TNiK is required for postsynaptic and nuclear signaling pathways and cognitive function.

Traf2 and NcK interacting kinase (TNiK) contains serine-threonine kinase and scaffold domains and has been implicated in cell proliferation and glutamate receptor regulation in vitro. Here we report its role in vivo using mice carrying a knock-out mutation. TNiK binds protein complexes in the synapse linking it to the NMDA receptor (NMDAR) via AKAP9. NMDAR and metabotropic receptors bidirectionally regulate TNiK phosphorylation and TNiK is required for AMPA expression and synaptic function. TNiK also organizes nuclear complexes and in the absence of TNiK, there was a marked elevation in GSK3ß and phosphorylation levels of its cognate phosphorylation sites on NeuroD1 with alterations in Wnt pathway signaling. We observed impairments in dentate gyrus neurogenesis in TNiK knock-out mice and cognitive testing using the touchscreen apparatus revealed impairments in pattern separation on a test of spatial discrimination. Object-location paired associate learning, which is dependent on glutamatergic signaling, was also impaired. Additionally, TNiK knock-out mice displayed hyperlocomotor behavior that could be rapidly reversed by GSK3ß inhibitors, indicating the potential for pharmacological rescue of a behavioral phenotype. These data establish TNiK as a critical regulator of cognitive functions and suggest it may play a regulatory role in diseases impacting on its interacting proteins and complexes.

Synapse-associated protein 102/dlgh3 couples the NMDA receptor to specific plasticity pathways and learning strategies.

Understanding the mechanisms whereby information encoded within patterns of action potentials is deciphered by neurons is central to cognitive psychology. The multiprotein complexes formed by NMDA receptors linked to synaptic membrane-associated guanylate kinase (MAGUK) proteins including synapse-associated protein 102 (SAP102) and other associated proteins are instrumental in these processes. Although humans with mutations in SAP102 show mental retardation, the physiological and biochemical mechanisms involved are unknown. Using SAP102 knock-out mice, we found specific impairments in synaptic plasticity induced by selective frequencies of stimulation that also required extracellular signal-regulated kinase signaling. This was paralleled by inflexibility and impairment in spatial learning. Improvement in spatial learning performance occurred with extra training despite continued use of a suboptimal search strategy, and, in a separate nonspatial task, the mutants again deployed a different strategy. Double-mutant analysis of postsynaptic density-95 and SAP102 mutants indicate overlapping and specific functions of the two MAGUKs. These in vivo data support the model that specific MAGUK proteins couple the NMDA receptor to distinct downstream signaling pathways. This provides a mechanism for discriminating patterns of synaptic activity that lead to long-lasting changes in synaptic strength as well as distinct aspects of cognition in the mammalian nervous system.

Maturation but not survival of dopaminergic nigrostriatal neurons is affected in developing and aging BDNF-deficient mice.

Brain-derived neurotrophic factor (BDNF) promotes survival of injured dopaminergic nigrostriatal neurons of the adult rodent substantia nigra pars compacta, as well their development in vitro. BDNF deficiency may play a role in Parkinson's disease, as the surviving dopaminergic nigrostriatal neurons have reduced levels of BDNF, and a BDNF gene polymorphism is present in a subpopulation of patients. Here, we investigated whether a lack of BDNF in early postnatal BDNF-/- mice or a chronic 50% reduction in BDNF levels in aging BDNF+/- mice would affect the survival of the dopaminergic nigrostriatal neurons. In general terms, BDNF-/- and BDNF+/- mice had morphologically and quantitatively normal nigrostriatal neurons at any time between postnatal day 14 (P14) and 18 months, when compared to their wild-type littermates. BDNF-/- mice (P14 and P21 only) had fewer dopaminergic dendrites in the substantia nigra, suggesting that BDNF plays a role in phenotypic maturation, but not in neuronal birth or survival. BDNF-/- mice also had aberrant tyrosine hydroxylase (TH) positive cell bodies in the pars reticulata. During adulthood and aging, BDNF+/- mice performed equally well as their wild-type littermates in tests of motor coordination, and both showed aging-related decreases in the size of the dopaminergic neurons as well as in motor coordination. These results suggest that chronic deficits in BDNF alone do not affect survival or function of dopaminergic nigrostriatal neurons during aging or potentially even in Parkinson's disease.

Evolution of GluN2A/B cytoplasmic domains diversified vertebrate synaptic plasticity and behavior.

Two genome duplications early in the vertebrate lineage expanded gene families, including GluN2 subunits of the NMDA receptor. Diversification between the four mammalian GluN2 proteins occurred primarily at their intracellular C-terminal domains (CTDs). To identify shared ancestral functions and diversified subunit-specific functions, we exchanged the exons encoding the GluN2A (also known as Grin2a) and GluN2B (also known as Grin2b) CTDs in two knock-in mice and analyzed the mice's biochemistry, synaptic physiology, and multiple learned and innate behaviors. The eight behaviors were genetically separated into four groups, including one group comprising three types of learning linked to conserved GluN2A/B regions. In contrast, the remaining five behaviors exhibited subunit-specific regulation. GluN2A/B CTD diversification conferred differential binding to cytoplasmic MAGUK proteins and differential forms of long-term potentiation. These data indicate that vertebrate behavior and synaptic signaling acquired increased complexity from the duplication and diversification of ancestral GluN2 genes.