발생중인 Mongolian gerbil의 전뇌에서 NGF, GFAP 및 CNTF의 분포

Immunohistochemical Localization of Nerve Growth Factor, Glial Fibrillary Acidic Protein and Ciliary Neurotrophic Factor in the Forebrain of the Developing Mongolian Gerbil

  • Park, Il-Kwon (College of Veterinary Medicine, Chungnam National University) ;
  • Lee, Kyoug-Youl (College of Veterinary Medicine, Chungnam National University) ;
  • Song, Chi-Won (College of Veterinary Medicine, Chungnam National University) ;
  • Kwon, Hyo-Jung (College of Veterinary Medicine, Chungnam National University) ;
  • Park, Mi-Sun (College of Veterinary Medicine, Chungnam National University) ;
  • Lee, Mi-Young (College of Veterinary Medicine, Chungnam National University) ;
  • Jeong, Young-Gil (College of Medicine, Konyang University) ;
  • Lee, Chul-Ho (Korean Research Institute of bioscience and biotechnology (KRIBB)) ;
  • Ha, Kwon-Soo (College of Medicine, Kangwon National University) ;
  • Lee, Kang-Yi (College of Oriental Medicine, Daejeon University) ;
  • Kim, Moo-Kang (College of Veterinary Medicine, Chungnam National University)
  • 심사 : 2002.06.10
  • 발행 : 2002.06.29

초록

중추신경계는 신경원(neuron)과 이를 지지해주는 신경아교세포(neuroglia)로 이루어져 있다. 발생과정중 신경원의 발달은 NGF(neruv growht factor)에 의해 관찰이 가능하고, 아교세포들중 별모양아교세포는 GFAP(Glial fibrillary acidic protein)항체로 밝혀낼 수 있다. CNTF(Cillary neurotrophic factor)는 이전에는 운동신경원의 발생 및 유지에 있어 중요한 역할을 하는 인자로 알려져 왔으며, 최근에는 발생과정 중 NGF와 GFAP의 역할에 도움을 주는 것으로 알려졌다. 본 연구에서는 이러한 NGF, GFAP, CNTF를 발생과정 중(임신 15,17,19,21일, 출생 1,2,3,일, 1,2,3주)의 몽골리안 저빌의 전뇌에서 시간에 따른 분포를 광학현미경, 형광염색을 통한 공초점현미경 및 전자현미경을 통해 알아보고자 하였다. 전뇌에서 NGF는 발생 19일령에서부터 대뇌피질에서 관찰되기 시작해서 후각망울, 해마체 및 diagonal band에 관찰되었고, 출생 3일령에서 가장 강한 염색성을 보였으며 그 반응은 출생 3주까지도 관찰되었다. GFAP는 출생17일령에서 뇌실로부터 아교세포가 관찰되는 형태가 보였으며, 외측뇌실과 제3뇌실에서부터 피질로 이동하는 형태로 관찰되었다. 또한 후각망울의 과립층, 대뇌피질 및 해마체에서 관찰되었으며, 출생 2일령에서 가장 강한 반응을 나타내었다. CNTF는 신경원과 신경아교세포에서 관찰되었으며, NGF와 GFAP와는 달리 출생 전에서는 관찰되지 않다가 출생 1일령부터 후각망울 및 대뇌피질에서 약하게 관찰되기 시작되었으며, 이러한 반응은 출생 2주령에서 잘 관찰되었다. 전자현미경상에서는 신경원과 신경아교세포에서 특징적인 구조는 관찰되지 않았으나, 각각의 항체에 대한 반응이 나타난 세포에서는 사립체와 형질내세망과 같은 세포소기관들이 많이 관찰되었다. 이러한 Mongolian gerbil 전뇌에서의 NGF, GFAP 및 CNTF의 분포는 비슷한 임신일령의 설치류와 거의 유사하게 관찰되었으며, 배아과정 및 출생 후 발달에 따른 전뇌에서의 신경원과 신경아교세포에서의 분포를 볼 수 있었다.

The immunohistochemical localization of the nerve growth factor (NGF), glial fibrillary acidic protein (GFAP) and ciliary neurotrophic factor (CNIF) in the developing Mongolian gerbil forebrain was investigated by the immunohistochemical and electron microscopy methods. Generally, the NGF specifically recognizes the neurons, the GFAP does the glia, and the CNIF does the motor neurons. This study demonstrates the location of the NGF, GFAP and CNTF in the developing Mongolian gerbil from the embryonic days 17 (E17) to the postnatal weeks 3 (PNW 3). The NGF was localized at E19 in the olfactocy bulb and the cerebral cortex, expanded to the hippocampus, and the diagonal bond from the late prenatal period to PNW 3. GFAP was observed in the lateral ventricle and the third ventricle at E17, projected into the cerebral cortex at E19. The GFAP was observed to have the largest numbers in several parts of the forebrain at the postnatal days 2 (PND2), while the most numerous CNTF was observed at PNW 2. The CNTF-IR cells were observed only in the postnatal days and were found in the olfactory bulb, cerebral cortex both neuron and neuroglia at PND3. Electron microscopy showed that the NGF, GFAP and CNTF were not related to any connections with any particular subcellular structure. These results suggest that NGF, GFAP and CNTF be related to the neuron and neuroglia at the prenatal and postnatal stages in the developing Mongolian gerbil.

키워드

참고문헌

  1. Mowla SJ, Sangeeta P, Homan FF, et al. Differencial sorting of nerve growth factor and brain-derived neurooophic factor in hippocampal neurnos. J Neurosci,19:2069-2080, 1999
  2. Barde YA. Trophic factors and neuronal survival. Neuron, 2:1525-1534. 1989
  3. Benowitz LI, Shashoua YE. Immunoreactive sites for nerve growth factor (NGF) in the goldfish brain. Brain Res, 172:561-565, 1979
  4. Conner JM, Muir D, Varon S, et at. The localization of nerve growth factor-like immnuoreacrivity in the adult rat basal forebrain and tippocampal formation. J Comp Neurol, 319: 454-462. 1992
  5. Finn PJ, Ferguson IA, Wilson PA, et at. Immuno histochemical evidence for the distribution of nerve growth factor in the embryonic mouse. J Neurocytol, 16:639-647, 1987
  6. Rush RA. Immunohistochemical localization of endogenous nerve growth factor. Nature (London), 312:364-367, 1984
  7. Williams R, Rush RA. Electron microscopic immunocytochemical localization of nerve growth factor in developing mouse olfactory neurons. Brain Res, 463:21-27, 1988
  8. Yan Q, Eugene M, Johnson Jr. Immunohistochemical localization and biochemical characterization of nerve growth factor receptor in adult rat brain. J Comp Neurol, 290:585-598, 1989
  9. Yan Q, Eugene M, Johnson Jr. An immunohistochemical study of the nerve growth factor receptor in developing rats. J Neurosci, 8:3481-3498, 1988
  10. Yu C, Crutcher KA. Nerve growth factor immumoreactivity and sympathetic sprouting in the rat hippocampal formation. Brain Res, 672:55-67, 1995
  11. Semokova I, Krieglstein J. Ciliary neurotrophic factor enhances the expression of NGF and p75 low-affivity NGF receptor in astrocytes. Brain Res, 838:184-19, 1999
  12. Gnahn H, Hefti F, Heumaim R, et al. NGF-mediated increase of choline acetyltiansferase (ChAT) in the neonatal rat forebrain: Evidence for a physiological role of NGF in the brain?. Dev Brain Res, 9:45-52, 1983
  13. Koh S, Oyler GA, Higgins GA. Localization of nerve growth factor receptor messenger RNA and protein in the adult rat brain Exp Neurol, 106:209-221, 1989
  14. Bigmnami A, Eng LF, Dahl D, et al. Localization of the glial fibrillary acidic protein in astrocytes by immunofluorescence, Brain Res, 43: 429-435, 1972
  15. Roessmann U, Velasco ME, Sindely SD, et al. Glial fibrillary acidic protein (GFAP) in ependymaI cells during development. An immunocytochemical study.Brain Res, 200:13-21, 1980
  16. Schiffer D, Giordana Mr, Migheli A, et al. Glial fibrillary protein and vimentin in the experimental glial reaction of the rat brain. Brain Res, 374:110-118, 1986
  17. Kalman M, Szekely AD, Csillag A. Distribution of glial fibrillary acidic protein and vimentin-immunopositive elements in the developing chicken brain hatch to adulthood. Anat Embryol, 198:213-235, 1998
  18. Kalman M, Hajas F. Distribution of glial fibrillary acidic protein(GFAP)-immunoreactive astrocytes in the rat brain. I. Forebrain. Exp Brain Res, 78:147-163, 1989
  19. Stockli KA, Lillien LE, Naher-Noe M, et al. Regional distribution, developmental changes, and cellular localization of CNTF-mRNA and protein in the rat brain. J Cell BioI, 115:447-459, 1991
  20. Sendtner M, Kreutzberg GW, Thoenen H Ciliary neurotrophic factor prevents the degeneration of motor neurons after axotorny. Nature, 345:440-441, 1990
  21. Jean-Claude L, Ella M, Seiji T, et at. CNTF-protection of oligodendrocytes against natural and tumor necrosis factor-induced death. Science, 259:689-692, 1993
  22. Margot M, Kishore B, Mark N. Ciliary neurotrophic factor and leukemia inhibitory factor promote the generation, maturation and survival of oligodendrocytesin vitro. Development, 120:143-153, 1994
  23. Piotr M, Haoxing L, Czeslaw R, et al. Recombinant human and rat ciliary neurotrophic factors. J Neurochem, 57:1003-1012, 1991
  24. Aloe L, Micera A, Bracci-Laudiero L, et al. Presence of nerve growth factor in the thymus of prenatal, paitnatal and pregnant rats. Thymus, 24:221-31, 1997
  25. Frisen J, Risling M, Korhonen L, et al. Nerve growth factor induces process formation in menigeal cells: Implications for scar formation in the injured CNS. JNeurosci, 18:5714-572, 1998
  26. Juanes JA, Riesco JM, Sanchez F, et al. Glial fibrillary acidic protein-like immunoreactive ependymal elements in the third ventricle of the rat, Acta Anat, 145:364-369, 1992
  27. Rickmatm M, Amaral DG, Cowan WM. Organization of radial glial cells during the development of the rat dentate gyrus. J Comp Neurol, 264:449-79, 1987
  28. Stockli KA, Lottspeich F, Sendtner M, et al. Cloning, expression and regional distribution of rat ciliary neurotrophic factor. Nature, 342:920-923, 1989
  29. Bajetto A, Schettini G, Chimini G. Nuclear localization of ciliary neurotrophic factor in glial cells. Brain Res, 818:565-569, 1999
  30. Hendeson JT, Seniuk NA, Roder JC. Localization of CNTF immunoreactivity to neurons and astroglia in the CNS, Mol Brain Res, 22:151-165, 1994
  31. Seniuk-Tatton NA, Henderson JT, and Roder JC. Neurons express ciliary neurotrophic factor mRNA in the early postnatal and adult rat brain. J Neurosci Res, 41:663-676, 1995