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H Minakata

Publications and source records attributed to H Minakata.

At least 37 records · Page 2Linked to original sources
PubMed1999

Characterization of a cDNA encoding a precursor of Carassius RFamide, structurally related to a mammalian prolactin-releasing peptide.

We have characterized the cDNA encoding Carassius RFamide (C-RFa), which is structurally related to mammalian prolactin-releasing peptides (PrRPs), from the brain of the crucian carp. The deduced C-RFa precursor has been shown to comprise 117 amino acids, encoding a single C-RFa sequence. A comparative study of amino acid sequences has revealed that several sequences conserved in preproPrRPs are also found in the C-RFa precursor. Furthermore, the abundant presence of the C-RFa mRNA in the telencephalon, optic tectum, medulla oblongata, and proximal half eye ball was demonstrated by Southern blot analysis of RT-PCR products.

Amino Acid Sequence↗
PubMed1999

Evidence for conservation of the vasopressin/oxytocin superfamily in Annelida.

Annetocin is a structurally and functionally oxytocin-related peptide isolated from the earthworm Eisenia foetida. We present the characterization of the annetocin cDNA. Sequence analyses of the deduced precursor polypeptide revealed that the annetocin precursor is composed of three segments: a signal peptide, an annetocin sequence flanked by a Gly C-terminal amidation signal and a Lys-Arg dibasic processing site, and a neurophysin domain, similar to other oxytocin family precursors. The proannetocin showed 37.4-45.8% amino acid homology to other prohormones. In the neurophysin domain, 14 cysteines and amino acid residues essential for association of a neurophysin with a vasopressin/oxytocin superfamily peptide were conserved, suggesting that the Eisenia neurophysin can bind to annetocin. Furthermore, in situ hybridization experiments demonstrated that the annetocin gene is expressed exclusively in neurons of the central nervous system predicted to be involved in regulation of reproductive behavior. These findings confirm that annetocin is a member of the vasopressin/oxytocin superfamily. This is the first identification of the cDNA encoding the precursor of an invertebrate oxytocin-related peptide and also the first report of the identification of an annelid vasopressin/oxytocin-related precursor.

Amino Acid Sequence↗
PubMed1999

Localization of leech excitatory peptide, a member of the GGNG peptides, in the central nervous system of a leech (Whitmania pigra) by immunohistochemistry and in situ hybridization.

We have recently isolated a myoactive peptide, called leech excitatory peptide, belonging to the GGNG peptide family from two species of leeches, Hirudo nipponia and Whitmania pigra. Immunohistochemistry and in situ hybridization were employed to localize leech excitatory peptide-like peptide(s) and its gene expression in the central nervous system of W. pigra. A pair of neuronal somata were stained by both immunohistochemistry and in situ hybridization in the supraesophageal, subesophageal, and segmental ganglia. In addition, several other neurons showed positive signals by either immunohistochemistry or in situ hybridization in these ganglia. An immunoreactive fiber was observed to run in the anterior root of segmental ganglion 6, which is known to send axons to the sexual organs, though we failed to detect immunoreactivity in possible target tissues. Antiserum specificity was established by enzyme-linked immunosorbent assay using different leech excitatory peptide-related peptides. Leech excitatory peptide elicited muscular contraction of isolated preparations of penis and intestine at concentrations of 10(-8 )M. These results suggest that leech excitatory peptide is a neuropeptide modulating neuromuscular transmission in multiple systems, including regulation of reproductive behavior.

Amino Acid Sequence↗
PubMed1999

Characterization of cDNA and expression of mRNA encoding an Achatina cardioexcitatory RFamide peptide.

Achatina cardioexcitatory peptide-1 (ACEP-1) is an RFamide family peptide isolated from the atria of the African giant snail, Achatina fulica. In this report, we describe an identification of the ACEP-1 cDNA sequence and localizations of the ACEP-1 mRNA. Southern blot analysis revealed that the ACEP-1 mRNA was present in the atrium as well as in the central nervous system. Furthermore, in situ hybridization revealed the localizations of the ACEP-1 mRNA in small neurons of the cerebral and pedal ganglia and a few large neurons of the right parietal and visceral ganglia.

Amino Acid Sequence↗
PubMed1999

Characterization of a cDNA encoding a precursor polypeptide of a D-amino acid-containing peptide, achatin-I and localized expression of the achatin-I and fulicin genes.

Achatin-I and fulicin, isolated from the ganglia and atria of the giant land snail Achatina fulica, are a tetrapeptide and pentapeptide containing a d-Phe and d-Asn at position 2, respectively. We succeeded in cloning a cDNA encoding a precursor of achatin-I from the Achatina ganglia, revealing that the d-Phe present in achatin-I is coded by a common l-Phe codon, TTT or TTC. The deduced polypeptide was found to comprise seven repeats of the achatin sequence GFAD and one analog GFGD flanked on both sides by the typical endoproteolytic site KR. Northern blot analysis of transcripts and Southern blot analysis of reverse transcription (RT)-PCR products demonstrated that achatin-I mRNA was localized in the subesophageal ganglia, whereas expression of fulicin mRNA was detected in the atrium as well as in the subesophageal ganglia. Furthermore, localization of the achatin gene transcript in the right and left pedal ganglia compartments was shown by in situ hybridization on sections of subesophageal ganglia, whereas the fulicin transcript was observed in the right and left parietal ganglia. These data suggested that achatin-I plays an essential role in the regulation of the heart as a neurotransmitter or neurohormone through production in the pedal ganglia and transport to the atrium, whereas fulicin serves not only as a neurotransmitter or neurohormone but also as a novel atrial hormone.

Amino Acid Sequence↗
PubMed1999

Comparison of precursor structures of the GGNG peptides derived from the earthworm Eisenia foetida and the leech Hirudo nipponia.

Earthworm and leech cDNAs encoding the GGNG peptides, a family of myotropic peptides, were cloned and examined in this study. Both of the predicted precursor proteins are of polyprotein structure and contain several putative peptides distinct from the GGNG peptides. However, the precursors show organizations distinct from each other and no sequence similarity except for the GGNG peptides.

Amino Acid Sequence↗
PubMed1998

Peptides controlling stifness of connective tissue in sea cucumbers.

We present the first evidence of a system of four bioactive peptides that affect the stiffness of sea cucumber dermis. The body wall dermis of sea cucumbers consists of catch connective tissue that is characterized by quick and drastic stiffness changes under nervous control. The peptides were isolated from the body wall, their amino acid sequences determined, and identical peptides synthesized. Two peptides, which we named holokinins, are homologous with bradykinin. We tested the effect of the peptides on the mechanical properties of sea cucumber dermis. Both of the holokinins softened the dermis, and a pentapeptide that we designated as NGIWYamide stiffened it. Both effects were reversibly suppressed by anesthesia with menthol. We called the fourth peptide stichopin; it had no direct effect on the stiffness of the dermis but suppressed action of the neurotransmitter acetylcholine reversibly. The results suggest that the peptides are neuropeptides and are part of a sophisticated system of neurotransmitters and neuromodulators that controls the connective tissue stiffness of sea cucumber dermis.

Amino Acid Sequence↗
PubMed1997

The leech excitatory peptide, a member of the GGNG peptide family: isolation and comparison with the earthworm GGNG peptides.

A member of the GGNG peptide family was isolated from Hirudo nipponia (leech). GGNG peptides had only been isolated previously from earthworms. The C-terminus structure of the leech peptide, LEP (leech excitatory peptide), was -Gly-Gly-Asn-amide, while that of the earthworm peptides, EEP (earthworm excitatory peptide), was -Gly-Gly-Asn-Gly. LEP exerted 1000-fold more potent activities on leech gut than did EEP-2. On the other hand, EEP-2 was 1000-fold more potent than LEP on the crop-gizzard of the earthworm. Analog peptides of LEP and EEP-2 were synthesized, and the myoactive potency of each analog on the leech and earthworm tissues was compared.

Amino Acid Sequence↗
PubMed1997

Demonstration, localization, and development of galanin receptors in the quail oviduct.

We recently isolated an oviposition-inducing peptide that was identified as avian galanin from the oviducts of the Japanese quail. Avian galanin was localized in neural fibers distributed in muscle layers in the uterine and vaginal oviduct regions, and potentiated spontaneous contractions of the uterus and vagina. To elucidate whether an oviposition-inducing effect of avian galanin is due to the direct action on the oviduct, therefore, a specific binding site for avian galanin was determined in the functional quail oviduct in this study. The binding of [125I]iodoavian galanin was primarily located in the oviduct as well as the brain. The galanin binding was specifically inhibited as a function of the concentrations of both avian and rat galanins. The specific binding of avian galanin to the quail oviduct was temperature dependent and reached the maximum level for 1 h at 20 degrees C. In several regions of the oviduct, a higher level of specific galanin binding was observed only in the uterus and vagina. In contrast, the specific binding was low in the isthmus and negligible in the magnum. A similar localization was evident in the functional chicken oviduct. The Scatchard plot analysis of the binding of avian galanin to the uterine preparation revealed that the dissociation constant (Kd) was 0.249 (95% confidence interval, 0.192-0.356) nM, and the number of binding sites was 1.13 (0.99-1.36) fmol per mg tissue, respectively. During development, the galanin-binding sites were apparent in the quail oviduct at 3 weeks of age and the number of binding sites markedly increased between 3 weeks and 3 months of age. However, there was no significant change in the Kd value in the developing quail oviduct. This is the first demonstration of the presence of galanin receptors in the reproductive tract, such as the uterine and vaginal oviduct. The present results suggest that the number of galanin receptors in the oviduct increases during development and that galanin acts directly on the mature uterus and vagina to induce their contractions. This mechanism may be essential to the avian oviposition.

Aging↗
PubMed1997

Immunochemical demonstration of Eisenia tetradecapeptide, a bioactive peptide isolated from the gut of the earthworm Eisenia foetida, in tissues of the earthworm.

The quantity and localization of Eisenia tetradecapeptide which was isolated from the gut of the earthworm Eisenia foetida were examined in tissues of the same species by enzyme-linked immunosorbent assay and immunohistochemistry. Analysis by enzyme-linked immunosorbent assay showed that Eisenia-tetradecapeptide-like immunoreactivity was present in both the central nervous system (cerebral ganglion, subesophageal ganglion, ventral ganglia, and ventral nerve cord) and the gut (esophagus, crop, gizzard, and intestine). The central nervous system contained a higher amount of Eisenia-tetradecapeptide-like immunoreactivity (1.3 pmol/mg wet weight) than the gut (0.2-0.6 pmol/mg wet weight). Eisenia-tetradecapeptide-like immunoreactivity was scarcely detected in the body-wall muscle, nephridia, and sexual organs (testis, ovary, seminal vesicle, and ovisac). Immunohistochemical analysis demonstrated that intense Eisenia-tetradecapeptide-like immunopositive cells and nerve fibers were present in the central nervous system. Immunoreactivity was found in the epithelial cells lining the esophagus and in the submucous plexus in various parts of the gut. Thus, the present study suggests that Eisenia tetradecapeptide is a neuropeptide and/or peptide hormone present in both the central nervous system and the gut of the earthworm and that its role involves the regulation of gut motility.

Amino Acid Sequence↗
PubMed1997

A novel D-amino acid-containing peptide, fulyal, coexists with fulicin gene-related peptides in Achatina atria.

Fulicin (Phe-D-Asn-Glu-Phe-Val-NH2) is a neuropeptide from ganglia of the African giant snail (Achatina fulica). Previously, the sequences of nine fulicin gene-related peptides (FGRP-1 to -9) have been predicted from the cDNA encoding the ganglia fulicin precursor and the transcripts have been detectable in the heart. We synthesized twenty peptides related to fulicin and FGRPs containing either an L- or a D-amino acid at position 2 and used them to identify FGRPs in atrial extracts. We identified ten alpha-amidated peptides, including fulicin and confirmed their structures as follows: Tyr-Ala-Glu-Phe-Leu-NH2 (FGRP-9), [D-Ala2]FGRP-9 (fulyal), [L-Asn2]fulicin, fulicin, Ser-Tyr-Asp-Phe-Val-NH2 (FGRP-2), Thr-Tyr-Asp-Phe-Leu-NH2 (FGRP-3), Tyr-Asp-Phe-Ile-NH2 (FGRP-5), Ser-Pro-Tyr-Asp-Phe-Ile-NH2 (FGRP-6), Asn-Tyr-Asp-Phe-Val-NH2 (FGRP-7) and Ser-Pro-Tyr-Asp-Phe-Val-NH2 (FGRP-8). We analyzed the biological activities of synthetic FGRPs using the snail penis retractor muscle. The results revealed that fulyal remarkably potentiated the tetanic contraction at concentrations as low as 10(-12) M. FGRP-9 was about 10,000-fold less potent. Fulyal, like fulicin, seems to undergo preferential maturation to participate in the penis retractor muscle contraction as a neuropeptide containing a D-amino acid.

Amino Acid Sequence↗
PubMed1996

Annetocin, an annelid oxytocin-related peptide, induces egg-laying behavior in the earthworm, Eisenia foetida.

Annetocin, an oxytocin-related peptide which we isolated from the earthworm Eisenia foetida, induced a series of egg-laying-related behaviors in the earthworms. These stereotyped behaviors consisted of well-defined rotatory movements, characteristic body-shape changes, and mucous secretion from the clitellum. Each of these behaviors is known to be associated with formation of the cocoon in which eggs are deposited. In fact, some of the earthworms injected with annetocin (> 5 nmol) laid eggs. Such egg-laying-related behaviors except for oviposition were also induced by oxytocin, but not by Arg-vasopressin and some other bioactive peptides isolated from E. foetida. Furthermore, annetocin also induced these egg-laying-like behaviors in the leech Whitmania pigra, but not in the polychaete Perinereis vancaurica. These results suggest that annetocin plays some key role in triggering stereotyped egg-laying behaviors in terrestrial or fresh-water annelids that have the clitella.

Amino Acid Sequence↗
PubMed1996

An oviposition-inducing peptide: isolation, localization, and function of avian galanin in the quail oviduct.

It is well established that avian oviposition is regulated, at least partly, by a neurohypophysial hormone, arginine vasotocin, and ovarian hormones, prostaglandins, are regulated through mechanisms of the induction of uterine contractions. Although abundant nerves are terminated in the musculature in the uterine and vaginal oviduct regions, limited information is available on the neuronal control mechanism of avian oviposition. To identify the oviduct factor that plays an important role in the induction of oviposition as a neurotransmitter or a neuromodulator, a study was conducted to isolate the bioactive substance involved in the vaginal and uterine contractions from mature oviducts of the Japanese quail. Acetic acid extracts of 200 quail oviducts were forced through disposable C-18 reversed-phase cartridges, and then the retained material was subjected to the reversed-phase and cation-exchange HPLC purifications. A purified bioactive substance showed a single peak on the reversed-phase HPLC and was further subjected to amino acid sequence analysis and molecular weight presumption. The substance was identified as avian galanin, which was previously isolated from chicken intestine. The synthetic peptide enhanced contractions of both the vagina and the uterus in a manner similar to the native peptide, and the threshold concentrations were 10(-9) - 10(-8) M in the vagina and 10(-10) - 10(-9) M in the uterus. An intraperitoneal injection of the synthetic peptide significantly evoked quail oviposition during 5 min after injection. Immunohistochemical analysis with the anti-galanin serum revealed that abundant immunoreactive fibers were distributed in muscle layers of the vagina and the uterus. The immunoreaction examined in these regions was completely inhibited by preincubation of the antibody with synthetic peptide. These results suggest that avian galanin in the oviduct evokes oviposition through mechanisms of the induction of uterine and vaginal contractions. This peptide may contribute as a neurotransmitter or a neuromodulator to avian oviposition.

Amino Acid Sequence↗
PubMed1995

The GGNG peptides: novel myoactive peptides isolated from the gut and the whole body of the earthworms.

Three novel bioactive peptides, which were structurally related to each other, were purified and chemically identified from two species of the earthworms, Eisenia foetida and Pheretima vittata. One peptide was isolated from the gut tissue of E. foetida, and the other two were purified from the whole body of E. foetida and P. vittata, respectively. These peptides consisted of 17 or 18 amino acid residues and were named GGNG peptides after their unique, common structure of the C-termini. These GGNG peptides augmented both tension and frequency of spontaneous contractions of isolated gut preparations of E. foetida, and also elicited contractions of other annelidan tissues such as the polychaete esophagus and the leech vagina. However, they showed no activity on molluscan or arthropodan tissues, suggesting that GGNG peptides may be peculiar to annelids. No peptides homologous to GGNG peptides have been known so far in any living organisms.

Amino Acid Sequence↗