• Title/Summary/Keyword: mud loach

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An efficient strategy for blocking the 1st mitotic cleavage of fish zygote using combined thermal treatment, exemplified by mud loach (Misgurnus mizolepis)

  • Nam, Yoon-Kwon;Park, Geyong-Cheol;Kim, Dong-Soo
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.38-38
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    • 2003
  • Blocking the first mitotic cleavage of the zygote is a key tool for chromosome-set manipulations in fish. We developed an improved method for inducing tetraploidy by blocking the mitosis with a combination of heat shock at 40.5$^{\circ}C$ for 1, 2 or 3 min followed by cold shock at $1.5^{\circ}C$ for 30, 45 or 60 min. When applied during the first cleavage metaphase of mud loach (Misgurnus mizolepis) zygotes, the optimal combination was heat for 2 min followed by cold for 45 min. At 1 month, the frequency of 4N survivors and the yield from total eggs fertilized was 55.7% and 14.4%, respectively, compared to heat shock alone with 20.0% efficiency and 3.6% yield. The effectiveness of the procedure was confirmed by diploid mitotic gynogenesis using transgenic markers. The overall yield of homozygous diploids, 34.0%, was better than that for single heat shock, 17.3%. The tetraploids and homozygous diploids had higher early mortality than normal diploid controls. However at 1 month, the viability of the tetraploids was the same as normal diploids. For gynogenetic diploids, the survival was similar to normal diploids after 3 months. The high efficiency of this new protocol extends the opportunity to study polyploidy in basic and applied research.

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Ploidy status of progeny from the crosses between tetraploid males and diploid females in mud loach (Misgurnus mizolepis)

  • Nam, Yoon-Kwon;Kim, Dong-Soo
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.40-40
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    • 2003
  • DNA content of the sperm of tetraploid mud loach (Misgurnus mizolepis) males and the ploidy status of progenies generated by crossing tetraploid males with diploid females are described. Reproductive performance of the induced adult tetraploid males ranged from sterility to fertility. Of 48 tetraploid males tested, 12 were sterile but the other 36 produced functional sperm. Of these 36, 26 produced haploid sperm, which on fertilizing the haploid eggs, generated diploid progenies. Seven tetraploid males were mosaics in their sperm, as indicated by the production of diploid, aneuploid and/or triploid offspring. Only 3 males produced diploid sperm rendering the production of all-triploid progenies. The DNA content of sperm of a tested tetraploid male was consistent throughout the 3 progeny tests, i.e. the haploid sperm-producing 4n males persisted to produce the haploid sperm only likewise the diploid sperm producing4n males consistently produced the diploid sperm only, when progeny testing was extended to 3 successive but alternate years. Hence, a careful and direct examination of the DNA profile of sperm from tetraploid males is a pre-requisite for reproductive containment of genetically modified fish.

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Early Gonadogenesis in Diploid and Triploid Mud Loach, Misgurnus mizolepis (2배체와 3배체 미꾸라지(Misgurnus mizolepis)의 원시생식소 형성과정)

  • Kim Bong-Seok;Kim Dong Soo
    • Journal of Aquaculture
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    • v.8 no.3
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    • pp.231-240
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    • 1995
  • This study was conducted to examine early gonadogenesis by using a histological method for the appearance of primordial germ cells (PGCs), protrude of genital ridge, and formation of primitive gonads in diploid and triploid mud loach, Misgurnus mizolepis. The pattern of early gonadogenesis including appearance of PGCs, formation of genital ridge, and development of primitive gonad in both diploid and triploid were not different histologically. Characteristics of PGCs of triploid were also the same as those of diploid. However, gonadal length of diploid was significantly longer than that of triploid (P<0.05).

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Transcriptional Alteration of Two Metallothionein Isoforms in Mud Loach (Misgurnus mizolepis) Fry during Acute Heavy Metal Exposure

  • Lee, Sang-Yoon;Stoliar, Oksana;Nam, Yoon-Kwon
    • Fisheries and Aquatic Sciences
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    • v.13 no.2
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    • pp.112-117
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    • 2010
  • Altered mRNA expression of two metallothionein isoforms (MT-IA and MT-IB) in response to acute heavy metal exposure was examined in mud loach, Misgurnus mizolepis, fry using a real-time RTPCR assay. Sublethal exposure (1 or 5 ${\mu}M$) to Cd, Cr, Fe, Mn, Ni, and Zn resulted in highly variable transcriptional responses of the two MT isoforms to the heavy metal ions, including upregulation, a steady state, and downregulation. Overall, the most potent inducer of both MT isoforms was Cd (up to 6-fold). Another exposure experiment using a series of doses of Cu revealed that the stimulation patterns of the two MT isoforms differed: MT-IA transcription was soon saturated at higher concentrations (about 2-fold at 1-4 ${\mu}M$ of Cu), whereas the activation of MT-IB was more dependent on the treatment dose (increased up to 5-fold at 3 ${\mu}M$). The isoform-specific allotment of constitutive and inducible functions was not as clear in fry as in adult tissues. Coordinated interaction between the MT-IA and MT-IB isoforms was hypothesized based on the finding that MT-IA represented a primary action under 'less stressful' or 'sublethal' conditions, whereas the activation of MT-IB became important under 'more stressful' or 'lethal' circumstances in this species.

Change of Skin Mucus Cells Related to Aerial Exposure of Misgurnus mizolepis (Cobitidae) Dwelling in a Rice Field (논에 서식하는 미꾸라지, Misgurnus mizolepis의 공기노출에 의한 피부 점액세포의 변화)

  • Oh, Min-Ki;Park, Jong-Young
    • Korean Journal of Ichthyology
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    • v.23 no.1
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    • pp.70-74
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    • 2011
  • During the winter, the rice field-dwelling muddy loach Misgurnus mizolepis is buried in burrows constructed of mud and are subjected to exposure to air at times of shortage of water. To investigate the environmental factors that lead to changes of the skin mucus cells of the muddy loach in rice fields, we carried out an experiment where we artificially exposure the fish to air, duplicating as close as possible winter conditions in nature. During the summer, a water tank containing M. mizolepis was filled with mud, and the water was allowed to evaporate. After a month of evaporation, the loach constructed burrows similar to those in a winter rice field. The epidermis in the experimental fish was mostly occupied by large elongated mucus cells, whose numbers drastically increased in all observed regions of the dorsum, lateral region, and the occiput. Such features are typically seen in fishes in wild habitats during the winter season.

Intraspecific Androgenesis in Mud Loach (Misgurnus mizolepis): II. Diploid Restoration and Viability Assessment (미꾸라지(Misgurnus mizolepis)의 웅성발생성 처녀생식: II. 웅성발생성 이배체 유도 및 생존능력 평가)

  • Nam, Yoon-Kwon;Bang, In-Chul;Noh, Choong-Hwan;Cho, Young-Sun;Kim, Dong-Soo
    • Journal of Aquaculture
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    • v.19 no.4
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    • pp.281-287
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    • 2006
  • Intraspecific diploid androgenesis was achieved in mud loach (Misgurnus mizolepis) by the inhibition of the first mitotic division using combined thermal treatment. A combined thermal treatment (heat shock at $40.5\;^{\circ}C$ for 120 sec followed by cold treatment at $1\;^{\circ}C$ for 45 min) applied to the 1st metaphase of cell division (28 min post insemination at $25\;^{\circ}C$) successfully recovered viable androgenetic diploidy. Mean hatching success of the androgenetic diploid group was 29.6%, and the average yield out of total eggs taken was about 7% assessed at 1 week of age. However, relatively large variations in the yield of diploid androgenesis were observed among different egg batches used as cytoplasmic donors. Successful diploidization was confirmed by flow cytometric analysis, and parthenogenic reproduction in a paternal exclusive manner was verified with transgene dosage. Significant mortality was found in most androgenetic groups especially from hatch to 1 month of age, although such mortality was stabilized later.