Nitric oxide synthase in ventral forebrain grafts and in early ventral forebrain development

Embryonic ventral forebrain (VFB) grafts to cortex contain neurons that synthesize acetylcholine and partially ameliorate behavioral deficits caused by excitotoxic damage to the nucleus basalis magnocelullaris in rats. An additional neurotransmitter, nitric oxide (NO), is synthesized by a subset of cholinergic neurons in rat ventral forebrain. If this neurotransmitter is expressed also by grafted cholinergic neurons (which include the embryonic medial septum and diagonal band), its functional contribution should be considered. Six to twelve months after transplantation of embryonic VFB tissue rats were sacrificed. Brain tissue was processed either for in situ hybridization of nNOS and neuropeptide Y (NPY) or for immunohistochemistry of choline acetyltransferase (ChAT) and neuronal nitric oxide synthase (nNOS). Quantification of messenger ribonucleic acid (mRNA) for nNOS was performed with radioactively labeled probes (silver grains were counted) and a preliminary comparison was made of graft sections to sections of the ventral forebrain of developing rats. Plots of silver grain counts against cell size revealed similar patterns in the grafts and in the ventral forebrain of developing rats. The rates of expression of mRNA for nNOS in the grafts were intermediate between those of the ventral forebrain of postnatal day 19 and those of postnatal day 12. Double immunohistochemical labeling revealed that 45.87 + 8.26% of cells expressing ChAT also expressed nNOS in the grafts, significantly higher than 33.16 + 3.9% which was the rate of co-expression observed in the adult ventral forebrain. This study suggests that possible contribution of NO to graft-associated modulation of behavior should be examined.