Co-expression of tyrosine hydroxylase, dopamine beta-hydroxylase and neuropeptide Y in the sympathetic neurons projecting to the submandibular gland in the sheep

Sympathetic neurons projecting to the ovine submandibular gland (SMG) from the superior cervical ganglion (SCG) and middle cervical ganglion (MCG) were identified using retrograde tracing with fluorescent dye (Fast Blue). Antibodies to tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DH) and neuropeptide Y (NPY) were used to determine the immunochemical characteristics of SMG-innervating sympathetic neurons. Immunohistochemistry combined with the retrograde tracing revealed that the population of SMG-projecting neurons consist of four distinct sub-populations, but taking into account their possible different physiological properties only three major sub-populations can be distinguished. The vast majority of neurons in both ganglia are noradrenergic in nature (co-express TH and DH). All examined TH-immunoreactive (IR) neurons also show immunoreactivity to DH. Sub-population of noradrenergic neurons can be divided into NPY-IR and non-NPY-IR. Noradrenergic neurons expressing NPY may act as vasoconstrictors. The second sub-population of SMG projecting neurons in the ganglia studied consists of non-noradrenergic neurons (containing NPY, but not TH). It is known that these kinds of neurons may play a vasodilatory role. In both examined ganglia the third sub-population consists of non-TH-IR and non-NPY-IR neurons of unknown physiological function. Since no DH immunoreactivity was found in any of TH– neurons these nerve cells can also be regarded as non-noradrenergic. It is possible that some neurons of the second as well as the third sub-population are cholinergic and some of them are non-noradrenergic/non-cholinergic in character.