• Korean Journal of Food Science and Technology
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• v.28 no.5
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• pp.910-915
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• 1996

Ascidian muscle was blackened during the frozen storage, so the prevention against blackening was investigated. Low storage temperature and packaging in polyethylene bags delayed the blackening of ascidian muscle during the frozen storage. The blackening was prevented by dipping for $3{\sim}5$ minutes in 3% brine solution containing 0.3% citric acid, packaging in the polyethylene bag, freezing at $-45^{\circ}C$ for 5 hours and storing at $-20^{\circ}C$. Under this condition, the color and the quality of frozen ascidian muscle were nearly not changed for 200 days.

• Korean Journal of Food Science and Technology
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• v.28 no.6
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• pp.1021-1025
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• 1996

The experiments were carried out to find how the processing of ascidian (Halocynthia roretzi) was affected by the frozen storage. The quality of ascidian which takes the shell-on was changed quickly in the frozen storage. The causes of the change were as follows: 1. the damage caused by the ice crystal in the muscle, 2. a lot of drips after thawing, 3. the discoloration of the muscle after thawing. On the contrary, the quality of ascidian which takes the shell-off was even better in color, texture, yield and drips after 60 days of the frozen storage. But the muscle was blackened after that.

• Development and Reproduction
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• v.25 no.4
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• pp.299-303
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• 2021

The ascidian Halocynthia aurantium (sea peach), a marine invertebrate, belongs to the same genus of the phylum Chordata along with the ascidian Halocynthia roretzi (sea pineapple), which is one of the model animals in the field of developmental biology. The characteristics of development and reproduction of H. aurantium are not yet known in detail. In order to find out the spawning period of H. aurantium, we investigated development of the gonads during the annual reproductive cycle. Testis and ovary were both in the bisexual gonads (ovotestes) of H. aurantium, which is a hermaphrodite like H. roretzi. In H. aurantium, the right gonad was longer and slightly larger than the left gonad throughout the year. In each gonad, the number of the testis gonoducts was slightly higher than that of the ovary gonoducts. These features were similarly observed in H. roretzi. However, the number of the testis gonoducts and the ovary gonoducts in each gonad of H. aurantium was about half that of H. roretzi. The gonads of H. aurantium contracted during the winter and summer seasons. The gonads decreased to the smallest size around February, and then started to increase again in March. The gonads were most developed in September of the year. Therefore, it is estimated that the spawning of H. aurantium begins around this period.

• Journal of fish pathology
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• v.22 no.3
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• pp.229-237
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• 2009

The causative agent for the tunic softness syndrome of the cultured ascidian Halocynthia roretzi from Jan 1999 to Feb 2009 was identified using virus isolation and polymerase chain reaction (PCR). The pathogenicity of the isolated virus MABV UR-1 strain was determined by experimental infection trials. The cytopathic effects was observed in CHSE-214 cell line at a level 5.1% (4/78) in normal ascidian and 1.8% in abnormal ascidian showing tunic softness syndrome signs. MABV gene was detected in 16.8% (18/107) of normal and 13.1% (5/38) of abnormal organisms by PCR. The ratio of MABV isolation and gene detection was similar level in normal and soft tunic diseased ascidian. Based on the VP2/NS junction region sequences, eight strains of virus isolated from ascidian, were included in the same genogroup with MABV which is originally isolated in wide ranges of marine fish and shellfish species. The UR-1 strain caused 60% mortality (36.5% mortality in control group) by immersion infection and 37% mortality (same mortality in control group) in injection infection indicating no significant differences in infected and control groups. These results suggest that ascidian can act as reservoir of the MABV, and this virus is not directly related with the ascidian mortality.

• Fisheries and Aquatic Sciences
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• v.12 no.1
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• pp.44-48
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• 2009

Nucleotide sequences of about 500 bp from the 5' end of mitochondrial (mt) DNA Cytochrome Oxidase I (COI) were analyzed to estimate the genetic variation between wild and cultured populations of the ascidian Halocynthia roretzi from two sites along the coast of Korea. A total of 25 haplotypes were defined by 21 variable nucleotide sites in the examined COI region. Genetic diversity (haplotype diversity and nucleotide divergence) of wild populations was higher than that of the cultured population. These data suggest that reduced genetic variation in the cultured population may have results from bottleneck effect caused by the use of a limited number of parental stock and pooling of gametes for fertilization. Pairwise population $F_{ST}$ estimates inferred that wild and cultured populations were genetically distinct. The combined results suggest that sequence polymorphism in the COI region would be preferable for estimating the genetic diversity of ascidian populations.

• Development and Reproduction
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• v.15 no.4
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• pp.359-364
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• 2011

The seven selected primers OPA-02, OPA-04, OPA-18, OPD-07, OPD-08, OPD-15 and OPD-16 were used to generate unique shared loci to each species and shared loci by the two species. The hierarchical dendrogram indicates three main branches: cluster 1 (RORETZI 01~RORETZI 11) and cluster 2 (HILGENDORF 12~HILGENDORF 22) from two geographic populations of ascidians, Halocynthia roretzi and H. hilgendorfi. The shortest genetic distance displaying significant molecular difference was between individuals' HILGENDORF no. 14~HILGENDORF no. 19 (genetic distance =0.008). Ultimately, individual no. 02 of the RORETZI ascidian was most distantly related to HILGENDORF no. 21 (genetic distance=0.781). These results demonstrate that the H. roretzi population is genetically different from the H. hilgendorfi population. From what has been said above, the potential of PCR analysis to identify diagnostic markers for the identification of two ascidian populations has been demonstrated. Generally speaking, using a variety of decamer primers, this PCR method has been applied to identify specific markers particular to line, species and geographical population, as well as genetic diversity/polymorphism in diverse species of organisms.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.3
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• pp.283-291
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• 2006

This study examined the development of a low-salt fermented seafood product using an ascidian (Halocynthia roretzi), and the optimum processing conditions and quality characteristics of the low-salt fermented ascidian (LSA). The optimum processing conditions for the LSA were as follows. The ascidian was shelled and its muscle sliced into 5 mm widths. This was soaked in a 10% salt and 1% sodium erythorbate solution for 20 min. The solution was drained and then the muscle was soaked in 0.1% sodium bisulfite solution for 1 min. To this was added a 1:1 mixture of anchovy sauce and rice gruel, and it was fermented at $5^{\circ}C$ for 15 days. The moisture content and salinity of the LSA were 75.0-75.4% and 8.0-8.5%, respectively. During salt-fermentation at $5^{\circ}C$ for 20 days, the amino-N content of the LSA increased, and the texture softened gradually. The viable cell counts in early salt-fermentation were $4.2-4.5{\times}10^4CFU/g$, and this decreased gradually. The ratio of saturated fatty acids tended to increase in early salt-fermentation, while that of polyunsaturated fatty acids decreased slightly. Chemical experiments and sensory evaluation showed that the dipping treatment in 1% sodium erythorbate solution and 0.1% sodium bisulfite solution resulted in a good color and prevented browning of the salt-fermented ascidian meat. Moreover, adding anchovy sauce and rice gruel mixture improved the flavor of the LSA.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.2
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• pp.94-99
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• 2006

The tadpole larvae of most ascidians have two sensory pigment cells in their brain vesicle. The anterior otolith pigment cell is sensitive to gravity, whereas the posterior ocellus pigment cell responds to light. Besides these two sensory cells, the larvae also possess another type of sensory receptor cell: hydrostatic pressure receptor (Hpr) cells. The Hpr cells have been presumed to sense hydrostatic water pressure, although no functional analysis has been performed. In larvae of the ascidian Halocynthia reretzi, the development of the Hpr cells and their structure in the brain vesicle are poorly understood. To investigate the morphology and formation of the Hpr cells, we established a monoclonal antibody, Hpr-1, that specifically recognizes Hpr cells. The Hpr-1 antigens became detectable in the brain vesicle at the late tailbud stage. Each Hpr cell projected a small globular body, connected by a short stalk, into the lumen of the brain vesicle. The brain vesicle showed remarkable left-right asymmetry. Pigment cells were located on the right side in the lumen of the brain vesicle, whereas Hpr cells were present in the left side. After metamorphosis, the Hpr cells were observed near the rudimental siphons of the juvenile.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.6
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• pp.913-919
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• 2015

In vertebrates, the vitamin D receptor (VDR), a member of the nuclear receptor superfamily, binds the biologically active ligand $1{\alpha},25-(OH)_2$-vitamin $D_3$ (1,25 $D_3$). Nearly all vertebrates, including Agnatha, possess a VDR with high ligand selectivity for 1,25 $D_3$ and related metabolites. Although a putative ancestral VDR gene is present in the genome of the chordate invertebrate Ciona intestinalis, the functional characteristics of marine invertebrate VDR are still obscure. To elucidate the ascidian Halocynthia roretzi VDR (HrVDR), we cloned full-length HrVDR cDNA and investigated the transcriptional activity of HrVDR in HEK293 cells. HrVDR consists of 1,680 nucleotides (559 amino acids [aa]), including a short N-terminal region (A/B domain; 26 aa), DNA-binding domain (C domain; 72 aa), hinge region (D domain; 272 aa), and C-terminal ligand-binding domain (E domain; 161 aa). The amino acid sequence identity of HrVDR was greatest to that of C. intestinalis VDR (56%). In the luciferase reporter assays, the transcriptional activity of HrVDR was not significantly increased by 1,25 $D_3$, whereas the farnesoid X receptor agonist GW4064 increased the transactivation of HrVDR. These results suggest the presence of a novel ligand for and a distinct ligand-binding domain in ascidian VDR.

• Development and Reproduction
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• v.21 no.4
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• pp.467-473
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• 2017

Ascidian embryos have become an important model for embryological studies, offering a simple example for mechanisms of cytoplasmic components segregation. It is a well-known example that the asymmetric segregation of mitochondria into muscle lineage cells occurs during ascidian embryogenesis. However, it is still unclear which signaling pathway is involved in this process. To obtain molecular markers for studying mechanisms involved in the asymmetric distribution of mitochondria, we have produced monoclonal antibodies, Mito-1, Mito-2 and Mito-3, that specifically recognize mitochondria-rich cytoplasm in cells of the ascidian Halocynthia roretzi embryos. These antibodies stained cytoplasm like reticular structure in epidermis cells, except for nuclei, at the early tailbud stage. Similar pattern was observed in vital staining of mitochondria with DiOC2, a fluorescent probe of mitochondria. Immunostaining with these antibodies showed that mitochondria are evenly distributed in the animal hemisphere blastomeres at cleavage stages, whereas not in the vegetal hemisphere blastomeres. Mitochondria were transferred to the presumptive muscle and nerve cord lineage cells of the marginal zone in the vegetal hemisphere more than to the presumptive mesenchyme, notochord and endoderm lineage of the central zone. Therefore, it is suggested that these antibodies will be useful markers for studying mechanisms involved in the polarized distribution of mitochondria during ascidian embryogenesis.