The extent of intrauterine growth restriction determines the severity of cerebral injury and neurobehavioural deficits in rodents

PLoS One. 2017 Sep 21;12(9):e0184653. doi: 10.1371/journal.pone.0184653. eCollection 2017.

Abstract

Background: Cerebral Palsy (CP) is the most common physical pediatric neurodevelopmental disorder and spastic diplegic injury is its most frequent subtype. CP results in substantial neuromotor and cognitive impairments that have significant socioeconomic impact. Despite this, its underlying pathophysiological mechanisms and etiology remain incompletely understood. Furthermore, there is a need for clinically relevant injury models, which a) reflect the heterogeneity of the condition and b) can be used to evaluate new translational therapies. To address these key knowledge gaps, we characterized a chronic placental insufficiency (PI) model, using bilateral uterine artery ligation (BUAL) of dams. This injury model results in intrauterine growth restriction (IUGR) in pups, and animals recapitulate the human phenotype both in terms of neurobehavioural and anatomical deficits.

Methods: Effects of BUAL were studied using luxol fast blue (LFB)/hematoxylin & eosin (H&E) staining, immunohistochemistry, quantitative Magnetic Resonance Imaging (MRI), and Catwalk neurobehavioural tests.

Results: Neuroanatomical analysis revealed regional ventricular enlargement and corpus callosum thinning in IUGR animals, which was correlated with the extent of growth restriction. Olig2 staining revealed reductions in oligodendrocyte density in white and grey matter structures, including the corpus callosum, optic chiasm, and nucleus accumbens. The caudate nucleus, along with other brain structures such as the optic chiasm, internal capsule, septofimbrial and lateral septal nuclei, exhibited reduced size in animals with IUGR. The size of the pretectal nucleus was reduced only in moderately injured animals. MAG/NF200 staining demonstrated reduced myelination and axonal counts in the corpus callosum of IUGR animals. NeuN staining revealed changes in neuronal density in the hippocampus and in the thickness of hippocampal CA2 and CA3 regions. Diffusion weighted imaging (DWI) revealed regional white and grey matter changes at 3 weeks of age. Furthermore, neurobehavioural testing demonstrated neuromotor impairments in animals with IUGR in paw intensities, swing speed, relative print positions, and phase dispersions.

Conclusions: We have characterized a rodent model of IUGR and have demonstrated that the neuroanatomical and neurobehavioural deficits mirror the severity of the IUGR injury. This model has the potential to be applied to examine the pathobiology of and potential therapeutic strategies for IUGR-related brain injury. Thus, this work has potential translational relevance for the study of CP.

MeSH terms

  • Animals
  • Animals, Newborn
  • Behavior, Animal*
  • Brain / diagnostic imaging
  • Brain / growth & development
  • Brain / pathology
  • Cell Death
  • Diffusion Tensor Imaging
  • Disease Models, Animal*
  • Female
  • Fetal Growth Retardation / diagnostic imaging
  • Fetal Growth Retardation / pathology*
  • Fetal Growth Retardation / physiopathology*
  • Fetal Growth Retardation / psychology
  • Ligation
  • Magnetic Resonance Imaging
  • Motor Activity
  • Placental Insufficiency
  • Pregnancy
  • Rats, Long-Evans
  • Severity of Illness Index
  • Uterine Artery

Grant support

This project received financial support from the Kids Brain Health Network as well as the Ontario Brain Institute. C.A.R. was supported by postdoctoral research grants from the Jeremy and Judith Freedman Family Foundation and the Ontario Stem Cell Initiative. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.