ABSTRACT
The neuropeptides vasoactive intestinal peptide (VIP) and pituitary adenylyl cyclase-activating peptide (PACAP) are induced strongly in neurons after several types of injury, and exhibit neuroprotective actions in vitro and in vivo. It is thought that changes in expression of neuropeptides and other molecules in injured neurons are mediated by new factors produced in Schwann and immune cells at the injury site, a loss of target-derived factors, or a combination of mediators. To begin to determine the role of the inflammatory mediators, we investigated axotomy-induced changes in VIP and PACAP gene expression in the facial motor nucleus in severe combined immunodeficient (SCID) mice, and in mice with targeted mutations in specific cytokine genes. In normal mice, VIP and PACAP mRNA was induced strongly in facial motor neurons 4 days after axotomy. The increase in PACAP mRNA was blocked selectively in SCID mice, indicating that mechanisms responsible for VIP and PACAP gene induction are not identical. The loss of PACAP gene expression in SCID mice after axotomy was fully reversed by an infusion of normal splenocytes, suggesting that PACAP mRNA induction requires inflammatory mediators. PACAP and VIP mRNA inductions, however, were maintained in mice lacking leukemia inhibitory factor (LIF) and interleukin-6 (IL-6), and in mice lacking both receptors for tumor necrosis factor alpha (TNFalpha). The data suggest that an inflammatory response, most likely involving T lymphocytes, is necessary for the axotomy-induced increase in PACAP but not in VIP. LIF, IL-6, and TNFalpha, however, are not required for this response to injury.
Subject(s)
Facial Nerve Injuries/immunology , Gene Expression Regulation/immunology , Motor Neurons/metabolism , Neuropeptides/biosynthesis , T-Lymphocytes/immunology , Animals , Axotomy , Cytokines/deficiency , Cytokines/genetics , Facial Nerve/immunology , Facial Nerve/metabolism , Facial Nerve/physiopathology , Facial Nerve Injuries/genetics , Facial Nerve Injuries/physiopathology , Inflammation/genetics , Inflammation/immunology , Inflammation/physiopathology , Male , Mice , Mice, Knockout , Mice, SCID , Motor Neurons/pathology , Mutation/genetics , Neuropeptides/genetics , Neuropeptides/metabolism , Pituitary Adenylate Cyclase-Activating Polypeptide , RNA, Messenger/metabolism , Rats , Rats, Wistar , Retrograde Degeneration/genetics , Retrograde Degeneration/immunology , Retrograde Degeneration/physiopathology , Transcriptional Activation , Up-Regulation/genetics , Vasoactive Intestinal Peptide/metabolismABSTRACT
The detailed mRNA distributions of pituitary adenylyl cyclase-activating polypeptide (PACAP) and its selective type I receptor (PAC(1)) were systematically compared in the brain of the frog Xenopus laevis. PACAP mRNA expression overlapped with that of PAC(1) in many brain areas such as the pallium, hypothalamic preoptic area, ventral hypothalamic nuclei, habenular nucleus, most thalamic nuclei, the cerebellular nucleus, and nuclei of isthmi. In some structures, PACAP and PAC(1) gene transcripts were present in anatomically distinct cell layers. For example, in the olfactory bulb, PACAP mRNA was present in the mitral cell layer, whereas gene transcripts for the receptor were observed in the granule layer. In a number of regions, expression showed no obvious overlap. PAC(1) but not PACAP mRNA was present at moderate levels in the Purkinje cell layer of the cerebellum and distal lobe of the pituitary. Conversely, PAC(1) gene expression was absent in the spinal cord while PACAP mRNA signals were observed in the medial portion of the ventral horn and deep portion of the dorsal horn. The granule and molecular cell layer of the cerebellum, alpha-motor neurons in the spinal cord, and reticular nucleus of isthmi showed neither PACAP nor PAC(1) gene transcripts. These localized patterns of ligand and receptor gene expression suggest possible PACAP projection and target fields in the frog brain.
