リケラボ論文検索は、全国の大学リポジトリにある学位論文・教授論文を一括検索できる論文検索サービスです。

リケラボ 全国の大学リポジトリにある学位論文・教授論文を一括検索するならリケラボ論文検索大学・研究所にある論文を検索できる

リケラボ 全国の大学リポジトリにある学位論文・教授論文を一括検索するならリケラボ論文検索大学・研究所にある論文を検索できる

大学・研究所にある論文を検索できる 「Absence of thyroid hormone induced delayed dendritic arborization in mouse primary hippocampal neurons through insufficient expression of brain-derived neurotrophic factor.」の論文概要。リケラボ論文検索は、全国の大学リポジトリにある学位論文・教授論文を一括検索できる論文検索サービスです。
リケラボ

コピーが完了しました

URLをコピーしました

論文の公開元へ論文の公開元へ
書き出し

Absence of thyroid hormone induced delayed dendritic arborization in mouse primary hippocampal neurons through insufficient expression of brain-derived neurotrophic factor.

矢島, 弘之 ヤジマ, ヒロユキ Yajima, Hiroyuki 群馬大学

2021.03.23

概要

1．研究の背景と目的
甲状腺ホルモンは、中枢神経系を含む多くの臓器の発達や機能維持に重要な役割を果たしている。周産期の甲状腺ホルモン欠乏症は、ヒトではクレチン症と呼ばれ、不可逆的な中枢神経障害が生じる。甲状腺ホルモンの脳発達への影響を評価するため、齧歯類がモデルとして広く用いられてきた。そして、行動学的実験・電気生理学的実験などから、甲状腺ホルモンが強い影響を及ぼす領域の一つに海馬があることが明らかとなった。先行研究において、周産期甲状腺ホルモン低下モデル動物で、海馬錐体細胞の樹状突起の分枝数の低下が報告されている。しかし、海馬の発達過程における甲状腺ホルモンの作用メカニズムは依然として不明な点が多い。そこで本研究では、海馬初代培養神経細胞を用い、ニューロンの成熟過程における甲状腺ホルモンの作用を明らかにすることを目的とした。

2. 実験方法
C57BL/6Jマウスの胎生15/16日の脳から、海馬を単離しパパイン処理にて分散培養した。海馬 初代培養神経細胞に使用される無血清サプリメントB27®には甲状腺ホルモンが含まれている。そ こで、Chenら (J Neurosci Methods.,2008) の組成を基に活性型の甲状腺ホルモンであるトリヨ ードサイロニン (T3) を含まない無血清培養液を作成し、T3添加群とT3未添加群で培養を行った。培養7、10、14日目において免疫細胞化学染色を行ない、顕微鏡下で観察した。Microtubule- associated protein 2 (MAP2)、Glial fibrillary acidic protein (GFAP) を染色し、神経細胞 数および、グリア細胞数を計測した。また、神経細胞の形態学的解析として、Sholl解析で細胞 体の中心から10 µm間隔で引かれた同心円と樹状突起との交点数を計測し、さらに各神経細胞に おける分枝数を計測した。培養14日目においてSynpatophysinを免疫細胞化学染色し、シナプス 数計測を行った。また、培養7、10、14日目において細胞からTotal RNAを抽出し甲状腺ホルモン 標的遺伝子やシナプス関連遺伝子の定量リアルタイムPCRを行った。培養10日目でBDNF受容体であるNeurotrophic tyrosine kinase receptor type 2 (Ntrk2, TrkB) のリン酸化レベルを Western blotで評価した。さらに、培養8、9日目に培地中にT3未添加群へBDNFを添加し、培養10 日目にSholl解析、分枝数の計測を行った。

3. 結果
細胞数を計測したところ、神経細胞数、グリア細胞数においてT3添加群とT3非添加群の間に有意な差はなかった。形態解析の結果、Sholl解析において、培養10日目にT3添加群と比較しT3未添加群の有意な交点数の低下が見られ、分枝数の計測においても、培養10日目にT3添加群と比較し T3未添加群の第二・第三分枝数の有意な減少が見られた。定量リアルタイムPCRにおいて、甲状腺ホルモン標的遺伝子Bdnfは培養10日目にT3添加群と比較し、T3未添加群の有意なmRNA量の減少が認められた。培養14日目におけるシナプス数解析の結果に差はなかった。培養10日目にBDNFの分泌量を評価するため、BDNFの受容体であるTrkBのリン酸化レベルを調べたところ、T3未添加群と比較しT3添加群は有意にリン酸化レベルが増加した。形態解析において、培養10日目にT3未添加群で樹状突起の形態変化が生じ、同時期において樹状突起の成長に関与している神経栄養因子であるBdnfの転写量が低下し、さらにBDNFの受容体であるTrkBのリン酸化レベルがT3未添加群において低下したことから、T3未添加群へBDNFの添加により樹状突起の形態変化が正常化するか解析した。培養8、9日目にT3未添加群に対しBDNFを外部から添加したBDNF添加群では、培養10日目におけるT3未添加群のSholl解析での交点数の低下、並びに分枝数の計測での第二・第三分枝の低下が正常化した。

4. 考察・まとめ
先行研究においてBDNFノックアウト動物で神経細胞の類似の形態変化が報告されており、T3未添加によるBdnfの発現低下がより形態的変化の一因になっている可能性を支持している。本研究の結果から、甲状腺ホルモンの低下により、海馬初代培養神経細胞では一時的な発達遅滞が生じることが明らかになった。さらに、発達期の海馬神経細胞において甲状腺ホルモンを介したBdnfの発現変化により神経細胞の形態変化が生じている可能性が示唆された。

論文の公開元へ

この論文で使われている画像

関連論文

関連論文をもっと見る

参考文献

1. Cabello G, Wrutniak C. Thyroid hormone and growth: Relationships with growth hormone effects and regulation. Reprod Nutr Dev (1989) 29:387–402. doi: 10.1051/rnd:19890401

2. Louzada RA, Carvalho DP. Similarities and differences in the peripheral actions of thyroid hormones and their metabolites. Front Endocrinol (Lausanne) (2018) 9:394:394. doi: 10.3389/fendo.2018.00394

3. Chen ZP, Hetzel BS. Cretinism revisited. Best Pract Res Clin Endocrinol Metab (2010) 24:39–50. doi: 10.1016/j.beem.2009.08.014

4. Glinoer D. Clinical and biological consequences of iodine deficiency during pregnancy. Endocr Dev (2007) 10:62–85. doi: 10.1159/000106820

5. Williams GR. Neurodevelopmental and neurophysiological actions of thyroid hormone. J Neuroendocrinol (2008) 20:784–94. doi: 10.1111/j.1365- 2826.2008.01733.x

6. Zoeller RT, Rovet J. Timing of thyroid hormone action in the developing brain: Clinical observations and experimental findings. J Neuroendocrinol (2004) 16:809–18. doi: 10.1111/j.1365-2826.2004.01243.x

7. Gilbert ME, Lasley SM. Developmental thyroid hormone insufficiency and brain development: A role for brain-derived neurotrophic factor (BDNF)? Neuroscience (2013) 239:253–70. doi: 10.1016/j.neuroscience.2012.11.022

8. Reid RE, Kim EM, Page D, O’Mara SM, O’Hare E. Thyroxine replacement in an animal model of congenital hypothyroidism. Physiol Behav (2007) 91:299–303. doi: 10.1016/j.physbeh.2007.03.005

9. Squire LR. Memory and the Hippocampuss: A Synthesis From Findings With Rats, Monkeys, and Humans. Psychol Rev (1992) 99:195–231. doi: 10.1037/ 0033-295X.99.2.195

10. Amano I, Takatsuru Y, Khairinisa MA, Kokubo M, Haijima A, Koibuchi N. Effects of Mild Perinatal Hypothyroidism on Cognitive Function of Adult Male Offspring. Endocrinology (2018) 159:1910–21. doi: 10.1210/en.2017- 03125

11. Gilbert ME, Sui L. Dose-dependent reductions in spatial learning and synaptic function in the dentate gyrus of adult rats following developmental thyroid hormone insufficiency. Brain Res (2006) 1069:10–22. doi: 10.1016/ j.brainres.2005.10.049

12. Khairinisa MA, Takatsuru Y, Amano I, Kokubo M, Haijima A, Miyazaki W, et al. In utero and postnatal propylthiouracil-induced mild hypothyroidism impairs maternal behavior in mice. Front Endocrinol (Lausanne) (2018) 9:228. doi: 10.3389/fendo.2018.00228

13. Sui L, Gilbert ME. Pre- and postnatal propylthiouracil-induced hypothyroidism impairs synaptic transmission and plasticity in area CA1 of the neonatal rat hippocampus. Endocrinology (2003) 144:4195–203. doi: 10.1210/en.2003-0395

14. Sánchez-Huerta KB, Montes S, Pérez-Severiano F, Alva-Sánchez C, Rıós C, Pacheco-Rosado J. Hypothyroidism reduces glutamate-synaptic release by ouabain depolarization in rat CA3-hippocampal region. J Neurosci Res (2012) 90:905–12. doi: 10.1002/jnr.22806

15. Vara H, Martıńez B, Santos A, Colino A. Thyroid hormone regulates neurotransmitter release in neonatal rat hippocampus. Neuroscience (2002) 110:19–28. doi: 10.1016/S0306-4522(01)00541-3

16. Vara H, Muñoz-Cuevas J, Colino A. Age-dependent alterations of long-term synaptic plasticity in thyroid-deficient rats. Hippocampus (2003) 13:816–25. doi: 10.1002/hipo.10132

17. Madeira MD, Cadete-Leite A, Andrade JP, Paula-Barbosa MM. Effects of hypothyroidism upon the granular layer of the dentate gyrus in male and female adult rats: A morphometric study. J Comp Neurol (1991) 314:171–86. doi: 10.1002/cne.903140116

18. Madeira MD, Sousa N, Lima-Andrade MT, Calheiros F, Cadete-Leite A, Paula-Barbosa MM. Selective vulnerability of the hippocampal pyramidal neurons to hypothyroidism in male and female rats. J Comp Neurol (1992) 322:501–18. doi: 10.1002/cne.903220405

19. Rami A, RabiéA, Patel AJ. Thyroid hormone and development of the rat hippocampus: Cell acquisition in the dentate gyrus. Neuroscience (1986) 19:1207–16. doi: 10.1016/0306-4522(86)90134-X

20. Rami A, Patel AJ, RabiéA. Thyroid hormone and development of the rat hippocampus: Morphological alterations in granule and pyramidal cells. Neuroscience (1986) 19:1217–26. doi: 10.1016/0306-4522(86)90135-1

21. Jan YN, Jan LY. Branching out: Mechanisms of dendritic arborization. Nat Rev Neurosci (2010) 11:316–28. doi: 10.1038/nrn2836

22. Cayrou C, Denver RJ, Puymirat J. Suppression of the basic transcription element-binding protein in brain neuronal cultures inhibits thyroid hormone- induced neurite branching. Endocrinology (2002) 143:2242–9. doi: 10.1210/ endo.143.6.8856

23. Kimura-Kuroda J, Nagata I, Negishi-Kato M, Kuroda Y. Thyroid hormone- dependent development of mouse cerebellar Purkinje cells in vitro. Dev Brain Res (2002) 137:55–65. doi: 10.1016/S0165-3806(02)00408-X

24. Amano I, Takatsuru Y, Toya S, Haijima A, Iwasaki T, Grasberger H, et al. Aberrant Cerebellar Development in Mice Lacking Dual Oxidase Maturation Factors. Thyroid (2016) 26:741–52. doi: 10.1089/thy.2015.0034

25. Koibuchi N, Chin WW. Thyroid hormone action and brain development. Trends Endocrinol Metab (2000) 11:123–8. doi: 10.1016/S1043-2760(00) 00238-1

26. Banker GA, Cowan WM. Rat hippocampal neurons in dispersed cell culture. Brain Res (1977) 126:397–425. doi: 10.1016/0006-8993(77)90594-7

27. Thompson CC, Potter GB. Thyroid Hormone Action in Neural Development. Cereb Cortex (2000) 10:939–45. doi: 10.1093/cercor/10.10.939

28. Koibuchi N. The role of thyroid hormone on cerebellar development. Cerebellum (2008) 7:530–3. doi: 10.1007/s12311-008-0069-1

29. Harada A, Teng J, Takei Y, Oguchi K, Hirokawa N. MAP2 is required for dendrite elongation, PKA anchoring in dendrites, and proper PKA signal transduction. J Cell Biol (2002) 158:541–9. doi: 10.1083/jcb.200110134

30. Chen Y, Stevens B, Chang J, Milbrandt J, Barres BA, Hell JW. NS21: Re- defined and modified supplement B27 for neuronal cultures. J Neurosci Methods (2008) 171:239–47. doi: 10.1016/j.jneumeth.2008.03.013

31. Ohkawa T, Satake S, Yokoi N, Miyazaki Y, Ohshita T, Sobue G, et al. Identification and characterization of GABAA receptor autoantibodies in autoimmune encephalitis. J Neurosci (2014) 34:8151–63. doi: 10.1523/ JNEUROSCI.4415-13.2014

32. Zagrebelsky M, Gödecke N, Remus A, Korte M. Cell type-specific effects of BDNF in modulating dendritic architecture of hippocampal neurons. Brain Struct Funct (2018) 223:3689–709. doi: 10.1007/s00429-018-1715-0

33. Cazzin C, Mion S, Caldara F, Rimland JM, Domenici E. Microarray analysis of cultured rat hippocampal neurons treated with brain derived neurotrophic factor. Mol Biol Rep (2011) 38:983–90. doi: 10.1007/s11033-010-0193-0

34. Patel MN, McNamara JO. Selective enhancement of axonal branching of cultured dentate gyrus neurons by neurotrophic factors. Neuroscience (1995) 69:763–70. doi: 10.1016/0306-4522(95)00281-M

35. Balkowiec A, Katz DM. Cellular mechanisms regulating activity-dependent release of native brain-derived neurotrophic factor from hippocampal neurons. J Neurosci (2002) 22:10399–407. doi: 10.1523/jneurosci.22-23-10399.2002

36. Ariyani W, Iwasaki T, Miyazaki W, Khongorzul E, Nakajima T, Kameo S, et al. Effects of gadolinium-based contrast agents on thyroid hormone receptor action and thyroid hormone-induced cerebellar purkinje cell morphogenesis. Front Endocrinol (Lausanne) (2016) 7:115:115. doi: 10.3389/fendo.2016.00115

37. Jacobs S, Cheng C, Doering LC. Hippocampal neuronal subtypes develop abnormal dendritic arbors in the presence of Fragile X astrocytes. Neuroscience (2016) 324:202–17. doi: 10.1016/j.neuroscience.2016.03.011

38. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, et al. Fiji: An open-source platform for biological-image analysis. Nat Methods (2012) 9:676–82. doi: 10.1038/nmeth.2019

39. Sholl DA. Dendritic organization in the neurons of the visual and motor cortices of the cat. J Anat (1953) 87:387–406. doi: 10.1038/171387a0

40. Paper O. Erratum: Iron deficiency impairs developing hippocampal neuron gene expression, energy metabolism, and dendrite complexity (Developmental Neuroscience (2016) 38 (264-276) DOI: 10.1159/000448514). Dev Neurosci (2018) 40:288. doi: 10.1159/000492448

41. Yajima H, Haijima A, Khairinisa MA, Shimokawa N, Amano I, Takatsuru Y. Early-life stress induces cognitive disorder in middle-aged mice. Neurobiol Aging (2018) 64:139–46. doi: 10.1016/j.neurobiolaging.2017.12.021

42. Xie C, Markesbery WR, Lovell MA. Survival of hippocampal and cortical neurons in a mixture of MEM+ and B27-supplemented neurobasal medium. Free Radic Biol Med (2000) 28:665–72. doi: 10.1016/S0891-5849(99)00268-3

43. Impey S, Davare M, Lasiek A, Fortin D, Ando H, Varlamova O, et al. An activity-induced microRNA controls dendritic spine formation by regulating Rac1-PAK signaling. Mol Cell Neurosci (2010) 43:146–56. doi: 10.1016/ j.mcn.2009.10.005

44. Molnár E. Long-term potentiation in cultured hippocampal neurons. Semin Cell Dev Biol (2011) 22:506–13. doi: 10.1016/j.semcdb.2011.07.017

45. Begni V, Riva MA, Cattaneo A. Cellular and molecular mechanisms of the brain-derived neurotrophic factor in physiological and pathological conditions. Clin Sci (2017) 131:123–38. doi: 10.1042/CS20160009

46. Gupta VK, You Y, Gupta VB, Klistorner A, Graham SL. TrkB receptor signalling: Implications in neurodegenerative, psychiatric and proliferative disorders. Int J Mol Sci (2013) 14:10122–42. doi: 10.3390/ijms140510122

47. Gil-Ibañez P, Garcıá-GarcıáF, Dopazo J, Bernal J, Morte B. Global transcriptome analysis of primary cerebrocortical cells: Identification of genes regulated by triiodothyronine in specific cell types. Cereb Cortex (2017) 27:706–17. doi: 10.1093/cercor/bhv273

48. Royland JE, Parker JS, Gilbert ME. A genomic analysis of subclinical hypothyroidism in hippocampus and neocortex of the developing rat brain. J Neuroendocrinol (2008) 20:1319–38. doi: 10.1111/j.1365-2826.2008.01793.x

49. Takahashi M, Negishi T, Tashiro T. Identification of genes mediating thyroid hormone action in the developing mouse cerebellum. J Neurochem (2008) 104:640–52. doi: 10.1111/j.1471-4159.2007.05049.x

50. Gilbert ME, Sanchez-Huerta K, Wood C. Mild thyroid hormone insufficiency during development compromises activity-dependent neuroplasticity in the hippocampus of adult male rats. Endocrinology (2016) 157:774–87. doi: 10.1210/en.2015-1643

51. Sui L, Li BM. Effects of perinatal hypothyroidism on regulation of reelin and brain-derived neurotrophic factor gene expression in rat hippocampus: Role of DNA methylation and histone acetylation. Steroids (2010) 75:988–97. doi: 10.1016/j.steroids.2010.06.005

52. Vella KR, Hollenberg AN. The actions of thyroid hormone signaling in the nucleus. Mol Cell Endocrinol (2017) 458:127–35. doi: 10.1016/ j.mce.2017.03.001

53. Binder DK, Scharfman HE. Brain-derived neurotrophic factor. Growth Factors (2004) 22:123–31. doi: 10.1080/08977190410001723308

54. McAllister AK, Lo DC, Katz LC. Neurotrophins regulate dendritic growth in developing visual cortex. Neuron (1995) 15:791–803. doi: 10.1016/0896-6273 (95)90171-X

55. Rauskolb S, Zagrebelsky M, Dreznjak A, Deogracias R, Matsumoto T, Wiese S, et al. Global deprivation of brain-derived neurotrophic factor in the CNS reveals an area-specific requirement for dendritic growth. J Neurosci (2010) 30:1739–49. doi: 10.1523/JNEUROSCI.5100-09.2010

56. Aid T, Kazantseva A, Piirsoo M, Palm K, Timmusk T. Mouse and rat BDNF gene structure and expression revisited. J Neurosci Res (2007) 85:525–35. doi: 10.1002/jnr.21139

57. Labelle C, Leclerc N. Exogenous BDNF, NT-3 and NT-4 differentially regulate neurite outgrowth in cultured hippocampal neurons. Dev Brain Res (2000) 123:1–11. doi: 10.1016/S0165-3806(00)00069-9

58. Yan Q, Radeke MJ, Matheson CR, Talvenheimo J, Welcher AA, Feinstein SC. Immunocytochemical localization of TrkB in the central nervous system of the adult rat. J Comp Neurol (1997) 378:135–57. doi: 10.1002/(SICI)1096-9861 (19970203)378:1<135::AID-CNE8>3.0.CO;2-5

59. Ledda F, Paratcha G. Mechanisms regulating dendritic arbor patterning. Cell Mol Life Sci (2017) 74:4511–37. doi: 10.1007/s00018-017-2588-8

60. Sawano E, Takahashi M, Negishi T, Tashiro T. Thyroid hormone-dependent development of the GABAergic pre- and post-synaptic components in the rat hippocampus. Int J Dev Neurosci (2013) 31:751–61. doi: 10.1016/ j.ijdevneu.2013.09.007

61. Zucker RS, Regehr WG. Short-term synaptic plasticity. Annu Rev Physiol (2002) 64:355–405. doi: 10.1146/annurev.physiol.64.092501.114547

62. Hajós F, Patel AJ, Balázs R. Effect of thyroid deficiency on the synaptic organization of the rat cerebellar cortex. Brain Res (1973) 50:387–401. doi: 10.1016/0006-8993(73)90740-3

63. Nicholson JL, Altman J. The effects of early hypo- and hyperthyroidism on the development of the rat cerebellar cortex. II. Synaptogenesis in the molecular layer. Brain Res (1972) 44:25–36. doi: 10.1016/0006-8993(72)90363-0

64. Nicholson JL, Altman J. Synaptogenesis in the rat cerebellum: effects of early hypo- and hyperthyroidism. Science (1972) 176:530–2. doi: 10.1126/ science.176.4034.530

65. Matteoli M, Verderio C, Krawzeski K, Mundigl O, Coco S, Fumagalli G, et al. Mechanisms of synaptogenesis in hippocampal neurons in primary culture. J Physiol - Paris (1995) 89:51–5. doi: 10.1016/0928-4257(96)80551-1

66. Kuczewski N, Porcher C, Lessmann V, Medina I, Gaiarsa JL. Activity- dependent dendritic release of BDNF and biological consequences. Mol Neurobiol (2009) 39:37–49. doi: 10.1007/s12035-009-8050-7

67. Matsuda N, Lu H, Fukata Y, Noritake J, Gao H, Mukherjee S, et al. Differential activity-dependent secretion of brain-derived neurotrophic factor from axon and dendrite. J Neurosci (2009) 29:14185–98. doi: 10.1523/JNEUROSCI.1863- 09.2009

68. Andreska T, Aufmkolk S, Sauer M, Blum R. High abundance of BDNF within glutamatergic presynapses of cultured hippocampal neurons. Front Cell Neurosci (2014) 8:107. doi: 10.3389/fncel.2014.00107

参考文献をもっと見る

全国の大学の
卒論・修論・学位論文
を
一発検索！

この論文の関連論文を見る