Prenatal protein malnutrition in rats alters the c-Fos response of neurons in the anterior cingulate and medial prefrontal region to behavioral stress

Prenatal protein malnutrition affects brain development and behavior despite dietary rehabilitation from birth. Behavioral alterations include abnormal responses to stressors. To explore what brain regions mediate this altered response, we used immunocytochemistry to c-Fos protein, a transcription factor marking neuronal activation. Controls (25% casein diet) and prenatally malnourished (6% casein) adult rats were subjected to 20 min of restraint stress or were unstressed. Plasma corticosterone levels were monitored before and after stress. Paired comparisons of corticosterone levels confirmed that both groups showed a significant post-stress increase. Three hours after onset of stress, rats were perfused with paraformaldehyde. Brain sections were immuno-stained together for c-Fos. Since anterior cingulate and medial prefrontal cortex modulate stress responses, labeled neurons in this region were quantified using unbiased stereology. A 2-way ANOVA of neuron numbers demonstrated a strong effect of stress and a stress by nutrition interaction. Post-hoc comparisons showed that stress significantly increased the number of c-Fos labeled neurons in both nutrition groups. Within the stress condition, prenatally malnourished rats showed a significantly greater number of c-Fos positive neurons than well-nourished rats. These results suggest that neurons in anterior cingulate and medial prefrontal regions respond excessively to restraint stress in prenatally malnourished rats.