Undernutrition during early life increases the level of apoptosis in the dentate gyrus but not in the CA2 + CA3 region of the hippocampal formation

Abstract

We have previously shown that undernutrition during early life causes a permanent deficit in the total number of dentate granule cells. However, it is unknown whether this deficit is due to neuronal cell death and/or to fewer cells being born during the period of neurogenesis. We have therefore used stereological methods combined with specific labeling techniques to examine the numbers of apoptotic cells in specific regions of the hippocampal formation. Rats were undernourished by restricting their daily food intake to about half that eaten by well-fed controls. Control and undernourished rats were killed on postnatal day 21, and their brains fixed in 4% paraformaldehyde. Serial sections through the hippocampal formation were labeled with the TUNEL technique to distinguish apoptotic cells. All care and animal handling procedures were approved by the institutional Animal Ethics Committee in line with Australian NHMRC procedures. There were about 21,500 and 57,000 TUNEL-positive cells in the dentate gyrus granule cell layer of control and undernourished rats, respectively. The difference between these values was statistically significant. In the CA3 + CA2 region, there were about 22,000 and 19,500 TUNEL-positive cells in control and undernourished rats, respectively. The difference between these values was not statistically significant. Furthermore, it was observed that the majority of the TUNEL-positive cells in the dentate gyrus were located close to the border between the dentate gyrus granule cells and hilus of the hippocampal formation. Our results show that undernutrition during gestation and lactation can result in an increase in the level of TUNEL-positive apoptotic cells in the rat dentate gyrus.