지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2016.1
- 수록면
- 120 - 129 (10page)
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초록· 키워드
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Assessing the cell-type expression pattern of a certain gene can be achieved by using cell-type-specific gene manipulation. Recently, cre-recombinase-dependent gene-silencing tool, pSico has become popular in neuroscientific research. However, pSico has a critical limitation that gene-silenced cell cannot be identified by fluorescence, due to an excision of the reporter gene for green fluorescence protein (GFP). To overcome this limitation, we newly developed pSico-Red, with mCherry gene as a reporter outside two loxP sites, so that red mCherry signal is detected in all transfected cells. When a cell expresses cre, GFP is excised and shRNA is enabled, resulting in disappearance of GFP. This feature of pSico-Red provides not only cell-type-specific gene-silencing but also identification of cre expressing cells. Using this system, we demonstrated for the first time the neuronal expression of the Bestrophin-1 (Best1) in thalamic reticular nucleus (TRN) and TRN-neuron-specific gene-silencing of Best1. We combined adeno-associated virus (AAV) carrying Best1-shRNA in pSico-Red vector and transgenic mouse expressing cre under the promoter of distal-less homeobox 5/6 (DLX5/6), a marker for inhibitory neurons. Firstly, we found that almost all of inhibitory neurons in TRN express Best1 by immunohistochemistry. Using pSico-Red virus, we found that 80% of infected TRN neurons were DLX5/6-cre positive but parvalbumin negative. Finally, we found that Best1 in DLX5/6-cre positive neurons were significantly reduced by Best1-shRNA. Our study demonstrates that TRN neurons strongly express Best1 and that pSico-Red is a valuable tool for cell-type-specific gene manipulation and identification of specific cell population.
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등록된 정보가 없습니다.
참고문헌 (29)
참고문헌 신청
Hartzell HC / 2008 / Molecular physiology of bestrophins: multifunctional membrane proteins linked to best disease and other retinopathies / Physiol Rev 88:639~672
Petrukhin K / 1998 / Identification of the gene responsible for Best macular dystrophy / Nat Genet 19:241~247
Jo S / 2014 / GABA from reactive astrocytes impairs memory in mouse models of Alzheimer’s disease / Nat Med 20:886~896
Lee S / 2010 / Channel-mediated tonic GABA release from glia / Science 330:790~796
Park H / 2009 / Bestrophin-1 encodes for the Ca2+-activated anion channel in hippocampal astrocytes / J Neurosci 29:13063~13073
Petrukhin K / 1998 / Identification of the gene responsible for Best macular dystrophy / Nat Genet 19:241~247
Jo S / 2014 / GABA from reactive astrocytes impairs memory in mouse models of Alzheimer’s disease / Nat Med 20:886~896
Lee S / 2010 / Channel-mediated tonic GABA release from glia / Science 330:790~796
Park H / 2009 / Bestrophin-1 encodes for the Ca2+-activated anion channel in hippocampal astrocytes / J Neurosci 29:13063~13073
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