Biology subjects

Roth, J. R.

Publications and source records attributed to Roth, J. R..

3 recordsLinked to original sources

bioRxiv2024

A suite of enhancer AAVs and transgenic mouse lines for genetic access to cortical cell types

The mammalian cortex is comprised of cells classified into types according to shared properties. Defining the contribution of each cell type to the processes guided by the cortex is essential for understanding its function in health and disease. We used transcriptomic and epigenomic cortical cell type taxonomies from mouse and human to define marker genes and putative enhancers and created a large toolkit of transgenic lines and enhancer AAVs for selective targeting of cortical cell populations. We report evaluation of fifteen new transgenic driver lines, two new reporter lines, and >800 different enhancer AAVs covering most subclasses of cortical cells. The tools reported here as well as the scaled process of tool creation and modification enable diverse experimental strategies towards understanding mammalian cortex and brain function.

molecular biology↗

bioRxiv2020

Alzheimer's disease risk gene BIN1 induces Tau-dependent network hyperexcitability

Genome-wide association studies identified the BIN1 locus as a leading modulator of genetic risk in Alzheimers disease (AD). One limitation in understanding BIN1s contribution to AD is its unknown function in the brain. AD-associated BIN1 variants are generally noncoding and likely change expression. Here, we determined the effects of increasing expression of the major neuronal isoform of human BIN1 in cultured hippocampal neurons. Higher BIN1 induced network hyperexcitability on multielectrode arrays, increased frequency of synaptic transmission, and elevated calcium transients, indicating that increasing BIN1 drives greater neuronal activity. In exploring the mechanism of these effects on neuronal physiology, we found that BIN1 interacted with L-type voltage-gated calcium channels (LVGCCs) and that BIN1-LVGCCs interactions were modulated by Tau in vitro and in vivo. Finally, Tau reduction prevented BIN1-induced network hyperexcitability. These data shed light on BIN1s neuronal function and suggest that it may contribute to Tau-dependent hyperexcitability in AD.

neuroscience↗

bioRxiv2019

A peptide inhibitor of Tau-SH3 interactions ameliorates amyloid-β toxicity

The microtubule-associated protein Tau is strongly implicated in Alzheimers disease (AD) and aggregates into neurofibrillary tangles in AD. Genetic reduction of Tau is protective in several animal models of AD and cell culture models of amyloid-{beta} (A{beta}) toxicity, making it an exciting therapeutic target for treating AD. A variety of evidence indicates that Taus interactions with Fyn kinase and other SH3 domain-containing proteins, which bind to PxxP motifs in Taus proline-rich domain, may contribute to AD deficits and A{beta} toxicity. Thus, we sought to determine if inhibiting Tau-SH3 interactions ameliorates A{beta} toxicity. We developed a peptide inhibitor of Tau-SH3 interactions and a proximity ligation assay (PLA)-based target engagement assay. Then, we used membrane trafficking and neurite degeneration assays to determine if inhibiting Tau-SH3 interactions ameliorated A{beta} oligomer (A{beta}o)-induced toxicity in primary hippocampal neurons from rats. We verified that Tau reduction ameliorated A{beta}o toxicity in neurons. Using PLA, we identified a peptide inhibitor that reduced Tau-SH3 interactions in HEK-293 cells and primary neurons. This peptide reduced Tau phosphorylation by Fyn without affecting Fyns kinase activity state. In primary neurons, endogenous Tau-Fyn interaction was present primarily in neurites and was reduced by the peptide inhibitor, from which we inferred target engagement. Reducing Tau-SH3 interactions in neurons ameliorated A{beta}o toxicity by multiple outcome measures, namely A{beta}o-induced membrane trafficking abnormalities and neurite degeneration. Our results indicate that Tau-SH3 interactions are critical for A{beta}o toxicity and that inhibiting them is a promising therapeutic target for AD.

neuroscience↗