Prenatal cerebral ischemia disrupts MRI-defined cortical microstructure through disturbances in neuronal arborization.

Children who survive preterm birth exhibit persistent unexplained disturbances in cerebral cortical growth with associated cognitive and learning disabilities. The mechanisms underlying these deficits remain elusive. We used ex vivo diffusion magnetic resonance imaging to demonstrate in a preterm large-animal model that cerebral ischemia impairs cortical growth and the normal maturational decline in cortical fractional anisotropy (FA). Analysis of pyramidal neurons revealed that cortical deficits were associated with impaired expansion of the dendritic arbor and reduced synaptic density. Together, these findings suggest a link between abnormal cortical FA and disturbances of neuronal morphological development. To experimentally investigate this possibility, we measured the orientation distribution of dendritic branches and observed that it corresponds with the theoretically predicted pattern of increased anisotropy within cases that exhibited elevated cortical FA after ischemia. We conclude that cortical growth impairments are associated with diffuse disturbances in the dendritic arbor and synapse formation of cortical neurons, which may underlie the cognitive and learning disabilities in survivors of preterm birth. Further, measurement of cortical FA may be useful for noninvasively detecting neurological disorders affecting cortical development.

GABAergic protection of hippocampal pyramidal neurons against glutamate insult: deficit in young animals compared to adults.

Hypoxia-ischemia (HI) injury in neonatal animals leads to selective regional loss of neurons including CA1 and CA3 pyramidal neurons of the hippocampus as well as nonlethal pathologies. Glutamate-receptor over-activation and Ca2+ influx are involved in these neonatal changes. We examined glutamate-evoked Ca2+ responses in neonatal (PN 7-13) and young adult (PN 21-27) CA1 pyramidal neurons in acute slices from rats. In neonates, transient exposure to glutamate produced large Ca2+ increases throughout neurons, including distal dendrites (primary Ca2+ responses). Repeated exposures resulted in sustained Ca2+ increases in apical dendrites (secondary Ca2+ responses) that were independent of continued glutamate exposure. These responses propagated and invaded the soma, resulting in irrecoverably high Ca2+ elevations. In neurons from adults, identical glutamate exposure evoked primary Ca2+ responses only in somata and proximal apical dendrites. Repeated glutamate exposures in the adult neurons also led to secondary Ca2+ responses, but they arose in the peri-somatic region and then spread outward to distal apical dendrites, again without recovery. More stimuli were required to initiate secondary responses in neurons from adult versus neonates. Block of GABAA receptors in adults caused the primary and secondary responses to revert to the spatial pattern seen in the neonates and greatly increased their vulnerability to glutamate. These findings suggest that neurodegenerative secondary Ca2+ events may be important determinants of susceptibility to HI injury in the developing CNS and that immature CA1 neurons may be more susceptible to excitotoxic injury due at least in part to insufficient development of GABAergic inputs to their dendrites.

Caffeine releasable stores of Ca2+ show depletion prior to the final steps in delayed CA1 neuronal death.

In addition to their role in signaling, Ca2+ ions in the endoplasmic reticulum also regulate important steps in protein processing and trafficking that are critical for normal cell function. Chronic depletion of Ca2+ in the endoplasmic reticulum has been shown to lead to cell degeneration and has been proposed as a mechanism underlying delayed neuronal death following ischemic insults to the CNS. Experiments here have assessed the relative content of ryanodine receptor-gated stores in CA1 neurons by measuring cytoplasmic Ca2+ increases induced by caffeine. These measurements were performed on CA1 neurons, in slice, from normal gerbils, and compared with responses from this same population of neurons 54-60 h after animals had undergone a standard ischemic insult: 5-min bilateral occlusion of the carotid arteries. The mean amplitude of responses in the postischemic population were less than one-third of those in control or sham-operated animals, and 35% of the neurons from postischemic animals showed very small responses that were approximately 10% of the control population mean. Refilling of these stores after caffeine challenges was also impaired in postischemic neurons. These observations are consistent with our earlier finding that voltage-gated influx is sharply reduced in postischemic in CA1 neurons and the hypothesis that the resulting depletion in endosomal Ca2+ is an important cause of delayed neuronal death.