Neuronal models for studying lipid metabolism and transport.
New methods have been developed for studying lipid metabolism and transport in primary cultures of neurons. Sympathetic neurons from rats and mice, as well as retinal ganglion neurons from rats, can be cultured in three-compartmented culture dishes in which the cell bodies reside in a compartment separate from that housing the distal axons. In addition, the three compartments contain completely independent fluid environments. Consequently, these neuronal cultures represent an excellent model for studying the intra-neuronal transport of lipids and proteins between cell bodies and distal axons. In addition, compartmented neuron cultures are particularly appropriate for investigating factors that regulate axonal growth and neuronal survival. The application of the compartmented culture model for use with murine neurons has opened up many new possibilities for studying lipid metabolism in neurons derived from genetically modified mice. Examples are given in which compartmented cultures of primary neurons have been used in studies on (i) lipid analysis of distal axons and cell bodies/proximal axons, (ii) immunoblotting of neuronal proteins involved in lipid metabolism, (iii) the compartmentalization of lipid metabolism, (iv) the role of lipids in axonal growth and survival, and (v) intracellular lipid transport.
Karten
Barbara
B
Group on Molecular and Cell Biology of Lipids, University of Alberta, Edmonton, Alta., Canada.
Hayashi
Hideki
H
Campenot
Robert B
RB
Vance
Dennis E
DE
Vance
Jean E
JE
D000818:Animals; D017209:Apoptosis; D001369:Axons / Q000378:metabolism; D001692:Biological Transport; D001695:Biology / Q000379:methods*; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism; D003470:Culture Media / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D005724:Ganglia / Q000378:metabolism; D015151:Immunoblotting; D047908:Intracellular Signaling Peptides and Proteins; D050356:Lipid Metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000378:metabolism*; D000077296:Niemann-Pick C1 Protein; D010743:Phospholipids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D012160:Retina / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors
D002352:Carrier Proteins; D003470:Culture Media; D018076:DNA, Complementary; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; C106881:NPC1 protein, human; D000077296:Niemann-Pick C1 Protein; D010743:Phospholipids; D011994:Recombinant Proteins; D002784:Cholesterol