During the development of sweat gland innervation, interactions with the target tissue induce a change from noradrenergic to cholinergic and peptidergic properties. To determine whether the change in neurotransmitter properties that occurs in the sweat gland innervation occurs more generally in sympathetic neurons, we identified a new target of cholinergic sympathetic neurons in rat, the periosteum, which is the connective tissue covering of bone, and characterized the development of periosteal innervation of the sternum. During development, sympathetic axons grow from thoracic sympathetic ganglia along rib periosteum to reach the sternum. All sympathetic axons displayed catecholaminergic properties when they reached the sternum, but these properties subsequently disappeared. Many axons lacked detectable immunoreactivities for vesicular acetylcholine transporter and vasoactive intestinal peptide when they reached the sternum and acquired them after arrival. To determine whether periosteum could direct changes in the neurotransmitter properties of sympathetic neurons that innervate it, we transplanted periosteum to the hairy skin, a noradrenergic sympathetic target. We found that the sympathetic innervation of the transplant underwent a noradrenergic to cholinergic and peptidergic change. These results suggest that periosteum, in addition to sweat glands, regulates the neurotransmitter properties of the sympathetic neurons that innervate it.

Understanding the development of cortical interneuron phenotypic diversity is critical because interneuron dysfunction has been implicated in several neurodevelopmental disorders. Here, tyrosine hydroxylase (TH)-immunoreactive neurons in the developing and adult rat cortex were characterized in light of findings regarding interneuron neurochemistry and development. Cortical THimmunoreactive neurons were first observed two weeks postnatally and peaked in number three weeks after birth. At subsequent ages, the number of these cell profiles was gradually reduced, and they were seen less frequently in adults. No DNA fragmentation or active caspase 3 was observed in cortical TH cells at any age examined, eliminating cell death as an explanation for the decrease in cell number. Although cortical TH cells reportedly fail to produce subsequent catecholaminergic enzymes, we found that the majority of these cells at all ages contained phosphorylated TH, suggesting that the enzyme may be active and producing L-DOPA as an end-product. Morphological criteria and colocalization of some TH cells with glutamic acid decarboxylase suggests that these cells are interneurons. Previously, parvalbumin, somatostatin, and calretinin were demonstrated in nonoverlapping subsets of interneurons. Cortical TH neurons colocalized with calretinin but not with parvalbumin or somatostatin. These findings suggest that the transitory increase in TH cell number is not due to cell death but possibly due to alterations in the amount of detectable TH present in these cells, and that at least some cortical TH-producing interneurons belong to the calretinin-containing subset of interneurons that originate developmentally in the caudal ganglionic eminence.

During development, the sympathetic innervation of two targets, sweat glands and periosteum, changes the neurotransmitters it expresses from noradrenaline to acetylcholine and vasoactive intestinal peptide (VIP). The target-derived molecules that induce, these changes have not been identified. Neuropoietic cytokines, including ciliary neurotrophic factor (CNTF) and leukemia inhibitory factor (LIF), induce the same phenotypic changes in sympathetic neurons in vitro as sweat glands and periosteum do in vivo, raising the possibility that one of these factors mediates induction of cholinergic traits and VIP by these target tissues. Because CNTF and LIF have overlapping functions and signalling pathways, they could act interchangeably or in concert to influence neurotransmitter expression. To determine whether CNTF or CNTF and LIF together are responsible for the induction of cholinergic and peptidergic function in vivo, we analyzed the neurotransmitter properties of sweat gland innervation in mice lacking CNTF or CNTF and LIF. We find that, as in wild-type mice, gland innervation in mice lacking one or both molecules appropriately expresses cholinergic properties and VIP immunoreactivity. Furthermore, footpads of mice lacking one or both genes contain choline acetyltransferase activity comparable to that of wild-type mice, and CNTF- or CNTF/LIF-deficient mice possess the normal complement of active sweat glands. We analyzed the innervation of a second, recently identified cholinergic sympathetic target, the periosteum, which is the connective tissue surrounding bone. Periosteal innervation of mice lacking CNTF, LIF, or both, like that of wild-type mice, is immunoreactive for the vesicular acetylcholine transporter, a recently identified cholinergic marker, and VIP. These results provide evidence that neither CNTF, LIF, nor a combination of the two are required for the developmental change from noradrenergic to cholinergic function that occurs in sympathetic innervation of sweat glands and periosteum.