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The dystrophin glycoprotein complex (DGC) is a membrane-associated protein complex binding extracellular matrix (ECM) molecules, such as laminin and forming a bridge towards the cytoskeleton. The molecular composition of the DGC is cell type dependent and it is involved in cell adhesion and motility. Here we present immunocytochemical localization of beta-dystroglycan, the central member of the DGC, utrophin and Dp71f, the spliced 71 kDa dystrophin protein product of the DMD gene, in cultured retinal Muller glial cells. It is shown that beta-dystroglycan and utrophin are colocalized in clusters in all parts of Muller cells including the lamellipodium and leading edge of migrating cells. As a contrast, Dp71f labels are distinct from beta-dystroglycan and confined to the perinuclear cytoplasm of Muller cells indicating that Dp71f is not a member of the DGC in cultured Muller cells.

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Bernocchi, G., Vignola, C., Scherini, E., Necchi, D., Pisu, M. B. (1998) Bioactive peptides and serotonin immunocytochemistry in the cerebral ganglia of hibernating Helix aspersa . J. Exp. Zool. 280 , 354–367. Pisu

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Acta Veterinaria Hungarica
Authors: Urszula Jankowska, Dariusz Jagielski, Michał Czopowicz, and Rafał Sapierzyński

, R. ( 2010 ): Practical aspects of immunocytochemistry in canine lymphomas . Pol. J. Vet. Sci. 13 , 661 – 668 . Schmidt , S. , Rainer , J. , Ploner , C. , Presul

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Amyotrophic lateral sclerosis (ALS) is a devastating neurological disorder affecting upper and lower motoneurons. Since immune disbalance is known to be an important manifestation of the disease, working with the familial ALS rat model, hSODG93A (containing multiple copies of the human SOD1 G93A mutation), we were particularly interested in following by live magnetic resonance imaging (MRI) the immune cells labeled by ultra small paramagnetic iron oxide (USPIO) nanoparticles. In addition, microglial activation was studied by immunocytochemistry. MRI of USPIO labeled Tcells revealed CD4+ lymphocyte infiltration in the midbraininterbrain region while the CD8+ cells were more confined to the brainstem region. By way of gadolinium (Gd) contrast it was also confirmed that the bloodbrain barrier (BBB) was compromised. Moreover, it was revealed that the regions of BBB breakthrough were congruent with the MRI foci of Tcell infiltration. Immunocytochemistry revealed microglial activation and fusion, possibly phagocytic interactions with neurons in the hippocampus and brainstem. These observations prove the existence of an elaborate inflammatory process in the brain of hSODG93A rats, and also demonstrates the complexity and multifocality of ALS as having its inflammatory manifestations also in the central nervous system (hippocampus) distinct from clinically described motor foci of degeneration.

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We investigated the presence and distribution of the D1 dopamine receptor in the CNS of Lymnaea stagnalis applying immunobloting and immunocytochemistry. We also investigated the effect of dopamine as well as the specific D1 receptor blocker, SCH23390, on the firing activity of the feeding modulator serotonergic neuron, CGC, which displayed D1 immunoreactivity. Immunoblot experiments showed one specifically labeled band with 62 kDa mw which is close to that of the mammalian D1 receptor. Neurons displaying D1-like immunoreactivity can be observed in each ganglion of the CNS but particularly in the pedal ganglia which are the center for locomotion. Dopamine regularly evokes burst activity in the serotonergic CGC at 1 mM and this effect could be antagonized by SCH23390. These observations suggest that a D1-like receptor molecule is present in the CNS of Lymnaea.

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Adrenocorticotropic hormone (ACTH), endorphin, progesterone and triiodothyronine (T 3 ) content of immune cells (lymphocytes, monocyte-macrophage-granulocyte group and mast cells of peritoneal fluid, blood lymphocytes and thymic lymphocytes) was studied by immunocytochemistry and flow cytometric analysis and compared in young (four weeks old) and sexually matured (two and a half months old) male and female rats. In most of cases, cells of female animals contained significantly more hormones, however, in the young group ACTH content of blood lymphocytes was double in males than that of females. In adults only T3 content of mast cells was higher in males. Progesterone was present only in the cells of the young group. The results show that i) hormone content in the immune cells is gender-dependent with the adventage of females and ii) this difference is not influenced by the sexual maturation.

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Elekes, K., Hustert, R., Geffard, M. (1987) Serotonin-immunoreactive and dopamine-immunoreactive neurons in the terminal ganglion of the cricket, Acheta domestica : Light-and electron-microscopic immunocytochemistry. Cell. Tissue Res. 250 , 167

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Embryogenesis of the histaminergic system in the pond snail, Lymnaea stagnalis, was investigated by means of immunocytochemistry and HPLC assay. From the earliest onset of the of histamine-immunoreactive (HA-IR) elements, the labelled neurons were confined to the pedal, cerebral and buccal ganglia, whereas no IR cells within the pleural, parietal and visceral ganglia were detectable during the embryogenesis. Peripheral projections of the embryonic HA-IR neurons were missing. No transient HA-IR neurons could be found either inside or outside the CNS. The first HA-IR elements appeared at about E55% of embryonic development, at the beginning of metamorphosis, and were represented by three pairs of neurons located in the cerebral ganglia. Following metamorphosis, four pairs of HA-IR neurons were added; two of them occurred in the pedal (E65% stage of development) and two in the buccal (E90% stage of development) ganglia. During embryogenesis, HA-IR fibers were present in the cerebro-pedal connectives and in the cerebral, pedal and buccal commissures, whereas only little arborization could be observed in the neuropil of the ganglia. HPLC measurements revealed a gradual increase of HA content in the embryos during development, corresponding well to the course of the appearance of immunolabeled elements. It is suggested that the developing HAergic system plays a specific role in the process of gangliogenesis and CNS plasticity of embryonic Lymnaea.

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We investigated the effect of elevated levels of humoral 5HT and DA on the feeding latency of Helix pomatia , 1 day, 3 days and 10 days following satiation, by injecting monoamines into the haemocoel. HPLC assay of monoamines showed that both 5HT and DA are present in pmol/ml concentrations in the haemolymph of both starved and non-starved animals. Elevated levels of 5HT and DA were most effective at decreasing the feeding latency 10 days following satiation when DA decreased the feeding latency in a concentration dependent manner between 10 −7 and 10 −5 M whereas 5HT levels decreased the feeding latency only at 10 −6 M but increased it at 10 −5 M. Immunocytochemistry revealed that both 5HT3 and D1 receptor-like immuno-reactivity are present in cell bodies located in the same areas of the buccal ganglia. Our observations suggest that both humoral DA and 5HT mutually modulate the activity of the feeding CPG through neurons which have these receptors.

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Acta Biologica Hungarica
Authors: Julianna Jójárt, I. Jójárt, Krisztina Boda, Márta Gálfi, A. Mihály, Zs. B.-Baldauf, and M. Vecsernyés

We investigated the distribution of oxytocin in rat spinal cord using immunocytochemistry and radioimmunoassay (RIA). Each segment of the spinal cord from cervical to coccygeal contained oxytocin-immunoreactive fibers. The Rexed laminae I and II of the dorsal horn showed moderate to intense immunoreactivity. A dense network was found around the central canal where some fibers apposed the ependyma. The autonomic centers of the spinal cord at the thoracolumbar and sacral segments were heavily innervated. Few fibers were found around the motoneurons. In the white matter, the immunoreactivity was localized mainly in the dorsal part of the lateral funiculus, in the pars funicularis of the nucleus intermediolateralis and in a longitudinal network of the lateral funiculus below the spinal cord surface. Some fibers from this network entered the pia mater. RIA measurements revealed that the cervical spinal cord had lower oxytocin content than that found in either the thoracic, lumbar, sacral or coccygeal region. Our results show that the distribution of oxytocin-immunoreactive fibers in the spinal cord correlates with anatomic locations related to nociceptive, autonomic and motor functions. We assume that oxytocin-containing axons play a role in secreting oxytocin directly into the liquor space of the spinal cord.

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