Effect of caloric restriction on myenteric neuroplasticity in the rat duodenum during aging

Abstract

The objective of this study was to evaluate the effects of caloric restriction (CR) on myenteric neurons in the duodenum of Wistar rats during aging. Thirty rats were divided into three groups: the C group (six-month-old animals that were fed a normal diet from weaning until six months of age), the SR group (18-month-old animals that were fed a normal diet from weaning until 18 months of age) and the CR group (18-month-old animals that were fed a 30% CR diet after six months of age). After 12 months, the animals were euthanized. Whole-mount preparations of the duodenums were either stained with Giemsa or underwent NADPH-diaphorase histochemistry to determine the general myenteric neuron population and the nitrergic neuron subpopulation (NADPH-d+), respectively. The NADPH-d-negative (NADPH-d-) neuron population was estimated based on the difference between the Giemsa-stained and NADPH-d+ neurons. The neurons were counted, and the cell body areas were measured. Aging was associated with neuronal loss in the SR group, which was minimized by caloric restriction in the CR group. The density (mm(2)) of the Giemsa-stained neurons was higher in the SR group (79.09 ± 6.25) than in the CR (92.37 ± 11.6) and C (111.68 ± 15.26) groups. The density of the NADPH-d+ neurons was higher in the SR group (44.90 ± 5.88) than in the C (35.75 ± 1.6) and RC (39.14 ± 7.02) groups. The density of NADPH-d- neurons was higher in the CR (49.73 ± 12.08) and C (75.64 ± 17.05) groups than in the SR group (33.82 ± 4.5). In the C group, 32% and 68% of the Giemsa-stained myenteric neurons were NADPH-d+ or NADPH-d-, respectively. With aging (SR group), the percentage of nitrergic neurons (56.77%) increased, whereas the percentage of NADPH-d- neurons (43.22%) decreased. In the CR group, the change in the percentage of nitrergic (42.37%) and NADPH-d- (57.62%) neurons was lower. As NADPH-d- neurons will be mostly cholinergic neurons, CR appears to reduce the loss of cholinergic neurons during aging. The cell body dimensions (μm(2)) were not altered by aging or CR. Thus, CR had a protective effect on myenteric neurons during aging.