Title: Dynamic label-free imaging of lipid droplets and their link to fatty acid and pyruvate oxidation in mouse eggs

Citation
Bradley JF, Pope I, Wang Y, et al. (2019). Dynamic label-free imaging of lipid droplets and their link to fatty acid and pyruvate oxidation in mouse eggs. Cardiff University. http://doi.org/10.17035/d.2019.0078592340


Access Rights: Data is provided under a Creative Commons Attribution (CC BY 4.0) licence
Access Method: Click to email a request for this data to opendata@cardiff.ac.uk

Dataset Details
Publisher: Cardiff University
Date (year) of data becoming publicly available: 2019
Data format: txt, tif
DOI: 10.17035/d.2019.0078592340

Description

Mammalian eggs generate most of their ATP by mitochondrial oxidation of pyruvate from the surrounding medium, or from fatty acids which are stored as triacylglycerols within lipid droplets. The balance between pyruvate and fatty acid oxidation in generating ATP is not established. We have combined coherent anti-Stokes Raman scattering (CARS) imaging with deuterium labelling of oleic acid to monitor turnover of fatty acids within lipid droplets of living eggs. We found that loss of labelled oleic acid is promoted by pyruvate removal, but minimised with inhibited β-oxidation. Pyruvate removal also causes a significant dispersion of lipid droplets, while inhibiting β-oxidation causes droplet clustering. Live imaging of luciferase, or FAD autofluorecence from mitochondria, suggest that inhibiting β-oxidation in mouse eggs only leads to a transient decrease in ATP because there are compensatory uptake of pyruvate into mitochondria. Inhibiting pyruvate uptake and then β-oxidation caused similar and successive declines in ATP. Our data suggest that β-oxidation and pyruvate oxidation contribute nearly equally to resting ATP production in mouse eggs and that reorganisation of lipid droplets occurs in response to metabolic demand.

The data sets consists of optical microscopy images and numerical data. Microscope images show eggs (as cross sections in two dimensions or 3D projections), obtained using Differential Interference Contrast microscopy (DIC) and CARS microscopy. Lipid droplets in eggs are specifically visualised in the CARS microscopy images. In some cases CARS images are from eggs that have been labelled with deuterium. Other data sets consists of autofluorescence or luciferase luminescence recordings from eggs using a CCD camera or photon counting imaging camera respectively.agg

Numerical data consist of the following groups:

1) Raman spectrum showing the vibrational resonances of the CD2 bond in deuterated oleic acid un-deuterated oleic acid. The data is in asci text format.

2) CARS and DIC images of eggs plus histograms of the occurrence of the aggregate size (number of lipid droplets per aggregate) in representative eggs. The data set is tif format and as ascii text files.

3) CARS and DIC images of eggs incubated with deuterated oleic acid. The data is in tif format.  

4) CARS and DIC images of eggs with loss of deuterated oleic acid label. The data is in tif format.  

5) DOA content of eggs with altered mitochondrial metabolism. Graph plots the mean DOA lipid volume (x-axis) against the total number of lipid droplets (y-axis).  The distribution of each variable is shown as the average value ± the standard error of the mean (error bars).The data is in asci text format.

6) Recordings of FAD autofluorescence from eggs with Y axis as fluorescence in arbitrary units and the X axis time in seconds. The data set is an ascii text format.

7)  Recordings of luciferase luminescence of ATP from eggs with Y axis as luminescence in arbitrary units and the X axis time in seconds. The data set is an ascii text file.

Research results based upon these data are published at http://doi.org/10.1242/jcs.228999



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Last updated on 2019-05-07 at 14:34