Applied Microscopy

Korean Society of Microscopy (한국현미경학회)
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Histochemical and Fine Structural Visualization of the Epithelial Apoptosis in the Anuran Tadpole during the Tail Regression Stages

무미양서류 꼬리 퇴화과정중 피부 상피조직의 세포사에 관한 조직화학적 및 미세구조적 연구

  • Lee, Hye-Won (Department of Biological Sciences & Institute of Basic Sciences, Dankook University) ;
  • Moon, Myung-Jin (Department of Biological Sciences & Institute of Basic Sciences, Dankook University)
  • 이혜원 (단국대학교 첨단과학대학 생명과학과 및 기초과학연구소) ;
  • 문명진 (단국대학교 첨단과학대학 생명과학과 및 기초과학연구소)
  • Received : 2009.04.14
  • Accepted : 2009.06.19
  • Published : 2009.06.30
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Abstract

The programed cell death of the cutaneous epithelial tissue during tail regression stages in anuran tadpoles of the blackspotted frog, Rana nigromaculata were visualized by the histochemical and transmission electron microscopic techniques. Metamorphotic changes in the tail regression during the period of the Shumway stage number 31 to 33 are characterized by the disappearance of mucous layer and formation of compound epithelium through cutaneous thickening. Following the TUNEL (terminal deoxynucleotidyl transferase-mediated biotinylated d-uridine triphosphate nick end labeling) staining technique, the apoptotic cells were detected at the distal region of the tail skin initially, but they can be seen at the proximal region according to their following development. It has been also revealed that the number of the TUNEL-positive cells gradually increased from apical to basal direction of the epithelial layers during the tail regressing stages. Following the TEM observation, the early apoptotic cells shown in the epithelium demonstrated condensation and margination of the chromatin material at the nuclear periphery. Another epithelial apoptotic cells were shown nuclear fragmentation, membrane blebbing and cytoplasmic condensation. Following the process of the apoptotic degradation, well preserved organelles and nuclear fragments can be identified in the cytoplasm of lysosome-rich cells, however they soon reduced to lysosomal residual bodies through the progressive degradation.

참개구리(Rana nigromaculata) 꼬리 퇴화과정에 따른 피부 상피조직의 분화와 세포자멸사에 의해 초래된 상피의 미세구조적 변화를 TUNEL 조직화학적 염색 및 고배율의 투과전자현미경 관찰을 통해 확인하였다. 꼬리의 퇴화가 시작되는 Shumway 31단계의 올챙이 피부에 발달되어 있던 점액층은 발생 32 단계를 거쳐 사라지고 상피는 비후되어 꼬리의 퇴화가 완료되는 발생 33 단계에 도달하면 성체인 개구리의 다층상피로 전환되었다. TUNEL 염색결과, 꼬리 퇴화과정중 apoptosis 세포의 출현은 원위부(distal region)의 꼬리에서 시작되어 몸통 쪽의 근위부(proximal region)로 이행되었고, 동일 상피층에서는 내강부로부터 기저부로 전이되는 특성을 보였다. 투과 전자현미경 관찰 결과, 세포자멸사에 의한 초기의 변형은 인접세포와의 세포간극이 사라지고 밀착된 후, 핵의 모양이 불규칙해지고 염색질이 응축되었다. 이어서 세포질에 다수의 자유 리보솜이 나타나고, 핵의 분절이 일어난 후, apoptotic body가 출현하며, 이들은 최종적으로 라멜라 구조를 지닌 잔여소체(residual body)를 형성하였다

Keywords

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