MAP2 provides reliable early assessment of neural injury in the newborn piglet model of birth asphyxia

Structural remodeling of gray matter astrocytes in the neonatal pig brain after hypoxia/ischemia

“…This model has been used in previous studies in our laboratories (Bj€ orkman et al, 2006;Foster et al, 2001;Lingwood et al, 2008;Pow et al, 2004;Sullivan et al, 2007a,b), and animals undergoing the H/I insult have a variety of outcomes (no damage; mild, moderate, or severe brain damage; and death), just like human infants exposed to H/I. Furthermore, H/I brain damage occurring in the pig mimics the patterns of brain damage seen in human babies: There is regional variation within the brain in terms of vulnerability to H/I damage, with forebrain regions such as cortex and hippocampus being most vulnerable (Nuñez and McCarthy, 2004).…”

Rapid loss of glutamine synthetase from astrocytes in response to hypoxia: Implications for excitotoxicity

“…This time line accords with that observed in rodent studies (Torp et al, 1995;Bruhn et al, 2000), and would imply that loss of transporters would lead to a moderately slow onset of glutamate dysregulation. Similarly loss of the neuronal marker MAP2 (expression of which is influenced by the exposure of neurons to glutamate) is first evident by about 6 h post-insult (Lingwood et al, 2008). However the overall problem of glutamate homeostasis is more complex: A critical component of the glutamate detoxification process is the conversion of toxic ammonia plus glutamate to form the non-neuroactive compound glutamine using the enzyme glutamine synthetase (GS).…”

“…We have now re-evaluated the question of whether GS levels are perturbed in response to hypoxia in our pig model of neonatal hypoxia, using a specific antibody against GS, in concert with double labelling for the neuronal marker MAP 2 which is a sensitive marker of neuronal perturbation in the hypoxic pig brain (Lingwood et al, 2008) and the glial glutamate transporter GLAST.…”

Hydrogen is neuroprotective and preserves neurovascular reactivity following asphyxia in newborn pigs

“…Evaluation of immunohistochemistry slides were performed according previously described methods [104]; briefly, images of whole labelled sections were scanned with automatic digital slide scanner (Panoramic MIDI, 3DHistech, Budapest, Hungary) and imported into Image Pro Plus 6.0 (Media Cybernetics, Bethesda, USA). Images were then converted to greyscale and thresholded at the same level in each image to produce images consisting only of black (labelled) or white (unlabelled) pixels.…”

“…Indeed, the recovery of the brain electrical activity on the aEEG and the mild neuronal damage detected with histopathology confirmed this notion. We performed immunohistochemistry of the neuronal cytoskeletal protein microtubule associated protein (MAP-2) in order to determine the so called MAP-2 score established by Lingwood et al [104]. MAP-2 is involved in maintaining the structural integrity of neuronal cytoskeleton.…”