Effect of central injection of tumor-necrosis factor-like cytokine 1A and interferons on food intake in chicks.

In mammals, anorexia accompanying infection is thought to be mediated via cytokines including interleukins, interferons (IFNs), and tumor necrosis factor (TNF). However, there is a lack of related knowledge on birds. Therefore, the purpose of the present study was to determine if cytokines are associated with reduced food intake in chicks (Gallus gallus). Specifically, we evaluated the effects of TNF-like cytokine 1A (TL1A), a member of the TNF family, interferon-α (IFN-α), and interferon-γ (IFN-γ) on food intake. Additionally, the effect of lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C) on cytokine mRNA expression in the diencephalon and spleen was also measured. Intracerebroventricular (ICV) injection of 0.05 or 0.5 µg TL1A, IFN-α, and IFN-γ had no effect on food intake. However, when 1.0 µg each of these factors was evaluated, TL1A significantly decreased food intake at 180 and 240 min after the injection, but IFN-α and IFN-γ had no effect. When chicks received intraperitoneal (IP) injections of 100 µg LPS or 400 µg poly I:C, their food intake was reduced. Diencephalic mRNA expression of TL1A was significantly decreased following IP injection of LPS or poly I:C. Additionally, diencephalic mRNA expression of IFN-γ mRNA was significantly increased by IP injection of LPS but decreased by IP injection of poly I:C. For the spleen, IP injection of LPS and poly I:C both significantly increased TL1A and IFN-γ mRNA expression. In sum, we have provided evidence that central TL1A but not IFN-α or IFN-γ are related to reduction of food intake in chicks, but the role of these cytokines for mediating anorexia associated with infections may differ from mammals.

Mice repeatedly exposed to Group-A ß-Haemolytic Streptococcus show perseverative behaviors, impaired sensorimotor gating, and immune activation in rostral diencephalon.

Repeated exposure to Group-A ß-Haemolytic Streptococcus (GAS) may constitute a vulnerability factor in the onset and course of pediatric motor disturbances. GAS infections/colonization can stimulate the production of antibodies, which may cross the blood brain barrier, target selected brain areas (e.g. basal ganglia), and exacerbate motor alterations. Here, we exposed developing SJL male mice to four injections with a GAS homogenate and evaluated the following domains: motor coordination; general locomotion; repetitive behaviors; perseverative responses; and sensorimotor gating (pre-pulse inhibition, PPI). To demonstrate that behavioral changes were associated with immune-mediated brain alterations, we analyzed, in selected brain areas, the presence of infiltrates and microglial activation (immunohistochemistry), monoamines (HPLC), and brain metabolites (in vivo Magnetic Resonance Spectroscopy). GAS-exposed mice showed increased repetitive and perseverative behaviors, impaired PPI, and reduced concentrations of serotonin in prefrontal cortex, a brain area linked to the behavioral domains investigated, wherein they also showed remarkable elevations in lactate. Active inflammatory processes were substantiated by the observation of infiltrates and microglial activation in the white matter of the anterior diencephalon. These data support the hypothesis that repeated GAS exposure may elicit inflammatory responses in brain areas involved in motor control and perseverative behavior, and result in phenotypic abnormalities.

Gaze palsy, hypogeusia and a probable association with miscarriage of pregnancy--the expanding clinical spectrum of non-opticospinal neuromyelitis optica spectrum disorders: a case report.

BACKGROUND: Neuromyelitis optica is characterised by optic neuritis, longitudinally-extensive transverse myelitis and presence of anti-aquaporin-4 antibodies in the serum. However, non-opticospinal central nervous system manifestations have been increasingly recognised. Awareness of the widening clinical spectrum of neuromyelitis optica (unified within the nosology of 'neuromyelitis optica spectrum disorders') is key to earlier diagnosis and appropriate therapy. We report 2 patients to illustrate the varied clinical manifestations of neuromyelitis optica spectrum disorders while postulating an effect of anti-aquaporin-4 antibodies on the miscarriage of pregnancy. This is the first report of horizontal gaze palsy as a presenting symptom of neuromyelitis optica spectrum disorders. CASE PRESENTATION: Patient 1: A 17-year-old Sri Lankan female presented with hypersomnolence, lateral gaze palsy and loss of taste of 1 week duration. Two years previously she had presented with intractable hiccups and vomiting followed by a brainstem syndrome. Magnetic resonance imaging showed a lesion in the left cerebellum extending into the pons while lesions in bilateral hypothalami and medulla noted 2 years ago had resolved. Autoimmune, vasculitis and infection screens were negative. Anti-aquaporin-4 antibodies were detected in serum. All her symptoms resolved with immunosuppressive therapy. Patient 2: A 47-Year-old Sri Lankan female presented with persistent vomiting lasting over 3 weeks. Three years previously, at 25-weeks of her 4(th) pregnancy, she had presented with quadriparesis and was found to have a longitudinally extensive transverse myelitis from C2 to T2 vertebral levels, which gradually improved following intravenous steroid therapy. Magnetic resonance imaging showed a hyper-intense lesion in the area postrema and longitudinally extensive atrophy of the cord corresponding to her previous myelitis. Autoimmune, vasculitis and infection screens were negative. Anti-aquaporin-4 antibodies were detected in serum. Her vomiting subsided with immunosuppressive therapy. Her second pregnancy had resulted in a first-trimester miscarriage. CONCLUSION: The clinical spectrum of neuromyelitis optica spectrum disorders has expanded beyond optic neuritis and myelitis to include non-opticospinal syndromes involving the diencephalon, brainstem and cerebrum. Our report highlights the varied central nervous system manifestations of neuromyelitis optica spectrum disorders and miscarriage of pregnancy possibly related to anti-aquaporin-4 antibodies.

Distribution of beacon immunoreactivity in the rat brain.

Beacon is a novel peptide isolated from the hypothalamus of Israeli sand rat. In the present study, we determined the distribution of beacon in the rat brain using immunohistochemical approach with a polyclonal antiserum directed against the synthetic C-terminal peptide fragment (47-73). The hypothalamus represented the major site of beacon-immunoreactive (IR) cell bodies that were concentrated in the paraventricular nucleus (PVN) and the supraoptic nucleus (SON). Additional immunostained cells were found in the septum, bed nucleus of the stria terminalis, subfornical organ and subcommissural organ. Beacon-IR fibers were seen with high density in the internal layer of the median eminence and low to moderate density in the external layer. Significant beacon-IR fibers were also seen in the nucleus of the solitary tract and lateral reticular formation. The beacon neurons found in the PVN were further characterized by double label immunohistochemistry. Several beacon-IR neurons that resided in the medial PVN were shown to coexpress corticotrophin-releasing hormone (CRH) and most labeled beacon fibers in the external layer of median eminence coexist with CRH. The topographical distribution of beacon-IR in the brain suggests multiple biological activities for beacon in addition to its proposed roles in modulating feeding behaviors and pituitary hormone release.

Acute stress increases permeability of the blood-brain-barrier through activation of brain mast cells.

Disruption of the blood-brain-barrier (BBB) is important in the pathophysiology of various inflammatory conditions of the central nervous system (CNS), such as multiple sclerosis (MS), in which breakdown of the BBB precedes any clinical or pathological findings. There is some evidence that relapsing-remitting MS attacks may be correlated with certain types of acute stressful episodes. Stress typically activates the hypothalamic-pituitary-adrenal (HPA) axis through the release of corticotropin releasing hormone (CRH), leading to production of glucocorticoids that down regulate immune responses. However, acute stress also has pro-inflammatory effects that appear to be mediated through activation of mast cells. Here we show that acute stress by immobilization increased permeability of rat BBB to intravenous 99Technetium gluceptate (99Tc). This effect was statistically significant in the diencephalon and the cerebellum, while it was absent in the cerebral cortex where there are not mast cells. Immobilization stress also induced activation of mast cells in diencephalon, the site where most mast cells are found in the rat brain. Both BBB permeability and mast cell activation were inhibited by the 'mast cell stabilizer' disodium cromoglycate (cromolyn). These results expand the pathophysiology of mast cells and implicate them in CNS disorders, that may possibly be induced or exacerbated by stress.

Distribution and regulation of telencephalic aromatase expression in the zebra finch revealed with a specific antibody.

In songbirds, aromatase (estrogen synthase) activity and mRNA are readily detectable in the brain. This neural aromatization presumably provides estrogen to steroid-sensitive targets via autocrine, paracrine, and synaptic mechanisms. The location of immunoreactive protein, however, has been difficult to describe completely, particularly in distal dendrites, axons, and terminals of the forebrain. Here we describe the neuroanatomical distribution of aromatase in the zebra finch by using a novel antibody raised specifically against zebra finch aromatase. The distribution of aromatase-positive somata in the zebra finch brain is in excellent agreement with previous reports. Additionally, this antibody reveals elaborate, spinous dendritic arbors, fine-beaded axons, and punctate terminals of telencephalic neurons that may synthesize estrogen. Some of these axon-like fibers extend into the high vocal center (HVC) and the robust nucleus of the archistriatum (RA) in males and females, suggesting a role for presynaptic aromatization in cellular processes within these loci. Adult males have more aromatase-positive fibers in the caudomedial neostriatum (NCM) and the preoptic area (POA) compared to females, despite the lack of detectable sex differences in the number of immunoreactive somata at these loci. Thus, the compartmentalization of aromatase in dendrites and axons may serve a sexually dimorphic function in the songbird. Finally, in adult males, aromatase expression is down-regulated by circulating estradiol in the hippocampus, but not in the NCM or POA. The distribution of aromatase suggests a role for aromatization in the regulation of pre- and postsynaptic function in steroid sensitive areas of the songbird forebrain.

Differential expression of inflammatory mediators in rat microglia cultured from different brain regions.

Microglial cells show a rather uniform distribution of cell numbers throughout the brain with only minor prevalences in some brain regions. Their in situ morphologies, however, may vary markedly from elongated forms observed in apposition with neuronal fibers to spherical cell bodies with sometimes extremely elaborated branching. This heterogeneity gave rise to the hypothesis that these cells are differentially conditioned by their microenvironment and, therefore, also display specific patterns of differential gene expression. In this study, microglia were isolated from 2-4 week-old mixed CNS cultures that had been prepared from neonatal rat diencephalon, tegmentum, hippocampus, cerebellum and cerebral cortex, and were investigated 24 h later. Messenger RNA levels of proteins involved in crucial immune functions of this cell type (TNF-alpha, CD4, Fcgamma receptor II, and IL-3 receptor beta-subunit) have been determined by semi-quantitative RT-PCR. The results clearly show, that three of these mRNAs (TNF-alpha, CD4, Fcgamma receptor II) are differentially expressed in microglia with hippocampal microglia displaying the highest levels of these mRNAs. The data strongly support the notion that the status of microglial gene expression depends on their localization in brain and on specific interactions with other neural cell types. Consequently, it is hypothesized that their responsiveness to signals arising in injury or disease may vary from one brain region to another.

Immunocytochemical localization of growth-associated protein GAP-43 in early human development.

Fibers labelled with antibody to the growth associated protein (GAP-43) were observed as early as 4 gestational weeks (g.w.) in the nervous system of human embryos. At 6 g.w. these fibers could be traced throughout the brainstem and the diencephalon. None of the immunolabeled fibers entered the telencephalic wall at that point, but 2 weeks later at 8 g.w., GAP-43 positive fibers were observed below the newly formed cortical plate of the cerebral cortex. GAP-43 positive fiber bundles had the same distribution as those previously labeled with tyrosine hydroxylase antibodies at the same age. These results strongly suggest that this growth associated protein is localized in the early growing dopaminergic fibers.

Effect of an antidiencephalon immune serum on pain and sleep in primary fibromyalgia.

The results of a double-blind, randomized, therapeutical trial with SER282, an antidiencephalon immune serum (Serolab, Lausanne, Switzerland), in 36 women, aged 24-56 years, with primary fibromyalgia are presented. Treatment was ambulatory and consisted of either SER282 (20 mg/ml) or amitryptiline (AMI, 50 mg) or placebo (PL) over an 8-week treatment course. Clinical and sleep EEG polygraphic data were obtained at baseline and after 4 and/or 8 weeks of therapy. Compared to an important PL response and moderate analgesia with AMI, pain and associated symptoms improved moderately with SER282. In contrast, polysomnographic recordings showed that SER282 tended to promote stage 4 sleep, while AMI and PL had few--if any--effect on sleep. These results are discussed together with the clinical characteristics of the patients and the relations between pain, associated symptoms, and sleep parameters in our patient population.

Distribution of corticotropin-releasing factor immunoreactive neurons in the brain of the tigerfrog, Rana tigrina.

The distribution of immunoreactive (ir) neurons containing corticotropin-releasing factor (CRF) is described in the brain of the tigerfrog, Rana tigrina. The olfactory bulb, medial pallium, nucleus of the diagonal band of Broca and medial area of the amygdala of the telencephalon showed ir-CRF perikarya. The anterior and ventromedial thalamic nuclei, and the magnocellular nucleus preopticus (NPO) revealed several ir cells; a few NPO neurons were cerebrospinal fluid contacting in nature. The nucleus hypothalamicus ventromedialis contained a few cells, but the nucleus infundibularis ventralis of the infundibulum revealed several diffusely distributed perikarya. Individual ir-CRF perikarya were visualized in the optic tectum and interpeduncular nucleus. Extensive fiber terminals were present in the median eminence, but no fibers were discerned either in the neural lobe or in the pituitary gland.

Immunohistochemical localization of argininosuccinate synthetase in the rat brain in relation to nitric oxide synthase-containing neurons.

The distribution of the urea cycle enzyme, argininosuccinate synthetase, in the rat brain was determined using immunohistochemistry. This enzyme participates in the only known metabolic pathway for citrulline, its condensation with aspartate to form argininosuccinate, which can then be cleaved to fumarate and arginine. It may thus provide a mechanism to recycle citrulline, formed in the nervous system via nitric oxide synthase activity, back to the nitric oxide precursor, L-arginine. Argininosuccinate synthetase immunoreactivity was detected in discrete populations of neurons throughout the brain. Double-staining with nicotinamide adenine dinucleotide phosphate (reduced form)-diaphorase histochemistry for the localization of nitric oxide synthase demonstrated that argininosuccinate synthetase coexists with nitric oxide synthase in some brain regions. However, many neurons were found that contained one of these two enzymes, but not the other. Thus some nitric oxide synthase-containing neurons appear able to recycle citrulline via argininosuccinate, while others do not. Additional roles for argininosuccinate synthetase in the brain are discussed.

Distribution of enkephalin-like immunoreactivity in the central nervous system of the rainbow trout: an immunocytochemical study.

The distribution of enkephalin-like immunoreactive (ELI) cell bodies and fibres in the brain of the teleost Salmo gairdneri L. was demonstrated with the indirect peroxidase-antiperoxidase immunocytochemical technique using a highly specific antiserum. In the telencephalon, ELI cell bodies were located in the area ventralis. In the diencephalon, they were found in the nucleus ventromedialis of the thalamus, nucleus lateralis tuberis, nucleus recessus lateralis, and nucleus recessus posterioris. In the mesencephalic tegmentum, ELI cell bodies were found in the nucleus of the rostral mesencephalic tegmentum, and in a group of neurons which was located dorsal to the nucleus of the rostral mesencephalic tegmentum. In the medial torus semicircularis, small numbers of immunoreactive cell bodies were found. In the cerebellum, numerous cell bodies were observed in the granule cell layer and at the border between the granular and molecular layer. ELI cell bodies were also seen in the nucleus tegmenti dorsalis lateralis and nucleus fasciculi solitarii. ELI fibres were widely distributed in the rainbow trout brain. The highest density of immunoreactive fibres was found in the area ventralis telencephali, the mesencephalic tegmentum, the stratum opticum of the optic tectum, the central gray of the brainstem, the caudal part of the fasciculi solitarii and the dorsal horn of the spinal cord. In the stratum fibrosum et griseum superficiale, stratum griseum centrale and stratum album centrale of the optic tectum, a moderate number of immunoreactive fibres was observed. In the olfactory bulb only a few immunoreactive fibres were present. No effect in the labelling was found after colchicine injections. These results provide the first complete mapping of the ELI in a fish brain. It is clear that enkephalins show a similar distribution pattern in Salmo gairdneri to that in other vertebrates; however, the number of ELI cell bodies in the fish brain is smaller than in land vertebrates. The distribution of enkephalins in specific hypothalamic nuclei, visual areas, and in the brainstem of the rainbow trout brain, suggests that these peptides are involved in the modulation of neuroendocrine and as well in visual and somatosensory functions.

Neuropeptide Y-like immunoreactivity in the diencephalon of the little brown bat (Myotis lucifugus): localized variations with physiological state.

The current study used light microscopic immunocytochemistry to demonstrate and compare neuropeptide Y-like immunoreactivity (NPY-IR) in the diencephalon of the little brown bat (Myotis lucifugus) at different stages in its annual cycle of activity and hibernation. Animals were sacrificed in each of three discrete physiological states: euthermic, hypothermic, and hibernating. In general, NPY-IR was abundant in the hypothalamus and sparse in other diencephalic areas. Immunoreactivity was present in a number of pathways which project to or originate from diencephalic nuclei; these include the ansa peduncularis, medial forebrain bundle, inferior thalamic peduncle, stria terminalis, stria medullaris, mammillary peduncle, and dorsal longitudinal fasiculus. Dense fiber plexuses were present throughout the hypothalamus; however, NPY-IR was conspicuously absent from the suprachiasmatic nucleus. Immunoreactive perikarya were located in the supraoptic, dorsomedial, ventromedial, and arcuate nuclei, in the external division of the ventral lateral geniculate nucleus, and in the pineal gland. Localized changes in density and/or distribution of NPY-IR were correlated with changes in physiological state.

Expression of nerve growth factor (NGF) receptors in the brain and retina of chick embryos: comparison with cholinergic development.

The expression of nerve growth factor receptor (NGFR) transcripts was investigated with in situ hybridization techniques in the CNS of chick embryos from 3 days of incubation (E3) to 14 days posthatch (P14). The time course and distribution of NGFR expression was compared with the development of the cholinergic phenotype. Cholinergic properties were assessed by immunolabeling for choline acetyltransferase (ChAT) and histochemistry for acetylcholinesterase (AchE) activity. NGFR transcripts are expressed transiently in the inner plexiform layer and ganglion cell layer of the retina (E4-P1), neostriatum and hippocampus (E18), infundibular hypothalamus (E7-18), spiriform complex (E9-15), layers 2, 3 (E9-18), and 10 (E11-18) of the optic tectum, nucleus mesencephalicus profundus, pars ventralis (E9-18), parvicellular isthmic nucleus (E7-P1), magnocellular isthmic nucleus (E9-E18), nucleus semilunaris (E7-18), isthmo-optic nucleus (E7-P14), rostral motor nuclei (E5-18), developing cerebellum (E7-15), internal granule cell layer (E11-18) and Purkinje cell layer (E15-P14) of the cerebellar cortex, and the inferior olivary nucleus (E9-15). A small number of neuronal populations with embryonic expression of NGFR remain strongly NGFR-positive in the posthatch animal:habenular nuclei (labeled after E5), nucleus subrotundus (after E9), mesencephalic trigeminal nucleus (after E5), caudal parts of locus ceruleus and nucleus subceruleus (after E7), medullar reticular nuclei (after E11), and motor nuclei IX, X, and XII (after E9). The majority of neuronal populations with NGFR expression show cholinergic properties in development, and NGFR expression always precedes the onset of ChAT immunoreactivity. Postnatal expression of growth factor receptors is largely confined to neurons of the reticular type. NGFR expression in avian CNS nuclei differs from that in mammals. Early loss of NGFR expression in the cholinergic basal forebrain (which remains strongly NGFR positive in mammals) and persistent NGFR expression in parts of the avian locus ceruleus indicate changes of growth factor receptor expression and growth factor requirements in phylogeny. Knowledge of the time and distribution of NGFR expression in the chick embryo will facilitate the assessment of specific functions of NGF and NGF-like molecules in an embryonic model with easy access for experimental manipulations.(ABSTRACT TRUNCATED AT 400 WORDS)

Distribution of galanin-like immunoreactivity in the brain of Rana esculenta and Xenopus laevis.

The immunocytochemical distribution of galanin-containing perikarya and nerve terminals in the brain of Rana esculenta and Xenopus laevis was determined with antisera directed toward either porcine or rat galanin. The pattern of galanin-like immunoreactivity appeared to be identical with antisera directed toward either target antigen. The distribution of galanin-like immunoreactivity was similar in Rana esculenta and Xenopus laevis except for the absence of a distinct laminar distribution of immunoreactivity in the optic tectum of Xenopus laevis. Galanin-containing perikarya were located in all major subdivisions of the brain except the metencephalon. In the telencephalon, immunoreactive perikarya were detected in the pars medialis of the amygdala and the preoptic area. In the diencephalon, immunoreactive perikarya were detected in the caudal half of the suprachiasmatic nucleus, the nucleus of the periventricular organ, the ventral hypothalamus, and the median eminence. In the mesencephalon, immunoreactive perikarya were detected near the midline of the rostroventral tegmentum, in the torus semicircularis and, occasionally, in lamina A and layer 6 of the optic tectum. In the myelencephalon, labelled perikarya were detected only in the caudal half of the nucleus of the solitary tract. Immunoreactive nerve fibers of varying density were observed in all subdivisions of the brain with the densest accumulations of fibers occurring in the pars lateralis of the amygdala and the preoptic area. Dense accumulations of nerve fibers were also found in the lateral septum, the medial forebrain bundle, the periventricular region of the diencephalon, the ventral hypothalamus, the median eminence, the mesencephalic central gray, the laminar nucleus of the torus semicircularis, several laminae of the optic tectum, the interpeduncular nucleus, the isthmic nucleus, the central gray of the rhombencephalon, and the dorsolateral caudal medulla. The extensive system of galanin-containing perikarya and nerve fibers in the brain of representatives of two families of anurans showed many similarities to the distribution of galanin-containing perikarya and nerve fibers previously described for the mammalian brain.

Distribution of the 3-fucosyl-N-acetyl-lactosamine (FAL) epitope in the adult mouse brain.

The distribution of the 3-fucosyl-N-acetyl-lactosamine (FAL) epitope within the adult mouse brain was studied by immunohistochemistry using the monoclonal antibody Leu-M1. Leu-M1-positive elements comprised astrocytes and neurons. FAL-positive astrocytes were particularly abundant in barrier structures of the brain, but were also prominent at the periphery of most medullated fiber tracts. Their intracerebral distribution led to a distinct pattern of organization, which in some locations, including the cerebral cortex, could be used for an extended regional architectonic description. Since only some FAL-positive astrocytes were also positive for glial fibrillary acid protein (GFAP), the emerging topography of the FAL-positive astrocytes often differed from the GFAP-distribution. In the cerebellum, Bergmann glia cells expressed the FAL epitope and, in the vermis, their arrangement had a band-like appearance. Positive oligodendrocytes could not be identified. The common ependymal cells were negative, whereas tanycytes were highly immunoreactive. The Leu-M1 antibody also stained some neurons. These occurred in selected neocortical regions, within the dorsal and ventral striatum, in the globus pallidus, the nucleus basalis of Meynert, the nucleus diagonalis and some hypothalamic areas. In some instances, their morphology and location indicated an association with neurochemically specified cell groups.

Age-related changes in serotonin-immunoreactivity in the telencephalon and diencephalon of rats.

Comparison of the appearance and density of the serotoninergic fibers and terminals in some tel- and diencephalon areas of young adult (3 months) and aged (28 months) rats was made immunohistochemically using an antibody to serotonin. In young adult rats a characteristic arrangement of the serotonin-immunoreactive (SER-IR) fibers and terminals in the tel- and diencephalon was observed. In aged rats the distribution pattern was the same but changes in the appearance and density of the SER-IR fibers were demonstrated. Many swollen and folded fibers forming cluster-like structures as well as reduced fiber networks were seen in the neocortex, hippocampal formation and striatum of aged rats. Aberrant fibers and decreased fiber density were also observed in the bed nucleus of the stria terminalis, lateral septum, nucleus accumbens and thalamic nuclei. Reduced fine granular immunostaining and scattered swollen varicosities were found in some amygdaloid and hypothalamic nuclei of aged rats. These results provide further morphological evidence for changes in the serotoninergic innervation during aging.

Serotonin-immunoreactive neurons in the brain of Eigenmannia lineata (Gymnotiformes, Teleostei).

The distribution of serotonin-immunoreactive (5-HTi) neurons was determined by the peroxidase-antiperoxidase technique. 5-HT-immunoreactive nerve cells (or perikarya) occurred in the diencephalic nucleus ventromedialis (nvm), in the so-called pre-pacemaker nucleus (ppm), in a laterally located (unnamed) nucleus (G1), in the nucleus posterior periventricularis (nppv), the nucleus recessus lateralis (nrl) and the nucleus recessus posterior (nrp). 5-HT-immunoreactivity could be localized in the pars intermedia and the proximal pars distalis of the pituitary gland. Large 5-HTi perikarya were found in the lamina VI of the mesencephalic torus semicircularis (ts). Intensely immunostained 5-HTi perikarya occurred in the raphe region. 5-HT-immunoreactivity was also located in the medullary pacemaker nucleus of the specialized electric organ (pm). Only a few 5-HTi fibres were observed in the telencephalon. Ventral to the 3rd ventricle of the diencephalon between the caudal hypothalamic nuclei most of these were fibre bundles dorsal to the nuc. recessus lateralis. Some 5-HTi fibres traversed the pituitary stalk to the proximal pars distalis of the pituitary gland. Scattered 5-HTi fibres were seen in the torus semicircularis, lobus inferior (li) and cerebellum, while more fibres appeared in the medulla oblongata (mo) and below the pacemaker nucleus. The results are compared to those in other fish.

[Effects of anti-tissue immune sera on the contractile response of cells isolated from smooth muscle of the pyloric antrum]. / Die Auswirkungen der Antigewebsimmunseren auf die Kontraktion von aus der glatten Muskulatur des Antrum pyloricum isolierten Zellen.

The effect of anti-tissue antisera on the contractile response of isolated smooth muscle cells from rabbit antrum has been investigated. Anti-stomach (SAE), anti-colon (SAC) and anti-diencephale (SAD) antisera induced a dose-dependent contraction is isolated cells, whereas both non-immune serum and antisera obtained by immunization with other foetal tissues were ineffective. Proglumide, the well known gastrin antagonist, similarly inhibited both gastrin- and SAE, SAC, SAD antisera-induced contraction of isolated smooth muscle cells.

Identification and immunocytochemical localization of a new thyrotropin-releasing hormone precursor in rat brain.

Antisera were raised to a tridecapeptide, Ser-Asp-Val-Thr-Lys-Arg-Gln-His-Pro-Gly-Arg-Arg-Phe, that was synthesized based on the sequence (residues 166-178) of a proposed cDNA for pro-TRH reported by Lechan et al. With this antiserum, immunostaining of Western blots of rat brain extracts revealed two major proteins with mol wt (Mr = 39,000 and 52,000) considerably larger than that of the largest protein (Mr = 29,000) that could be encoded by the cDNA of Lechan et al. Because these observations suggested the possibility of novel TRH precursors, we studied the immunocytochemical distribution of pro-TRH (39-52K) in rat brain. Our anatomical findings were 4-fold. 1) The distributions of 29K pro-TRH and 39-52K pro-TRH are not identical. 2) TRH is found only in regions containing 29K pro-TRH, 39-52K pro-TRH, or both. 3) There are regions that contain both 29K pro-TRH and 39-52K pro-TRH, but no TRH. 4) Regions containing only 39-52K pro-TRH do not contain 29K pro-TRH mRNA as mapped by Segerson et al. From these electrophoretic and anatomical observations, we postulate the existence of at least one and possibly two additional precursors that can be processed to TRH in rat brain.