Development of choline acetyltransferase (CAT) in the sympathetic innervation of rat sweat glands. (1/2)

It has been postulated that the developing sympathetic innervation of rat eccrine sweat glands changes from adrenergic to cholinergic under the influence of its target. In agreement with previous evidence that the sympathetic innervation of adult rat sweat glands is cholinergic, we found that choline acetyltransferase (CAT)-immunoreactive nerve fibers are present in adult glands, and that gland-rich chunks of adult footpads contain CAT enzyme activity. We were therefore interested in determining when CAT activity is first expressed in the developing gland innervation. Low levels of acetylating activity were observed in rat footpads as early as postnatal day 4, when sympathetic fibers first contact the glands. A greater than fourfold increase in CAT specific activity occurred between postnatal days 11 and 21. Neonatal treatment of rats with the adrenergic neurotoxin 6-hydroxydopamine (6-OHDA) eliminated most of the CAT activity in 14 and 19 d footpads. In contrast, the acetylating activity observed prior to day 11 was unaffected by neonatal 6-OHDA treatment, and only slightly reduced by the selective CAT inhibitor, naphthylvinylpyridine. These results indicate that the sympathetic fibers that innervate rat sweat glands do not acquire detectable levels of CAT activity until a full week after they contact the glands.  (+info)

Acetylcholine concentration and its role in ionic transport by the corneal epithelium. (2/2)

Corneal epithelium is known to have a high acetylcholine (ACh) concentration, but its role remains uncertain. Furthermore, rabbit corneal epithelium is devoid of cholinergic receptors. ACh concentration in calf, rabbit, and frog corneal epithelium was determined with Fellman's fluorometric assay to be 16.9, 11.1, and 21.8 micrograms of ACh per gram of epithelium, respectively. The isolated frog cornea was used to examine a possible role of the cholinergic system on active ionic transport. A 2 mM concentration of exogenous ACh has a moderate inhibitory effect on Na transport but no effect on Cl transport. A 10(-4)M concentration of 4-(1-naphthylvinyl) pyridine (NVP), a choline acetyltransferase inhibitor, reversibly inhibited both Na and Cl transport by about 70%. NVP also reduced ACh content of frog corneal epithelium by 51%. Thus endogenous ACh, but not exogenous ACh, seems to be stimulatory of active ionic transport. Of several muscarinic or nicotinic blockers tested, 10(-3)M atropine inhibited Na transport by 55% and Cl transport by 83%; 10(-3)M nicotine inhibited Na transport by 33% and Cl transport by 17%. If frog cornea (like rabbit cornea) contains no cholinergic receptors, the effects of ACh, nicotine, and atropine on ionic transport may be mediated through a nonspecific pathway.  (+info)