Functional significance of neuronal activity-dependent transcriptional regulation in the nervous system

The ability of extrinsic environmental cues to modify the nervous system is critical both for the appropriate maturation of the nervous system, as well as for important adaptive functions of the mature brain, such as learning and memory. The discovery that, in response to sensory experience, neurotransmitter release at synapses and subsequent calcium influx into postsynaptic neurons lead to the synthesis of new gene products suggested a compelling mechanism by which long-lasting, use-dependent changes occur in the nervous system. Despite considerable progress in our understanding of the program of neuronal activity-regulated gene expression, direct evidence that the activity-dependent component of transcription per se is specifically important for nervous system development or function has been elusive. The first part of this thesis addresses this question through the development of a mutant mouse model in which the activity-dependent component of Bdnf expression is specifically disrupted. We find that mutation of the CaRE3/CRE (CREm) at endogenous Bdnf promoter IV by gene targeting results in an animal in which the neuronal activity-dependent component of Bdnf transcription in the cortex is selectively disrupted. CREm knock-in mice exhibit a reduction in the number of inhibitory synapses formed by cortical neurons in culture, a reduction in spontaneous inhibitory quantal transmission measured in acute brain slices, and a reduction in the level of inhibitory presynaptic markers in the cortex. These results indicate a specific requirement for activity-dependent Bdnf expression in the development of inhibition in the cortex and demonstrate that the activation of gene expression in response to experience-driven neuronal activity has important biological consequences in the nervous system. The second part of my thesis investigates the functional significance of the calcium-dependent regulation of MeCP2, a transcriptional regulator that has been implicated in the activity-dependent expression of Bdnf, and the protein that is mutated in the neurodevelopmental disorder Rett syndrome. We find that MeCP2 becomes phosphorylated at a specific amino acid residue, Serine 421 (S421), selectively in the nervous system in response to neuronal activity via a CaMKII-dependent mechanism. Mutation of MeCP2 at S421 disrupts the function of MeCP2 in regulating dendritic growth, spine morphogenesis, and activity-dependent Bdnf transcription in an in vitro over-expression model of RTT. These findings suggest that, by triggering MeCP2 phosphorylation, neuronal activity regulates a program of gene expression that mediates neuronal connectivity in the nervous system. The disruption of this process in individuals with mutations in MeCP2 may underlie the neural-specific pathology of RTT. Together, these studies demonstrate how an understanding of the molecular mechanisms by which neuronal activity regulates gene transcription allows one to specifically isolate and examine the significance of the neuronal activity-dependent component of the process under study.