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Tadaaki Moritomo

Publications and source records attributed to Tadaaki Moritomo.

25 records · Page 2Linked to original sources
PubMed2003

Molecular cloning and sequencing of the banded dogfish (Triakis scyllia) interleukin-8 cDNA.

The dogfish (Triakis scyllia) interleukin-8 (IL-8) cDNA was isolated from mitogen-stimulated peripheral white blood cells (WBCs) utilising the polymerase chain reaction (PCR). The cDNA sequence showed that the dogfish IL-8 clones contained an open reading frame encoding 101 amino acids. A short 5' untranslated region (UTR) of 70 nucleotides and a long 3' UTR of 893 nucleotides were also present in this 1.2-kb cDNA. Furthermore, the 3' UTR of the mRNA contained the AUUUA sequence that has been implicated in shortening of the half-life of several cytokines and growth factors. The predicted IL-8 peptide had one potential N-linked glycosylation site (Asn-72-Thr-74) that is not conserved in other vertebrates. It also contained four cysteine residues (Cys-34, 36, 61 and 77), which are characteristic of CXC subfamily cytokines and found in all vertebrates, to date. The dogfish IL-8 lacked an ELR motif as found in the lamprey and trout. Comparison of the deduced amino acids showed that the dogfish IL-8 sequence shared 50.5, 41.2, 37.1 and 40.4-45.5% identity with the chicken, lamprey, trout and mammalian IL-8 sequences, respectively.

Amino Acid Sequence↗
PubMed2003

Flow cytometric analysis of the neutrophil respiratory burst of ayu, Plecoglossus altivelis: comparison with other fresh water fish.

Neutrophils of vertebrates undergo respiratory burst activity (RBA) as a defense mechanism against bacterial infections. We report here that ayu (Plecoglossus altivelis) have unusually high RBAs even when they are in a healthy condition. Kidney and blood leukocytes were obtained from ayu, rainbow trout (Oncorhynchus mykiss), carp (Cyprinus carpio), eel (Anguilla japonica), and pond smelt (Hypomesus nipponensis). Neutrophil RBA was measured by flow cytometry using dihydrorhodamine after stimulation with phorbol myristate acetate. The amount of RBA of neutrophils from both blood and kidney was significantly higher in ayu than in the other species (e.g. the fluorescence intensity of ayu blood neutrophils was about 3-7 times higher than that from trout and carp, and that of ayu kidney neutrophils was 2-19 times higher than that of rainbow trout, carp, eel, and pond smelt). This unique character of ayu neutrophils was invariable even at different ages, locations, and sex-maturation stages.

Anguilla↗
PubMed2002

Characterization of immunoglobulin light chain isotypes in the common carp.

Two or three isotypes of immunoglobulin (Ig) light (L) chain have been demonstrated in several teleost species thus far. A reverse transcription-polymerase chain reaction (RT-PCR) strategy was used with the aim of isolating cDNA clones encoding IgL chain from the common carp ( Cyprinus carpio. L.) different from the previously isolated isotype, designated L1A. cDNA clones representing two novel isotypes, designated L1B and L3, were obtained in this study. For CL segments, L1B sequences were similar to type L1/G in various teleost species, as well as L1A, while L3 sequences were closely related with type L3/F in catfish and zebrafish. For VL segments, however, L1A sequences demonstrated higher similarity to L3 rather than L1B. These results were also supported by the phylogenetic study. The similarity found between L1A and L3 VL sequences was based on the very high degree of nucleotide identity in FR1, and/or FR2 to FR3, strongly suggesting the involvement of interlocus recombination between them. Southern blot analyses suggested that the locus of L1B has a cluster-like organization, but that those of L3 and L1A are currently unknown due to cross-hybridization between those VL probes. Northern blot analyses showed that CL mRNA of each isotype was expressed in lymphoid tissues examined, particularly strongly in pronephros, and was predominantly composed of approximately 1-kb and 0.6-kb transcripts, possibly corresponding to fully spliced (LVJC) and truncated (JC/J-C) forms, respectively.

Amino Acid Sequence↗
PubMed2002

A new method for fish leucocyte counting and partial differentiation by flow cytometry.

A simple and rapid method for analysis of fish blood cells is presented. Carp (Cyprinus carpio) blood was diluted 200 times with Hanks' solution containing 1 microg/ml of DiOC6(3) which is a fluorescent, lipophilic dye. After staining for 10 min, the blood cells were measured by a flow cytometer (FACS). Several blood cell populations were identified by different FL-1 (green fluorescence), FSC (forward scatter), and SSC (side scatter) properties. FL-1 v. SSC or FSC v. SSC dot-plot of stained blood cells displayed five separate cell populations: erythrocytes: a mixture of thrombocytes plus lymphocytes; monocytes; neutrophils; and basophils. The number of each type of blood cell counted by the FACS was in good agreement with those counted microscopically.

Animals↗
PubMed2002

Granulocyte responses to experimental injection of live and formalin-killed bacteria in carp (Cyprinus carpio).

The objective of the study was to investigate the dynamics of changes in number of granulocytes in bacterial infections of carp (Cyprinus carpio). Carp were inoculated with non-pathogenic or pathogenic bacteria and changes in type I (neutrophils) and type II granulocyte (basophils/eosinophils) counts in kidney, circulating blood and peritoneal cavity were assessed. After the injection of non-pathogenic bacteria (Escherichia coli), the number of type I and II cells in blood increased after 6-12h, but returned to the control level after 24-48 h. In contrast, after the injection of pathogenic bacteria (Aeromonas hydrophila), the number of type I cells initially increased followed by an increase in the number of type II cells. The peak counts of type I and II cells were at 12 and 24h after the injection, respectively. When the fish were given serial-injections of formalin-killed bacteria at 12-h intervals, the type II cells also predominantly increased and remained at high levels, following the peak count of type I cells.

Aeromonas hydrophila↗
PubMed2002

A new method for counting of quail leukocytes by flow cytometry.

An automatic counting method was developed for fish blood cells using a fluorescent dye, 3, 3-dihexyloxacarbocyanine (DiOC (6)(3)), that selectively stain lipid bilayers in living cells. In the present study, the DiOC(6)(3) method was applied to quail (Cotumix cotumix japonica) blood cells. After quail blood cells were stained with DiOC(6)(3), absolute counts and relative proportions of erythrocytes, granulocytes, monocytes, and lymphocytes plus thrombocytes in whole blood were obtained by means of flow cytometry (FC). The number of each cell types by the FC was in good agreement with those counted microscopically. This method will offer new possibilities for routine blood cell counting for avian medicine.

Animals↗