Project summary
The clinical manifestations of autism spectrum disorder (ASD) and related neurodevelopmental disorders
(NDDs) are thought to be caused by an imbalance of excitatory and inhibitory neuronal activity. We have
discovered that changes in activity cause long-term changes in the intracellular architecture of neurons. The
long-term changes occur through the formation of novel organelles, called Golgi satellites (GSats), that have
many of the functions of the Golgi apparatus (GA) in the soma. Preliminary data demonstrate that activity
changes cause GSats to position at synapses, particularly in dendrites at spine heads. Preliminary data also
show that the ASD- and NDD-related protein Ube3a localizes to GSats, and that this association is increased in
response to neuronal stimulation. At dendritic synaptic sites, GSats are the “missing” organelles needed for
proper glycosylation of locally translated membrane and secreted proteins important for synaptic plasticity. GSat
formation transforms local secretory pathways at postsynaptic sites so that locally translated membrane and
secreted proteins are properly glycosylated. In addition, multiple glycoproteins which are both associated with
ASD and involved in synaptic function are endocytosed into early endosomes and then trafficked into GSats
where glycans can again be processed. Through these functions, GSats can remodel the neuronal surface
glycoproteome and mediate rapid changes in the sialic acid content of synaptic glycoproteins. These processes
can lead to changes in protein function which can then contribute to the altered synaptic function observed in
ASD and NDDs.
In this proposal we will examine how GSats interact with the products of the ASD- and NDD-risk gene UBE3A.
The association of Ube3a with GSats is hypothesized to regulate GSat acidification, which in turn influences the
activity of resident sialyltransfereases and their ability to remodel the neuronal surface glycoproteome and
rapidly alter synaptic protein sialic contents. We will determine how changes in Ube3a expression alters sialic
acid content at synapses and on a set of synaptic and ASD-related glycoproteins. We will also characterize how
changes in Ube3a expression regulate activity-induced modulation of GSat formation and localization in relative
to synapse, how these changes correlate with synaptic plasticity. We have developed a series of techniques,
assays and preparations that will allow us to perform these experiments in both primary neuronal cultures and
in ex vivo mouse hippocampal preparations.