온도; pH, 염분 내성 및 충격이 개불, Urechis unicinctus 유생에 미치는 영향

최상덕,김호진,라성주,정형택,윤호섭,신종암,이원교 國立 麗水大學校 環境問題硏究所 1999 環境硏究論文集 Vol.2 No.-

The effect of different temperature, salinity and pH on survival rate of Urechis unicinctus's swimming embryo was examined to develop the method for the efficient seed production. In addition to, this study was designed to investigate the possibility of pollutant marker diagnosis of marine pollutions by the change in different temperature, salinity and pH. Treatment was carried out whit different temperature (5˚C, 10˚C, 17˚C, 20˚C, 25˚C,), salinity(0‰, 10‰, 20‰, 30‰, 34‰, 40‰) and pH(4, 6, 7.2, 8.2, 9.2, 10). All the swimming embryo of the temperature experiment was survived within all experiment section, but the others experiment were not. At the temperature-shock and tolerance experiment ranging from 5˚C to 25˚C the survival rate was occurred: 75%, 95%, 100%, 100%, 50%, at the ph-Shock experiment ranging from 4 to 10, the survival rate was occurred: 0%, 50%, 85%, 100%, 60%, 0%, at the pH-tolerance experiment ranging 4 to 10 the survival rate was occurred : 0%, 75% 90%, 100%, 80%, 0%, and at the salinity-shock experiment ranging from 0% to 40% the survival rate was occurred : 0% 10% 90% 95% 100% 0% , at the salinity-tolerance experiment ranging from 0% to 40% , the survival rate was occurred : 0% 0% 90% 100% 100% 0%. These results suggest, that the survival rate of larvae of Urechis unicinctus was the highest at 17˚C, 34%. pH8.2 and this Urechis unicinctus could be live at the zone of euryceious and zone of euryhalines

아미노산 6종에 대한 개불, Urechis unicinctus 유생의 착저반응

최태봉,라성주,윤순기,유태식,최상덕 한국수산해양교육학회 2017 水産海洋敎育硏究 Vol.29 No.6

Settlement and metamorphosis of marine invertebrate larvae are induced by substratum-specific biochemical signals from the environment. In several species, a basic similarity between the biochemical signals required for induction of larval settlement and metamorphosis has been found. These inducers are protein or peptide associated from amino acids. So we thought that larvae and adult were closely connected. We analyzed amino acids of the Urechis unicinctus larvae, mucus, egg and sperm. And investigation was carried out on the effects of six different amino acids on larval settlement of the Urechis unicinctus. Among them, the most effective amino acid was leucine which had a lower threshold of settlement of 1×10-7M and caused settlement rate was more than 20%. Glycyn, alanine, lysine, aspartic acid and glutamic acid had thresholds between 1×10-6M and 1×10−2M. On the other hand, lysine induced a high percentage of larval settlement at 1×10-5M, but became toxic at high concentration.

개불 (Urechis unicinctus) 체강에서의 단위집단 정자형성(Solitary Spermatogenesis)에 관한 미세구조 연구

신길상,김완종,Shin, Kil-Sang,Kim, Wan-Jong 한국현미경학회 2002 Applied microscopy Vol.32 No.2

U. unicinctus의 초기 정자형성 과정은 체액 부유성 웅성 생식세포 집단(cluster)에서 수행되는 것으로 관찰되었다. 이들 웅성 생식세포 집단은 체강의 체액 속에 산재하고 다른 체세포와 특정 구조 또는 조직을 형성하지 않았으므로 분산형 정자형성(diffuse spermatogenesis) 또는 단위집단 정자형성 (solitary spermatogenesis) 과정을 거치는 것으로 볼 수 있었다. 단위집단 정자 형성과정은 생식세포들이 체세포와 함께 조직을 형성하는 포유동물의 집단 정자형성(localized spermatogenesis)과 비교하여 미세구조 측면에서 여러 가지 특징을 갖는 것으로 관찰되었다. U. unicinctus의 단위집단 정자형성에서 웅성 생식세포 발생단계는 핵의 크기, 핵질(nuclear matrices)의 위상 그리고 핵 응축도 및 웅성 생식세포 기관의 발달 상태를 고려하여 따로 분류하였다. 정원세포와 제1 정모세포의 차이는 핵질의 위상에서 분명하였고, 정모세포와 정세포는 핵 응축도에서 차이를 볼 수 있었다. 단위집단을 형성하는 시기는 정원세포기로서 각 정원세포 돌기들이 서로 융합한다는 증거를 볼 수 있었다. 제1, 제2 정모세포 그리고 정세포들은 단위집단의 외곽에 부착하고 있었으며 내부에는 웅성 생식세포 응축 과정에 수반되어 분리된 또는 제거된 세포질 성분(cytoplasmic mass)이 채워지는 것으로 볼 수 있었다. 성숙 후 단위집단에서 체강으로 분리되는 시기는 정세포기이며, 체강에서 관찰된 성숙정자는 물질농도에서 구획이 형성된 원반형 첨체, 미토콘드리아와 편모의 구조로 보아서 운동성과 수정력이 있는 것으로 보인다. 이와 같은 단위집단 생식세포 형성은 체액을 통한 간접 방법에 의해 대사 작용에 필요한 정보와 물질을 교환하기 때문에 가능한 것으로 추측하였으며, 이를 응용하면 적절한 양의 체액을 공급하여 in vitro에서 생식세포 배양이 가능할 것으로 사료되었다. Early spermatocytes of U. unicinctus are found in cluster floating in the coelomic fluid. The spermatocytes in a cluster form a syncytium or cytoplasmic mass, but there are no indications that the cytoplasmic mass is a component of a somatic cell. This work suggested that this type of spermatogenesis can be subordinated to solitary spermatogenesis in the sense excluding structural and functional support of a somatic cell for sperm developments. The solitary spermatogenesis in U. unicinctus is different in appearances and developmental details of sperm organelles and stage distributions from that of localized spermatogenesis. The acrosomal rudiments and centrioles can be observed in the early single cells of spermatogonia and clearly disclosed in the primary spermatocyte. In the stage of secondary spermatocyte, the acrosomal precursor and the centrioles begin to move to each cytoplasmic poles. The polarities of the organelles are attained at stage of spermatids. The spermatocytes and spermatids are arranged circumferentially along the cytoplasmic mass in which some amorphological cytoplasmic components are included. The spermatids reveal to be detached from the cytoplasmic mass into coelomic fluid. It suggests that the spermatogenesis are progressed in support of coelomic fluid, and the fact take into consideration that the spermatogenic cells can be in vitro cultured without somatic cells and with supplements of coelomic fluid.

한국산 개불, Urechis unicinctus의 서식생태에 관한 연구

최상덕,김호진,이원교,서호영,이우범 國立 麗水大學校 環境問題硏究所 2000 環境硏究論文集 Vol.3 No.-

For the commercial culture of marine echiuran worm Urechis unicinctus, a series of basic ecological study has been conducted for fishery period(November, 1998~April, 1999) in the laboratory and southern coast of Korea. The environmental factors of echiurans' habitat such as water temperature, salinity and dissolved oxygen were ranged 8.0~14.9 °C, 30.1~33.1 psu 7.0~8.9 ㎎/ℓ, respectively. The habitat sediments were composed of coarse sand, sand silt and silty sand. The highest densities and biomass of U. unicinctus were found in areas of the sand silt. In the laboratory, all echiurans performed burrowing success in coarse sand and sand within 24 hours, but none did in silt and clay within 10 days. In sand silt, echiurans enabled to dig 0.7~2.8 burrows/day/ind. And U. unicinctus made a burrow of U shape.

Attribution of Cortical Granules to Formation of Fertilization Envelopes and Polyspermy Block in Urechis unicinctus

Shin, Kil-Sang,Kwon, Hyuk-Jae,Kim, Wan-Jong The Korean Society for Integrative Biology 2005 Integrative biosciences Vol.9 No.2

Cortical reaction and polyspermy block are well defined in most marine invertebrates. In Urechis species, the function of cortical granules (CGs) is not yet known, and there is controversy on whether the cortical reaction occurs, or the fertilization envelope (FE) is attributed to CG releases or functions to prevent polyspermy. This study was carried out to determine the cortical reactions and functions of the FE in Urechis unicinctus. Artificial insemination of the eggs revealed that CG release occurred to give rise to perivitelline space (PS) into the final FE. Both PS and final FE effectively blocked polyspermy. The final FE was accomplished within 10 min after sperm-egg initial binding. No massive release of CGs occurred within the early phase of 5 min after the initial binding, initially and the PS seemed to playa role to prevent polyspermy. The CG massively released its content into the PS in late phase of FE formation, and differentiated PS into five intermediate layers. The layers opened into each other by anastomosis, so that the final FE consisted of two layers, the inner layer ($15{\mu}m$ in thickness) and the outer layer ($1{\mu}m$ in thickness). The outer layer derived from vitelline layer and the inner layer consisted of PS and CG secretions. Immunofluorescence and confocal laser microscopy revealed that the spermatozoon took up residence in the egg cortex during FE formation and successive meioses of the fertilized egg. These results suggest that both PS and final FE of U. unicinctus were equivalent to the early and late block, respectively, of other marine animals.

개불의 체벽으로부터 i-type 라이소자임의 정제

오혜영(Hye Young Oh),박남규(Nam Gyu Park) 한국생명과학회 2018 생명과학회지 Vol.28 No.3

라이소자임은 선천성 면역 물질로 개불을 포함하는 여러 무척추동물의 병원균에 대한 방어에 주요하게 작용한다. 본 논문은 개불(Urechis unicinctus)의 체벽 조직 추출물로부터 무척추형 라이소자임의 정제와 그 특성에 관한 분석을 기술하고 있다. 체벽 추출물은 우선적으로 Sep-Pak C18 cartridge를 사용하여 부분적으로 분리되었으며, 분리된 분획 중 60% 메탄올에 용출된 분획이 Bacillus subtilis KCTC 1021에서 강한 항균활성을 나타내었다. 그후 여러 단계의 역상과 이온교환 고속액체크로마토그래피(High Performance Liquid Chromatography, HPLC)를 사용하여 항균성 물질이 정제되었으며, 분자량은 약 14 kDa이었다. 이 단백질의 일차서열은 LC-MS/MS를 통해 분석되었으며, 얻은 부분적 아미노산 서열을 NCBI BLAST를 통하여 분석해 본 결과, 이 항균성 물질은 다른 동물들로부터 동정된 무척추동물형 라이소자임(invertebrate-type lysozyme)의 서열과 유사도를 가지고 있어, 체벽으로부터 정제된 개불 라이소자임(Urechis unicinctus invertebrate-type lysozyme from body wall, Uu-iLysb)으로 명명하였다. 개불 라이소자임의 활성을 확인하기 위하여 항균활성 및 라이소자임 효소 활성실험을 진행한 결과, 개불 라이소자임이 항균활성과 라이소자임 효소활성 모두 강하게 가지고 있는 것을 확인하였다. Lysozymes are innate immune factors that play a critical role in the defense against pathogens in various invertebrate animals including spoon worms. In this study, an invertebrate-type lysozyme was isolated from the body wall of spoon worm, Urechis unicinctus. The acidified body wall extract was partially separated using a Sep-Pak C18 cartridge. Among the fractions, the materials that were eluted with 60% methanol/0.1% trifluoroacetic acid showed the most potent antimicrobial activity against Bacillus subtilis KCTC 1021. A series of high performance liquid chromatography (HPLC) steps were then utilized to isolate a single antimicrobial absorbance peak. The molecular weight of the antimicrobial peak was approximated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which was approximately 13 to 14 kDa. The partial primary structure of this antimicrobial protein that was analyzed, using LC-MS/MS, was CTGGRPPTCEDYAK (1611.69 Da). Homology search of these fourteen residues, using the National Center for Biotechnology Information Basic Local Alignment Search Tool (NCBI BLAST), revealed that the isolated protein was similar to the invertebrate-type lysozymes described in other animals. Then, the antimicrobial and lysozyme enzymatic (muramidase) activities of this protein were assessed. The isolated protein possessed antimicrobial activity and potent muramidase activity, which were comparable to those of hen egg white lysozyme. Therefore, the isolated protein was designated as Urechis unicinctus invertebrate-type lysozyme from the body wall, Uu-iLysb.

개불과 넙치의 혼합 사육시 성장과 사육환경의 변화

김용구,박일웅,안윤근,배애란,최상덕 國立麗水大學校 環境問題硏究所 2004 環境硏究論文集 Vol.6 No.-

This present study was conducted to determine the effects of sea water quality and sediment quality on growth and body composition in poly culture system, Paralichthys olivaceus and Urechis unicinctus. The seawater temperature of the experimental groups was ranges of 17.1~23.8℃, the lowest in 0 day and the highest in 70 day. The concentration of COD was 1.2~1.6㎎/ℓ, DIN 6.19~28.89㎎/ℓ in the D group. Mean IL concentration was 5.42~5.07% and it was maximum in 0 day and relatively minimum 70day. The concentration of AVS was 0.36~0.22㎎/g-dry, COD 4.36~4.08㎎/g-dry in the D group. Growth and feed intake of fish were affected by diets and feeding frequencies(P<0.05). There were differences at all experimental groups especially C group and control group but A, B group and C, D group were not difference. The lowest increased body weight was observed 4.7g in the D experimental group and the highest experimental group was observed 8.6g in the C group.

Improvement of erectile dysfunction by the active pepide from <i>Urechis unicinctus</i> by high temperature/pressure and ultra - wave assisted lysis in Streptozotocin Induced Diabetic Rats

Kim, Kang Sup,Bae, Woong Jin,Kim, Su Jin,Kang, Kyong-Hwa,Kim, Se-Kwon,Cho, Hyuk Jin,Hong, Sung-Hoo,Lee, Ji Youl,Kim, Sae Woong Sociedade Brasileira de Urologia 2016 International Braz j urol Vol.42 No.4

<P><B>ABSTRACT</B></P><P><B>Introduction:</B></P><P>We investigate the effect of active peptide from Urechis unicinctus (UU) by high temperature/pressure and ultra-wave assisted lysis on erectile dysfunction in streptozotocin-induced diabetic rats.</P><P><B>Materials and Methods:</B></P><P>Forty 12-week-old Sprague-Dawley rats were used in this study. Diabetes was induced by a one-time intraperitoneal injection of streptozotocin (50mg/kg). One week later, the diabetic rats were randomly divided into four groups: normal control, untreated diabetes control, and groups treated with 100 or 500mg/kg/d UU peptide. Rats were fed with UU peptide by intragastric administration for 8 weeks. After 8 weeks, penile hemodynamic function was evaluated in all groups by measuring the intracavernosal pressure after electrostimulating the cavernous nerve. Nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) activities were measured and endothelial nitric oxide synthase (eNOS) and neuronal NOS (nNOS) protein expression was determined by Western blot.</P><P><B>Results:</B></P><P>Maximum intracavernosal pressure in diabetic control rats decreased significantly compared to normal control rats, and was increased significantly compared to untreated diabetic rats after UU peptide supplementation. Treatment with the higher dose of UU peptide significantly increased the NO and cGMP levels compared with the diabetic control group. Decreased activity and expression eNOS and nNOS were found in the diabetic rats compared with the normal control group. Decreased eNOS and nNOS in diabetic rats were improved by UU peptide administration.</P><P><B>Conclusions:</B></P><P>Active peptide from UU ameliorates erectile function in a streptozotocin induced diabetic rat model of erectile dysfunction.</P>

Hypoglycemic Effects of Glycosaminoglycan from Urechis unicinctus in Diabetic Mice

Chunying Yuan,Ping Liu,Xu Han,Qingman Cui 한국식품영양과학회 2015 Journal of medicinal food Vol.18 No.2

To further utilize glycosaminoglycan from Urechis unicinctus, the hypoglycemic effect and possible mechanism of glycosaminoglycan on diabetic mice were evaluated. Diabetes was induced in mice by intraperitoneal injections of streptozotocin for 3 consecutive days and fed with high-sugar and high-lipid fodder. After diabetes was confirmed, the hypoglycemic effect of glycosaminoglycan from U. unicinctus was investigated in the diabetic mice. Results demonstrated that glycosaminoglycan could significantly decrease blood glucose concentrations, HOMA-IR, AUG, and liver MDA content in diabetic mice. In addition, it significantly enhanced liver SOD and GSH-Px activity, as well as liver GCK activity and hepatic glycogen levels. Glycosaminoglycan from U. unicinctus exhibited efficacy against diabetes, suggesting its potential use as a natural intervention against diabetes.

Cytochemical Localization of Nuclear Actin of Sperm and Spermatids in Urechis unicinctus

Shin, Kil-Sang,Kim, Ho-Jin,Kwon, Hyuk-Jae,Kim, Wan-Jong The Korean Society for Integrative Biology 2005 Integrative biosciences Vol.9 No.2

In this study, we found that sperm ball of Urechis unicinctus consisted of a somatic cell and spermatogenic cells. After separation from the sperm ball, individual spermatid floated freely in the coelomic fluid and differentiated into a mature sperm. Because of many nuclear vacuoles, spermatid nucleus was observed to be heterogeneous. Later, the spermatid nucleus condensed into the homogeneous round nucleus of the mature sperm. Perinuclear microtubules could be seen but did not seem to be organized into manchette microtubules. To understand the nature of nuclear condensation during spermiogenesis, the sperm and spermatids (spermiogenic cells) were treated with FITC-phalloidin, or anti-actin-FITC, or labeled with antiactin immunogold particles (AAIP; 10 nm) followed by transmission electron microscopy or confocal laser scanning microscopy. The anti-actin-FITC and FITC-phalloidin reactions occurred distinctly in the nuclei of both spermiogenic cells. FITC-phalloidin reacted more intensely with acrosomes. The AAIP were incorporated mainly into nuclei of both cells sometimes showing local distribution in the nucleus. Nuclear vacuoles of spermatids disappeared progressively with condensation of the nucleus, as the number of incorporated $AAIP/{\mu}m^2$ increased. These results suggest that nuclear actin microfilaments might be closely related to nuclear condensation.