BDNF-TrkB Signaling in Single-Spine Structural Plasticity

Multiple lines of evidence reveal that activation of the tropomyosin related kinase B (TrkB) receptor is a critical molecular mechanism underlying status epilepticus (SE) induced epilepsy development. However, the cellular consequences of such signaling remain unknown. To this point, localization of SE-induced TrkB activation to CA1 apical dendritic spines provides an anatomic clue pointing to Schaffer collateral-CA1 synaptic plasticity as one potential cellular consequence of TrkB activation. Here, we combine two-photon glutamate uncaging with two photon fluorescence lifetime imaging microscopy (2pFLIM) of fluorescence resonance energy transfer (FRET)-based sensors to specifically investigate the roles of TrkB and its canonical ligand brain derived neurotrophic factor (BDNF) in dendritic spine structural plasticity (sLTP) of CA1 pyramidal neurons in cultured hippocampal slices of rodents. To begin, we demonstrate a critical role for post-synaptic TrkB and post-synaptic BDNF in sLTP. Building on these findings, we develop a novel FRET-based sensor for TrkB activation that can report both BDNF and non-BDNF activation in a specific and reversible manner. Using this sensor, we monitor the spatiotemporal dynamics of TrkB activity during single-spine sLTP. In response to glutamate uncaging, we report a rapid (onset less than 1 minute) and sustained (lasting at least 20 minutes) activation of TrkB in the stimulated spine that depends on N-methyl-D-aspartate receptor (NMDAR)-Ca2+/Calmodulin dependent kinase II (CaMKII) signaling as well as post-synaptically synthesized BDNF. Consistent with these findings, we also demonstrate rapid, glutamate uncaging-evoked, time-locked release of BDNF from single dendritic spines using BDNF fused to superecliptic pHluorin (SEP). Finally, to elucidate the molecular mechanisms by which TrkB activation leads to sLTP, we examined the dependence of Rho GTPase activity - known mediators of sLTP - on BDNF-TrkB signaling. Through the use of previously described FRET-based sensors, we find that the activities of ras-related C3 botulinum toxin substrate 1 (Rac1) and cell division control protein 42 (Cdc42) require BDNF-TrkB signaling. Taken together, these findings reveal a spine-autonomous, autocrine signaling mechanism involving NMDAR-CaMKII dependent BDNF release from stimulated dendritic spines leading to TrkB activation and subsequent activation of the downstream molecules Rac1 and Cdc42 in these same spines that proves critical for sLTP. In conclusion, these results highlight structural plasticity as one cellular consequence of CA1 dendritic spine TrkB activation that may potentially contribute to larger, circuit-level changes underlying SE-induced epilepsy.

Full of Hot Air? Three Examinations of Climate Change in the American Political Information Environment

Climate change is thought to be one of the most pressing environmental problems facing humanity. However, due in part to failures in political communication and how the issue has been historically defined in American politics, discussions of climate change remain gridlocked and polarized. In this dissertation, I explore how climate change has been historically constructed as a political issue, how conflicts between climate advocates and skeptics have been communicated, and what effects polarization has had on political communication, particularly on the communication of climate change to skeptical audiences. I use a variety of methodological tools to consider these questions, including evolutionary frame analysis, which uses textual data to show how issues are framed and constructed over time; Kullback-Leibler divergence content analysis, which allows for comparison of advocate and skeptical framing over time; and experimental framing methods to test how audiences react to and process different presentations of climate change. I identify six major portrayals of climate change from 1988 to 2012, but find that no single construction of the issue has dominated the public discourse defining the problem. In addition, the construction of climate change may be associated with changes in public political sentiment, such as greater pessimism about climate action when the electorate becomes more conservative. As the issue of climate change has become more polarized in American politics, one proposed causal pathway for the observed polarization is that advocate and skeptic framing of climate change focuses on different facets of the issue and ignores rival arguments, a practice known as “talking past.” However, I find no evidence of increased talking past in 25 years of popular newsmedia reporting on the issue, suggesting both that talking past has not driven public polarization or that polarization is occurring in venues outside of the mainstream public discourse, such as blogs. To examine how polarization affects political communication on climate change, I test the cognitive processing of a variety of messages and sources that promote action against climate change among Republican individuals. Rather than identifying frames that are powerful enough to overcome polarization, I find that Republicans exhibit telltale signs of motivated skepticism on the issue, that is, they reject framing that runs counter to their party line and political identity. This result suggests that polarization constrains political communication on polarized issues, overshadowing traditional message and source effects of framing and increasing the difficulty communicators experience in reaching skeptical audiences.