From the Université René Descartes, Paris, France.
Correspondence to C. Charriaut-Marlangue, INSERM U29, 123 Boulevard de Port-Royal, 75014 Paris, France. E-mail cm{at}u29.cochin.inserm.fr
Background and Purpose—The
mechanisms leading to delayed cell death after hypoxic-ischemic
injury in the developing brain remain to be elucidated. The aim of this
study was to develop a model of transient focal ischemia in the
neonatal rat in an attempt to create a reperfusion phase since in the
filament model of reversible middle cerebral artery occlusion, size
limitations precluded performing this procedure before 14 to 18 days.
We then analyze whether apoptosis or necrosis occurs in
this model.
Methods—Seven-day-old Wistar rat pups (n=96) underwent permanent
left middle cerebral artery occlusion in association with 1-hour
occlusion of the left common carotid artery. Evolution of the brain
infarction was studied from 24 hours to 3 months on cresyl
violet–stained coronal sections. Infarct volume was determined with
the use of the mitochondrial stain
2,3,5-triphenyltetrazolium chloride.
Neuronal death was demonstrated by the silver staining method of
Gallyas et al (1980). Chromatin condensation was shown by DNA
fragmentation assessed with the use of terminal
deoxynucleotidyl transferase–mediated dUTP-biotin
nick end-labeling (TUNEL) assay in cryostat sections and electron
microscopic analysis.
Results—Almost all of the animals who survived had reproducible
cortical infarcts. The mean infarct volume was 31±7
mm3 (mean±SD). The ipsilateral hemisphere showed a
well-delineated lesion in the frontoparietal cortex at 3-month
recovery. Argyrophilic (dying) neurons were observed a few hours after
reperfusion and increased with time. Cells exhibiting DNA fragmentation
were shown as early as 6 hours, increased up to and peaked at 24 to 96
hours, then progressively decreased and persisted for several days,
suggesting an ongoing process. Electron microscopy analysis
demonstrated high condensation and clumping of chromatin beneath
nuclear membrane in shrunken neurons.
Conclusions—Our study demonstrates the feasibility of performing
ischemia-reperfusion in 7-day-old rats that develop progressive
neuronal death with features characteristic of apoptosis. The
reperfusion phase mimics events that occur during neonatal human
hypoxic-ischemic encephalopathy at birth, since perinatal
intensive care most often permits recirculation.
Anesthesiology/Critical
Care Medicine,
Johns Hopkins Medical Institutes,
Baltimore, Maryland
© 1998 American Heart Association, Inc.
Original Contributions
A Model of Transient Unilateral Focal Ischemia With Reperfusion in the P7 Neonatal Rat
Morphological Changes Indicative of Apoptosis
Editorial Comment
Morphological Changes Indicative of Apoptosis
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