Immunohistochemical study on the development of cholinergic and nitrergic nerve structures in the bovine esophageal groove

The proper functioning of the perinatal sucking reflex in calves is essential for the prevention of milk leakage into the rumen. The complex process behind its regulation is mediated at the gut level via multiple excitatory and inhibitory neurotransmitters, of which acetylcholine and nitric oxide are of fundamental importance. The aim of our study was to depict age-related alterations in the cholinergic and nitrergic innervation of the esophageal groove (EG) using immunohistochemistry and Real-Time PCR methods. We found out that the highest number of cholinergic nerve cells was present in the second trimester fetuses. From this developmental stage onward, their amount was gradually decreasing and reached the lowest value in 4-year-old cows. The same developmental pattern was observed for nitrergic nerve structures with the highest percentage of nitrergic neurons in the third trimester fetuses. Our observations prove that both neuronal populations are crucial for a proper closure of EG in calves. Therefore, their contribution to a general neuronal activity in the ENS diminishes with age as the high motility of a gastric groove is not necessarily required in older cattle