Dysregulation of brown adipocyte metabolism have potential consequences for development of metabolic disease. This raises interest in understanding the individual cellular components and their role in brown adipocyte activation in response to thermogenic activation. Long non-coding RNAs (lncRNAs) have been increasingly recognized as potent regulators of adipose tissue function. We have previously reported that LINC00473, a novel human specific lncRNA, contributes to increased metabolic activity by translocating from the nucleus to connect mitochondria and lipid droplets in human brown adipocytes (1). However, localization of most lncRNAs in different cellular compartments and how they control organelle activity is to date unexplored.
Here we use smRNA FISH and a high-content imaging screening of highly expressed lncRNAs in human brown and white adipocytes, to screen for novel lncRNAs regulating adipose metabolism in response to norepinephrine stimulation. This approach allows us to trace lncRNA localisation and interaction with specific organelles and proteins. Furthermore, we are able to explore translocations of individual RNAs within single cells and thus uncover possible regulatory mechanisms specific to various cell subpopulations. Interestingly, we have identified several lncRNAs that changed cellular location upon adrenergic activation of brown and white adipocytes some of which changed specifically in cellular subtypes. To gain further insight in which RNA sequences that determines subcellular location, we are now combining the identification of lncRNA location with full length Iso-sequencing.