생쥐 뇌의 발생과정에 따른 맥락총에서의 coxsackievirus and adenovirus receptor 발현

Abstract

맥락상피세포 사이의 밀착결합들은 혈액뇌척수액관문을 둘러싸고있다. 혈액뇌척수액관문에서 발생단계에 따라 변화하는 coxsackievirus and adenovirus receptor (CAR) 발현을 알아보기 위하여 생쥐 뇌의 맥락총에서 발생단계별로 실험을 하였다. Total CAR mRNA 와 단백질은 태아, 신생아, 태어난지 7일이 되는 생쥐의 뇌에서 비교적 높게 발현하였다. 하지만 성숙한 쥐의 뇌에서는 현저하게 감소하였다. CAR mRNA는 뇌하수체와 맥락총에서 비교적 높게 발현되었다. 맥락총에서 total CAR mRNA는 성숙한 뇌에서 현저하게 증가하였다. 맥락총에서의 multiplex PCR 결과, CAREX7 (long isoform) mRNA 발현이 태어난지 7일까지 현저하게 감소하였지만, 성숙한 뇌에서는 발현양이 증가하였다. 반면에 CAREX8 (short isoform) mRNA는 출생 후 맥락총의 발생과정동안 감소하였다. Western blot에서 MW 46kDa CAR 단백질은 성숙한 뇌의 맥락총에서 현저하게 증가하였다. CAR의 발현을 면역조직화학법으로 확인한 결과 성숙한 뇌의 뇌실막 세포와 맥락총의 상피세포에서 증가하였다. 이중면역형광법 결과 CAR는 ZO-1과 apical 밀착결합과 lateral peduncular 사이와 인접한 부분 그리고 floccular 맥락총의 상피세포에서 colocalization 되었다. 이러한 결과로 보았을때, CAR의 long isoform은 맥락총 상피세포의 밀착결합 뿐만 아니라 세포부착과 혈액뇌척수액관문의 성숙에도 관여할 것으로 사료된다. 뇌실막 세포에서의 CAR의 발현이 뇌가 성숙 할수록 증가하는 것은 아데노바이러스와 연관된 유전자 치료 방법에 잠재적인 가능성을 가지고 있다는 사실을 보여준다. | Tight junctions (TJs) between choroids plexus (CP) epithelial cell build up blood cerebrospinal fluid barrier (BCSFB). In an effort to elucidate developmental changes in the molecular architecture of BCSFB the expression of coxsackievirus and adenovirus receptor (CAR) were examined in the choroid plexus in developing mouse brain. Total CAR mRNA and protein were relatively high in fetal and immature brain until PND 7. CAR expression was markedly decreased in adult brain in which CAR mRNA levels were relative high in the pituitary and CP. In CP, total CAR mRNA was markedly increased in adult brain. In multiplex PCR using CP cDNA CAREx7(long isoform) mRNA levels were relatively low until PND7 but markedly increased in adult mice. In contrast, CAREx8(short isoform) mRNA was decreased during postnatal development of CP. On Western blot, MW 46 kDa CAR proteins were markedly increased in adult CP. CAR immunoreactivity was visibly increased in CP epithelial cells and ependymal cells in adult brain. CAR was colocalized with ZO-1 at apical TJs and lateral contacts between lateral peduncular and floccular CP epithelial cells. Together, long isoform of CAR may participate in the cell adhesion as well as TJs in CP epithelial cells and play a role in the maturation of BCSFB. Adulthood increase in CAR in the CP and ependyma may have potential implication in adenovrirus mediated gene therapy intraventricular route.; Tight junctions (TJs) between choroids plexus (CP) epithelial cell build up blood cerebrospinal fluid barrier (BCSFB). In an effort to elucidate developmental changes in the molecular architecture of BCSFB the expression of coxsackievirus and adenovirus receptor (CAR) were examined in the choroid plexus in developing mouse brain. Total CAR mRNA and protein were relatively high in fetal and immature brain until PND 7. CAR expression was markedly decreased in adult brain in which CAR mRNA levels were relative high in the pituitary and CP. In CP, total CAR mRNA was markedly increased in adult brain. In multiplex PCR using CP cDNA CAREx7(long isoform) mRNA levels were relatively low until PND7 but markedly increased in adult mice. In contrast, CAREx8(short isoform) mRNA was decreased during postnatal development of CP. On Western blot, MW 46 kDa CAR proteins were markedly increased in adult CP. CAR immunoreactivity was visibly increased in CP epithelial cells and ependymal cells in adult brain. CAR was colocalized with ZO-1 at apical TJs and lateral contacts between lateral peduncular and floccular CP epithelial cells. Together, long isoform of CAR may participate in the cell adhesion as well as TJs in CP epithelial cells and play a role in the maturation of BCSFB. Adulthood increase in CAR in the CP and ependyma may have potential implication in adenovrirus mediated gene therapy intraventricular route.