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P Panula

Publications and source records attributed to P Panula.

At least 73 records · Page 4Linked to original sources

Catechol-O-methyltransferase in rat sensory ganglia and spinal cord.

The localization of catechol-O-methyltransferase immunoreactivity in rat dorsal root ganglia and in the spinal cord and its co-existence with substance P, calcitonin gene-related peptide and fluoride-resistant acid phosphatase in dorsal root ganglion cells was examined with immunohistochemical and histochemical double-staining methods. Analysis of dorsal of dorsal root ganglia at both cervical and lumbar levels revealed catechol-O-methyltransferase immunoreactivity in numerous dorsal root ganglion cells. Double-staining studies showed that catechol-O-methyltransferase and substance P immunoreactivities were located in different cells with a few exceptions, whereas both catechol-O-methyltransferase and calcitonin gene-related peptide immunoreactivities were detected in about 10% of all labeled cells positive for one of the two markers at both levels studied. The great majority of fluoride-resistant alkaline phosphatase-positive cells were also immunoreactive for catechol-O-methyltransferase. Again, no difference was found between cervical and lumbar levels. Catechol-O-methyltransferase immunoreactivity was also found in the neuropil of the dorsal horn of the spinal cord. The staining was most intense in the superficial laminae (I-III) and overlapped partly with substance P and calcitonin gene-related peptide immunoreactivity. Western blotting analysis revealed that soluble catechol-O-methyltransferase was the clearly dominating form of the enzyme in dorsal root ganglia. The distribution pattern of catechol-O-methyltransferase in dorsal horn and sensory neurons suggests that the enzyme may modulate sensory neurotransmission.

Acid Phosphatase↗

Catechol-O-methyltransferase (COMT) in rat brain: immunoelectron microscopic study with an antiserum against rat recombinant COMT protein.

Localization of catechol-O-methyltransferase (COMT) in rat cerebral cortex, neostriatum and cerebellar cortex was studied with preembedding immunoelectron microscopy using a specific antiserum raised against rat recombinant COMT protein. In all areas, immunoreactivity was found both in astrocytes and in neuronal processes. Reaction product was seen in the cytoplasm and in association with tubular structures of dendritic processes. Immunoreactivity was also located postsynaptically in dendritic spines and associated with the postsynaptic membrane. Strong immunoreaction was also seen in the cytoplasm of ependymal cells lining the ventricles, and in tanycytes in median eminence. The results suggest that postsynaptic dendritic spines and astrocytic processes may be the sites of catecholamine inactivation by COMT in rat brain.

Animals↗

Gastric mucosal histamine storing cells. Evidence for different roles of mast cells and enterochromaffin-like cells in humans.

Gastric mucosal histamine content, enterochromaffin-like cell density, and mast cell density were studied in 13 subjects under omeprazole therapy, 13 partially gastrectomized subjects with a Billroth II reconstruction, 10 partially gastrectomized subjects with a Roux-en-Y reconstruction, and 9 control subjects. Histamine content was significantly greater both in the subjects with higher gastrinemic levels (omeprazole-treated subjects) and those with more abundant enterogastric reflux (Billroth II subjects) than in controls. Enterochromaffin-like cell density was significantly greater in the omeprazole subjects than in each of the other groups. Mast cell density was significantly greater in Billroth II subjects than in controls. Serum gastrin levels, mucosal histamine content, and enterochromaffin-like cell density were positively correlated. Gastrin was not correlated to mast cell density. These results support the existence of different control pathways for enterochromaffin-like and mast cells. Moreover, they suggest that enterochromaffin-like cells and mast cells are involved in the regulation of gastric secretion and in gastric mucosal injury-repair mechanisms, respectively, due to histamine release.

Aged↗

Neuropeptide FF-like immunoreactivity in human plasma.

In order to examine whether neuropeptide FF (NPFF), an octapeptide with pain-modulating and blood pressure-raising properties in the rat, is present in circulating human blood, a radioimmunoassay (RIA) was established. Using this highly specific and sensitive RIA, the mean concentration of NPFF in human plasma was 2.9 pg/ml +/- 1.1 (n = 111). The concentration did not correlate with age or sex. Reversed-phase high pressure liquid chromatography (HPLC) followed by RIA using two different antisera for NPFF showed that plasma NPFF eluted in a position identical to that of synthetic NPFF. In view of published Kd values (0.06 mM) for NPFF receptor, the concentrations detected of NPFF in human plasma may be too low for systemic actions. Thus, plasma NPFF may represent leakage of the peptide from nervous tissue.

Amino Acid Sequence↗

Neuropeptide FF-containing efferent projections from the medial hypothalamus of rat: a Phaseolus vulgaris leucoagglutinin study.

Neuropeptide FF (FMRFamide-like peptide, morphine-modulating peptide) is an octapeptide isolated from the bovine brain. There is evidence that neuropeptide FF participates in the modulation of nociceptive mechanisms. Neuropeptide FF acts through its own receptors which are distinct from the opiate receptors. In the rat brain neuropeptide FF is found in two major cell populations. We have studied the efferent connections of the hypothalamic neuropeptide FF-containing cell group, which is located in the medial hypothalamus between the dorsomedial, ventromedial and periventricular hypothalamic nuclei. By using an anterograde tracing method (Phaseolus vulgaris leucoagglutinin) combined with double-staining immunohistochemistry we characterized the connections of this cell group with the limbic system, certain hypothalamic nuclei, periaqueductal gray and with the solitary tract nucleus. In the limbic system, the major targets were the lateral septal nucleus, bed nucleus of stria terminalis and certain subnuclei in the amygdala. These connections suggest that neuropeptide FF may act, in addition to its well-characterized action in the sensory system, in limbic functions. Efferent connections to the periaqueductal gray suggest that neuropeptide FF may modulate the opiate mediated analgesia at this site. Good correlation between our results and receptor autoradiography support the idea that the terminal areas which our results show are target areas of the neuropeptide FF-containing system.

Animals↗

Neuronal and non-neuronal catechol-O-methyltransferase in primary cultures of rat brain cells.

Previous biochemical and histochemical studies have suggested that catechol-O-methyltransferase (COMT) is a predominantly glial enzyme in the brain. The aim of this work was to study its localization and molecular forms in primary cultures, where cell types can be easily distinguished with specific markers, COMT immunoreactivity was studied in primary astrocytic cultures from newborn rat cerebral cortex, and in neuronal cultures from rat brain from 18-day-old rat embryos using antisera against rat recombinant COMT made in guinea pig. Double-staining studies with specific cell markers to distinguish astrocytes, neurons and oligodendrocytes were performed. COMT immunoreactivity colocalized with a specific oligodendrocyte marker galactocerebroside in cells displaying oligodendrocyte morphology, flat cells displaying type-1 astrocyte morphology and glial fibrillary acidic protein, in branched cells displaying type-2 astrocyte morphology and in cell bodies of neurons, the processes of which displayed neurofilament immunoreactivity. Western blots detected both soluble 24 kDa and membrane-bound 28-kDa COMT proteins in neuronal and astrocyte cultures. The results suggest that COMT is synthesized by cultured astrocytes, oligodendrocytes and neurons.

Animals↗

Histamine-immunoreactive neurons in the brain of the teleost Gasterosteus aculeatus L. Correlation with hypothalamic tyrosine hydroxylase- and serotonin-immunoreactive neurons.

The distribution of putative histaminergic neurons in the brain of a teleost, the three-spined stickleback, was investigated by means of immunocytochemistry using specific antibodies against histamine (HA), and conventional microscopy as well as confocal laser scanning microscopy. Histamine-immunoreactive (HAir) neurons form discrete populations ventral to the nucleus of the posterior recess (NRP) and in the nucleus saccus vasculosus (NSV), which belong to the periventricular hypothalamic nuclei. The neuronal somata are subependymally located, and do not possess apical neurites contacting the cerebrospinal fluid. They give rise to both long-range and local axonal projections. The local projections give rise to a field of dense punctate immunoreaction dorsal to the NRP and lateral to the NSV. Long-range projections are comprised of ascending projections to the thalamus, habenula, preoptic area and dorsal telencephalon; and descending projections via the posterior tuberal nucleus, ventrally to the nucleus interpeduncularis, and dorsally into the central gray. HAir neurons occur together with serotoninergic cerebrospinal fluid-contacting (CSFc) neurons in the NRP, and with tyrosine hydroxylase-immunoreactive (THir) neurons in the NSV. Although HAir elements occur together with THir ones in many brain areas, direct contacts between the two neurotransmitter systems are rare. The putative histaminergic neurons in the brain of the three-spined stickleback constitute a very discrete neuronal system, with a major projection area in the dorsal telencephalon in a region which is considered homologous with the dorsal pallium of land vertebrates.

Animals↗

The antiopioid peptide, neuropeptide FF, enhances the effects of acute morphine on the cerebral monoamines in rats.

The effects of neuropeptide FF (NPFF) on the changes induced by acute morphine in cerebral monoamines were studied in male Wistar rats. We measured the concentrations of noradrenaline (NA), 5-hydroxytryptamine (5-HT), dopamine and their main metabolites in the limbic area (LIM), hypothalamus (HYP) and lower brain stem (LBS). NPFF 10 micrograms x 3 at 20 min intervals, i.c.v., bilaterally) did not alter the concentrations of monoamines or their metabolites. Morphine (10 mg kg-1, s.c., 1 h) did not alter NA or 5-HT or their metabolites in any brain area studied, but elevated 3,4-dihydroxyphenylacetic acid (DOPAC) and/or homovanillic acid (HVA) in LIM and HYP. Combination of morphine and NPFF decreased NA in LIM, elevated 3-methoxy-4-hydroxyphenylethylene glycol in HYP and LBS as well as 5-hydroxyindoleacetic acid in LIM and HYP. These responses are typical to morphine at doses larger than 10 mg kg-1, and suggest a synergistic effect of the two compounds. Further, the morphine-induced elevations of DOPAC and HVA were potentiated by NPFF in LIM and HYP. In conclusion, our results suggest that NPFF does not attenuate but rather enhances the changes induced by acute morphine in the cerebral monoamines.

Animals↗

Expression of histidine decarboxylase and cellular histamine-like immunoreactivity in rat embryogenesis.

In this study we investigated the developmental expression of histidine decarboxylase (HDC) mRNA and the distribution of histamine-immunoreactive (histamine-ir) cells in the rat embryonic tissues. We applied Northern blot analysis, in situ hybridization with synthetic oligonucleotide probes complementary to the rat HDC cDNA, and indirect histamine immunocytochemistry. Northern blot analysis revealed the appearance of a major (2.6 KB) HDC mRNA species in liver on embryonic Day 14. Its hybridization level peaked on Day E18, when two minor (1.6 and 3.5 KB) mRNA species were also present. During the periparturition period, a rapid decrease in HDC RNA was apparent, as the 2.6 KB mRNA species was expressed at a low level on postnatal Day P1. The embryonic liver expressed HDC on days E14-E20. On days E18 and E20, the periosteum and the epiphyseal growth plates of the endochondrally ossificating bones, and some striated muscle cells, showed hybridization signal for HDC. Histamine immunoreactivity was detected in many epithelial and neuronal cell types during embryogenesis. An intense histamine immunoreaction appeared first in essentially all cells of the liver parenchyma on day E12. This parenchymal histamine immunoreactivity disappeared by birth, after which this immunofluorescence in liver was restricted to a few scattered mast cells until adulthood. Some neurons in the peripheral sensory, sympathetic and cranial nerve ganglia were histamine-immunoreactive from day E16 to birth. In addition, many immunoreactive nerve fibers were detected in the gastrointestinal muscularis externa, mesentery, salivary glands, kidney, lung, and muscle tissue. We conclude that during rat embryogenesis histamine is produced and stored transiently by cells in liver, developing bone, and a few striated muscle cells, in addition to previously reported neurons in rat brain. Many peripheral neurons, epithelial cells, and mast cells display histamine immunoreactivity during rat embryogenesis but are devoid of detectable HDC mRNA with the current method. It remains possible that histamine is formed by another enzyme or is taken up from the extracellular space. The results support the concept that a significant proportion of histamine is formed and stored by embryonic cells other than mast cells.

Animals↗

Developmental patterns of histamine-like immunoreactivity in the mouse.

We studied the appearance and distribution of histamine (HA) during mouse embryogenesis, neonatal period, and adulthood using a specific rabbit HA antiserum and indirect immunofluorescence. HA first appeared on the Embryonic Day 13 (E13) in scattered mast cells in the gastrointestinal (GI) muscularis externa and liver. The splenic primordium contained a dense population of intensely HA-immunoreactive (HA-ir) cells from E13 on. From E15 to the birth, HA was detected in many embryonic cell types. On E15, the first HA-ir epithelial endocrine cells appeared in the oxyntic mucosa. In addition to the HA-ir cells in GI tract and liver, some nerve cells in ganglia of the peripheral nervous system (PNS), some fibers in spinal and cranial nerves, nerve fibers in mesenterium, and nerve plexuses of the gastrointestinal muscularis externa were HA-ir from E15 on. Occasional HA-ir nerve fibers were detected within the glandular epithelium of the oxyntic mucosa, pancreas, and salivary glands during late embryogenesis. During the same period, bright fluorescence was observed in cells of the kidney convoluted tubules and pancreatic islet cells. From E14 on, mast cells exhibiting bright fluorescence were scattered throughout the connective tissue of the fetus, and their number increased rapidly with age. Their density was especially high in subcutaneous connective tissue. Embryonic epidermal cells showed faint HA immunoreactivity. In musculoskeletal tissues, developing bone and occasional striated muscle cells exhibited HA immunoreactivity. Interestingly, most cells in liver showed transiently weak HA immunoreactivity during embryogenesis. In adult mouse, HA was stored only by scattered mast cells, oxyntic epithelial cells, and neurons in the tuberomamillary nucleus of the brain. The other HA-containing embryonic cells were negative for HA in adult mouse. In conclusion, HA immunoreactivity is widely distributed in epithelial, neuronal, and mast cells in various organs during mouse embryogenesis.

Animals↗

Distribution of histamine-, 5-hydroxytryptamine-, and tyrosine hydroxylase-immunoreactive neurons and nerve fibers in developing rat brain.

Although the general patterns of the developing histaminergic system in the rat brain are known, no comparative studies between the development of the brain histaminergic system and the development of other neuroactive substances have yet been published. Interestingly, separate immunohistochemical studies on the development of the 5-HT system and on the catecholaminergic system in the rat imply common features in the different aminergic systems. Therefore, the spatial distribution of histamine-immunoreactive (HA-ir) neurons and nerve fibers was compared to the distribution of 5-hydroxytryptamine (5-HT)-, and tyrosine hydroxylase-immunoreactive (TH-ir) ones in the developing rat brain between embryonic days 12 (E12) and 20 (E20) by using a double-immunostaining method. The high-pressure liquid chromatography (HPLC) fluorometric method was used for determination of histamine concentration in different brain regions during the same period of development and synthetic oligonucleotide probes complementary to the rat histidine decarboxylase (HDC) to determine the origin of HA in the brain during the development with in situ hybridization. The immunohistochemical results revealed co-localization of HA and 5-HT within a subgroup of cells in the developing raphe nuclei between E14 and E18. From E18 onwards HA immunoreactivity started to gradually disappear from the rhombencephalon, and was totally abolished by E20, while 5-HT-ir cells continued to establish their adult positions. No significant colocalization of HA and TH immunoreactivities was detected. The biochemical results were in agreement with the immunohistochemical ones and confirmed that histamine detected in the early developing brain is authentic. A positive in situ hybridization signal for HDC was detected in a small area in the ventrolateral pons in the same areas as HA- and HDC-ir cell bodies at E16, suggesting that at least some HA may be synthesized locally. These results confirm that HA is one of the first neurotransmitters to appear in the developing brain. In addition, the transient co-localization of HA and 5-HT immunoreactivities and the transient HDC expression at E16 within the developing pontine raphe nuclei may imply an interesting and a more general role for HA in modification of brain development.

Animals↗

Distribution of neuropeptide FF in porcine spinal cord in comparison with other neuropeptides and serotonin.

A large number of neurotransmitters and neuropeptides are concentrated in the dorsal horn of the spinal cord, where they interact in a complex manner and modulate sensory mechanisms. Most studies are carried out in the rat, and little is known of other species. It is relevant to study mammals with a more complex central nervous system, because pain mechanisms are central in both human and veterinary medicine. Immunoreactivity for neuropeptide FF, an amidated octapeptide originally isolated from bovine brain, was found immunocytochemically at all levels of porcine spinal cord. In contrast to other species studied so far, the peptide immunoreactivity in porcine spinal cord was confined to the intermediolateral gray matter, especially to the intermediolateral cell column and lamina X of the gray matter. This distribution was remarkably different from that of substance P, proenkephalin A-derived peptides, thyrotropin-releasing hormone, serotonin, and neuropeptide Y. Pharmacologic administration of neuropeptide FF alters behavior in assays for analgesia. The distribution of neuropeptide FF immunoreactivity as revealed by this study suggests that there may be marked species differences in the distribution and function of the peptide.

Animals↗

gamma-Aminobutyric acid in the nervous system of a planarian.

The amino acid gamma-aminobutyric acid (GABA) is an important inhibitory neurotransmitter in both vertebrates and invertebrates. Despite this, no reports of GABA in flatworms have to date been published. We have studied the presence of GABA in the planarian Dugesia tigrina with immunocytochemical methods and high-pressure liquid chromatography. Fibers showing GABA-like immunoreactivity (GABA-IR) were present in abundance in the longitudinal nerve cords and lateral nerves. GABA-IR was revealed in fibers forming commissures in the brain. The ventral part of the subepidermal plexus showed GABA-IR. No cell somata containing GABA-IR could be identified with certainty. The chromatographic analysis showed that the average GABA concentration in D. tigrina is 533.6 pmol/mg protein. This is substantially higher than the concentrations of dopamine (62.87 pmol/mg) and serotonin (233.20 pmol/mg). An enzyme assay confirmed the capacity for GABA-synthesis in D. tigrina. The results indicate that GABA-containing neurons appeared earlier in evolution than was previously thought and that GABA may serve an important role already in the flatworms.

Animals↗

Expression of HB-GAM (heparin-binding growth-associated molecules) in the pathways of developing axonal processes in vivo and neurite outgrowth in vitro induced by HB-GAM.

HB-GAM (heparin-binding growth-associated molecule; p18) was previously isolated as a neurite outgrowth-promoting protein that is expressed at high levels in perinatal rat brain. cDNA cloning and expression revealed that HB-GAM is a novel secretory protein that is homologous with the retinoic acid-inducible MK protein. In the present paper we have used affinity-purified anti-peptide and anti-protein antibodies to study the expression of HB-GAM in the developing nervous system of the rat. In general, HB-GAM accumulates to extracellular structures that line growing axonal processes but is absent or only occurs at low levels in the axonal pathways after neurite extension has essentially ceased. During early stages of the nervous system development, HB-GAM is strongly expressed in the developing fiber tracts of the peripheral nervous system on embryonic days 12-14 (E12-E14). In the early central nervous system, HB-GAM is first expressed in a radial pattern along the neuroepithelial cells on E11-E12 and in early ascending neuron fibers in superficial layers of the brain vesicles on E12-E14. On E16-E18, HB-GAM is strongly expressed in the subplate and the marginal zone of the primordial neocortex. After this local expression in the primordial brain, HB-GAM is more widely expressed in the pathways of the developing axons during the late embryonic and early postnatal period. We have also extended in vitro studies on the interactions of HB-GAM with perinatal rat brain neurons by creating patterned substrates of HB-GAM upon culture wells and upon mixtures of extracellular matrix structures. These studies confirm the neurite-promoting effect of HB-GAM and suggest, together with the patterns of tissue localization, that HB-GAM may also guide axonal processes of brain neurons. The interactions of HB-GAM with brain neurons are specifically inhibited by heparin and its fragments and by incubation of the neurons with heparitinase. We suggest that in developing nervous tissues HB-GAM is deposited to an extracellular location in developing axon pathways and it interacts with heparin-like molecules of the neuron surface to promote formation of neural connections.

Animals↗

Distribution of catechol-O-methyltransferase enzyme in rat tissues.

In the present study we show the distribution of catechol-O-methyltransferase (COMT) in various rat tissues with a highly specific antiserum prepared against recombinant rat COMT. Immunoprecipitation and immunocytochemical controls confirmed the COMT-specificity of the antibodies. The antiserum detected both the 24 KD soluble and the 28 KD membrane-bound forms of the enzyme. By immunohistochemical staining the COMT enzyme was found in most rat tissues. Staining was most intense in the liver and in the kidney, in agreement with previous studies and our immunoblotting results. In the gastrointestinal tract, epithelial cells of the stomach, duodenum, and ileum were immunoreactive for COMT. In pancreas, COMT immunoreactivity was found in insulin-producing beta-cells and somatostatin-producing D-cells but not in glucagon-producing alpha-cells of the islets of Langerhans. In pituitary, COMT immunoreactivity was found in cleft cells, in pituicytes of the posterior lobe, and in the anterior lobe, partly in the same cells containing luteinizing hormone (LH). In other endocrine organs, COMT immunoreactivity was found in epithelial cells of the thyroid gland and in zona glomerulosa of the adrenal cortex. In the brain, brightest immunofluorescence was seen in ependymal cells of the cerebral ventricles and choroid plexus. Weak to moderate immunofluorescence was found in the neuropil of several brain areas, including striatum and cortex. Scattered small neurons in spinal sensory ganglia were also COMT immunoreactive. Previous immunocytochemical studies, enzyme activity determinations, and distribution of the COMT mRNA are in general agreement with the results presented here. The wide distribution of COMT in different tissues suggests an important role for this protein in inactivation of catechol compounds.

Animals↗

Endogenous histamine in cultured bovine adrenal chromaffin cells.

Histamine releases catecholamines and opioids in primary cultured bovine adrenal medullary (BAM) chromaffin cells. We have studied whether histamine is synthesized and localized in BAM cells, and whether it can be released upon activation with secretagogues. In BAM cells histamine is immunohistochemically co-localized with tyrosine hydroxylase in 45 +/- 8% of all cells. Only histamine immunoreactivity was observed in 8 +/- 2% of all BAM cells. No mast-cell-like cells were observed in our system. Histamine can be released from BAM cells by high potassium (56 mM K+) in a calcium-dependent manner. Compound 48/80 did not release histamine from BAM cells but nicotine caused a dose-dependent liberation of the amine. Cultured BAM cells have histidine decarboxylase activity which is inhibited by alpha-fluoromethylhistidine. These results indicate that endogenous histamine is synthesized, stored and released in BAM chromaffin cells in vitro.

Adrenal Medulla↗