Circadian regulation of c-fos expression in the suprachiasmatic pacemaker by light.

The primary objective of this research was to examine expression of the immediate-early gene c-fos within the suprachiasmatic nucleus (SCN) for evidence of circadian regulation by photic stimuli. In vivo and in vitro analyses demonstrate that photic signals have an inductive effect on c-fos expression in the SCN, but only at critical times when light is capable of phase-shifting circadian rhythms. This evidence for correlative relations between the effects of light signals in inducing c-fos gene expression in the SCN and modulating the circadian period of the SCN pacemaker suggests that immediate-early genes may be components of the signal transduction cascade by which light entrains circadian rhythms. In addition, dual-immunostaining methods were utilized to examine neurochemical identity of SCN cells that exhibit this circadian induction of c-fos expression in response to light. Within the ventrolateral SCN, the photic induction of Fos expression occurred in neurons expressing gastrin-releasing peptide (GRP), but not in those containing vasoactive intestinal polypeptide (VIP). This finding suggests that SCN neurons containing GRP may be involved in the transduction of photic signals mediating circadian entrainment.

Postnatal development of cholinergic neurotransmitter enzymes in the mouse cerebellum. Biochemical, light microscopic and electron microscopic cytochemical investigations.

The activity and distribution of the cholinergic neurotransmitter enzymes acetylcholinesterase (AChE) and choline acetyltransferase (ChAc) in the developing cerebellum of the mouse were investigated using biochemical assays and light microscopic histochemistry for AChE and ChAc, and electron microscopic histochemistry for AChE. Postnatal alterations in the levels of AChE and ChAc in the cerebellum of the mouse are characterized by a divergent pattern. During the first two postnatal weeks, AChE activity increases progressively, whereas ChAc activity remains low. Beyond day 14, when AChE activity is steady or gradually decreasing, ChAc increases sharply to reach a peak on day 32. Histochemically, AChE activity is associated with the glomeruli and the Golgi cells of the internal granular layer, the medullary layer, and the deep cerebellar nuclei. Purkinje cells exhibit transient staining between days 2 and 9. At the structural level, AChE staining is first demonstrated on day 6 within the Golgi cell soma, on day 9 at the mossy fiber-granule cell synapse, and on day 11 at the Golgi terminal-granule cell synapse. The staining intensity of these structures reaches that of the adult on day 19. The histochemical reaction for ChAc is localized to moderate number of presumed Bergmann glial cells, a few large cells of the deep cerebellar nuclei, small numbers of Golgi cells, and all immature Purkinje cells. The molecular distribution of ChAc sharply contrasts with the localization established by other methods previously and is interpreted in the light of the drawbacks of the histochemical procedure. The biochemical fluctuations in AChE activity, correlated with the histochemical and cytochemical data, suggest that the postnatal increases in the enzyme are related to the ingrowth of the mossy fibers and to the maturation of the AChE-positive Golgi cells. Histochemical evidence for the correlation between the ontogenetic increases in cerebellar ChAc activity and progressive mossy fiber innervation must await the application of the immunohistochemical method to the developing cerebellum.

The subnuclear distribution of substance P, cholecystokinin, vasoactive intestinal peptide, somatostatin, leu-enkephalin, dopamine-beta-hydroxylase, and serotonin in the rat interpeduncular nucleus.

The distribution of immunofluorescent somata and processes within the interpeduncular nucleus (IPN) containing substance P (SP), cholecystokinin (CCK), vasoactive intestinal peptide (VIP), somatostatin (SST), leu-enkephalin (L-ENK), dopamine beta hydroxylase (DBH), and serotonin (5HT) was examined in male rats treated with colchicine 48 hours prior to perfusion. Serial sections were examined for immunofluorescence and variations in the density of fluorescence rated 1 + (sparse) to 4 + (dense). The rostral subnucleus contained SP, SST, and L-ENK-positive somata and processes. Substance P and VIP processes were present throughout the rostral subnucleus but were concentrated in two ovoid areas located dorsally in the caudal region of this subnucleus. Cholecystokinin and L-ENK processes surrounded these ovoid areas. The entire width of the central subnucleus was crossed by SP and L-ENK processes oriented horizontally in narrow bands. Substance P processes were also aligned into vertical columns adjacent to the lateral margins of the central subnucleus. Leu-enkephalin and 5HT processes were distributed throughout this subnucleus, while VIP processes were present only caudally. Dopamine beta hydroxylase processes were evenly distributed but were restricted from the vertical columns laterally. The intermediate subnuclei contained a sparse density of SP and 5HT processes that were present in proximity to the major blood vessels penetrating this subnucleus. Only DBH processes were evenly distributed. The lateral subnuclei contained a dense concentration of SP processes. The medial edges of this subnucleus were distinguished by VIP, CCK, L-ENK, and 5HT processes. The dorsal subnucleus contained 5HT, L-ENK, and SST-positive somata and processes. Substance P, VIP, CCK, and DBH processes were also present. Dorsal-lateral subnuclei contained SP, SST, L-ENK, and DBH processes. Interstitial subnuclei contained SP, CCK, L-ENK, 5HT, and DBH processes. This study demonstrates that perikarya and processes containing peptides and monoamines are distributed within subnuclei of IPN in a topographic and heterogeneous pattern. New features of IPN organization are revealed.

Expression of ICAM-1, VCAM-1, L-selectin, and leukosialin in the mouse central nervous system during the induction and remission stages of experimental allergic encephalomyelitis.

Adhesion molecules facilitate infiltration of leukocytes into the central nervous system (CNS) of mice with experimental allergic encephalomyelitis (EAE). Expression of the adhesion molecules ICAM-1 (CD54), VCAM-1 (CD106), L-selectin (CD62L), and leukosialin (CD43) was analyzed via immunocytochemistry 4-28 days after the injection of encephalitogen into EAE-susceptible SWXJ mice. Constitutive ICAM-1 expression on large-diameter CNS vessels was upregulated on post-injection days 8, 11, 14 and 18 (concurrent with de novo expression on smaller capillaries and glial cells), partially downregulated by day 23, and back to control levels by day 28. Constitutive VCAM-1 expression was upregulated by day 14 and back to control levels by day 28. Upregulation of ICAM-1 temporally coincided with the immigration of CD4+ lymphocytes and L-selectin+ leukocytes into the CNS, while downregulation coincided with their emigration. The infiltration of CD43+ leukocytes also coincided with the upregulation of vascular adhesion molecules, but CD43+ cells remained in the CNS after ICAM-1 and VCAM-1 had returned to control levels. Cellular infiltration and adhesion-molecule expression preceded EAE clinical symptoms by a minimum of 3 days, suggesting a causal role of adhesion molecules in the initiation of CNS inflammation. However, prophylactic injections of monoclonal antibodies against either ICAM-1, L-selectin, or CD43, did not ameliorate the clinical severity of EAE in this model.

Enhanced glial activation and expression of specific CNS inflammation-related molecules in aged versus young rats following cortical stab injury.

Aging is associated with increased glial responsiveness that may enhance the brain's susceptibility to injury and disease. To determine whether unique age-related molecular responses occur in brain injury, we assessed mRNA levels of representative central nervous system (CNS) inflammation-related molecules in young (3 months) and aged (36 months) Fisher 344/Brown Norwegian F1 hybrid rats following cortical stab. Enhanced glial activation in older animals was accompanied by increased expression of a subset of inflammation-related mRNAs, including IL-1beta, TNFalpha, IL-6, ICAM-1, inducible nitric oxide synthase (iNOS), metalloproteinase-9 (MMP-9), and complement 3alpha-chain 1 (C3alpha1). Recognition of these age-specific differences may guide development of novel treatment regimes for older individuals.

TNF alpha and IL-1beta mediate intercellular adhesion molecule-1 induction via microglia-astrocyte interaction in CNS radiation injury.

Radiation injury to the central nervous system (CNS) results in glial activation accompanied by expression of pro-inflammatory cytokines and adhesion molecules. In this study we demonstrate intercellular adhesion molecule-1 (ICAM-1) induction in the irradiated mouse brain at the mRNA and protein levels. Immunocytochemical analysis revealed that ICAM-1 protein was primarily expressed in endothelial cells and microglia. In vitro, ionizing radiation significantly induces TNF alpha, IL-1beta and ICAM-1 mRNA in primary microglia cultures. Interestingly, although ionizing radiation activated primary astrocyte cultures, it did not induce ICAM-1 expression. However, exposure of astrocytes to conditioned medium collected from irradiated microglia resulted in ICAM-1 induction, which was abrogated when the conditioned medium was pre-incubated with neutralizing antibodies raised against murine TNF alpha and IL-1beta. These results indicate that pro-inflammatory cytokines may be necessary for ICAM-1 expression in astrocytes in CNS radiation injury.

A system of corticotropin releasing factor-containing amacrine cells in the rat retina.

Immunohistochemical processing of Long-Evans retina wholemounts using an antiserum directed against rat, human corticotropin releasing factor revealed a group of immunoreactive amacrine cells. Two subpopulations could be distinguished based primarily on the location of their cell bodies. One subpopulation had cell bodies situated along the junction of the inner nuclear layer and the inner plexiform layer. The other subpopulation had cell bodies in the ganglion cell layer. The latter was judged to be displaced amacrine cells since double-label experiments indicated that the pattern of corticotropin releasing factor-like immunoreactive staining in the ganglion cell layer did not coincide with that of ganglion cells labeled retrogradely with fluorogold. Corticotropin releasing factor-like immunoreactive amacrine cells on either side of the inner plexiform layer emitted processes which ramified extensively in sublamina 5 and, to a lesser degree, in sublamina 4. A minority of these cells also sent a single process to ramify in sublamina 1. Throughout the retina, corticotropin releasing factor-like immunoreactive cells were distributed relatively evenly, with a tendency to peak in the superior temporal region. Despite the anatomical classification into two subpopulations, it is proposed that the corticotropin releasing factor-like immunoreactive cells are functionally one system, influencing preferentially synaptic interactions associated with the inner half of the inner plexiform layer. The results of this study provide anatomical basis for further investigations of corticotropin releasing factor as a putative peptidergic neurotransmitter in the retina.

Ultrastructural demonstration of noradrenergic synapses in the rat central nervous system by dopamine-beta-hydroxylase immunocytochemistry.

Noradrenergic (NA) cell bodies and axonal processes were identified in the electron microscope by the immunocytochemical localization of the norepinephrine-synthesizing enzyme, dopamine-beta-hydroxylase (DBH). DBH immunoreactivity, visualized by the peroxidase-antiperoxidase method, was observed in the somata and proximal processes of locus coeruleus neurons and in the distal axons of several NA terminal fields. DBH immunoreactivity is distributed throughout the cytoplasm of the NA neuron, but demonstrates a selective association with endoplasmic reticulum, Golgi apparatus, mitochondrial outer membranes, large granular vesicles, and small, round synaptic vesicles. DBH-positive axonal profiles, typically interspersed between unlabeled dendrites, form two distinct populations: a) thin, unmyelinated intervaricose segments (ca. 0.28 micron) and b) spherical varicosities (ca. 1.00 micron). No DBH-positive varicosities were observed in contact with intracerebral capillaries. In order to determine whether or not NA axons typically form synaptic contacts, a quantitative analysis of selected areas of the diencephalon, cerebellum, and limbic cortex was carried out. More than half (58%) of all DBH-positive varicosities form axodendritic synapses characterized by specialized junctional appositions. The results suggest that NA neurons typically exert their influence on other neurons through highly restricted synaptic contacts.

Induction of interleukin-1 beta mRNA in adult rat brain.

The ability of the central nervous system to produce the cytokine interleukin-1 beta (IL-1 beta) in response to challenge by activators of the mononuclear phagocyte system has been examined in vivo. Unilateral injection of a mixture of gamma-interferon (IFN-gamma) and lipopolysaccharide (LPS) into the forebrain of adult rats induced expression of IL-1 beta mRNA. In situ hybridization of IL-1 beta mRNA showed a gradient of cellular hybridization, which was most intense at the site of IFN-gamma/LPS injection. The reverse transcription--polymerase chain reaction (RT-PCR) was used to demonstrate the presence of IL-1 beta mRNA in normal rat brain, and to confirm increases in IL-1 beta mRNA levels following IFN-gamma/LPS injection. These studies show that IL-1 beta can be induced to high levels within the CNS as a consequence of exposure to potent stimulators of macrophage activation.

The coexistence and release of bovine pancreatic polypeptide-like immunoreactivity from noradrenergic superior cervical ganglia neurons.

The coexistence of bovine pancreatic polypeptide-like (BPP) immunoreactivity within the cell bodies and axons of the superior cervical ganglia (SCG) was studied. Adjacent sections stained by the indirect immunofluorescence technique for either BPP or dopamine-beta-hydroxylase (DBH) revealed that virtually all (90-95%) SCG cells contained DBH and a subpopulation (30-50%) also contained BPP. Ligation of the pre- and postganglionic nerves of the SCG demonstrated BPP-positive fibers emanating from the SCG via both axonal trunks. BPP-containing fibers were also observed entering the SCG via the preganglionic nerve. The peptide was co-released with catecholamine by electrical stimulation of the preganglionic cervical sympathetic nerve trunk, but was not effected by reserpine. The extensive distribution and unique coexistence of a PP-like peptide with peripheral sympathetic nerves suggests a neuromodulatory role in autonomic functions.

Bovine pancreatic polypeptide-like immunoreactivity in brain and peripheral nervous system: coexistence with catecholaminergic nerves.

The rat central and peripheral nervous system contains a widespread distribution of BPP-like immunoreactive neurons. Some of these neurons coexist with a catecholamine, probably mostly NE. This peptide appears to be releaseable by nerve stimulation. Catecholamine releasing agents such as reserpine do not appear to deplete the BPP. This extensive distribution and unique coexistence of a peptide with peripheral sympathetic nerves suggests a neuromodulatory role of BPP in autonomic functions.

The distribution of bovine pancreatic polypeptide-like immunoreactive neurons in rat brain.

Using the indirect immunofluorescent technique, bovine pancreatic polypeptide (BPP)-like immunoreactive nerve fibers and cell bodies were observed widely distributed in rat brain. A detailed stereotaxic atlas of BPP-immunoreactive neurons was prepared. Large numbers of BPP-containing perikarya were observed in the acute nucleus, with scattered cells in the cerebral cortex, nucleus olfactorius anterior, nucleus tractus diagonalis, olfactory tubercle, nucleus accumbens, neostriatum, nucleus interstitialis stria terminalis, nucleus preopticus medialis, area retrochiasmatica, zona interna of the median eminence, substantia grisea centralis, locus coeruleus, nucleus tractus solitarius, and in the region of the nucleus reticularis lateralis. Large numbers of varicose BPP-like nerve fibers were observed in the following nuclei: accumbens, interstitialis stria terminalis, preopticus medialis, preopticus suprachiasmaticus, suprachiasmaticus, periventricular thalamic and hypothalamic, paraventricularis, dorsomedialis, ventromedialis, arcuatus, parabrachialis dorsalis, tractus solitarius and the substantia gelatinosa trigemini. The present findings suggest that a BPP-like peptide may be involved in significant neuronal circuitry, possibly in a neurotransmitter or neuromodulator role. However, the exact identity of this peptide and its physiological role remain to be determined.

The development of peptide-containing neurons within neocortical transplants in adult mice.

Transplantation of embryonic neocortex into adult host neocortex leads to the survival of many donor cells, with the subsequent differentiation of the cortical neurons within a loosely laminated cellular pattern. We wanted to know whether peptide-containing neurons that are known to exist in normal neocortex would survive in the transplants, and if so, whether they would differentiate into morphological cell types that normally contain these peptides in cortex. By 30 days after transplantation, the implants were well vascularized and the donor neurons appeared healthy in Nissl-stained preparations. AChE-positive axons grew across the interface and innervated the transplant in moderate densities. Immunocytochemical localization of peptides in the transplant revealed that processes containing the four peptides normally present in cortex also develop in the transplants. These were vasoactive intestinal polypeptide, cholecystokinin, pancreatic polypeptide and somatostatin. Other peptides not yet demonstrated in and presumably not present in neocortex, did not develop in the transplants. These included alpha-melanocyte stimulating hormone, arginine-vasopressin, corticotropin releasing factor, beta-endorphin and substance P. The results demonstrate that peptide-immunoreactive neurons survive in neural transplants, where they develop complicated patterns of axonal arborization. The conditions used in these experiments produced no evidence that peptidergic neurons within the transplant grow out of the transplant and into the host brain within six weeks. Similarly, host peptidergic axons were never seen crossing the interface zone and entering the transplant in any significant numbers.

Histochemical localization of FMRFamide-like immunoreactivity in the rat brain.

Molluscan cardioexcitatory neuropeptide or FMRFamide is present in the invertebrate central nervous system (CNS) and FMRFamide like peptide has been demonstrated in the mammalian CNS. In this study, the distribution of FMRFamide immunoreactivity was studied in rat brain using the indirect immunofluorescent method. The highest number of FMRFamide staining cell bodies was found in the nucleus (n) arcuatus. N. paraventricularis, n. hypothalamus, n. ventromedialis, n. dorsomedialis and n. tractus solitarii also contained high numbers. FMRFamide positive nerve fibers and terminals were widely distributed. The septal complex contained high densities, especially in n. interstitialis striae terminalis. N. paraventricularis hypothalami, n. paraventricularis, n. hypothalamicus, n. ventromedialis and n. dorsomedialis showed a high to very high degree of immunoreactivity. In myelencephalon, n. tractus solitarii had the densest innervation. Spinal cord had a dense band of FMRFamide positive fibers in lamina I and II of the dorsal horn. The present findings support a neurotransmitter role for a FMRFamide like peptide in the mammalian brain, possibly related to endocrine and autonomic regulation as well as pain modulation.

Behavioral investigation of the coexistence of substance P, corticotropin releasing factor, and acetylcholinesterase in lateral dorsal tegmental neurons projecting to the medial frontal cortex of the rat.

Colocalization of substance P (SP), corticotropin releasing factor (CRF), and acetylcholinesterase (AChE) was detected by retrograde tracing and immunocytochemical staining in the nucleus tegmentalis dorsalis lateralis (ntdl) projecting to the medial frontal cortex (MFC), septum, and thalamus of the rat. The histochemical results suggest that SP and CRF coexist within a subpopulation of ntdl cholinergic neurons that project to a number of forebrain regions including the MFC. Behavioral studies of the effects of SP, CRF, and the cholinergic agonist, carbachol, employed microinjections into the MFC of rats. SP and CRF did not elicit any behavioral effects when administered alone. Carbachol (1-5 micrograms/side) produced a stereotyped motor behavior, consisting of rapid forepaw treading while in an upright posture, resembling "boxing." SP (1 micrograms/side) increased carbachol-induced "boxing." CRF (1-10 ng/side) decreased carbachol-induced "boxing." One possible functional significance of the coexistence of SP, CRF, and acetylcholinesterase, in neurons projecting to the medial frontal cortex in rats, appears to be a modulatory potentiation of cholinergic response by SP, and a modulatory inhibition of the cholinergic response by CRF.

The distribution of corticotropin releasing factor-like immunoreactive neurons in rat brain.

Using the indirect immunofluorescent technique, corticotropin releasing factor (CRF)-like immunoreactive nerve fibers and cell bodies were observed to be widely distributed in rat brain. A detailed stereotaxic atlas of CRF-like immunoreactive neurons was prepared. Large numbers of CRF-containing perikarya were observed in the nucleus paraventricularis, with scattered cells in the following nuclei: accumbens, interstitialis stria terminalis, preopticus medialis, supraopticus, periventricularis hypothalami, amygdaloideus centralis, dorsomedialis, substantia grisea centralis, parabrachialis dorsalis and ventralis, tegmenti dorsalis lateralis, vestibularis medialis, tractus solitarius and reticularis lateralis. The most intense staining of CRF-containing fibers was observed in the external lamina of the median eminence. Moderate numbers of CRF-like fibers were observed in the following nuclei: lateralis and medialis septi, tractus diagonalis, interstitialis stria terminalis, preopticus medialis, supraopticus, periventricularis thalami and hypothalami, paraventricularis, anterior ventralis and medialis thalami, rhomboideus, amygdaloideus centralis, habenulae lateralis, dorsomedialis, ventromedialis, substantia grisea centralis, cuneiformis, parabrachialis dorsalis and ventralis, tegmenti dorsalis lateralis, cerebellum, vestibularis medialis, reticularis lateralis, substantia gelatinosa trigemini and lamina I and II of the dorsal horn of the spinal cord. The present findings suggest that a CRF-like peptide may be involved in a neurotransmitter or neuromodulator role, as well as a hypophysiotropic role.

Dehydration induces Fos, but not increased vasopressin mRNA in the supraoptic nucleus of aged rats.

Dehydration induces Fos expression and increases the length of the vasopressin (VP) mRNA poly-A tail and the content of VP mRNA in the supraoptic (SON) and paraventricular nuclei (PVN) of the hypothalamus. The current studies were performed to evaluate the effect of aging on these responses. Fischer 344 rats of 4, 14, and 28-30 months of age were either water deprived for 72 h or allowed ad libitum access to water. Fos induction in the SON and PVN was examined by immunocytochemistry in order to provide an index of cellular activation. VP mRNA content and size was examined in SON by Northern analysis as an index of VP synthetic capacity. Dehydration induced the expected increase in plasma osmolality in all three ages, however, serum VP was only increased in the 4- and 14-month-old rats. The increase in serum VP was accompanied by a decrease in VP content of the posterior pituitary (PP) in the dehydrated 4- and 14-month-old rats. PP VP content was reduced in both the hydrated and dehydrated old rats relative to the other ages (P = 0.0007). Fos was induced in both SON and PVN of all water deprived rats regardless of age. The density of Fos staining was increased in both nuclei following dehydration (SON, P = 0.002; PVN, P = 0.0001). There was also a significant increase in the number of cells expressing Fos in both nuclei in the dehydrated animals (SON, P = 0.002; PVN, P = 0.0056). There was no significant effect of age on the density of Fos staining. In contrast, dehydration failed to elicit the expected increase in VP mRNA size and content in the SON of the aged dehydrated rats although both of these parameters were increased in the 4- and 14-month-old rats (P < 0.05). Thus, the inability of old Fischer rats to increase serum VP during chronic dehydration is not caused by decreased activation of the neurons (as indicated by Fos induction), but apparently reflects depletion of PP stores of VP due to an inability to increase the amount of VP mRNA available for translation.

Light induces expression of fos-related proteins within gastrin-releasing peptide neurons in the rat suprachiasmatic nucleus.

The present study was conducted to determine whether the photic induction of c-fos expression in the rat suprachiasmatic nucleus (SCN) occurs within neurons containing gastrin-releasing peptide (GRP) and/or vasoactive intestinal polypeptide (VIP) because these peptidergic cells are closely associated with retinal projections to the ventrolateral subfield. Using dual immunostaining and thin sectioning techniques, the ventrolateral SCN of light-exposed rats was examined for evidence of individual neurons coexpressing nuclear immunostaining for c-fos proteins (Fos) with cytoplasmic immunoreactivity for GRP or VIP. In all animals, the photic induction of Fos expression in the SCN was mainly confined to cells segregated within the ventrolateral subfield and was evident in approximately 40% of the SCN neurons with cytoplasmic immunoreactivity for GRP. However, neurons coexpressing Fos and GRP comprised only a small fraction of the total number of cells within the ventrolateral SCN exhibiting light-induced Fos immunoreactivity. No sign of Fos expression was detected within VIP-immunoreactive perikarya in the ventrolateral SCN of light-treated rats. These results demonstrate that light induces Fos expression in a number of GRP-containing neurons within the SCN, suggesting that these peptidergic cells may process photic information mediating circadian entrainment.

Increase of interleukin-1beta mRNA and protein in the spinal cord following experimental traumatic injury in the rat.

Interleukin-1 beta (IL-1beta) is a major mediator of inflammation and a growth promoter for many cell types that could play an important role in the consequences of traumatic spinal cord injury (SCI). In the present study, the expression of IL-1beta and its mRNA was determined in the rat spinal cord following a standardized contusion injury. IL-1beta mRNA, measured with quantitative RT-PCR, was significantly increased in the lesion site by 1 h after SCI (35.2 +/- 5.9 vs. 9.1 +/- 2.1 pg/mg RNA, n = 3, P < 0.05) and remained significantly higher than in the normal spinal cord for at least 72 h post-injury (p.i.). IL-1beta mRNA levels in tissue immediately caudal to the lesion site did not change after the injury. IL-1beta protein levels, measured by an ELISA, were determined at the lesion site and in cerebrospinal fluid (CSF) and serum samples. IL-1beta levels in the CSF and serum were much lower than in the spinal cord. At the lesion site, IL-1beta was increased significantly by 1 h p.i., peaked at 8 h (32.3 +/- 0.1 vs. 7.6 +/- 1.9, ng/g tissue, n = 5, P < 0.05) and remained significantly higher than normal through at least 7 days p.i. These results suggest that the increased IL-1beta mRNA and protein levels are an early and local response at the lesion site that could trigger other, later, responses to traumatic SCI.

Interaction of opioid peptide-containing terminals with dopaminergic perikarya in the rat hypothalamus.

Both direct pituitary and indirect CNS mechanisms have been postulated for the influence of opiate agonists on prolactin secretion. By examining the interactions between terminals of neurons containing opioid peptides and hypothalamic TH-positive cell bodies, this paper addressed the anatomical basis for the latter mechanism. Initial electron microscopic studies directly demonstrated contact between opioid peptide terminals and dopaminergic cell bodies and provided some visual criteria for assessing opioid-dopamine interactions at the light microscopic level. Using these guidelines, we examined the rates of contact on both A12 and A14 neurons of each of the three opioid peptide families: pro-enkephalin, pro-dynorphin, and pro-opiomelanocortin (POMC). For A14 neurons, many of which project to the posterior pituitary, contact rates were estimated at 15, 20, and 5% for dynorphin, Met-enkephalin, and ACTH (a POMC derivative), respectively. In contrast, the A12 dopamine neurons, which regulate prolactin secretion by inhibition, showed a roughly 70% contact rate with dynorphin axons (P less than 0.001) with Met-enkephalin and ACTH contact rates remaining low at 20 and 5% respectively. Contact frequency varied significantly during the estrus cycle only with dynorphin contacts on A12 neurons. Proestrus and diestrus (less so) showed a small but significant (P less than 0.05) elevation in contact rates versus estrus, male, lactating and pregnant groups. No other significant difference emerged among these groups. On the basis of these observations, we conclude that dynorphin represents a significant and specific factor in the innervation of A12 dopamine neurons. This relationship may account for some if not most of the influence of opiate agonists and antagonists on prolactin secretion.