Conditional gene targeting using UCP1-Cre mice directly targets the central nervous system beyond thermogenic adipose tissues.
Conditional gene targeting using UCP1-Cre mice directly targets the central nervous system beyond thermogenic adipose tissues.
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DOI：
10.1016/j.molmet.2021.101405
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发表时间：
2022-01
影响因子：
8.1
通讯作者：
Potthoff MJ
中科院分区：
文献类型：
--
作者：
Claflin KE;Flippo KH;Sullivan AI;Naber MC;Zhou B;Neff TJ;Jensen-Cody SO;Potthoff MJ
解偶联蛋白1（UCP 1）是脂肪组织中对适应性产热至关重要的线粒体蛋白，并且通常认为其仅限于产热脂肪组织。UCP 1-Cre转基因小鼠在许多研究中用于提供“棕色脂肪特异性”条件基因靶向。在这里，我们研究了Cre和UCP 1在UCP 1-Cre报告小鼠体内的分布。利用UCP 1-Cre小鼠与Ai 14-tdTomato和Ai 9-tdTomato报告基因小鼠杂交，研究Cre重组酶和Ucp 1 mRNA在不同组织中的组织分布。UCP 1-Cre小鼠独立感染Cre依赖性PHP.eB-tdTomato病毒或Cre依赖性AAV-tdTomato病毒，以确定UCP 1是否在成人中枢神经系统中活跃表达以及在何处表达。原位分析沉积的单细胞RNA测序数据被用来评估Ucp 1在下丘脑中的表达。正如预期的那样，在棕色和腹股沟脂肪组织中均检测到Ucp 1表达。在肾脏、肾上腺、胸腺和下丘脑中也检测到Ucp 1表达。与可检测的Ucp 1表达一致，通过荧光成像和qPCR，在雄性和雌性UCP 1-Cre; Ai 14-tdTomato和UCP 1-Cre; Ai 9-tdTomato小鼠的棕色脂肪组织、腹股沟白色脂肪组织、肾脏、肾上腺和下丘脑中也观察到tdTomato表达。重要的是，在中枢神经系统内观察到tdTomato的表达，从而在对能量稳态调节至关重要的大脑区域中观察到UCP 1的表达，包括腹内侧下丘脑（VMH）。UCP 1-Cre; tdTomato小鼠中的TdTomato表达不限于产热脂肪组织。TdTomato还在肾脏、肾上腺和整个大脑中表达，包括对能量稳态的中枢调节的多个方面至关重要的大脑区域和细胞类型。总的来说，这些数据具有重要的意义，UCP 1-Cre小鼠作为遗传工具，专门在棕色脂肪组织中研究基因功能的效用。UCP 1-Cre转基因小鼠在产热脂肪组织以外的组织中表达Cre。UCP 1-Cre; tdTomato报告小鼠显示Cre也在整个脑、肾和肾上腺中表达。Ucp 1 mRNA在成年小鼠的中枢神经系统中表达活跃。
Uncoupling protein 1 (UCP1) is a mitochondrial protein critical for adaptive thermogenesis in adipose tissues, and it is typically believed to be restricted to thermogenic adipose tissues. UCP1-Cre transgenic mice are utilized in numerous studies to provide “brown adipose-specific” conditional gene targeting. Here, we examined the distribution of Cre and UCP1 throughout the body in UCP1-Cre reporter mice. UCP1-Cre mice crossed to Ai14-tdTomato and Ai9-tdTomato reporter mice were used to explore the tissue distribution of Cre recombinase and Ucp1 mRNA in various tissues. UCP1-Cre mice were independently infected with either a Cre-dependent PHP.eB-tdTomato virus or a Cre-dependent AAV-tdTomato virus to determine whether and where UCP1 is actively expressed in the adult central nervous system. In situ analysis of the deposited single cell RNA sequencing data was used to evaluate Ucp1 expression in the hypothalamus. As expected, Ucp1 expression was detected in both brown and inguinal adipose tissues. Ucp1 expression was also detected in the kidney, adrenal glands, thymus, and hypothalamus. Consistent with detectable Ucp1 expression, tdTomato expression was also observed in brown adipose tissue, inguinal white adipose tissue, kidney, adrenal glands, and hypothalamus of both male and female UCP1-Cre; Ai14-tdTomato and UCP1-Cre; Ai9-tdTomato mice by fluorescent imaging and qPCR. Critically, expression of tdTomato, and thus UCP1, within the central nervous system was observed in regions of the brain critical for the regulation of energy homeostasis, including the ventromedial hypothalamus (VMH). TdTomato expression in UCP1-Cre; tdTomato mice is not restricted to thermogenic adipose tissues. TdTomato was also expressed in the kidneys, adrenal glands, and throughout the brain, including brain regions and cell types that are critical for multiple aspects of central regulation of energy homeostasis. Collectively, these data have important implications for the utility of UCP1-Cre mice as genetic tools to investigate gene function specifically in brown adipose tissue. UCP1-Cre transgenic mice express Cre in tissues beyond thermogenic adipose tissues. UCP1-Cre; tdTomato reporter mice show that Cre is also expressed throughout the brain, kidney, and adrenal glands. Ucp1 mRNA is actively expressed in the central nervous system of adult mice.
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