Summary
Lipid droplets (LDs) provide an “on demand” source of
fatty acids (FAs) that can be mobilized in response to fluctuations in nutrient
abundance. Surprisingly, the amount of LDs increases during prolonged periods of
nutrient deprivation. Why cells store FAs in LDs during an energy crisis is
unknown. Our data demonstrate that mTORC1-regulated autophagy is necessary and
sufficient for starvation-induced LD biogenesis. The ER-resident diacylglycerol
acyltransferase 1 (DGAT1) selectively channels autophagy-liberated FAs into new,
clustered LDs that are in close proximity to mitochondria and are lipolytically
degraded. However, LDs are not required for FA delivery to mitochondria, but
instead function to prevent acylcarnitine accumulation and lipotoxic
dysregulation of mitochondria. Our data support a model in which LDs provide a
lipid buffering system that sequesters FAs released during the autophagic
degradation of membranous organelles, reducing lipotoxicity. These findings
reveal an unrecognized aspect of the cellular adaptive response to starvation
mediated by LDs.
Imaging of nucleic acids is important for studying cellular processes such as cell division and apoptosis. A noninvasive label-free technique is attractive. Raman spectroscopy provides rich chemical information based on specific vibrational peaks. However, the signal from spontaneous Raman scattering is weak and long integration times are required, which drastically limits the imaging speed when used for microscopy. Coherent Raman scattering techniques, comprising coherent anti-Stokes Raman scattering (CARS) and stimulated Raman scattering (SRS) microscopy, overcome this problem by enhancing the signal level by up to five orders of magnitude. CARS microscopy suffers from a nonresonant background signal, which distorts Raman spectra and limits sensitivity. This makes CARS imaging of weak transitions in spectrally congested regions challenging. This is especially the case in the fingerprint region, where nucleic acids show characteristic peaks. The recently developed SRS microscopy is free from these limitations; excitation spectra are identical to those of spontaneous Raman and sensitivity is close to shot-noise limited. Here we demonstrate the use of SRS imaging in the fingerprint region to map the distribution of nucleic acids in addition to proteins and lipids in single salivary gland cells of Drosophila larvae, and in single mammalian cells. This allows the imaging of DNA condensation associated with cell division and opens up possibilities of imaging such processes in vivo.
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