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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳示國 | |
| dc.contributor.author | Yi-Ting Chang | en |
| dc.contributor.author | 張宜婷 | zh_TW |
| dc.date.accessioned | 2021-06-15T13:55:55Z | - |
| dc.date.available | 2018-09-22 | |
| dc.date.copyright | 2015-09-30 | |
| dc.date.issued | 2015 | |
| dc.date.submitted | 2015-08-27 | |
| dc.identifier.citation | Abrahamson, E.E., Leak, R.K., and Moore, R.Y. (2001). The suprachiasmatic nucleus projects to posterior hypothalamic arousal systems. Neuroreport 12, 435-440.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/51895 | - |
| dc.description.abstract | 在哺乳類視網膜中，自主感光視神經細胞傳遞光訊息至腦區，以調控非視覺相關的功能，如生理時鐘和瞳孔反射。目前研究顯示，自主感光視神經細胞會直接投射至生理時鐘的中樞-視叉上核。但現階段對此神經迴路的了解，仍無法解釋光如何造成多樣的生理時鐘變化。這是由於我們的認知侷限於組織層次，究竟單一自主感光視神經細胞如何與視叉上核的細胞連結，光訊息又是如何經由這複雜的神經網路所傳遞和整合，目前仍未知。故本研究欲剖析視網膜和視叉上核間的神經迴路，我們結合藥理與基因轉殖鼠的技術，成功地追蹤單一自主感光視神經細胞，並分析其在視網膜中的樹突型態，與腦中的軸突分佈。我們的研究發現，單一自主感光視神經細胞可同時投射至多個腦區，主要投射目標為–視叉上核(SCN) 和膝狀間區小葉(IGL)。令人驚訝的是，即使異側對同側自主感光視神經細胞的比例為9:1，它們可透過非典型的軸突同時連至兩側的視叉上核，最終使得兩側的視叉上核接收等量的單眼光訊息。此外，每個自主感光視神經細胞會傾向投射至視叉上核中的特定區域，代表可能與單一種視叉上核的細胞連結，也就是光可能透過不同的神經迴路來影響生理時鐘。總結來說，建構單一自主感光視神經細胞至視叉上核的神經連結，是解開此複雜神經迴路突破性的一步，並奠定日後研究光如何調控生理時鐘的基礎。 | zh_TW |
| dc.description.abstract | In the mammalian retina, intrinsically photosensitive retinal ganglion cells (ipRGCs) convey photic inputs to several brain regions for non-image forming functions, such as circadian photoentrainment and pupil light reflex. Recent studies have shown that ipRGCs directly project to the suprachiasmatic nucleus (SCN), the mammalian master clock, through the retinohypothalamic tract (RHT). However, our understandings of retinal -SCN circuit are still limited at the tissue level, which cannot explain the variety of light effects on circadian photo-entrainment. To dissect out the complicated retinal-SCN connectivity by tracing single ipRGC is definitely a key breakthrough for revealing how light information is processed in the SCN. Here we applied a conditional and inducible Cre-LoxP system to randomly label a single ipRGC in mice, and reconstructed their dendritic structure and axonal architecture from the retina to the brain. Our findings reveal that a single ipRGC projects to multiple brain nuclei, especially the predominant targets- the SCN and IGL. Strikingly, we also found that individual ipRGCs, despite with the 9:1 contralateral to ipsilateral ratio, can bilaterally project to both ipsilateral and contralateral sides of the SCN. As a result, the photic inputs from monocular ipRGCs to both sides of the SCN are close to equal. Finally, our results show that individual ipRGCs preferentially target specific regions in the SCN, suggesting that a single ipRGC may form synapses with specific SCN neurons. These distinct neuronal circuits may explain the diverse light responses of SCN in circadian photo-entrainment. Taken together, these detailed morphological features and innervation patterns in a single-cell resolution provide us valuable insight into the complexity of retinal-SCN circuit. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T13:55:55Z (GMT). No. of bitstreams: 1 ntu-104-R02b21007-1.pdf: 262627643 bytes, checksum: 7d738a3df3ed4f599c0abdbffa72a9bb (MD5) Previous issue date: 2015 | en |
| dc.description.tableofcontents | 口試委員審定書…………………………………………………..……………………i
謝誌….......................................……………………………………...……………ii 摘要…………………………………………….……………………...……………....iv Abstract …………………………………………..…………………………..…....….v Abbreviations …………………………………..………………………………….…vii Contents ……………………………………………...………………………..….….ix Chapter I Introduction…..…...…………………………...………………………….1 1.1 Retina structure and visual processing in the retina……………...…….…1 1.2 Visual system and binocular vision……..……………......………………….2 1.3 Intrinsically photosensitive retinal ganglion cells…..…….……….……….3 1.3.1 Properties of melanopsin photopigment………...……...…..………..4 1.3.2 Projections and physiological functions of ipRGCs…………………5 1.3.3 Subtypes of ipRGCs………………….……………….…………….……5 1.4 Circadian rhythm………………………………….……………………………6 1.5 Suprachiasmatic nucleus……………………………………………….........7 1.5.1 Structures of the SCN…………………………………………...…….7 1.5.2 Inputs of the SCN………………………………………………...……...8 1.5.3 Synchronization of SCN neurons……………………………..………9 1.5.4 Outputs of the SCN………………..…………..………………………10 Statement of Purpose…………………………………………………..………...12 Chapter II Materials and methods…..………………………….....…………….13 2.1 Animals…………………………………………………..…..……………...…13 2.2 Genotyping …………………………..……………...…….………………….13 2.2.1 DNA isolation……….….....……………………...…….……………….13 2.2.2 Polymerase chain reaction (PCR)…….……...…….………………….14 2.2.3 X-gal staining……….……………...……………...…….………………14 2.3 Enucleation…………………………………………………..……………...…14 2.4 Tamoxifen injection……….………………………...…….…………………14 2.5 Alkaline phosphatase staining……….……………………...…….………15 2.6 Immunofluorescence staining………………………………………………15 2.6.1 Preparation of frozen-tissue……….………………………....…….…16 2.6.2 Immunohistochemistry…………..……………………...…….………16 2.7 3D reconstructions and analysis………..………………..……………...…17 2.7.1 3D reconstruction …….....……..……………...….…………………...17 2.7.2 Analysis ……………........……..……………...…….………………...17 2.7.3 Statistics ………………………………………………………………..18 2.8 In situ hybridization (ISH) ………………………………………………….18 2.8.1 Retina RNA extraction ………………………………………………...18 2.8.2 cDNA synthesis ………………………………………………………..19 2.8.3 Cloning for preparation of probe template…………………………19 2.8.4 Probe Synthesis …………………………………………………….…..21 2.8.5 Preparation of whole retina and frozen sections...…………………21 2.8.6 Sample pretreatment …………………………….....…………….…..22 2.8.7 Hybridization and antibody staining ………………………………...22 Chapter III Results…..…...………..…………………...…………...……………...24 3.1 A single M1 ipRGC projects to multiple brain nuclei with collateral axons…...24 3.2 The bilateral innervation of single ipRGCs contributes to the symmetric input from the retina to the SCN…………………..…………………………….25 3.3 Unilateral and bilateral ipRGCs have similar dendritic morphologies..27 3.4 Individual ipRGC prefers specific region of the SCN……………………27 3.5 A single ipRGC prefer to connect to one type of SCN neurons…….....29 3.6 The dendritic- axonal correlation of ipRGCs…………………………...…30 Chapter IV Discussion…..…...…………..…………...………….....…….……...31 4.1 M1 ipRGCs send collateral axon to multiple brain regions……………31 4.2 A single ipRGC bilaterally innervates both sides of the SCN…………33 4.3 A single ipRGC prefers to innervate a specific region in the SCN and to have a certain synaptic partners………...………………………………………36 4.4 The correlation between dendritic morphology and axonal properties.38 4.5 Limitations and future directions of current technology of single cell tracing…39 Chapter V Significance of work…..…...…………..…………...…….....………41 Chapter VI Reference……………..…...…………..…………...……....………….42 Figures……………………………………………………………………………....54 Figure 1. Schematic representation of the mouse genetic lines……………54 Figure 2. Representative images of AP staining and 3D reconstruction from a single ipRGC in the mouse………………………………………………………55 Figure 3. A single ipRGC innervates multiple brain targets……………………56 Figure 4. Overall innervation patterns of single ipRGCs……………………….58 Figure 5. Axonal architectures of a single ipRGC in the IGL…………………59 Figure 6. Axonal architectures of a single ipRGC in the SCN…………………60 Figure 7. The spatial distribution of contralateral and ipsilateral ipRGCs in the retina..61 Figure 8. Innervation patterns of single ipRGCs in the SCN……………...….62 Figure 9. The axon branch right at the optic chiasm…………………………63 Figure 10. Bilateral indexes of contralateral and ipsilateral ipRGCs.………64 Figure 11. Axonal distribution of contralateral and ipsilateral ipRGCs in both sides of the SCN………………………………………………………………...65 Figure 12. Unilateral and bilateral projecting ipRGCs have similar somato-dendritic features………………………………………………………………….66 Figure 13. A single ipRGC preferentially innervates a specific region of the SCN……68 Figure 14. Scheme of axonal Sholl analysis ….…………………………………69 Figure 15. Axonal Sholl analysis of ventral, middle and dorsal targeting ipRGCs……70 Figure 16. Merged images of axonal fields of ventral, middle and dorsal targeting ipRGCs…………………………………………………………………...71 Figure 17. Ventral, middle and dorsal targeting ipRGCs have similar somato-dendritic features but different soma spatial distribution……………………72 Figure 18. A single ipRGC connects to VIP neurons in the SCN……………74 Figure 19. A single ipRGC connects to AVP neurons in the SCN……………75 Figure 20. A single ipRGC connects to GRP neurons in the SCN……………76 Figure 21. Dendritic-axonal correlation of ipRGCs……..………………………77 Figure 22. Expression of robo3 in the P4 mouse retinas………………………78 Figure 23. Models of neuronal circuits of single M1 ipRGCs in the mouse brain……..79 Tables………………………………………………………………………………...80 Table 1. List of primers for genotyping…………………………………………...80 Table 2. List of primary antibodies used in this study…………...…..………...81 Table 3. List of secondary antibodies used in this study……………..……...82 Table 4. Summary of brain targets from single ipRGCs.………………………83 Table 5. Dendritic-axonal correlation of ipRGCs………………………………84 Appendix……………………………………………………………………………...85 Appendix I pCRRII-TOPO vector used in ISH………..………………………85 Appendix II Light promotes hair follicle regeneration via ipRGCs………….86 Appendix III Abstract and Poster…….…………………………………………101 | |
| dc.language.iso | en | |
| dc.subject | 神經迴路 | zh_TW |
| dc.subject | 生理時鐘 | zh_TW |
| dc.subject | 單一神經元追蹤技術 | zh_TW |
| dc.subject | 視叉上核 | zh_TW |
| dc.subject | 視神經細胞 | zh_TW |
| dc.subject | circadian rhythm | en |
| dc.subject | suprachiasmatic nucleus | en |
| dc.subject | neuronal circuit | en |
| dc.subject | single cell tracing | en |
| dc.subject | retinal ganglion cells | en |
| dc.title | 建構單一M1 自主感光視神經細胞之神經迴路及探討其下游之生理功能 | zh_TW |
| dc.title | Determine the Neuronal Circuits of M1 Intrinsically Photosensitive Retinal Ganglion Cells (ipRGCs) at the Single-Cell Level and Their Downstream Functions | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 103-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 王致恬,林頌然,周申如 | |
| dc.subject.keyword | 視神經細胞,視叉上核,生理時鐘,神經迴路,單一神經元追蹤技術, | zh_TW |
| dc.subject.keyword | retinal ganglion cells,suprachiasmatic nucleus,circadian rhythm,neuronal circuit,single cell tracing, | en |
| dc.relation.page | 105 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2015-08-28 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生命科學系 | zh_TW |
| 顯示於系所單位： | 生命科學系 | |
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