RESUMEN
Metabolism and mRNA translation represent critical steps involved in modulating gene expression and cellular physiology. Being the most energy-consuming process in the cell, mRNA translation is strictly linked to cellular metabolism and in synchrony with it. Indeed, several mRNAs for metabolic pathways are regulated at the translational level, resulting in translation being a coordinator of metabolism. On the other hand, there is a growing appreciation for how metabolism impacts several aspects of RNA biology. For example, metabolic pathways and metabolites directly control the selectivity and efficiency of the translational machinery, as well as post-transcriptional modifications of RNA to fine-tune protein synthesis. Consistently, alterations in the intricate interplay between translational control and cellular metabolism have emerged as a critical axis underlying human diseases. A better understanding of such events will foresee innovative therapeutic strategies in human disease states.
Asunto(s)
Biosíntesis de Proteínas , ARN Mensajero , Humanos , Animales , ARN Mensajero/metabolismo , ARN Mensajero/genética , Redes y Vías Metabólicas , Procesamiento Postranscripcional del ARNRESUMEN
Hind limb ischemia is a useful model to assess metabolic and cellular responses. Here, we present a protocol for evaluating post-natal angiogenesis in a mouse hind limb ischemia model. We describe steps to induce a severe restriction of blood supply of the femoral artery and vein that mimics the real-life scenario observed in clinical settings. We then detail procedures for follow-up laser Doppler imaging to compare post-ischemic responses of four different mouse strains in their capacity to trigger compensatory arteriogenesis. For complete details on the use and execution of this protocol, please refer to Oberkersch et al. (2022).1.
RESUMEN
The DNA polymerase gamma (Polg) is a nuclear-encoded enzyme involved in DNA replication in animal mitochondria. In humans, mutations in the POLG gene underlie a set of mitochondrial diseases characterized by mitochondrial DNA (mtDNA) depletion or deletion and multiorgan defects, named POLG disorders, for which an effective therapy is still needed. By applying antisense strategies, ENU- and CRISPR/Cas9-based mutagenesis, we have generated embryonic, larval-lethal and adult-viable zebrafish Polg models. Morphological and functional characterizations detected a set of phenotypes remarkably associated to POLG disorders, including cardiac, skeletal muscle, hepatic and gonadal defects, as well as mitochondrial dysfunctions and, notably, a perturbed mitochondria-to-nucleus retrograde signaling (CREB and Hypoxia pathways). Next, taking advantage of preliminary evidence on the candidate molecule Clofilium tosylate (CLO), we tested CLO toxicity and then its efficacy in our zebrafish lines. Interestingly, at well tolerated doses, the CLO drug could successfully rescue mtDNA and Complex I respiratory activity to normal levels, even in mutant phenotypes worsened by treatment with Ethidium Bromide. In addition, the CLO drug could efficiently restore cardio-skeletal parameters and mitochondrial mass back to normal values. Altogether, these evidences point to zebrafish as a valuable vertebrate organism to faithfully phenocopy multiple defects detected in POLG patients. Moreover, this model represents an excellent platform to screen, at the whole-animal level, candidate molecules with therapeutic effects in POLG disorders.
Asunto(s)
Enfermedades Mitocondriales/genética , Compuestos de Amonio Cuaternario/metabolismo , Animales , Modelos Animales de Enfermedad , Fenotipo , Pez CebraRESUMEN
A key question in vascular biology is how the diversity of origin of vascular mural cells, namely smooth muscle cells (SMCs) and pericytes influences vessel properties, in particular the regional propensity to vascular diseases. This review therefore first describes the role and regulation of mural cells during vascular formation, with a focus on embryonic origin. We then consider the evidence that connects heterogeneities in SMC and pericyte origins with disease. Since this idea has major implications for understanding and modelling human disease, then there is a pressing need for new model systems to investigate mural cell development and the consequences of heterogeneity. Recent advances arising from in vitro strategies for deriving mural cells from human pluripotent stem cells as well as from the zebrafish model will be discussed and the medical relevance of these discoveries will be highlighted.
Asunto(s)
Linaje de la Célula , Células Madre Embrionarias/patología , Células Madre Pluripotentes Inducidas/patología , Miocitos del Músculo Liso/patología , Enfermedades Vasculares/patología , Animales , Comunicación Celular , Células Madre Embrionarias/metabolismo , Regulación del Desarrollo de la Expresión Génica , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Miocitos del Músculo Liso/metabolismo , Neovascularización Fisiológica , Fenotipo , Transducción de Señal , Enfermedades Vasculares/genética , Enfermedades Vasculares/metabolismo , Enfermedades Vasculares/fisiopatología , Pez Cebra/genética , Pez Cebra/metabolismoRESUMEN
Formation and maturation of a functional vascular network is a process called angiogenesis. This is a crucial biological event in all vertebrates. Precise morphogenetic and cellular mechanisms act in endothelial cells (ECs) to drive angiogenesis during growth and throughout adulthood. Reactive oxygen species (ROS) and their metabolism are proving to be crucial participants in the shaping and stabilizing of blood vessels. Often, the same mechanisms are responsible for the insurgence of vascular-associated pathologies. In this review we discuss how ROS-mediated signalling events and distinctive metabolic pathways drive the biology of endothelial cells. We support the use of alternative anti-angiogenic therapy based on the manipulation of ROS signalling and metabolism to solve angiogenesis-related diseases.
Asunto(s)
Células Endoteliales/metabolismo , Neovascularización Patológica/metabolismo , Neovascularización Fisiológica/fisiología , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/fisiología , Animales , Humanos , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Feline leukemia virus subgroup C receptor 1 (Flvcr1) encodes two heme exporters: FLVCR1a, which localizes to the plasma membrane, and FLVCR1b, which localizes to mitochondria. Here, we investigated the role of the two Flvcr1 isoforms during erythropoiesis. We showed that, in mice and zebrafish, Flvcr1a is required for the expansion of committed erythroid progenitors but cannot drive their terminal differentiation, while Flvcr1b contributes to the expansion phase and is required for differentiation. FLVCR1a-down-regulated K562 cells have defective proliferation, enhanced differentiation, and heme loading in the cytosol, while FLVCR1a/1b-deficient K562 cells show impairment in both proliferation and differentiation, and accumulate heme in mitochondria. These data support a model in which the coordinated expression of Flvcr1a and Flvcr1b contributes to control the size of the cytosolic heme pool required to sustain metabolic activity during the expansion of erythroid progenitors and to allow hemoglobinization during their terminal maturation. Consistently, reduction or increase of the cytosolic heme rescued the erythroid defects in zebrafish deficient in Flvcr1a or Flvcr1b, respectively. Thus, heme export represents a tightly regulated process that controls erythropoiesis.
Asunto(s)
Diferenciación Celular/fisiología , Eritropoyesis/fisiología , Hemo/metabolismo , Líquido Intracelular/metabolismo , Proteínas de Transporte de Membrana/fisiología , Receptores Virales/fisiología , Secuencia de Aminoácidos , Animales , Humanos , Células K562 , Ratones , Ratones Noqueados , Ratones Transgénicos , Datos de Secuencia Molecular , Pez CebraRESUMEN
CD (cathepsin D) is a ubiquitous lysosomal hydrolase involved in a variety of pathophysiological functions, including protein turnover, activation of pro-hormones, cell death and embryo development. CD-mediated proteolysis plays a pivotal role in tissue and organ homoeostasis. Altered expression and compartmentalization of CD have been observed in diseased muscle fibres. Whether CD is actively involved in muscle development, homoeostasis and dystrophy remains to be demonstrated. Zebrafish (Danio rerio) is emerging as a valuable 'in vivo' vertebrate model for muscular degeneration and congenital myopathies. In this work, we report on the perturbance of the somitic musculature development in zebrafish larvae caused by MPO (morpholino)-mediated silencing of CD in oocytes at the time of fertilization. Restoring CD expression, using an MPO-non-matching mutated mRNA, partially rescued the normal phenotype, confirming the indispensable role of CD in the correct development and integrity of the somitic musculature. This is the first report showing a congenital myopathy caused by CD deficiency in a vertebrate experimental animal model.
Asunto(s)
Catepsina D/genética , Miopatías Estructurales Congénitas/genética , Pez Cebra/genética , Cigoto/metabolismo , Animales , Catepsina D/biosíntesis , Modelos Animales de Enfermedad , Regulación del Desarrollo de la Expresión Génica/genética , Técnicas de Silenciamiento del Gen , Silenciador del Gen , Humanos , Miopatías Estructurales Congénitas/etiología , Miopatías Estructurales Congénitas/patología , ARN Mensajero/genética , Pez Cebra/crecimiento & desarrollo , Cigoto/patologíaRESUMEN
MicroRNAs are small non-coding RNAs endogenously expressed by all tissues during development and adulthood. They regulate gene expression by controlling the stability of targeted messenger RNA. In cardiovascular tissues microRNAs play a role by modulating essential genes involved in heart and blood vessel development and homeostasis. The zebrafish (Danio rerio) system is a recognized vertebrate model system useful to study cardiovascular biology; recently, it has been used to investigate microRNA functions during natural and pathological states. In this review, we will illustrate the advantages of the zebrafish model in the study of microRNAs in heart and vascular cells, providing an update on recent discoveries using the zebrafish to identify new microRNAs and their targeted genes in cardiovascular tissues. Lastly, we will provide evidence that the zebrafish is an optimal model system to undercover new microRNA functions in vertebrates and to improve microRNA-based therapeutic approaches.