Haploinsufficiency of the autism-associated Shank3 gene leads to deficits in synaptic function, social interaction, and social communication
Authors: Bozdagi Ozlem , Sakurai Takeshi , Papapetrou Danae , Wang Xiaobin , Dickstein Dara L , Takahashi Nagahide , Kajiwara Yuji , Yang Mu , Katz Adam M , Scattoni Maria Luisa , Harris Mark J , Saxena Roheeni , Silverman Jill L , Crawley Jacqueline N , Zhou Qiang , Hof Patrick R , Buxbaum Joseph D 
Published: January 2011

AbstractBackgroundSHANK3 is a protein in the core of the postsynaptic density (PSD) and has a critical role in recruiting many key functional elements to the PSD and to the synapse, including components of α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionic acid (AMPA), metabotropic glutamate (mGlu) andN-methyl-D-aspartic acid (NMDA) glutamate receptors, as well as cytoskeletal elements. Loss of a functional copy of theSHANK3gene leads to the neurobehavioral manifestations of 22q13 deletion syndrome and/or to autism spectrum disorders. The goal of this study was to examine the effects of haploinsufficiency of full-lengthShank3in mice, focusing on synaptic development, transmission and plasticity, as well as on social behaviors, as a model for understandingSHANK3haploinsufficiency in humans.MethodsWe used mice with a targeted disruption ofShank3in which exons coding for the ankyrin repeat domain were deleted and expression of full-length Shank3 was disrupted. We studied synaptic transmission and plasticity by multiple methods, including patch-clamp whole cell recording, two-photon time-lapse imaging and extracellular recordings of field excitatory postsynaptic potentials. We also studied the density of GluR1-immunoreactive puncta in the CA1 stratum radiatum and carried out assessments of social behaviors.ResultsInShank3heterozygous mice, there was reduced amplitude of miniature excitatory postsynaptic currents from hippocampal CA1 pyramidal neurons and the input-output (I/O) relationship at Schaffer collateral-CA1 synapses in acute hippocampal slices was significantly depressed; both of these findings indicate a reduction in basal neurotransmission. Studies with specific inhibitors demonstrated that the decrease in basal transmission reflected reduced AMPA receptor-mediated transmission. This was further supported by the observation of reduced numbers of GluR1-immunoreactive puncta in the stratum radiatum. Long-term potentiation (LTP), induced either with θ-burst pairing (TBP) or high-frequency stimulation, was impaired inShank3heterozygous mice, with no significant change in long-term depression (LTD). In concordance with the LTP results, persistent expansion of spines was observed in control mice after TBP-induced LTP; however, only transient spine expansion was observed inShank3heterozygous mice. MaleShank3heterozygotes displayed less social sniffing and emitted fewer ultrasonic vocalizations during interactions with estrus female mice, as compared to wild-type littermate controls.ConclusionsWe documented specific deficits in synaptic function and plasticity, along with reduced reciprocal social interactions inShank3heterozygous mice. Our results are consistent with altered synaptic development and function inShank3haploinsufficiency, highlighting the importance of Shank3 in synaptic function and supporting a link between deficits in synapse function and neurodevelopmental disorders. The reduced glutamatergic transmission that we observed in theShank3heterozygous mice represents an interesting therapeutic target inShank3-haploinsufficiency syndromes.