A new model of repeat mTBI in adolescent rats

Abstract

Sports-related injury is frequently associated with repeated diffuse and mild traumatic brain injury (mTBI). We combined two existing models for inducing TBI in rats, the Impact Acceleration and Controlled Cortical Impact models, to create a new method relevant to the study of cognitive sequelae of repeat mTBI in adolescent athletes. Repeated mTBI, such as those incurred in sports, can result in a wide range of outcomes, with many individuals experiencing no chronic sequela while others develop profound cognitive and behavioral impairments, typically in the absence of lasting motor symptoms or gross tissue loss appreciable antemortem. It is critical to develop models of mTBI and repeat mTBI that have the flexibility to assess multiple parameters related to injury (e.g. number and magnitude of impacts, inter-injury interval, etc) that are associated with brain vulnerability compared to normal recovery. We designed a 3D-printed plastic implant to permanently secure a metal disc to the skull of adolescent rats in order to induce multiple injuries without performing multiple survival surgeries and also to minimize pre-injury anesthesia time. Rats were randomly assigned to sham injury (n = 12), single injury (n = 12; injury on P41), or repeat injury (n = 14; injuries on P35, P38, and P41) groups. Compared to single injury and sham injury, repeat injuries caused increased toe pinch reflex latency (F(2,34) = 4.126, p < .05) and diminished weight gain (F(2, 34) = 4.767, p < .05). Spatial navigation was tested using Morris water maze, beginning one week after the final injury (P48). While there were no differences between groups during acquisition, both single and repeat injuries resulted in deficits on probe trial performance (p < .01 and p < .05 respectively). Single injury animals also exhibited a deficit in working memory deficit across three days of testing (p < .05). Neither injury group had neuronal loss in the hilus or CA3, according to stereological quantification of NeuN. Therefore, by implanting a helmet we have created a relevant model of sports-related injury and repeated mTBI that results in subtle but significant changes in cognitive outcome in the absence of significant hippocampal cell death.