A Novel In Vivo Model for Multiplexed Analysis of Callosal Connections upon Cortical Damage.

Autores de CIPF

• 

  Mª Carmen Navarro González

  Autor

• Jose Manuel Saborit Torres

  Autor

• 

  Pietro Fazzari

  Autor

Participantes ajenos a CIPF

• González-Manteiga A
• Sebestyén VE
• Talhada D
• Vayá MI
• Ruscher K

Grupos de Investigación

• Laboratorio de Circuitos Corticales en Salud y Enfermedad

  

  Jefe/a de Grupo

  Pietro Fazzari

• Laboratorio de Obesidad, Diabetes y Cormobilidades

  

  Jefe/a de Grupo

  Stefania Carobbio

• Unidad Mixta CIPF-FISABIO de Imagen Biomédica

Abstract

Brain damage is the major cause of permanent disability and it is particularly relevant in the elderly. While most studies focused on the immediate phase of neuronal loss upon injury, much less is known about the process of axonal regeneration after damage. The development of new refined preclinical models to investigate neuronal regeneration and the recovery of brain tissue upon injury is a major unmet challenge. Here, we present a novel experimental paradigm in mice that entails the (i) tracing of cortico-callosal connections, (ii) a mechanical lesion of the motor cortex, (iii) the stereological and histological analysis of the damaged tissue, and (iv) the functional characterization of motor deficits. By combining conventional microscopy with semi-automated 3D reconstruction, this approach allows the analysis of fine subcellular structures, such as axonal terminals, with the tridimensional overview of the connectivity and tissue integrity around the lesioned area. Since this 3D reconstruction is performed in serial sections, multiple labeling can be performed by combining diverse histological markers. We provide an example of how this methodology can be used to study cellular interactions. Namely, we show the correlation between active microglial cells and the perineuronal nets that envelop parvalbumin interneurons. In conclusion, this novel experimental paradigm will contribute to a better understanding of the molecular and cellular interactions underpinning the process of cortical regeneration upon brain damage.

Keywords

• brain damage, callosal neurons, cortico-cortical connections, microglia, neuronal regeneration, perineuronal network, pre-clinical models, stroke, traumatic injury