lucifer-yellow and Cognition-Disorders

In humans, mutations in the gene encoding ATRX, a chromatin remodeling protein of the sucrose-nonfermenting 2 family, cause several mental retardation disorders, including α-thalassemia X-linked mental retardation syndrome. We generated ATRX mutant mice lacking exon 2 (ATRX(ΔE2) mice), a mutation that mimics exon 2 mutations seen in human patients and associated with milder forms of retardation. ATRX(ΔE2) mice exhibited abnormal dendritic spine formation in the medial prefrontal cortex (mPFC). Consistent with other mouse models of mental retardation, ATRX(ΔE2) mice exhibited longer and thinner dendritic spines compared with wild-type mice without changes in spine number. Interestingly, aberrant increased calcium/calmodulin-dependent protein kinase II (CaMKII) activity was observed in the mPFC of ATRX(ΔE2) mice. Increased CaMKII autophosphorylation and activity were associated with increased phosphorylation of the Rac1-guanine nucleotide exchange factors (GEFs) T-cell lymphoma invasion and metastasis 1 (Tiam1) and kalirin-7, known substrates of CaMKII. We confirmed increased phosphorylation of p21-activated kinases (PAKs) in mPFC extracts. Furthermore, reduced protein expression and activity of protein phosphatase 1 (PP1) was evident in the mPFC of ATRX(ΔE2) mice. In cultured cortical neurons, PP1 inhibition by okadaic acid increased CaMKII-dependent Tiam1 and kalirin-7 phosphorylation. Together, our data strongly suggest that aberrant CaMKII activation likely mediates abnormal spine formation in the mPFC. Such morphological changes plus elevated Rac1-GEF/PAK signaling seen in ATRX(ΔE2) mice may contribute to mental retardation syndromes seen in human patients.

Topics: Adaptation, Ocular; Analysis of Variance; Animals; Animals, Newborn; Astrocytes; Benzylamines; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Cell Count; Cells, Cultured; Cognition Disorders; Conditioning, Classical; Dendritic Spines; Disease Models, Animal; DNA Helicases; Exons; Exploratory Behavior; Fear; Gene Expression Regulation; Green Fluorescent Proteins; Guanine Nucleotide Exchange Factors; Humans; Immunoprecipitation; Isoquinolines; Learning Disabilities; Maze Learning; Mice; Mice, Transgenic; Motor Activity; Mutation; Neurons; Nuclear Proteins; Phosphopyruvate Hydratase; Phosphorylation; Prefrontal Cortex; Protein Kinase Inhibitors; Protein Phosphatase 1; Protein Phosphatase 2; RNA, Messenger; Sulfonamides; T-Lymphoma Invasion and Metastasis-inducing Protein 1; X-linked Nuclear Protein

Alterations in neuronal morphology occur in primate cerebral cortex during normal aging, vary depending on the neuronal type, region and cortical layer, and have been related to memory and cognitive impairment. We analyzed how such changes affect a specific subpopulation of cortical neurons forming long corticocortical projections from the superior temporal cortex to prefrontal area 46. These neurons were identified by retrograde transport in young and old macaque monkeys. Dendritic arbors of retrogradely labeled neurons were visualized in brain slices by intracellular injection of Lucifer Yellow, and reconstructed three-dimensionally using computer-assisted morphometry. Total dendritic length, numbers of segments, numbers of spines, and spine density were analyzed in layer III pyramidal neurons forming the projection considered. Sholl analysis was used to determine potential age-related changes in dendritic complexity. We observed statistically significant age-related decreases in spine numbers and density on both apical and basal dendritic arbors in these projection neurons. On apical dendrites, changes in spine numbers occurred mainly on the proximal dendrites but spine density decreased uniformly among the different branch orders. On basal dendrites, spine numbers and density decreased preferentially on distal branches. Regressive dendritic changes were observed only in one particular portion of the apical dendrites, with the general dendritic morphology and extent otherwise unaffected by aging. In view of the fact that there is no neuronal loss in neocortex and hippocampus in old macaque monkeys, it is possible that the memory and cognitive decline known to occur in these animals is related to rather subtle changes in the morphological and molecular integrity of neurons subserving identifiable neocortical association circuits that play a critical role in cognition.

Topics: Aging; Animals; Cerebral Cortex; Cognition Disorders; Dendrites; Female; Fluorescent Dyes; Isoquinolines; Macaca fascicularis; Macaca mulatta; Male; Memory Disorders; Neural Pathways; Neurons; Prefrontal Cortex; Pyramidal Cells; Temporal Lobe