내동보존이 진주조개(Pinctada fucata martensii) 유생의 미세구조에 미치는 영향
The Effects of Cryopreservation on Fine Structure of Pearl Oyster(Pinctada fucata martensii) Larvae
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냉동보존이 진주조개(Pincata fucata martensii) 유생의 형태 및 구조에 미치는 영향을 알아보기 위하여 냉동전후 D형 및 각정기 유생을 광학 및 전자현미경으로 조사하였다. 동해방지제는 0.2 M sucrose를 첨가한 2.0 M DMSO를 사용하였다. 냉동후 유생은 일부 패각이 손상되긴 했지만 hinge와 prodissoconch가 뚜렷하게 나타났으며, 소포체, 지질 과립, 미토콘드리아, 핵 등을 포함한 세포내 소기관들이 고르게 분포되어 있었다. 또한 섬모가 규칙적으로 배열되어 있었고 섬모 아래 미토콘드리아와 지질과립이 위치해 있는 것이 관찰되었으나, 일부 해동된 유생에서 섬모의 불규칙적인 배열과 섬모환이 둥글게 뭉쳐져 있는 모습이 관찰되었다. 이러한 결과는 진주조개의 D형 유생과 각정기 유생이 냉동에 쉽게 영향을 받을 수 있다는 것을 보여준다. 따라서 냉동보존 시 세포의 손상을 감소시킬 수 있는 연구가 이루어져야 할 것으로 사료된다.
Abstract
The freezing susceptibilities of two larval stages (D-shaped and umbo) of the pearl oyster (Pinctada fucata martensii) were evaluated by the electron microscopy (light, transmission electron and scanning electron). The morphological shapes were examined from each pre-frozen or frozen-thawed stage of the cryopreserved larvae in liquid nitrogen by using the cryoprotectant, dimethyl sulfoxide mixed with sucrose. Although a portion of the shell was damaged, the hinge and prodissoconch were intact and clearly visible after preservation in liquid nitrogen. In addition, the cytoplasm of the frozen-thawed larvae maintained the normal organelle integrities, e.g., endoplasmic reticula, lipid droplets, mitochondria, nucleus and microvilli. However, some of the frozen-thawed larvae showed irregularly arranged cilia, rough shell surfaces and round-lumped cilium heads. These results indicate that P. fucata martensii larvae are susceptible to freezing, at least at those two critical developmental stages (D-shaped and umbo), and suggest a new industrial investigation including reduction method of cell injury for preserving microbial starter cultures need to be developed.
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