Digital tomosynthesis and high resolution computed tomography as clinical tools for vertebral endplate topography measurements: Comparison with microcomputed tomography.

Endplate morphology is understood to play an important role in the mechanical behavior of vertebral bone as well as degenerative processes in spinal tissues; however, the utility of clinical imaging modalities in assessment of the vertebral endplate has been limited. The objective of this study was to evaluate the ability of two clinical imaging modalities (digital tomosynthesis, DTS; high resolution computed tomography, HRCT) to assess endplate topography by correlating the measurements to a microcomputed tomography (µCT) standard. DTS, HRCT, and µCT images of 117 cadaveric thoracolumbar vertebrae (T10-L1; 23 male, 19 female; ages 36-100 years) were segmented, and inferior and superior endplate surface topographical distribution parameters were calculated. Both DTS and HRCT showed statistically significant correlations with µCT approaching a moderate level of correlation at the superior endplate for all measured parameters (R(2)Adj=0.19-0.57), including averages, variability, and higher order statistical moments. Correlation of average depths at the inferior endplate was comparable to the superior case for both DTS and HRCT (R(2)Adj=0.14-0.51), while correlations became weak or nonsignificant for higher moments of the topography distribution. DTS was able to capture variations in the endplate topography to a slightly better extent than HRCT, and taken together with the higher speed and lower radiation cost of DTS than HRCT, DTS appears preferable for endplate measurements.

System A transporter SAT2 mediates replenishment of dendritic glutamate pools controlling retrograde signaling by glutamate.

Glutamate mediates several modes of neurotransmission in the central nervous system including recently discovered retrograde signaling from neuronal dendrites. We have previously identified the system N transporter SN1 as being responsible for glutamine efflux from astroglia and proposed a system A transporter (SAT) in subsequent transport of glutamine into neurons for neurotransmitter regeneration. Here, we demonstrate that SAT2 expression is primarily confined to glutamatergic neurons in many brain regions with SAT2 being predominantly targeted to the somatodendritic compartments in these neurons. SAT2 containing dendrites accumulate high levels of glutamine. Upon electrical stimulation in vivo and depolarization in vitro, glutamine is readily converted to glutamate in activated dendritic subsegments, suggesting that glutamine sustains release of the excitatory neurotransmitter via exocytosis from dendrites. The system A inhibitor MeAIB (alpha-methylamino-iso-butyric acid) reduces neuronal uptake of glutamine with concomitant reduction in intracellular glutamate concentrations, indicating that SAT2-mediated glutamine uptake can be a prerequisite for the formation of glutamate. Furthermore, MeAIB inhibited retrograde signaling from pyramidal cells in layer 2/3 of the neocortex by suppressing inhibitory inputs from fast-spiking interneurons. In summary, we demonstrate that SAT2 maintains a key metabolic glutamine/glutamate balance underpinning retrograde signaling by dendritic release of the neurotransmitter glutamate.

Alpha-synuclein overexpression in PC12 and chromaffin cells impairs catecholamine release by interfering with a late step in exocytosis.

Alpha-synuclein (alpha-syn), a protein implicated in Parkinson's disease pathogenesis, is a presynaptic protein suggested to regulate transmitter release. We explored how alpha-syn overexpression in PC12 and chromaffin cells, which exhibit low endogenous alpha-syn levels relative to neurons, affects catecholamine release. Overexpression of wild-type or A30P mutant alpha-syn in PC12 cell lines inhibited evoked catecholamine release without altering calcium threshold or cooperativity of release. Electron micrographs revealed that vesicular pools were not reduced but that, on the contrary, a marked accumulation of morphologically "docked" vesicles was apparent in the alpha-syn-overexpressing lines. We used amperometric recordings from chromaffin cells derived from mice that overexpress A30P or wild-type (WT) alpha-syn, as well as chromaffin cells from control and alpha-syn null mice, to determine whether the filling of vesicles with the transmitter was altered. The quantal size and shape characteristics of amperometric events were identical for all mouse lines, suggesting that overexpression of WT or mutant alpha-syn did not affect vesicular transmitter accumulation or the kinetics of vesicle fusion. The frequency and number of exocytotic events per stimulus, however, was lower for both WT and A30P alpha-syn-overexpressing cells. The alpha-syn-overexpressing cells exhibited reduced depression of evoked release in response to repeated stimuli, consistent with a smaller population of readily releasable vesicles. We conclude that alpha-syn overexpression inhibits a vesicle "priming" step, after secretory vesicle trafficking to "docking" sites but before calcium-dependent vesicle membrane fusion.