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Biomedical subjects

C Yanaihara

Publications and source records attributed to C Yanaihara.

At least 55 records · Page 3Linked to original sources
PubMed1987

Effects of naloxone on basal and vagus nerve-induced secretions of GRP, gastrin, and somatostatin from the isolated perfused rat stomach.

The effects of naloxone, an opiate antagonist, on basal and vagus nerve-induced secretions of GRP, gastrin, and somatostatin were examined using the isolated perfused rat stomach prepared with vagal innervation. Naloxone (10(-6) M) significantly inhibited basal somatostatin secretion in the presence and absence of atropine and of hexamethonium, whereas basal GRP and gastrin secretion was not affected by naloxone. Electrical stimulation (10 Hz, lms duration, 10V) of the distal end of the subdiaphragmatic vagal trunks elicited a significant increase in both GRP and gastrin but a decrease in somatostatin. Naloxone (10(-6) M) failed to affect these responses in the presence or absence of atropine. On the other hand, when hexamethonium was infused, naloxone significantly inhibited both the GRP and gastrin responses to electrical vagal stimulation. Somatostatin secretion was unchanged by vagal stimulation during the infusion of hexamethonium with or without naloxone. These findings suggest that basal somatostatin secretion is under the control of an opiate neuron and that opioid peptides might be involved in vagal regulation of GRP and gastrin secretion.

Animals↗
PubMed1987

Immunocytochemical distribution of met-enkephalin-Arg6-Gly7-Leu8 in the rat lower brainstem.

The distribution of methionine-enkephalin-Arg6-Gly7-Leu8, a unique peptide derived from proenkephalin A in the rat brainstem, was studied immunocytochemically by using a highly specific antiserum to this octapeptide sequence. Immunoreactive perikarya with various shapes and sizes were detected in many regions of the rat brainstem. Dense accumulation of immunoreactive perikarya and fibers was seen in the nuclei associated with special sensory and visceral functions, such as the interpeduncular nucleus, the parabrachial nucleus, the nucleus of the solitary tract, and the nucleus of the spinal tract of the trigeminal nerve. Clusters of methionine-enkephalin-Arg6-Gly7-Leu8-like immunoreactive perikarya and fibers were observed in certain areas considered to play a role in nociception and analgesia, such as the central gray of the midbrain central gray and the raphe magnus nucleus. Some methionine-enkephalin-Arg6-Gly7-Leu8-like immunoreactive perikarya were distributed in the lateral reticular nucleus, the nucleus of the solitary tract, and the raphe magnus nucleus, where monoaminergic neurons were also detected. In addition to the previously reported enkephalinergic cells, we found many methionine-enkephalin-Arg6-Gly7-Leu8 containing neurons; the rostral and caudal linear nucleus of raphe, the median raphe nucleus, entire length of the raphe magnus nucleus, the medial longitudinal fasciculus, the cuneate nucleus, the external cuneate nucleus, the gracile nucleus, and the area postrema. The wide distribution of this octapeptide-like immunoreactivity reflected neurons expressing the preproenkephalin A gene distributed more widely than previously reported and that innervated many regions.

Animals↗
PubMed1987

An immunohistochemical study of methionine-enkephalin-Arg6-Gly7-Leu8-like immunoreactivity-containing neurons in the parasympathetic preganglionic regions of the rat spinal cord.

The localization of the methionine-enkephalin-Arg6-Gly7-Leu8 (Met-Enk-Arg-Gly-Leu)-like immunoreactivity-containing neurons in the rat lumbosacral spinal cord was immunohistochemically examined by an antiserum very specific to Met-Enk-Arg-Gly-Leu. The immunoreactive neurons occupied the positions corresponding to the parasympathetic preganglionic nuclei determined by the previous horseradish peroxidase (HRP)-tracing experiments. The present study suggests that the parasympathetic preganglionic neurons in the rat lumbosacral spinal cord produce preproenkephalin A and its related peptides.

Animals↗
PubMed1987

Corelease of PHI and VIP by vagal stimulation in the dog.

Electrical vagal stimulation increased immunoreactive peptide histidine isoleucine (IR-PHI) and vasoactive intestinal polypeptide (IR-VIP) levels in the portal plasma in anesthetized dogs, depending on the stimulation frequency. Atropine failed to suppress the vagal release of IR-PHI and IR-VIP, whereas hexamethonium abolished the increase of both peptide immunoreactivities. Gel filtration profiles of IR-PHI and IR-VIP in the portal plasma obtained during vagal stimulation revealed major peaks of IR-PHI and IR-VIP eluting in the same positions as synthetic PHI-27 and VIP-28, respectively. These data demonstrate that PHI and VIP are coreleased by vagal stimulation via a nicotinic ganglionic mechanism. An additional peak of larger molecular weight IR-PHI was observed in gel filtration of the portal plasma obtained under higher frequency vagal stimulation, when measured by the N-terminal-specific PHI antiserum. The presence of this larger form of IR-PHI together with a PHI-27-like component was also demonstrated in tissue extracts of the entire length of the gastrointestine, particularly in higher concentration in the stomach. The structure and the physiological significance of this larger form of IR-PHI remains to be elucidated.

Animals↗
PubMed1986

Inhibitory effects of galanin on the isolated spinal cord of the newborn rat.

The effects of galanin, a 29-amino-acid peptide, on spinal reflexes were studied. In the isolated hemisected spinal cord of the newborn rat, galanin (0.1-5 microM) depressed the monosynaptic reflex that was induced by dorsal root stimulation and recorded from the corresponding ventral root. In the isolated spinal cord-tail preparation of the newborn rat, galanin (0.3-0.6 microM) depressed the nociceptive reflex that was induced by application of capsaicin to the tail and recorded from a lumbar ventral root. In both preparations the inhibitory effects of galanin were reversible and the full recovery of the reflexes was observed within 3-20 min after removal of the peptide. The mechanisms of action of galanin on the spinal reflexes and the physiological role of the peptide in the spinal cord are discussed.

Animals↗
PubMed1986

An immunohistochemical study of methionine-enkephalin-Arg6-Gly7-Leu8-like immunoreactivity-containing liquor-contacting neurons (LCNs) in the rat spinal cord.

The localization of methionine-enkephalin-Arg6-Gly7-Leu8 (Met-Enk-Arg-Gly-Leu)-like immunoreactivity in the medullospinal liquor-contacting neurons (LCNs) of the rat was immunohistochemically investigated using a modified Sternberger's PAP method. In this study, we also examined the distribution of Met-Enk-Arg-Gly-Leu-like immunoreactive LCNs in the entire length of the medulla oblongata and spinal cord. The immunoreactive LCN possessed an oval, polygonal or rod-shaped perikaryon which protruded an axon-like process and a CSF-contacting process. In the entire length of the medulla oblongata and spinal cord, 60-70 immunoreactive LCNs were observed. They were sparsely distributed along the central canal. The ratio of the number of immunoreactive LCNs in each segment/total number of immunoreactive LCNs in the medulla oblongata and whole spinal cord was as follows: medulla oblongata, 0%; cervical segments, 28.1%; thoracic segments, 56.8%; lumbar segments, 12.0%; sacral segments, 3.0%; and coccygeal segments, 0%. When the immunoreactivity of perikarya of LCNs was weak, a few nerve terminals with a strong Met-Enk-Arg-Gly-Leu-like immunoreactivity were noticed on them. These findings suggest that preproenkephalin A-related opioid peptides including Met-Enk-Arg-Gly-Leu may be secreted into the cerebrospinal fluid (CSF) through the terminal portions of axon-like processes and/or CSF-contacting processes of LCNs. Enkephalin-containing nerve terminals on the LCNs may represent modulating afferents aiding the receptive and/or secretory functions of the neurons.

Animals↗
PubMed1986

Glucagon-related peptides in the rat hypothalamus.

Immunohistochemically, nerve fibers and terminals reacting with anti-N-terminal-specific but not with anti-C-terminal-specific glucagon antiserum were observed in the following rat hypothalamic regions: paraventricular nucleus, supraoptic nucleus, anterior hypothalamus, arcuate nucleus, ventromedial hypothalamic nucleus and median eminence. Few fibers and terminals were demonstrated in the lateral hypothalamic area and dorsomedial hypothalamic nucleus. Radioimmunoassay data indicated that the concentration of gut glucagon-like immunoreactivity was higher in the ventromedial nucleus than in the lateral hypothalamic area. In food-deprived conditions, this concentration increased in both these parts. This was also verified in immunostained preparations in which a marked enhancement of gut glucagon-like immunoreactivity-containing fibers and terminals was observed in many hypothalamic regions. Several immunoreactive cell bodies were found in the ventromedial and arcuate nuclei of starved rats. Both biochemical and morphological data suggest that glucagon-related peptides may act as neurotransmitters or neuromodulators in the hypothalamus and may be involved in the central regulatory mechanism related to feeding behavior and energy metabolism.

Animals↗
PubMed1986

Identification of two pro-VIP forms in a human neuroblastoma cell line.

The immunoreactivity of VIP and PHI standards, immobilized on a nitrocellulose membrane, was first assayed with various detection procedures. For VIP, the double bridge peroxidase-antiperoxidase (PAP) method was the most sensitive procedure, giving a detection limit of 0.1-0.3 pmol per mm2 with the 4 rabbit anti-VIP antisera tested. By contrast, the detection limit of immobilized PHI was 100 times higher with the 4 rabbit anti-PHI/PHM antisera tested presumably because major antigenic sites were masked in the immobilized peptide. With this information at hand, the VIP and PHI immunoreactivity of human neuroblastoma NB-OK-1 cells was tested after extraction, SDS-PAGE, electrotransfer, and PAP immunodetection. Two faint immunoreactive bands corresponding to two pro-VIP forms with an Mr of, respectively, 19 kDa and 18 kDa, were detected in undifferentiated cells. These distinct bands increased progressively and markedly during differentiation in the presence of dibutyryl cyclic AMP. In addition, two intermediary VIP forms of lower Mr (11 kDa and 6 kDa) and 3 kDa VIP itself were also present after 2 days of differentiation. The 19 kDa and 18 kDa pro-VIP forms were detected with a sensitivity several times higher than that of VIP and their staining was specific for VIP epitopes. By contrast, when using 4 rabbit anti-PHI/PHM antisera, we observed essentially the strong unspecific staining of a 17 kDa polypeptide. VIP immunoreactivity was also visualized by immunocytochemistry in neuroblastoma cells cultured on glass coverslips and fixed in situ. Specific VIP staining using the PAP method was present in 10 percent of the cells in the undifferentiated state.(ABSTRACT TRUNCATED AT 250 WORDS)

Bucladesine↗
PubMed1986

Interaction of synthetic N- and C-terminal fragments of helodermin with rat liver VIP receptors.

Synthetic helodermin was more potent than natural helodermin (purified from the venom of Gila monster) in activating adenylate cyclase in rat liver membranes. Two possible reasons for this discrepancy were discussed. Comparing adenylate cyclase activation to the binding of 125I-natural helodermin and 125I-VIP in the presence of six synthetic helodermin fragments (1-27, 7-35, 13-35, 17-35, 18-35 and 22-35), we conclude that the effects of both synthetic and natural helodermin were mediated through an interaction with VIP receptors. The C-terminal (28-35) extension of these peptides favored VIP receptor recognition. Their N-terminal extremity was not necessary for binding and for ensuing adenylate cyclase activation, illustrating again the atypical coupling of VIP receptors with the effector enzymatic system, in rat liver membranes.

Adenylyl Cyclases↗
PubMed1986

PHI structural requirements for potentiation of glucose-induced insulin release.

Immunoreactive PHI was detected in rat pancreas. The potentiating effect of 10(-9) M PHI upon insulin release from the isolated perfused rat pancreas was significant and most consistent when 250 mg% glucose was present in the perfusion medium. PHI(1-15) retained a substantial potentiating effect on insulin release, while PHI(14-27) was essentially inactive. Replacement of amino-terminal decapeptide portion of the PHI molecule with the corresponding part of VIP resulted in a drastic decrease of the potentiating effect of PHI on insulin release. 10(-8) M PHI(14-27) substantially diminished the potentiation by 10(-9) M PHI while PHI(1-15) was without an inhibitory effect. The present results indicate that the PHI active site for potentiation of glucose-induced insulin release resides in the amino-terminal segment of the molecule but requires the carboxyl terminal segment primarily for binding to exhibit full biological activity.

Animals↗
PubMed1986

Helodermin has a VIP-like effect upon canine blood flow.

The effect of helodermin on vascular physiology was studied in anesthetized dogs using a synthetic replicate of helodermin and helodermin related peptides. Intraarterial infusion of helodermin caused a dose-dependent increase in femoral blood flow. Helodermin was 16 times less potent than VIP and 5 times more potent than PHM (human PHI). The helodermin effect lasted significantly longer; the half-life of the helodermin effect was 6.5 times longer than VIP. Synthetic helodermin (Hd) N-terminal fragment Hd(1-27)NH2 retained substantial activity similar to the full helodermin molecule but the prolonged effect was lost. Hd(7-35) and Hd(22-35) were inactive in this system. Intravenous injection of synthetic helodermin produced prolonged systemic hypotension and tachycardia; and, similar to VIP, it increased the common carotid arterial blood flow while those of the superior mesenteric and femoral arteries were decreased. The results demonstrate the VIP-like vasodilating activity and cardiovascular effects of helodermin in anesthetized dogs.

Animals↗
PubMed1986

Inhibitory effects of rimorphin and dynorphin on insulin secretion from the isolated, perfused rat pancreas.

In order to settle the question about whether or not opioid peptides stimulate or inhibit insulin secretion, we studied effects of rimorphin and dynorphin, two members of the preproenkephalin B group, on glucose-induced insulin secretion in the isolated, perfused rat pancreas. These peptides (3.95 X 10(-8) M), like morphine (3.95 X 10(-8) M), significantly inhibited the glucose-induced insulin secretion. The inhibitory effect of rimorphin was attenuated by naloxone (1.2 X 10(-6) M) and phentolamine (10(-6) M), suggesting an involvement of adrenergic alpha receptors in the inhibition of glucose-induced insulin secretion mediated through specific opiate receptors. Rimorphin also inhibited glucose-induced insulin secretion even in the cysteamine-treated rat pancreas from which somatostatin had been depleted. Thus, somatostatin does not appear to play a major regulatory role in the insulin secretion in the pancreas.

Animals↗
PubMed1985

Distribution of human leumorphin-like immunoreactivity in the monkey spinal cord revealed by immunocytochemistry.

The regional distribution of human leumorphin (HL)-like immunoreactivity (HL-LI) in monkey (Macaca fuscata) spinal cord and dorsal root ganglia was investigated by peroxidase-anti-peroxidase immunocytochemistry using specific antiserum. HL-LI-positive fibers and terminals were distributed densely in laminae (Rexed) I and II, and sparsely in laminae III-VII and X, but no immunoreactive elements were observed in the ventral horn, the white matter or the dorsal root ganglia. Many immunoreactive neuronal perikarya were found in laminae I and II. Intrathecal injection of colchicine also revealed the presence of immunoreactive neuronal perikarya in laminae III-VII and X. These results suggest the presence of HL-LI, which represents HL and/or its C-terminal fragment, in the neuronal elements of the monkey spinal cord.

Animals↗
PubMed1985

The influence of serotonergic inputs on peptide neurons in the rat suprachiasmatic nucleus: an immunocytochemical study.

The influence of serotonin (5-hydroxytryptamine; 5-HT) innervation on peptide-containing neurons in the rat suprachiasmatic nucleus (SCN) was investigated by peroxidase-anti-peroxidase (PAP) immunocytochemistry. The 5-HT neuronal system was chemically severed by 5,6-dihydroxytryptamine (5,6-DHT) injection into the medial forebrain bundle bilaterally. After this treatment, a marked decrease of vasoactive intestinal peptide (VIP)-like immunoreactivity in neuronal perikarya occurred in the SCN corresponding to a decrease in number of 5-HT immunoreactive fibers and terminals. However, no alteration of arginine-vasopressin-like immunoreactivity was detected between 5,6-DHT-treated animals and the controls. It is speculated that VIP-like immunoreactive neurons play an important role in the SCN under the influence of strong 5-HT innervation.

5,6-Dihydroxytryptamine↗
PubMed1985

Presence of helodermin-like peptides of the VIP-secretin family in mammalian salivary glands and saliva.

Helodermin is a biologically active peptide isolated from the venom of the Gila monster lizard (Heloderma suspectum) whose structure is related to that of vasoactive intestinal peptide and secretin. Using a specific radioimmunoassay based on antisera prepared by immunizing rabbits with natural helodermin, we demonstrated the presence of helodermin-like material in mammalian salivary glands, including parotid, submaxillary and sublingual glands from rat and dog, and parotid and submaxillary glands from man. All helodermin-like materials had an apparent molecular mass of 4-12 kDa. Dog saliva, collected after pilocarpine stimulation, revealed similar immunoreactivity with a major component around 6 kDa.

Amino Acid Sequence↗
PubMed1985

Isolation and chemical characterization of glicentin C-terminal hexapeptide in porcine pancreas.

Using a radioimmunoassay specific for porcine glicentin C-terminal hexapeptide, we isolated a peptide from porcine pancreas and characterized it as the C-terminal 64-69 sequence of glicentin: H-Asn-Lys-Asn-Asn-Ile-Ala-OH. The purification steps included gel filtration, ion-exchange chromatography and HPLC. In each step, the recovery of the desired peptide, radioimmunologically estimated from the respective elution profile, was 71.4-91.7%. The final yield of the hexapeptide was 22 micrograms (4.3%) from 800 g pancreas. The pancreatic content of this peptide was estimated to be approximately equimolar to that of pancreatic glucagon. No hexapeptide-like component was detected in porcine intestinal extracts. The data confirmed that the processing of pancreatic proglucagon liberates the C-terminal hexapeptide of the intramolecular glicentin sequence in a tissue-specific manner during the production of glucagon.

Amino Acid Sequence↗