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.

Enteric neuronal development in canine small intestine - an immunohistochemical study.

The development of the enteric nervous system (ENS) is still a valid and intensely studied issue. However, literature in the field has no data on this topic in the dog. The present investigations were performed in three groups of fetuses from mongrel dogs - from the third, sixth- -seventh, and ninth week of pregnancy - and in 3-5-day-old puppies (3 specimens for each age group). The tissues (the medial parts of the duodenum, jejunum, and ileum with the cecum and a small portion of the adjacent ascending colon) were cut using a cryostat and the sections were processed for single- and double-labeling immunohistochemistry using antisera against acetylated tubulin (AcTub), vesicular acetylcholine transporter (VAChT), nitric oxide synthase (NOS), vasoactive intestinal polypeptide (VIP), galanin (GAL), neuropeptide Y (NPY), substance P (SP), and calcitonin gene-related peptide (CGRP). In the 3-week-old fetuses, some oval cells invading the gut wall were found. From the seventh week of pregnancy onwards, two different enteric ganglia were present: submucosal and myenteric. The estimated number of nerve elements in the 9-week-old fetuses was much higher than that observed in the 6-7-week-old individuals. There was no significant difference in the estimated number of nerve structures between the 9-week-old fetuses and the 3-5-day-old puppies. The colonization pattern and the development of the ENS in the canine small intestine are very similar to those observed in other mam- mals. However, a few exceptions have been confirmed, regarding the time of appearance of the VIP-, GAL-, and CGRP-immunoreactive neurons, and their distribution in different portions of the canine bowel during development.