Innervation of the endolymphatic sac.

Previous studies suggest that the endolymphatic sac plays an important role in the homeostasis of endolymph. Factors that influence blood flow in the sac may affect its function. This blood flow may be influenced by autonomic innervation; however, no such innervation has been demonstrated. The purpose of this study was to demonstrate catecholaminergic and cholinergic fibers on the endolymphatic sac. Endolymphatic sacs from Hartley guinea pigs were stained either immunocytochemically for tyrosine hydroxylase to reveal catecholaminergic fibers or histochemically for acetylcholinesterase to reveal cholinergic fibers. For tyrosine hydroxylase immunostaining, the endolymphatic sacs were treated with dilute hydrogen peroxide and then incubated in the primary antiserum. The tissue was further processed by the avidin-biotin immunoperoxidase method and reacted with diaminobenzidine. For acetylcholinesterase histochemistry, the tissue was processed by a modification of the direct thiocholine method. Light microscopy of the whole-mounted endolymphatic sacs revealed tyrosine hydroxylase-positive and acetylcholinesterase-positive fibers. Some of the acetylcholinesterase-positive fibers were clearly associated with vessels. This innervation, which has not been described previously, may significantly influence blood flow and function of the endolymphatic sac.

Cholinergic innervation of the guinea pig tympanic membrane.

While cholinergic nerve fibers of the parasympathetic system have been demonstrated in the middle ear mucosa, such innervation of the tympanic membrane has never been shown. Such fibers may prove important since the tympanic membrane may be one of the initial sites of effusion production, and since parasympathetic innervation is thought to be involved with middle ear effusion. To demonstrate cholinergic innervation, we have used modified direct thiocholine histochemical staining. Anesthetized Hartley guinea pigs were killed, and the tympanic bullae were removed intact, fixed in 4% paraformaldehyde, and then stained whole. Following staining, the tympanic membrane was dissected from each bulla and whole-mounted for light microscopy. Numerous acetylcholinesterase-positive fibers were observed on the tympanic membrane. Some fibers appeared to be vessel associated, although the majority did not. This suggests that these fibers may act on the mucosa or vasculature of the tympanic membrane and contribute to the pathogenesis of middle ear effusion.