Pattern recognition receptors involved in the immune system of hagfish (Eptatretus burgeri).

The initial defense against invading pathogenic microbes is the activation of innate immunity by binding of pattern recognition receptors (PRRs) to pathogen associated molecular patterns (PAMPs). To explain the action of PRRs from hagfish, one of the extant jawless vertebrates, we purified the GlcNAc recognition complex (GRC) from serum using GlcNAc-agarose. The GRC comprises four proteins of varying molecular masses: 19 kDa, 26 kDa, 27 kDa, and 31 kDa.

The Germline Marker Expressed in the Skin Layer of the Polychaete After Injury.

Nereidid polychaete is a homonomous metameric worm with a complete septum between each segment. Each segment has germ cells localized in the distal area of the parapodia. is also known to have high abilities of tissue regeneration; however, it is still unclear whether germ cells can regenerate in the healing tissue.

Germ Cell Development in Male and Gamete Spawning Mechanisms in Males and Females.

is a fully segmented worm with complete intersegmental septa. A previous study of females revealed that germ cells of this animal originate in the tail end segment, called the pygidium. Germ cells were duplicated in the pygidium, transferred to a newly generated segment, and then settled in the parapodia.

High concentrations of perfluorooctane sulfonate in mucus of tiger puffer fish Takifugu rubripes: a laboratory exposure study.

Distribution of perfluorooctane sulfonate (PFOS) was investigated in tissues (plasma, blood clot, mucus, skin, liver, muscle, and gonad) of tiger puffer fish Takifugu rubripes. A single dose of PFOS was intraperitoneally injected at 0.1 mg/kg body weight with samples taken over a 14-day period.

A model for germ cell development in a fully segmented worm.

Introduction: Polychaetes are segmented marine worms with body segments separated by a complete or incomplete septum. In most polychaetes the whole body cavity is filled with gametes during the breeding season. Platynereis dumerilii (Pl.

An oocyte-specific astacin family protease, alveolin, is released from cortical granules to trigger egg envelope hardening during fertilization in medaka (Oryzias latipes).

It has long been hypothesized that in fishes the contents of cortical granules are involved in the hardening of egg envelope following fertilization. We previously purified the egg envelope hardening initiation factor from the exudates released from activated medaka (Oryzias latipes) eggs and tentatively termed this protein alveolin. Alveolin is a member of the astacin metalloprotease family and was proposed to be a protease which hydrolyzes ZPB at one restricted position to allow starting cross-linking with ZPC.

A relaxin-like peptide purified from radial nerves induces oocyte maturation and ovulation in the starfish, Asterina pectinifera.

Gonad-stimulating substance (GSS) of starfish is the only known invertebrate peptide hormone responsible for final gamete maturation, rendering it functionally analogous to the vertebrate luteinizing hormone (LH). Here, we purified GSS of starfish, Asterina pectinifera, from radial nerves and determined its amino acid sequence. The purified GSS was a heterodimer composed of 2 different peptides, A and B chains, with disulfide cross-linkages.

Neuronal peptides induce oocyte maturation and gamete spawning of sea cucumber, Apostichopus japonicus.

Extracts prepared from tissues containing buccal ring nerve or longitudinal radial nerve of sea cucumber induce oocyte maturation and ovulation from ovarian tissues. We purified two small peptides, a pentapeptide and a heptapeptide, from the buccal tissues of Japanese common sea cucumber, Apostichopus japonicas. Both peptides induced oocyte maturation and gamete spawning.

Preliminary study on the receptor of gonad-stimulating substance (GSS) as a gonadotropin of starfish.

Previously, the gonad-stimulating substance (GSS) which acts as the gonadotropin was purified from the starfish (Asterina pectinifera) and subsequently, its structure was deciphered. In this study, artificial GSS was synthesized and its interaction with the receptors was examined further. According to competitive experiments using radioiodinated and radioinert GSS in various tissues of A.

Phosphorylation of the p34(cdc2) target site on goldfish germinal vesicle lamin B3 before oocyte maturation.

The nuclear membranes surrounding fish and frog oocyte germinal vesicles (GVs) are supported by the lamina, an internal, mesh-like structure that consists of the protein lamin B3. The mechanisms by which lamin B3 is transported into GVs and is assembled to form the nuclear lamina are not well understood. In this study, we developed a heterogeneous microinjection system in which wild-type or mutated goldfish GV lamin B3 (GFLB3) was expressed in Escherichia coli, biotinylated, and microinjected into Xenopus oocytes.

Purification and characterisation of blarinasin, a new tissue kallikrein-like protease from the short-tailed shrew Blarina brevicauda: comparative studies with blarina toxin.

A new tissue kallikrein-like protease, blarinasin, has been purified from the salivary glands of the short-tailed shrew Blarina brevicauda. Blarinasin is a 32-kDa N-glycosylated protease with isoelectric values ranging between 5.3 and 5.

Identification of a protein kinase which phosphorylates a subunit of the 26S proteasome and changes in its activity during meiotic cell cycle in goldfish oocytes.

The proteasome is involved in the progression of the meiotic cell cycle in fish oocytes. We reported that the alpha4 subunit of the 26S proteasome, which is a component of the outer rings of the 20S proteasome, is phosphorylated in immature oocytes and dephosphorylated in mature oocytes. To investigate the role of the phosphorylation, we purified the protein kinase from immature oocytes using a recombinant alpha4 subunit as substrate.

Blarina toxin, a mammalian lethal venom from the short-tailed shrew Blarina brevicauda: Isolation and characterization.

Venomous mammals are rare, and their venoms have not been characterized. We have purified and characterized the blarina toxin (BLTX), a lethal mammalian venom with a tissue kallikrein-like activity from the submaxillary and sublingual glands of the short-tailed shrew Blarina brevicauda. Mice administered BLTX i.

Inhibitory effect of a SALMFamide neuropeptide on secretion of gonad-stimulating substance from radial nerves in the starfish Asterina pectinifera.

In starfish, the peptide hormone gonad-stimulating substance (GSS) secreted from nervous tissue stimulates oocyte maturation to induce 1-methyladenine (1-MeAde) production by ovarian follicle cells. The SALMFamide family is also known to an echinoderm neuropeptide. The present study examined effect of SALMFamide 1 (S1) on oocyte maturation of starfish Asterina pectinifera.

A shift in steroidogenesis occurring in ovarian follicles prior to oocyte maturation.

Gonadotropins (GTHs; FSH and LH) require two major steroidal mediators, estradiol-17 beta (E(2)) and 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17 alpha,20 beta-DP) to act as critical hormones to execute oocyte growth and maturation, respectively. A two-cell type model has been proposed, where the theca cells provide the precursor steroids, and the granulosa cells produce the two steroidal mediators under the direct influence of FSH and LH. A distinct shift in steroidogenesis, i.

Molecular cloning of DAX1 and SHP cDNAs and their expression patterns in the Nile tilapia, Oreochromis niloticus.

Piscine DAX1 and SHP cDNAs with an open reading frame encoding 296 and 258 amino acid residues, respectively, as well as SHP partial gene fragment, were cloned from Nile tilapia. Phylogenetic analyses of DAX1s, SHPs, and homologous EST fragments indicate that DAX1 and SHP are conserved in gene structure and are present throughout vertebrates. A single band of approximately 1.

Characterization of ovarian membrane receptor for 17,20beta-dihydroxy-4-pregnen-3-one, a maturation-inducing hormone in yellowtail, Seriola quinqueradiata.

In our previous studies, we tentatively identified 17,20 beta-dihydroxy-4-pregnen-3-one (17,20 beta-P) as a maturation-inducing hormone (MIH) in yellowtail (Seriola quinqueradiata) through in vivo and in vitro experiments. In this study, we investigated the binding sites for radioactive 17,20 beta-P and characterized the receptor binding to the ovarian plasma membrane in yellowtail undergoing first stage of maturation (FSM). Equilibrium binding sites for 17,20 beta-P have been detected within 1h incubation and the binding dissociated completely within 50 min at 4 degrees C and was pH dependent (optimum pH 7.

Vitellogenin Incorporation into Oocytes of Rainbow Trout, Oncorhynchus mykiss, in Vitro: Effect of Hormones on Denuded Oocytes: (vitellogenin/oocyte growth/insulin/thyroxine/rainbow trout).

An in vitro culture procedure to measure vitellogenin (VTG) incorporation into oocytes without follicle cell layers was developed using oocytes of the rainbow trout, Oncorhynchus mykiss. Oocytes incorporated VTG specifically and linearly for up to 24 hr. The maximum incorporation observed was 314 μg/24 hr/oocyte, using vitellogenic (3.

A Monoclonal Antibody Against the PSTAIR Sequence of p34 , Catalytic Subunit of Maturation-Promoting Factor and Key Regulator of the Cell Cycle: (monoclonal antibody/PSTAIR sequence/oocyte maturation/cell cycle/cdc2).

A homolog of the serine/threonine protein kinase (p34 ), encoded by the cdc2 gene of the fission yeast (Schizosaccharomyces pombe), is a catalytic subunit of maturation-promoting factor and a key regulator of the cell cycle. We have raised a monoclonal antibody against the most conserved amino acid sequence, the PSTAIR sequence (EGVPSTAIREISLLKE) of p34 This antibody recognizes 31-34 kDa proteins by immunoblotting in all species examined so far. The proteins recognized by the anti-PSTAIR antibody are probably either p34 itself or proteins highly homologous to p34 in the given species, since, in all species studies to date, they are all precipitated with p13 , the fission yeast suc1 gene product, which binds to p34 with high specificity.

Involvement in Meiotic Prophase of H1 Histone Kinase and p34 Homologues in Lily (Lilium longiflorum) Microsporocytes: (lily microsporocyte/p34 kinase/meiosis/pachytene).

We have taken advantage of the synchrony of meiotic prophase I in Lilium microsporocytes to investigate the presence and involvement in four stages of meiotic prophase I (leptotene, zygotene, pachytene, and diplotene) of the p34 H1 histone kinase, a component of MPF and a key participant in division control in other eukaryotes. H1 kinase activity showed a peak pattern during meiotic prophase I with the highest kinase activity at pachytene. A monoclonal antibody directed against a highly conserved region of p34 (termed the 'PSTAIR') recognized three major protein forms by immunoblotting.

Inhibition of Starfish Oocyte Maturation by Tumor-Promoting Phorbol Esters*: (starfish oocyte maturation/tumor promoters/phorbol esters/teleocidin/retinoids).

Effect of tumor promoters including phorbol esters and teleocidin on 1-methyladenine (1-MeAde)-induced oocyte maturation was studied in the starfish. When isolated immature oocytes were treated with 1-MeAde and 12-O-tetradecanoylphorbol-13-acetate (TPA), 1-MeAde-induced maturation was completely inhibited at more than 2.5 μg/ml.