3in1 Activator Shank

3in1 Activator Shank

the results presented above are the first to show that knockdown of both shank2 and shank3 reduces levels of full-length and cleaved nrf-2. to further confirm our data, we transfected hippocampal neurons with a construct expressing a dominant-negative form of nrf-2. transfected neurons were challenged with a tnf/il-1-cytotoxic stimulus. as expected, wild-type nrf-2 protected against apoptosis induced by tnf/il-1 and the protection was reduced in neurons expressing a dominant-negative nrf-2 mutant. in contrast, tnf/il-1-induced neuronal apoptosis was not attenuated by a dominant-negative form of nrf-1. similar to the effects of shrnas, overexpression of dominant-negative nrf-2 completely abolished the protection against apoptosis induced by shank2 or shank3 overexpression, whereas overexpression of a dominant-negative nrf-1 mutant had no effect on neuronal survival (fig. 5). these data demonstrate that both shank2 and shank3 protect against tnf/il-1-induced neuronal death and that the protective effect requires endogenous nrf-2.

to determine the role of the endogenous nrf-2 in affecting tnf-induced neuronal apoptosis, we tested whether pharmacological activation of nrf-2 could protect against tnf/il-1-induced neurotoxicity. we found that treatment of hippocampal neurons with the nrf-2 activator tbhq attenuated tnf/il-1-induced neuronal apoptosis in a time-dependent fashion. the protection was complete within 24 h of treatment. the number of dopaminergic neurons in various conditions was expressed relative to the normal fv condition.

development of appropriate therapeutic modalities to treat the underlying molecular causes of autism is only possible through extensive investigation of the mechanistic underpinnings of asd and attendant co-morbid conditions. therefore, we established experimental paradigms in model organisms to investigate the aetiological perturbations that have been associated with autism. we developed and characterized precise transgenic mouse models for asd that harbour murine shank3 mutations (l68p, r12c) mimicking reported patient-derived mutations, as well as a novel, risk-associated mutation in the disc1 coding region, which leads to partial loss of protein expression (p533l). here, we describe a characterization of the phenotypes that emerged in these pre-clinical models of asd, including assessment of aberrant social behaviour, as well as endophenotypic assessment of social cognition and sensorimotor function in mice subjected to measures of context-dependent fear conditioning and prepulse inhibition, respectively. moreover, we provide an assessment of the utility of our novel mouse models as systems for furthering the understanding of the nature of the heterogeneous asds, and in particular, their conserved genetic underpinnings.

to determine whether or not nrf-1 is involved in the generation of reactive oxygen species (ros) following cell stimulation with tnf/il-1 in primary fibroblasts, we performed a ros assay. control lung fibroblasts were first stimulated with tnf/il-1 for 6 h, then immediately subjected to tem. panel a: ros levels visualized as tem deposits. isolated tem deposits were counted in the presence or absence of 30 nm nqdi-1, a known selective inhibitor of nrf-1. unstimulated cells have 3.36±.01 (sd) tem deposits (indicated by the horizontal line) compared to cells stimulated with tnf/il-1 (5.46±.06) (indicated by a bar). b: number of tem deposits in the presence or absence of nqdi-1. * denotes significant difference (p
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