title: A model for sealing plasmalemmal damage in neurons and other eukaryotic cells.
abstract: Plasmalemmal repair is necessary for survival of damaged eukaryotic cells. Ca(2+) influx through plasmalemmal disruptions activates calpain, vesicle accumulation at lesion sites, and membrane fusion proteins; Ca(2+) influx also initiates competing apoptotic pathways. Using the formation of a dye barrier (seal) to assess plasmalemmal repair, we now report that B104 hippocampal cells with neurites transected nearer (<50 μm) to the soma seal at a lower frequency and slower rate compared to cells with neurites transected farther (>50 μm) from the soma. Analogs of cAMP, including protein kinase A (PKA)-specific and Epac-specific cAMP, each increase the frequency and rate of sealing and can even initiate sealing in the absence of Ca(2+) influx at both transection distances. Furthermore, Epac activates a cAMP-dependent, PKA-independent, pathway involved in plasmalemmal sealing. The frequency and rate of plasmalemmal sealing are decreased by a small molecule inhibitor of PKA targeted to its catalytic subunit (KT5720), a peptide inhibitor targeted to its regulatory subunits (PKI), an inhibitor of a novel PKC (an nPKCη pseudosubstrate fragment), and an antioxidant (melatonin). Given these and other data, we propose a model for redundant parallel pathways of Ca(2+)-dependent plasmalemmal sealing of injured neurons mediated in part by nPKCs, cytosolic oxidation, and cAMP activation of PKA and Epac. We also propose that the evolutionary origin of these pathways and substances was to repair plasmalemmal damage in eukaryotic cells. Greater understanding of vesicle interactions, proteins, and pathways involved in plasmalemmal sealing should suggest novel neuroprotective treatments for traumatic nerve injuries and neurodegenerative disorders.
authors.affiliation: Institute for Cellular and Molecular Biology, University of Texas, Austin, TX 78712, USA.
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Spaeth
authors.forename: Elaine A
authors.initials: EA
authors.lastname: Boydston
authors.forename: Lauren R
authors.initials: LR
authors.lastname: Figard
authors.forename: Aleksej
authors.initials: A
authors.lastname: Zuzek
authors.forename: George D
authors.initials: GD
authors.lastname: Bittner
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000502:physiology*; D005057:Eukaryotic Cells / Q000473:pathology / Q000502:physiology*; D006624:Hippocampus / Q000166:cytology / Q000473:pathology / Q000502:physiology; D008959:Models, Neurological; D009474:Neurons / Q000473:pathology / Q000502:physiology*; D051381:Rats