돼지난소에서 난포폐쇄와 큰포식세포에 관한 형태학적 연구 = Follicular Atresia and Macrophages in the Porcine Ovary; Immunohistochemical and Transmission Electron Microscopic Study

국문 초록 (Abstract)

난포폐쇄가 과립층세포의 세포자멸사에 의해 이루어지고 이 과정에 큰포식세포가 직, 간접적으로 관여한다는 사실은 널리 알려져 있다. 그러나 난포내에서 일어나는 세포자멸사가 과립층의 어디에서부터 개시되고 어떻게 파급되는지, 세포자멸사 산물과 퇴화된 난모세포의 제거 방법 그리고 난포폐쇄의 완성은 어떻게 이루어지는가에 대해서는 아직 확실히 밝혀져 있지 않다. 이에 저자들은 가임기 돼지(Yorkshire-breed)를 실험동물로 난소내 난포의 광학현미경적 및 투과전자현미경적 관찰과 돼지 대식세포 단크론항체 4E9를 이용한 면역조직화학적 방법으로 본 연구를 실시하였다.
광학현미경적 관찰 결과, 난포폐쇄는 과립층세포의 세포자멸사로부터 개시되고 이어 난모세포의 퇴화와 난포막 속층 세포들의 세포자멸사가 일어나면서 난포폐쇄가 완성되는 것으로 보인다. 과립층세포의 세포자멸사는 핵농축으로부터 시작되고 과립층의 깊이에 관계없이 짧은 시간안에 모든 층으로 파급되고 최종적으로 난모세포의 투명대를 둘러싸고 있는 과립층세포의 세포자멸사로 이어지는 것으로 보인다. 투과전자현미경 관찰에서는 세포자멸사중에 있는 과립층세포들이 인접한 정상적인 과립층세포들과 큰포식세포들에 의해 포식되는 것을 확인할 수 있었고, 난포동내에 산재한 세포자멸사 소체들의 제거에도 큰포식세포들이 관여하고 있음을 알 수 있었다. 난포폐쇄와 큰포식세포와의 관계를 알아보기 위해 실시한 면역조직화학적 관찰에서는, 과립층세포의 세포자멸사 초기에는 큰포식세포가 과립층내에서만 발견되었으나, 세포자멸사의 진행과 함께 난포동과 난모세포를 둘러싸고 있는 투명대 주위에서 각각 순서대로 관찰되었고, 마지막으로 난포막 속층에서도 발견되어, 이들이 과립층세포들의 세포자멸사 소체들을 제거하는 것은 물론 최종적으로는 난포막 속층 세포들의 세포자멸사 소체들을 제거하여 난소 실질화에 관여하는 것으로 보인다. 이러한 본 연구의 결과는 난포폐쇄와 관련된 큰포식세포의 구조, 기능적 연구에 유용한 자료로 생각된다.

다국어 초록 (Multilingual Abstract)

Apoptosis of granulosa cells leads follicular atresia and macrophages have an important role during the apoptotic process. However, the propagation of apoptosis within the follicle, the ways of elimination of apoptotic bodies and degenerated oocyte, and the completion of follicular atresia are still controversial and unidentified clearly. Using adult porcine (Yorkshire-breed) ovary, in this morphological study, transmission electron microscopic observation and immunohistochemical study with pig macrophage monoclonal antibody 4E9 were performed.
In light microscopy, the follicular atresia initiated with apoptosis of granulosa cells, followed by degeneration of oocyte and apoptosis of theca interna cells. Apoptosis occured in random fashion among the granulosa cells and propagated multidirectionally, and finally to the granulosa cells surrounding zona pellucida of degenerating oocyte.
Pyknosis of granulosa cells was the first sign of apoptosis. In immunohistochemistry, macrophages were found only in the granulosa layer at the stage of beginning of apoptosis. With progression of apoptosis, they were proliferated greatly in number enough to eliminate all the apoptotic bodies, and found within the follicular antrum. In advanced stage of atresia, macrophages surrounded the zona pellucida of degenerating oocyte, and found also in the theca interna. In transmission electron microscopy, phagocytic granulosa cells maintained characteristic gap junctions with neighboring granulosa cells and contained several apoptotic bodies and lipid droplets within their cytoplasm. Macrophages kept many apoptotic bodies, vacuoles and autophagosomes in their cytoplasm. Apoptotic granulosa cells were ingested by intact granulosa cells and macrophages initially, but lately, all the apoptotic granulosa cells and degenerated oocyte were eliminated by macrophages. Ovarian follicular atresia completed with phagocytosis of apoptotic theca interna cells by macrophages, and the remnants of the atretic follicle became ovarian stroma.
It is well known that macrophages may play an important role during follicular atresia, such as elimination of
apoptotic granulosa cells, theca interna cells and degenerated oocytes, but, the valid action mechanisms of macrophages on the initiation of granulosa cell apoptosis and on the completion of atresia through the secretion of paracrine factors and autocrine factors still unclear.

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