RESUMEN
Here, we describe a method for inducing subcortical white matter stroke in mice, as well as tracking cellular proliferation through drinking water administration of EdU and ex vivo labeling.
Asunto(s)
Accidente Cerebrovascular , Sustancia Blanca , Ratones , Animales , Sustancia Blanca/patología , Accidente Cerebrovascular/patología , Proliferación Celular , Hiperplasia/patologíaRESUMEN
Ependyma have been proposed as adult neural stem cells that provide the majority of newly proliferated scar-forming astrocytes that protect tissue and function after spinal cord injury (SCI). This proposal was based on small, midline stab SCI. Here, we tested the generality of this proposal by using a genetic knock-in cell fate mapping strategy in different murine SCI models. After large crush injuries across the entire spinal cord, ependyma-derived progeny remained local, did not migrate and contributed few cells of any kind and less than 2%, if any, of the total newly proliferated and molecularly confirmed scar-forming astrocytes. Stab injuries that were near to but did not directly damage ependyma, contained no ependyma-derived cells. Our findings show that ependymal contribution of progeny after SCI is minimal, local and dependent on direct ependymal injury, indicating that ependyma are not a major source of endogenous neural stem cells or neuroprotective astrocytes after SCI.
Asunto(s)
Astrocitos/fisiología , Diferenciación Celular , Cicatriz/fisiopatología , Epéndimo/fisiopatología , Traumatismos de la Médula Espinal/fisiopatología , Animales , Modelos Animales de Enfermedad , Ratones , Ratones Transgénicos , Células-Madre Neurales/fisiologíaRESUMEN
Astrocytes respond to a variety of CNS injuries by cellular enlargement, process outgrowth, and upregulation of extracellular matrix proteins that function to prevent expansion of the injured region. This astrocytic response, though critical to the acute injury response, results in the formation of a glial scar that inhibits neural repair. Scar-forming cells (fibroblasts) in the heart can undergo mesenchymal-endothelial transition into endothelial cell fates following cardiac injury in a process dependent on p53 that can be modulated to augment cardiac repair. Here, we sought to determine whether astrocytes, as the primary scar-forming cell of the CNS, are able to undergo a similar cellular phenotypic transition and adopt endothelial cell fates. Serum deprivation of differentiated astrocytes resulted in a change in cellular morphology and upregulation of endothelial cell marker genes. In a tube formation assay, serum-deprived astrocytes showed a substantial increase in vessel-like morphology that was comparable to human umbilical vein endothelial cells and dependent on p53. RNA sequencing of serum-deprived astrocytes demonstrated an expression profile that mimicked an endothelial rather than astrocyte transcriptome and identified p53 and angiogenic pathways as specifically upregulated. Inhibition of p53 with genetic or pharmacologic strategies inhibited astrocyte-endothelial transition. Astrocyte-endothelial cell transition could also be modulated by miR-194, a microRNA downstream of p53 that affects expression of genes regulating angiogenesis. Together, these studies demonstrate that differentiated astrocytes retain a stimulus-dependent mechanism for cellular transition into an endothelial phenotype that may modulate formation of the glial scar and promote injury-induced angiogenesis.
Asunto(s)
Astrocitos/citología , Astrocitos/metabolismo , Células Endoteliales/citología , Células Endoteliales/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Animales , Cadherinas/metabolismo , Linaje de la Célula , Plasticidad de la Célula , Células Cultivadas , Análisis por Conglomerados , Medio de Cultivo Libre de Suero , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Ratones Endogámicos C57BL , MicroARNs/genética , MicroARNs/metabolismo , Neovascularización Fisiológica/genética , Células-Madre Neurales/citología , Células-Madre Neurales/metabolismo , Plasticidad Neuronal , Transcripción GenéticaRESUMEN
Angiogenesis is a key feature of central nervous system injury. A neovessel-derived signal mediated by prostacyclin triggers axonal sprouting and functional recovery in a mouse model of inflammatory spinal cord injury (pages 1658-1664). Are such angiocrine signals relevant to neurovascular remodeling and recovery in other neurological contexts?