Short Communication
Volume 2 Issue 5 - March 2017
DOI: 10.19080/OAJNN.2017.02.555600
Open Access J Neurol Neurosurg
Copyright © All rights are reserved by Kimberle M Jacobs
Neurophysiological Study of Traumatic Brain
Injury: To Slice or Not to Slice
Jianli Sun and Kimberle M Jacobs*
Department of Anatomy & Neurobiology, Virginia Commonwealth University, USA
Submission: March 04, 2017; Published: March 20, 2017
*Corresponding author: Kimberle M Jacobs, PhD Associate Professor, Department of Anatomy & Neurobiology Virginia Commonwealth
University, PO Box 980709 Richmond, VA 23298, USA, Tel: ; Fax: ; Email:
Introduction
One of the most critical neuropathological features of
traumatic brain injury (TBI) is axotomy [1-5]. Even in mild (m)
TBI, diffuse axotomy is easily and consistently observed as axons
that end in swellings and dislocation from the distal part of the
axon [6-8]. Integrity of axons can be measured in the clinical
population with diffusion tensor imaging (DTI) [9-12]. Patient
outcome has been correlated with DTI results, [13-16] although
subacute imaging may be most effective in this regard [17,18].
Correlations may also be stronger for severe and moderate as
opposed to mild injuries [19]. DTI results have also been shown
to correlate with the degree of diffuse axonal injury (DAI)
as measured with amyloid precursor protein (APP) staining
that accumulates in the axonal swelling [20,21]. Treatment
assessments have typically been focused on reduction of this
axonal injury [22-29], although it has also been recognized
that some measures of DAI improve over time while cognitive
dysfunction persists [30].
Because axotomy also occurs during preparation of ex vivo
brain slices, it could be argued that this methodology should
not be used for study of mechanisms associated with TBI.
Vibratome-axotomized neurons can certainly be identified at
the surface of acute slices (Figure 1A & 1B). This is one reason
most investigators performing recordings in slices choose
neurons that are at least one cell layer deep to surface neurons,
and typically 40-60 m into the tissue. Creation of ex vivo slices
for study of normal physiological function has been a staple
of neuroscience research for decades [31,32]. The benefits of
stability, visualized access to specific cell types and placement
of electrodes, as well as feasibility of drug application have been
previously described [33-35]. However, the idea that slicing
produces pathology has also been used to create a model of
post-traumatic epilepsy utilizing organotypic hippocampal
slices that survive for weeks [36-40]. In this model, interictal-
like epileptiform activity is first observed after 14 days in vitro,
while ictal-like activity is seen by 21 days [38]. The fact that this
abnormal activity develops in days and not hours suggests that
it may be due to synaptic reorganization that is secondary to
the axotomy and thus unlikely to play a role in the physiology
of acute slices.
We recently chose to use the acute slice to study physiological
effects of trauma, specifically in order to target the axotomized
neurons which are in the minority and located diffusely within
Open Access J Neurol Neurosurg 2(5): OAJNN.MS.ID.555600 (2017) 001
Abstract
Diffuse axotomy is characteristic of mild traumatic brain injury (TBI) both clinically and in experimental models. Axotomy also occurs during
preparation of acute brain slices for physiological recordings. Brain slices have been used for decades and have many advantages, including
the ability to select cells within particular locations or with specific morphologies for study. The question addressed here is whether creating
additional axotomy by preparing brain slices introduces additional injury that is already typical of TBI. We have previously demonstrated
alterations in both intrinsic membrane and synaptic properties recorded in brain slices after mTBI. Here we examined whether axotomized
neurons differ from intact neurons in slices made from naïve mouse pups. There was no significant difference in either excitatory synaptic
postsynaptic currents or in any measures of intrinsic membrane and cellular properties. These results suggest that creating the brain slices does
not induce alterations similar to that observed within two days of mTBI induced by fluid percussion injury. Thus ex vivo slices are an appropriate
methodology to study the effects of TBI.
Keywords: Ex vivo slice; Electrophysiology; Axotomy; EPSC; Membrane properties; Neurotrauma
Abbreviations: AHP: After Hyperpolarization; APP: Amyloid Precursor Protein; DAI: Diffuse Axonal Injury; DAP: Depolarizing After Potential;
DTI: Diffusion Tensor Imaging; TBI: Traumatic Brain Injury