Molecular and Cellular Neurobiology
We are
interested in both the mechanisms of neuronal plasticity and its impairment in
neurodegeneration. One facet of plasticity is the regulation of mRNA
translocation and translation in dendrites. RNAs are not uniformly distributed
in neurons, and a subset of mRNAs that extend into dendrites appear to
position their translation products strategically to implement the
morphological changes associated with activity-related changes in synapses.
Emerging work has attempted to link the translational regulation of these
dendritic mRNAs to synaptic activity. The population of RNAs which segregate
to the dendrite create a specialized locale possibly capable of implementing
activity-related structural changes, including dendritic spine morphogenesis
associated with enduring long-term potentiation. In dendrites, mRNAs are
present as granules. We have observed translocation of RNA granules in neurons
along microtubules and have engineered a nucleic acid-peptide complex capable
of directly visualizing in living neurons the translocation of a 5'
untranslated RNA sequence complexed to green fluoresecent protein.
Although
synaptic activation can induce translation, how activation is coupled to
translation of specific mRNAs is poorly understood. In contrast to local
translation control, the delivery of new mRNAs in granules to the dendrite is
a slower means of altering local protein composition. Using RNA sedimentation
techniques we have isolated and characterized the RNA granule as a
macromolecular control site where specific mRNAs are held in translational
arrest until stimulated. Available projects are directed at identification of
components within these granules, characterization of their interaction with
microtubules, identification of specific proteins that interact with highly
segregated neuronal RNAs, and the direct visualization of RNA transport. RNAs
that sediment with a somewhat lower mass than granules are polysomes. In this
fraction are a remarkable variety of microRNAs potentially capable of
regulating translation locally.
The basis for
experience-dependent modification of neural circuitry involves changes in both
the efficacy of existing synapses and the patterning of new anatomical
connections such as the elaboration of motile filopodial, new synapses and
spines. We cloned a protein called delta-catenin which is shared by both
adherens junctions and synapses, two structures with an intertwined function
and evolutionary history. Spine formation and filopodial elaboration must
involve adhesive changes for neurite outgrowths to penetrate the neuropil and
delimit portions of the newly extended membrane for a synapse of a defined
composition. In the adherens junction, delta-catenin binds to classical
cadherin as a neuronal specific Arm-repeat family member. Among the Arm-repeat
family members, a sub-family that includes delta-catenin and the prototypical
member, p120ctn, is distinguished by the presence of ten Arm-repeats and
binding to the juxtamembrane region of the classical cadherins. Through its
PDZ binding domain, delta-catenin is linked to the PDZ domains of several
synaptic proteins. Delta-catenin is ideally positioned to bridge and
coordinate activity-related changes in the synapse with changes in adhesion of
the post-synaptic membrane.
In parallel to
this work are studies directed at the underlying cellular mechanisms by which
plasticity is lost in the course of neurodegeneration. The
microtubule-associated protein tau is the focus of these studies and the
projects focus principally on the changes that neurons undergo as tau becomes
vulnerable to the formation of aggregates and inclusions. Most recently we
have completed a screen for compounds that are likely to inhibit the
phosphorylation of tau, the likely first step in its transition toward the
formation of intra-cellular aggregates.