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      Acyl-CoA synthetase 2 overexpression enhances fatty acid internalization and neurite outgrowth.

      The Journal of Biological Chemistry
      3T3-L1 Cells, Animals, Arachidonic Acid, metabolism, Cell Differentiation, Cell Line, Transformed, Cell Separation, Coenzyme A Ligases, biosynthesis, physiology, Docosahexaenoic Acids, Fatty Acids, Fatty Acids, Unsaturated, Flow Cytometry, Green Fluorescent Proteins, Luminescent Proteins, Mice, Neurons, Oleic Acid, PC12 Cells, RNA, Messenger, Rats, Retroviridae, genetics, Reverse Transcriptase Polymerase Chain Reaction, Time Factors

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          Abstract

          During neurodevelopment neurons increase phospholipid synthesis to generate additional plasma membrane that makes up the growing neurites. Compared with most cell types, neurons contain a high percentage of the polyunsaturated fatty acids (PUFAs) arachidonic acid (AA) and docosahexaenoic acid (DHA). By utilizing PC12 cell lines as a model neuronal cell line, we examined the internalization rate of AA, DHA, and non-essential oleic acid (OA), as well as their effects on neurite outgrowth. When wild type cells were differentiated, the rate of AA and DHA internalization increased 50% more than the rate of OA internalization. When media were supplemented with AA or DHA, the average neurite length was increased by approximately 40%, but supplementation with the same amount of OA had no effect. We also increased the levels of acyl-CoA synthetase-1 (ACS1) and ACS2 proteins to determine whether they contribute to PUFA internalization or neurite outgrowth. Overexpression of ACS1 increased the rate of OA internalization by 55%, and AA and DHA uptake was increased by 25%, but there was no significant change in neurite outgrowth. In ACS2-overexpressing cells, in contrast, the rate of OA internalization increased by 90%, AA by 115%, and DHA by 70%. The average aggregate neurite length in ACS2-overexpressing cells was increased by approximately 40% when the media were supplemented with PUFAs, but there was no change with OA supplementation. Taken together, these results support the hypotheses that ACSs are rate-limiting for fatty acid internalization and that ACS2 enhances neurite outgrowth by promoting PUFA internalization.

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