","doi":"10.3390/jdb13010003","DOIUrl":"10.3390/jdb13010003","url":null,"abstract":"<p><p>Human genome studies have suggested that strawberry notch homologue 1 (<i>SBNO1</i>) is crucial for normal brain development, with mutations potentially contributing to neurodevelopmental disorders. In a previous study, we observed significant developmental abnormalities in the neocortex of <i>Sbno1</i> as early as one week after birth. In the present study, we conducted an extensive analysis of Sbno1 postnatal expression in the brain of C57BL/6 mice using a newly developed in-house polyclonal antibody against Sbno1. We found that Sbno1 is expressed in all neurons, with certain neuronal populations exhibiting distinct dynamic changes (both temporal and spatial) in expression level. These findings suggest that the neuronal expression of Sbno1 is developmentally regulated after birth. They also indicate that while Sbno1 may play a general role across all neurons, it may also serve more specialized functions in certain neuronal types and/or for certain cellular activities related to particular neuronal pathways.</p>","PeriodicalId":15563,"journal":{"name":"Journal of Developmental Biology","volume":"13 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11843823/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143468270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Regeneration, Regengrow and Tissue Repair in Animals: Evolution Indicates That No Regeneration Occurs in Terrestrial Environments but Only Recovery Healing.","authors":"Lorenzo Alibardi","doi":"10.3390/jdb13010002","DOIUrl":"10.3390/jdb13010002","url":null,"abstract":"<p><p>The present, brief review paper summarizes previous studies on a new interpretation of the presence and absence of regeneration in invertebrates and vertebrates. Broad regeneration is considered exclusive of aquatic or amphibious animals with larval stages and metamorphosis, where also a patterning process is activated for whole-body regeneration or for epimorphosis. In contrast, terrestrial invertebrates and vertebrates can only repair injury or the loss of body parts through a variable \"recovery healing\" of tissues, regengrow or scarring. This loss of regeneration likely derives from the change in genomes during land adaptation, which included the elimination of larval stages and intense metamorphosis. The terrestrial conditions are incompatible with the formation of embryonic organs that are necessary for broad regeneration. In fact, no embryonic organ can survive desiccation, intense UV or ROS exposition on land, and rapid reparative processes without embryonic patterning, such as recovery healing and scarring, have replaced broad regeneration in terrestrial species. The loss of regeneration in land animals likely depends on the alteration of developmental gene pathways sustaining regeneration that occurred in progenitor marine animals. Terrestrial larval stages, like those present in insects among arthropods, only metamorphose using small body regions indicated as imaginal disks, a terrestrial adaptation, not from a large restructuring process like in aquatic-related animals. These invertebrates can reform body appendages only during molting, a process indicated as regengrow, not regeneration. Most amniotes only repair injuries through scarring or a variable recovery healing, occasionally through regengrow, the contemporaneous healing in conjunction with somatic growth, forming sometimes new heteromorphic organs.</p>","PeriodicalId":15563,"journal":{"name":"Journal of Developmental Biology","volume":"13 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11755470/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143023532","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mesenchymal Traits as an Intrinsic Feature of Undifferentiated Cells.","authors":"Mirco Galiè","doi":"10.3390/jdb13010001","DOIUrl":"10.3390/jdb13010001","url":null,"abstract":"<p><p>Since its first conceptualization over a century ago, the mesenchymal phenotype has traditionally been viewed as either a transient phase between successive epithelial stages or as a feature of cell types primarily devoted to structural support. However, recent findings in cancer research challenge this limited view, demonstrating that mesenchymal traits and hybrid mesenchymal/epithelial states can mark cancer cells with stem cell properties. By analyzing publicly available single-cell transcriptome datasets from early embryonic stages and adult tissues, this study aims to extend this concept beyond pathological contexts, suggesting that a partial or fully mesenchymal phenotype may represent the morphological expression of undifferentiated and multipotent states in both the developing embryo and adult organs.</p>","PeriodicalId":15563,"journal":{"name":"Journal of Developmental Biology","volume":"13 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11755434/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143023529","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genetics and Genomics of Gastroschisis, Elucidating a Potential Genetic Etiology for the Most Common Abdominal Defect: A Systematic Review.","authors":"John P Marquart, Qian Nie, Tessa Gonzalez, Angie C Jelin, Ulrich Broeckel, Amy J Wagner, Honey V Reddi","doi":"10.3390/jdb12040034","DOIUrl":"10.3390/jdb12040034","url":null,"abstract":"<p><p>(1) Background: The exact etiology for gastroschisis, the most common abdominal defect, is yet to be known, despite the rising prevalence of this condition. The leading theory suggests an increased familial risk, indicating a possible genetic component possibly in the context of environmental risk factors. This systematic review aims to summarize the studies focused on the identification of a potential genetic etiology for gastroschisis to elucidate the status of the field. (2) Methods: Following the PRISMA-ScR method, Pubmed and Google Scholar were searched, and eligible publications were mined for key data fields such as study aims, cohort demographics, technologies used, and outcomes in terms of genes identified. Data from 14 human studies, with varied cohort sizes from 40 to 1966 individuals for patient vs. healthy controls, respectively, were mined to delineate the technologies evaluated. (3) Results: Our results continue the theory that gastroschisis is likely caused by gene-environment interactions. The 14 studies utilized traditional methodologies that may not be adequate to identify genetic involvement in gastroschisis. (4) Conclusions: The etiology of gastroschisis continues to remain elusive. A combination of omics and epigenetic evaluation studies would help delineate a possible genetic etiology for gastroschisis.</p>","PeriodicalId":15563,"journal":{"name":"Journal of Developmental Biology","volume":"12 4","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11727865/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142894984","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comprehensive Predictions of Mef2-Mediated Chromatin Loops, Which May Inhibit Ubx Binding by Blocking Low-Affinity Binding Sites.","authors":"Katrin Domsch","doi":"10.3390/jdb12040033","DOIUrl":"10.3390/jdb12040033","url":null,"abstract":"<p><p>Gene regulation depends on the interaction between chromatin-associated factors, such as transcription factors (TFs), which promote chromatin loops to ensure tight contact between enhancer and promoter regions. So far, positive interactions that lead to gene activation have been the main focus of research, but regulations related to blocking or inhibiting factor binding are also essential to maintaining a defined cellular status. To understand these interactions in greater detail, I investigated the possibility of the muscle differentiation factor Mef2 to prevent early Hox factor binding, leading to the proper timing of regulatory processes and the activation of differentiation events. My investigations relied on a collection of publicly available genome-wide binding data sets of Mef2 and Ubx (as the Hox factor), Capture-C interactions, and ATAC-seq analysis in <i>Mef2</i> mutant cells. The analysis indicated that Mef2 can form possible chromatin loops to Ubx-bound regions. These regions contain low-affinity Ubx binding sites, and the chromatin architecture is independent of Mef2's function. High levels of Ubx may disrupt the loops and allow specific Ubx bindings to regulate defined targets. In summary, my investigations highlight that the use of many publicly available data sets enables computational approaches to make robust predictions and, for the first time, suggest a molecular function of Mef2 as a preventer of Hox binding, indicating that it may act as a timer for muscle differentiation.</p>","PeriodicalId":15563,"journal":{"name":"Journal of Developmental Biology","volume":"12 4","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11678716/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142894982","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Loss of Tafazzin Transacetylase Activity Is Sufficient to Drive Testicular Infertility.","authors":"Paige L Snider, Elizabeth A Sierra Potchanant, Catalina Matias, Donna M Edwards, Jeffrey J Brault, Simon J Conway","doi":"10.3390/jdb12040032","DOIUrl":"10.3390/jdb12040032","url":null,"abstract":"<p><p>Barth syndrome (BTHS) is a rare, infantile-onset, X-linked mitochondriopathy exhibiting a variable presentation of failure to thrive, growth insufficiency, skeletal myopathy, neutropenia, and heart anomalies due to mitochondrial dysfunction secondary to inherited TAFAZZIN transacetylase mutations. Although not reported in BTHS patients, male infertility is observed in several <i>Tafazzin</i> (<i>Taz</i>) mouse alleles and in a <i>Drosophila</i> mutant. Herein, we examined the male infertility phenotype in a BTHS-patient-derived <i>D75H</i> point-mutant knockin mouse (<i>Taz^PM</i>) allele that expresses a mutant protein lacking transacetylase activity. Neonatal and adult <i>Taz^PM</i> testes were hypoplastic, and their epididymis lacked sperm. Histology and biomarker analysis revealed <i>Taz^PM</i> spermatogenesis is arrested prior to sexual maturation due to an inability to undergo meiosis and the generation of haploid spermatids. Moreover, <i>Taz^PM</i> testicular mitochondria were found to be structurally abnormal, and there was an elevation of p53-dependent apoptosis within <i>Taz^PM</i> seminiferous tubules. Immunoblot analysis revealed that <i>Taz^PM</i> gamete genome integrity was compromised, and both histone γ-H2Ax and Nucleoside diphosphate kinase-5 protein expression were absent in juvenile <i>Taz^PM</i> testes when compared to controls. We demonstrate that Taz-mediated transacetylase activity is required within mitochondria for normal spermatogenesis, and its absence results in meiotic arrest. We hypothesize that elevated <i>Taz^PM</i> spermatogonial apoptosis causes azoospermia and complete infertility.</p>","PeriodicalId":15563,"journal":{"name":"Journal of Developmental Biology","volume":"12 4","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11677720/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142894985","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcriptomic Evidence for Cell-Autonomous Sex Differentiation of the Gynandromorphic Fat Body in the Silkworm, <i>Bombyx mori</i>.","authors":"Fumiko Yamamoto, Takeshi Yokoyama, Yan Su, Masataka G Suzuki","doi":"10.3390/jdb12040031","DOIUrl":"10.3390/jdb12040031","url":null,"abstract":"<p><p>The classic model of sex determination in insects suggests that they do not have sex hormones and that sex is determined in a cell-autonomous manner. On the other hand, there is accumulating evidence that the development of secondary sexual traits is controlled in a non-cell-autonomous manner through external factors. To evaluate the degrees of the cell-autonomous and non-cell-autonomous regulation of secondary sexual trait development, we analyzed the dynamics of the sexually dimorphic transcriptome in gynandromorphic individuals of the <i>mo</i> mutant strain in the silkworm <i>Bombyx mori</i>. The silkworm possesses a female heterogametic sex-determination system (ZZ = male/ZW = female), where the master regulatory gene for femaleness, <i>Feminizer</i> (<i>Fem</i>), is located in the W chromosome. As a secondary sexual trait, we focused on the fat body, which shows remarkable differences between the sexes during the last instar larval stage. A comparison of the transcriptomes between the fat bodies of male and female larvae identified 232 sex-differentially expressed genes (S-DEGs). The proportions of ZZ and ZW cells constituting the fat body of the gynandromorphic larvae were calculated according to the expression level of the <i>Fem</i>. Based on the obtained values, the expression level of each S-DEG was estimated, assuming that the levels of S-DEG expression were determined according to the proportion of ZZ and ZW cells. The estimated expression levels of 207 out of 232 S-DEGs were strongly correlated with the corresponding S-DEG expression level of the gynandromorphic fat body, determined by RNA-seq. These results strongly suggest that most of the sexually dimorphic transcriptome in the fat body is regulated in a cell-autonomous manner.</p>","PeriodicalId":15563,"journal":{"name":"Journal of Developmental Biology","volume":"12 4","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11587106/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710266","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Methyl-Beta-Cyclodextrin Restores Aberrant Bone Morphogenetic Protein 2-Signaling in Bone Marrow Stromal Cells Obtained from Aged C57BL/6 Mice.","authors":"Daniel Halloran, Venu Pandit, Kelechi Chukwuocha, Anja Nohe","doi":"10.3390/jdb12040030","DOIUrl":"10.3390/jdb12040030","url":null,"abstract":"<p><p>During aging, disruptions in various signaling pathways become more common. Some older patients will exhibit irregular bone morphogenetic protein (BMP) signaling, which can lead to osteoporosis (OP)-a debilitating bone disease resulting from an imbalance between osteoblasts and osteoclasts. In 2002, the Food and Drug Administration (FDA) approved recombinant human BMP-2 (rhBMP-2) for use in spinal fusion surgeries as it is required for bone formation. However, complications with rhBMP-2 arose and primary osteoblasts from OP patients often fail to respond to BMP-2. Although patient samples are available for study, previous medical histories can impact results. Consequently, the C57BL/6 mouse line serves as a valuable model for studying OP and aging. We find that BMP receptor type Ia (BMPRIa) is upregulated in the bone marrow stromal cells (BMSCs) of 15-month-old mice, consistent with prior data. Furthermore, conjugating BMP-2 with Quantum Dots (QDot^®s) allows effective binding to BMPRIa, creating a fluorescent tag for BMP-2. Furthermore, after treating BMSCs with methyl-β-cyclodextrin (MβCD), a disruptor of cellular endocytosis, BMP signaling is restored in 15-month-old mice, as shown by von Kossa assays. MβCD has the potential to restore BMPRIa function, and the BMP signaling pathway offers a promising avenue for future OP therapies.</p>","PeriodicalId":15563,"journal":{"name":"Journal of Developmental Biology","volume":"12 4","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11586967/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}