To evaluate the effects of protein and tryptophan restriction on cholinergic neurotransmission in terms of choline acetyltransferase (CAT) activity and its expression as well as muscarinic receptors number at cerebral cortex and hippocampus, Wistar rats were raised on a chronically protein and tryptophan restricted diets with 8% protein based on either Purina chow or corn. There was a significant decrease in both body and cerebral weight in the restricted animals compared with the control group fed with 23% protein diet. In cerebral cortex CAT activity and its expression were significantly increased in corn fed (C) versus protein restricted (HP) and control (T) animals, without no significant changes in muscarinic receptors number. On the other hand, hippocampus CAT activity and its expression were significantly lesser and muscarinic receptors significantly increased in C group and CAT activity in HP group was significantly increased without significant changes in muscarinic receptors related to T group. These results indicate that a reduction in 5-HT disponibility on brain induced by tryptophan restricted diet during development affects clearly the cholinergic system modifying probably the short term memory and learning.