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在哺乳期,细胞特异性和通用启动子在转导催产素神经元和监测其神经活动方面的效率。

Efficiency of cell-type specific and generic promoters in transducing oxytocin neurons and monitoring their neural activity during lactation.

机构信息

Department of Psychiatry, Icahn School of Medicine at Mount Sinai, 1 Gustave L. Levy Pl, New York, NY, 10029, USA.

Seaver Autism Center for Research and Treatment, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

出版信息

Sci Rep. 2021 Nov 18;11(1):22541. doi: 10.1038/s41598-021-01818-x.

Abstract

Hypothalamic oxytocin (OXT) and arginine-vasopressin (AVP) neurons have been at the center of several physiological and behavioral studies. Advances in viral vector biology and the development of transgenic rodent models have allowed for targeted gene expression to study the functions of specific cell populations and brain circuits. In this study, we compared the efficiency of various adeno-associated viral vectors in these cell populations and demonstrated that none of the widely used promoters were, on their own, effective at driving expression of a down-stream fluorescent protein in OXT or AVP neurons. As anticipated, the OXT promoter could efficiently drive gene expression in OXT neurons and this efficiency is solely attributed to the promoter and not the viral serotype. We also report that a dual virus approach using an OXT promoter driven Cre recombinase significantly improved the efficiency of viral transduction in OXT neurons. Finally, we demonstrate the utility of the OXT promoter for conducting functional studies on OXT neurons by using an OXT specific viral system to record neural activity of OXT neurons in lactating female rats across time. We conclude that extreme caution is needed when employing non-neuron-specific viral approaches/promoters to study neural populations within the paraventricular nucleus of the hypothalamus.

摘要

下丘脑催产素(OXT)和精氨酸加压素(AVP)神经元一直是许多生理和行为研究的中心。病毒载体生物学的进步和转基因啮齿动物模型的发展,使得针对特定细胞群体和脑回路的靶向基因表达成为可能。在这项研究中,我们比较了各种腺相关病毒载体在这些细胞群体中的效率,并证明在 OXT 或 AVP 神经元中,没有一种广泛使用的启动子能够有效地驱动下游荧光蛋白的表达。正如预期的那样,OXT 启动子可以有效地驱动 OXT 神经元中的基因表达,而这种效率完全归因于启动子,而不是病毒血清型。我们还报告说,使用 OXT 启动子驱动 Cre 重组酶的双重病毒方法显著提高了 OXT 神经元中病毒转导的效率。最后,我们通过使用 OXT 特异性病毒系统在哺乳期雌性大鼠中记录 OXT 神经元的神经活动,证明了 OXT 启动子在 OXT 神经元功能研究中的实用性。我们得出结论,在研究下丘脑室旁核内的神经群体时,需要极其谨慎地使用非神经元特异性的病毒方法/启动子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2175/8602291/9b7789622ad2/41598_2021_1818_Fig1_HTML.jpg

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