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Rapid lipolytic oscillations in ex vivo adipose tissue explants revealed through microfluidic droplet sampling at high temporal resolution.
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-03-23 , DOI: 10.1039/d0lc00103a Juan Hu 1 , Xiangpeng Li , Robert L Judd , Christopher J Easley
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-03-23 , DOI: 10.1039/d0lc00103a Juan Hu 1 , Xiangpeng Li , Robert L Judd , Christopher J Easley
Affiliation
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Our understanding of adipose tissue biology has steadily evolved. While structural and energy storage functionalities have been in the forefront, a key endocrine role for adipocytes was revealed only over the last few decades. In contrast to the wealth of information on dynamic function of other endocrine tissues, few studies have focused on dynamic adipose tissue function or on tool development toward that end. Here, we apply our unique droplet-based microfluidic devices to culture, perfuse, and sample secretions from primary murine epididymal white adipose tissue (eWAT), and from predifferentiated clusters of 3T3-L1 adipocytes. Through automated control, oil-segmented aqueous droplets (∼2.6 nL) were sampled from tissue or cells at 3.5 second temporal resolution (including sample and reference droplets), with integrated enzyme assays enabling real-time quantification of glycerol (down to 1.9 fmol per droplet). This high resolution revealed previously unreported oscillations in secreted glycerol at frequencies of 0.2 to 2.0 min-1 (∼30-300 s periods) present in the primary tissue but not in clustered cells. Low-level bursts (∼50 fmol) released in basal conditions were contrasted with larger bursts (∼300 fmol) during stimulation. Further, both fold changes and burst magnitudes were decreased in eWAT of aged and obese mice. These results, combined with immunostaining and photobleaching analyses, suggest that gap-junctional coupling or nerve cell innervation within the intact ex vivo tissue explants play important roles in this apparent tissue-level, lipolytic synchronization. High-resolution, quantitative sampling by droplet microfluidics thus permitted unique biological information to be observed, giving an analytical framework poised for future studies of dynamic oscillatory function of adipose and other tissues.
中文翻译：
通过以高时间分辨率的微流体液滴采样揭示了离体脂肪组织外植体中的快速脂解振荡。
我们对脂肪组织生物学的认识已经稳步发展。尽管结构和能量存储功能一直处于最前沿，但脂肪细胞的内分泌作用仅在最近几十年才被揭示。与其他内分泌组织的动态功能有关的大量信息相反，很少有研究集中于动态脂肪组织的功能或为此目的开发的工具。在这里，我们将我们独特的基于液滴的微流控设备应用于原代小鼠附睾白色脂肪组织（eWAT）以及3T3-L1脂肪细胞预分化簇的培养，灌注和样品分泌。通过自动控制，从组织或细胞中以3.5秒的时间分辨率（包括样品和参比液滴）对油状含水液滴（约2.6 nL）进行采样，集成的酶检测技术可实现甘油的实时定量（每滴低至1.9 fmol）。这种高分辨率揭示了原发组织中而不存在于簇状细胞中的，以0.2至2.0 min-1（约30-300 s周期）的频率存在的分泌甘油的先前未报道的振荡。在刺激期间，在基础条件下释放的低水平爆发（〜50 fmol）与较大爆发（〜300 fmol）形成对比。此外，在老年和肥胖小鼠的eWAT中，倍数变化和爆发强度均降低。这些结果与免疫染色和光漂白分析相结合，表明完整的离体组织外植体中的缝隙连接偶联或神经细胞神经支配在这种表观组织水平的脂解同步中起重要作用。高分辨率，
更新日期：2020-04-24
中文翻译：
通过以高时间分辨率的微流体液滴采样揭示了离体脂肪组织外植体中的快速脂解振荡。
我们对脂肪组织生物学的认识已经稳步发展。尽管结构和能量存储功能一直处于最前沿，但脂肪细胞的内分泌作用仅在最近几十年才被揭示。与其他内分泌组织的动态功能有关的大量信息相反，很少有研究集中于动态脂肪组织的功能或为此目的开发的工具。在这里，我们将我们独特的基于液滴的微流控设备应用于原代小鼠附睾白色脂肪组织（eWAT）以及3T3-L1脂肪细胞预分化簇的培养，灌注和样品分泌。通过自动控制，从组织或细胞中以3.5秒的时间分辨率（包括样品和参比液滴）对油状含水液滴（约2.6 nL）进行采样，集成的酶检测技术可实现甘油的实时定量（每滴低至1.9 fmol）。这种高分辨率揭示了原发组织中而不存在于簇状细胞中的，以0.2至2.0 min-1（约30-300 s周期）的频率存在的分泌甘油的先前未报道的振荡。在刺激期间，在基础条件下释放的低水平爆发（〜50 fmol）与较大爆发（〜300 fmol）形成对比。此外，在老年和肥胖小鼠的eWAT中，倍数变化和爆发强度均降低。这些结果与免疫染色和光漂白分析相结合，表明完整的离体组织外植体中的缝隙连接偶联或神经细胞神经支配在这种表观组织水平的脂解同步中起重要作用。高分辨率，
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