Experimental modelling of the consequences of brief late gestation asphyxia on newborn lamb behaviour and brain structure

PLoS One. 2013 Nov 6;8(11):e77377. doi: 10.1371/journal.pone.0077377. eCollection 2013.

Abstract

Brief but severe asphyxia in late gestation or at the time of birth may lead to neonatal hypoxic ischemic encephalopathy and is associated with long-term neurodevelopmental impairment. We undertook this study to examine the consequences of transient in utero asphyxia in late gestation fetal sheep, on the newborn lamb after birth. Surgery was undertaken at 125 days gestation for implantation of fetal catheters and placement of a silastic cuff around the umbilical cord. At 132 days gestation (0.89 term), the cuff was inflated to induce umbilical cord occlusion (UCO), or sham (control). Fetal arterial blood samples were collected for assessment of fetal wellbeing and the pregnancy continued until birth. At birth, behavioral milestones for newborn lambs were recorded over 24 h, after which the lambs were euthanased for brain collection and histopathology assessments. After birth, UCO lambs displayed significant latencies to (i) use all four legs, (ii) attain a standing position, (iii) find the udder, and (iv) successfully suckle--compared to control lambs. Brains of UCO lambs showed widespread pathologies including cell death, white matter disruption, intra-parenchymal hemorrhage and inflammation, which were not observed in full term control brains. UCO resulted in some preterm births, but comparison with age-matched preterm non-UCO control lambs showed that prematurity per se was not responsible for the behavioral delays and brain structural abnormalities resulting from the in utero asphyxia. These results demonstrate that a single, brief fetal asphyxic episode in late gestation results in significant grey and white matter disruption in the developing brain, and causes significant behavioral delay in newborn lambs. These data are consistent with clinical observations that antenatal asphyxia is causal in the development of neonatal encephalopathy and provide an experimental model to advance our understanding of neuroprotective therapies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Animals, Suckling
  • Apoptosis
  • Asphyxia / pathology*
  • Behavior, Animal
  • Brain / pathology*
  • Caspase 3 / metabolism
  • Female
  • Pregnancy
  • Prenatal Exposure Delayed Effects / pathology*
  • Sheep
  • Umbilical Cord / blood supply

Substances

  • Caspase 3

Grants and funding

This study was supported by grants to DWW and MCM from the National Health & Medical Research Council of Australia (NHMRC), Cerebral Palsy Alliance Australia, and the Network in Genes, Environment and Development (NGED), an initiative of the NHMRC and Australia Research Council (ARC). The authors also gratefully acknowledge support from the Victorian Government’s Operational Infrastucture Support Program to Monash Institute of Medical Research. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.