The effect of polyunsaturated fatty acids on the homeostasis of yolk lipoprotein in C. elegans examined by CARS and two-photon excitation fluorescence (TPE-F) microscopy

Wei-Wen Chen; Yung-Hsiang Yi; Cheng-Hao Chien; Kuei-Ching Hsiung; Yi-Chun Lin; Tian-Hsiang Ma; Szecheng J. Lo; Ta-Chau Chang

doi:10.1117/12.2210787

9 March 2016 The effect of polyunsaturated fatty acids on the homeostasis of yolk lipoprotein in C. elegans examined by CARS and two-photon excitation fluorescence (TPE-F) microscopy

Wei-Wen Chen, Yung-Hsiang Yi, Cheng-Hao Chien, Kuei-Ching Hsiung, Yi-Chun Lin, Tian-Hsiang Ma, Szecheng J. Lo, Ta-Chau Chang

Author Affiliations +

Proceedings Volume 9716, Optical Methods in Developmental Biology IV; 97160G (2016) https://doi.org/10.1117/12.2210787
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

Yolk lipoprotein constitutes the major source of energy and the materials for synthesizing signaling factors for the development of oocytes and embryos in C. elegans. Polyunsaturated fatty acids (PUFAs) packed in yolk lipoprotein have been recently recognized as critical molecules for fertilization and reproduction.¹ However, the relation between PUFAs and the homeostasis of yolk lipoprotein is not clear. Here we use coherent anti-Stokes Raman scattering (CARS) microscopy and two-photon excitation fluorescence (TPE-F) microscopy to examine the transportation of yolk lipoprotein. We demonstrate that CARS microscopy is a more sensitive method than the traditional Nile Red staining method in probing the abnormal accumulation of yolk lipoprotein in the body cavity of C. elegans. It is found that the accumulation of yolk lipoprotein is a time-dependent process. In addition, a negative correlation (r = -0.955) between reproductive aging and abnormal accumulation of yolk lipoprotein is established. We further examine wild-type, fat-1, and fat-2 worms with or without the expression of GFP-tagged yolk lipoprotein (VIT-2-GFP). Our data reveal that PUFAs have a positive effect on the synthesis and endocytosis of yolk lipoprotein, confirming the model proposed by Edmonds et al.²

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Wei-Wen Chen, Yung-Hsiang Yi, Cheng-Hao Chien, Kuei-Ching Hsiung, Yi-Chun Lin, Tian-Hsiang Ma, Szecheng J. Lo, and Ta-Chau Chang "The effect of polyunsaturated fatty acids on the homeostasis of yolk lipoprotein in C. elegans examined by CARS and two-photon excitation fluorescence (TPE-F) microscopy", Proc. SPIE 9716, Optical Methods in Developmental Biology IV, 97160G (9 March 2016); https://doi.org/10.1117/12.2210787

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KEYWORDS

Microscopy

Green fluorescent protein

Genetics

Proteins

CARS tomography

Intestine

Signal detection

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