노화 흰쥐 소뇌에서 세포사 조절인자들의 변화에 대한 연구
Differential Expression of Neuronal Death-related Factors in Aged Rat Cerebellum
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세포자멸사는 발생과정과 중추신경계의 항상성 유지에서 중요한 역할을 하는 것으로 알려져 있다. 본 연구에서는 흰쥐 소뇌에서 노화에 따른 세포사 조절인자들의 변화를 in situ hybridization법과 면역조직화학 법을 사용하여 관찰하였다. TUNEL 염색 결과에서는 염색된 세포핵이 노화 흰쥐 소뇌에서만 발견되었다. IGF-IR, NT 및 p53에 대한 면역 염색성은 성숙군의 소뇌에서는 거의 관찰되지 않은 반면, 노화군에서는 소뇌 겉질의 조롱박 세포에서 현저한 면 역염색성을 보였다. ICH-1 (caspase-2)의 면역염색성은 노화 쥐의 분자층과 조롱박세포층에서 관찰되었다. c-Fos는 성숙군의 과립층에서 진하게 염색되었고, 면역염색성은 노화군의 과립층에서 유의성 있게 감소하였다. Bcl-2는 조 롱박 세포 (Purkinje cell)층과 과립층에서 가장 강하게 발현되었고 노화에 따라 유의성 있게 감소하는 경향을 나 타냈다. Bax는 성숙군의 조롱박 세포에서 가장 진하게 염색되었고, Bax 면역염색성은 소뇌 겉질과 소뇌핵에서 노 화에 따른 유의성 있는 차이를 나타내지 않았다. 결론적으로, 본 연구는 흰쥐 소뇌에서 세포사 관련인자들의 노화에 따른 변화에 대한 최초의 형태학적 자료를 제공하였다. 이러한 연구는 노화를 비롯한 여러 신경퇴행성 질환 연구에 필요한 기본적인 형태학적 자료를 제공 할 수 있을 것이라 사료된다.
Abstract
In the present study, we investigated the expression of apoptosis-associated proteins in the cerebellum of aged rats: IGF-I receptor (IGF-IR), nitrotyrosine (NT), p53, key pro-apoptotic gene ICH-1 (caspase-2), c-Fos and Bcl-2 family members (Bcl-2 and Bax). Twelve adult (4~6 month old) and 15 aged (24~29 month old) Sprague-Dawley rats were examined in this study. We performed immunohistochemical staining, in situ hybridization and densitometric measurement using a NIH image program (Scion Image) to determine the staining density. In adult rats, there were no immunoreactivities for insulin-like growth factor-I receptor (IGF-IR), nitrotyrosine (NT) or p53 in any region of cerebellum. However, IGF-IR immunoreactivity was found in some Purkinje cells in aged rat cerebellum. The prominent staining of NT or p53 was also localized in the Purkinje cell layer in aged rats. A high density of ICH-1 (caspase-2) immunoreactivity was observed in the molecular and Purkinje cell layers in aged rats. Immunoreactivity for c-Fos was significantly decreased in the granule cells in aged rats. Positive signal for bcl-2 was significantly decreased in the Purkinje cells and granule cells of aged rats. The most intense staining for Bax was observed in the soma of Purkinje cells of adult rats. However, Bax immunoreactivity was not changed in any layers in the cerebellar cortex of aged rats. In conclusion, this study provides the first morphological data concerning the differential regulation of apoptosisrelated genes in rat cerebellum during aging.
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