Abstract
The overactivation of microglia leads to the excessive release of proinflammatory mediators, which is detrimental to brain health. Docosahexaenoic acid (DHA) has been shown to alleviate neuroinflammation by inhibiting the release of proinflammatory mediators from microglia. Therefore, the uptake of DHA into microglia is essential for reducing neuroinflammation.
Cytoplasmic carrier proteins, fatty acid-binding proteins (FABPs), are involved in DHA trafficking in other cell types. Whether FABPs are expressed and involved in DHA trafficking into microglia remains to be investigated. This study focused on screening whether various FABP isoforms are expressed in microglia, and whether they are involved in the uptake of DHA into microglia. Using immortalised mouse microglia (BV-2) cells, RT-qPCR and western blotting were used to quantitatively determine the mRNA and protein levels of the 10 known FABP isoforms. FABP3,FABP4, and FABP5 were expressed at both the mRNA and protein level in BV-2 cells. A genetic knockdown approached was taken to investigate the involvement of these FABP isoforms in the microglial uptake of DHA-d5, a surrogate of DHA. DHA-d5 uptake in BV-2 cells was quantified using an optimised LC-MS/MS technique. Following 77.5-92.3% (at mRNA level) and 45.4-81.7%(at protein level) knockdown of FABP3, FABP4, and FABP5 at 48 hours, no changes in DHA-d5uptake into microglia was observed at 2 min. This suggested the involvement of other microglial DHA uptake mechanisms, such as the possible involvement of membrane transporters like fatty acid transport proteins. Therefore, further DHA uptake mechanisms present in microglia will need to be further investigated.
Cytoplasmic carrier proteins, fatty acid-binding proteins (FABPs), are involved in DHA trafficking in other cell types. Whether FABPs are expressed and involved in DHA trafficking into microglia remains to be investigated. This study focused on screening whether various FABP isoforms are expressed in microglia, and whether they are involved in the uptake of DHA into microglia. Using immortalised mouse microglia (BV-2) cells, RT-qPCR and western blotting were used to quantitatively determine the mRNA and protein levels of the 10 known FABP isoforms. FABP3,FABP4, and FABP5 were expressed at both the mRNA and protein level in BV-2 cells. A genetic knockdown approached was taken to investigate the involvement of these FABP isoforms in the microglial uptake of DHA-d5, a surrogate of DHA. DHA-d5 uptake in BV-2 cells was quantified using an optimised LC-MS/MS technique. Following 77.5-92.3% (at mRNA level) and 45.4-81.7%(at protein level) knockdown of FABP3, FABP4, and FABP5 at 48 hours, no changes in DHA-d5uptake into microglia was observed at 2 min. This suggested the involvement of other microglial DHA uptake mechanisms, such as the possible involvement of membrane transporters like fatty acid transport proteins. Therefore, further DHA uptake mechanisms present in microglia will need to be further investigated.
| Original language | English |
|---|---|
| Pages | 62-62 |
| Number of pages | 1 |
| Publication status | Published - 2020 |
| Event | Macquarie Neurodegeneration Meeting - Webinar, Sydney, Australia Duration: 28 Oct 2020 → 29 Oct 2020 Conference number: 3rd https://www.mq.edu.au/research/research-centres-groups-and-facilities/healthy-people/centres/macquarie-university-centre-for-motor-neuron-disease-research/conference |
Conference
| Conference | Macquarie Neurodegeneration Meeting |
|---|---|
| Country/Territory | Australia |
| City | Sydney |
| Period | 28/10/20 → 29/10/20 |
| Other | The Macquarie Neurodegeneration Meeting is an annual event hosted by the Centre for Motor Neuron Disease Research, Macquarie University. The aim of this event is for Australian neuroscientists to showcase their research and to stimulate conversation and foster collaboration to develop treatments for diseases including motor neuron disease, Alzheimer’s disease, frontotemporal dementia, Parkinson’s disease and other degenerative brain disorders. |
| Internet address |