생명과학회지 (Journal of Life Science)

  • 제23권7호
  • /
  • Pages.941-946
  • /
  • 2013
  • /
  • 1225-9918(pISSN)
  • /
  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
권호 표지
DOI QR코드

DOI QR Code

발생단계별 해마신경세포에서 eIF4E 및 eIF4EBP1의 표현

Developmental Expression of Eukaryotic Initiation Factor 4E (eIF4E) and eIF4E-binding Protein 1 (eIF4EBP1) in Rat Hippocampal Neurons

  • 박재완 (동국대학교 의과대학 해부학교실) ;
  • 문일수 (동국대학교 의과대학 해부학교실)
  • Park, Jaewan (Department of Anatomy, Dongguk University, College of Medicine, and Medical Institute of Dongguk University) ;
  • Moon, Il Soo (Department of Anatomy, Dongguk University, College of Medicine, and Medical Institute of Dongguk University)
  • 투고 : 2013.07.18
  • 심사 : 2013.07.26
  • 발행 : 2013.07.30
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

신경세포의 가지돌기 내 단백질합성은 필요한 단백질을 실시간으로 제공할 수 있는 이점을 제공한다. 본 연구에서는 단백질합성인자 eIF4E와 그 억제 단백질인 eIF4EBP1의 발생단계별 표현을 배양한 해마신경세포를 면역 염색하여 조사하였다. eIF4E는 가지돌기에 점박이 모양으로 표현되었으며, 핵에는 표현되지 않았다. 그러나 eIF4EBP1는 가지돌기 뿐 아니라 발생초기(DIV 0.5)부터 핵에서 표현되었으며 성숙한 세포에서 핵에 더욱 뚜렷이 표현되었다. eIF4E 혹은 eIF4EBP1의 PSD95과의 colocalization은 $39.1{\pm}9.6%$ 및 $70.5{\pm}5.2%$ (DIV 7), $57.7{\pm}8.2%$ 및 $36.0{\pm}3.1%$ (DIV 10), $29.9{\pm}2.9%$ 및 $40.2{\pm}11.7%$ (DIV 20)이었다. eIF4E와 eIF4EBP1의 colocalizatin은 $18.5{\pm}2.6%$ (DIV 7), $11.1{\pm}3.9%$ (DIV 10), $38.6{\pm}5.6%$ (DIV 20)이었다. 이 결과는 eIF4E 및 eIF4EBP1의 많은 부분이 연접후에 위치하며, 발생초기에는 eIF4E가 활동적인 형태로 존재하지만, 성숙 신경세포에서는 eIF4EBP1과 결합하여 비활성적인 형태로 존재함을 의미한다.

Local protein synthesis at subsynaptic sites plays a key role in the regulation of the protein composition in local domains. In this study, we carried out immunocytochemistry of cultured rat hippocampal neurons in various developmental stages to investigate the expression of eIF4E and its binding protein, eIF4EBP1. Both proteins were distributed in dendrites. In addition, eIF4EBP1 was highly expressed in the nucleus throughout the development, whereas eIF4E was not expressed in the nucleus. Punctate expression of eIF4E and eIF4EBP1 was evident in DIV 3. The colocalization rates of eIF4E or eIF4EBP1 puncta with PSD95 were higher in the dendrogenic than in the mature stages. In contrast, the colocalization rates of eIF4E and eIF4EBP1 puncta were higher in the mature than in the dendrogenic stages. As eIF4E is inactive when it is bound to eIF4EBP1, these data indicate that most dendritic eIF4E's are active during development but that they are mostly under inhibition in mature neurons.

키워드

참고문헌

  1. Asaki, C., Usuda, N., Nakazawa, A., Kametani, K. and Suzuki, T. 2003. Localization of translational components at the ultramicroscopic level at postsynaptic sites of the rat brain. Brain Res 972, 168-176. https://doi.org/10.1016/S0006-8993(03)02523-X
  2. Brewer, G. J., Torricelli, J. R., Evege, E. K. and Price, P. J. 1993. Optimized survival of hippocampal neurons in B27-supplemented Neurobasal, a new serum-free medium combination. J Neurosci Res 35, 567-576. https://doi.org/10.1002/jnr.490350513
  3. Choi, M.-K., Park, S. D., Park, I. S. and Moon, I. S. 2011. Localization of translation initiation factors to the postsynaptic site. J Life Sci 21, 1526-1531. https://doi.org/10.5352/JLS.2011.21.11.1526
  4. Frey, U., Krug, M., Reymann, K. G. and Matthies, H. 1988. Anisomycin an inhibitor of protein synthesis blocks late phases of LTP phenomena in the hippocampal CA region in vitro. Brain Res 452, 57-65. https://doi.org/10.1016/0006-8993(88)90008-X
  5. Gingras, A. C., Gygi, S. P., Raught, B., Polakiewicz, R. D., Abraham, R. T., Hoekstra, M. F., Aebersold, R. and Sonenberg, N. 1999. Regulation of 4E-BP1 phosphorylation: A novel two-step mechanism. Genes Dev 13, 1422-1437. https://doi.org/10.1101/gad.13.11.1422
  6. Goslin, K., Assmussen, H. and Banker, G. 1998. Rat hippocampal neurons in low density culture, pp. 339-370. In Banker, G. and K. Goslin (eds.), Culturing Nerve Cells, 2nd eds., MIT Press, Cambridge, MA.
  7. Haghighat, A., Mader, S., Pause, A. and Sonenberg, N. 1995. Repression of cap-dependent translation by 4E-binding protein 1: competition with p220 for binding to eukaryotic initiation factor-4E. EMBO J 14, 5701-5709.
  8. Kang, H. and Schuman, E. M. 1996. A requirement for local protein synthesis in neurotrophin-induced synaptic plasticity. Science 273, 1402-1406. https://doi.org/10.1126/science.273.5280.1402
  9. Moon, I. S, Lee, H. J and Park, I. S. 2012. Dendritic eIF4E-binding protein 1 (eIF4E-BP1) mRNA is upregulated by neuronal activation. J Korean Med Sci 27, 1241-1247. https://doi.org/10.3346/jkms.2012.27.10.1241
  10. Moon, I. S., Cho, S. J., Jin, I. and Walikonis, R. 2007. A simple method for combined fluorescence in situ hybridization and immunocytochemistry. Mol Cells 24, 76-82.
  11. Moon, I. S., Cho, S. J., Seog, D. H. and Walikonis, R. 2009. Neuronal activation increases the density of eukaryotic translation initiation factor 4E mRNA clusters in dendrites of cultured hippocampal neurons. Exp Mol Med 41, 601-610. https://doi.org/10.3858/emm.2009.41.8.066
  12. Murphy, J. A., Jensen, O. N., and Walikonis, R. 2007. BRAG1, a sec7 domain-containing protein, is a component of the postsynaptic density of excitatory synapses. Brain Res 1120, 35-45.
  13. Nguyen, P. V., Abel, T. and Kandel, E. R. 1994. Requirement of a critical period of transcription for induction of a late phase of LTP. Science 265, 1104-1107. https://doi.org/10.1126/science.8066450
  14. Richter, J. D. and Sonenberg, N. 2005. Regulation of cap-de-pendent translation by eIF4E inhibitory proteins. Nature 433, 477-480. https://doi.org/10.1038/nature03205
  15. Rong, L., Livingstone, M., Sukarieh, R., Petroulakis, E., Gingras, A. C., Crosby, K., Smith, B., Polakiewicz, R. D., Pelletier, J., Ferraiuolo, M. A. and Sonenberg, N. 2008. Control of eIF4E cellular localization by eIF4E-binding proteins, 4E-BPs. RNA 14, 1318-1327. https://doi.org/10.1261/rna.950608
  16. Tang, S. J., Reis, G., Kang, H., Gingras, A. C., Sonenberg, N. and Schuman, E. 2002. A rapamycin-sensitive signaling pathway contributes to long-term synaptic plasticity in the hippocampus. Proc Natl Acad Sci USA 99, 467-472. https://doi.org/10.1073/pnas.012605299
  17. von der Harr, T., Gross, J. D., Wagner, G. and McCarthy, J. E. 2004. The mRNA cap-binding protein eIF4E in post-transcriptional gene expression. Nat Struct Mol Biol 11, 503-511. https://doi.org/10.1038/nsmb779