Regulated interactions of the norepineprhine transporter by the actin and microtubule cytoskeletons.
One role of the actin cytoskeleton is to maintain the structural morphology and activity of the pre-synaptic terminal. We sought to determine if the actin cytoskeleton plays a role in regulating interactions between the norepinephrine transporter (NET) and alpha-Synuclein (alpha-Syn), two proteins expressed in the pre-synaptic terminal. In cells transfected with either 0.5 microg/mL or 3 microg/mL of alpha-Syn and 1 microg/mL of NET DNA, treatment with cytochalasin D, an actin depolymerizing agent, caused a dose-dependent decrease and increase, respectively, in [3H]-NE uptake. Protein interactions between NET, beta-actin, and alpha-Syn were modified, along with levels of surface transporters. Treatment of primary brainstem neurons and frontal cortex synaptosomes with cytochalasin D caused a 115% and 28% increase, respectively, in NET activity. Depolymerization of both actin and microtubules did not alter NET activity in cells with 0.5 microg/mL alpha-Syn, but caused an increase in [3H]-NE uptake in cells transfected with 3 microg/mL of alpha-Syn and primary neurons. This is the first direct demonstration of NET activity being regulated via actin and modulated by interactions with alpha-Syn.
Jeannotte
Alexis M
AM
Interdisciplinary Program in Neuroscience, Department of Biochemistry and Molecular and Cell Biology, Georgetown University, Washington, DC 20007, USA.
Sidhu
Anita
A
D000199:Actins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D008870:Microtubules / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D009474:Neurons / Q000378:metabolism; D050484:Norepinephrine Plasma Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism
D000199:Actins; D050484:Norepinephrine Plasma Membrane Transport Proteins; D051844:alpha-Synuclein