Search

Tens of thousands of chemicals with poorly understood biological properties are released into the environment each day. High-throughput screening (HTS) is potentially a more efficient and cost-effective alternative to traditional toxicity tests. Using HTS, one can profile chemicals for potential adverse effects and prioritize a manageable number for more in-depth testing. Importantly, it can provide clues to mechanism of toxicity. The Tox21 program has generated >50 million quantitative high-throughput screening (qHTS) data points. A library of several thousands of compounds, including environmental chemicals and drugs, is screened against a panel of nuclear receptor and stress response pathway assays. The National Center for Advancing Translational Sciences (NCATS) has organized an International data challenge in order to crowd-source data and build predictive toxicity models. This Challenge asks a crowd of researchers to use these data to elucidate the extent to which the interference of biochemical and cellular pathways by compounds can be inferred from chemical structure data. The data generated against the Tox21 library served as the training set for this modeling Challenge. The competition attracted participants from 18 different countries to develop computational models aimed at better predicting chemical toxicity. The winning models from nearly 400 model submissions all achieved >80% accuracy. Several models exceeded 90% accuracy, which was measured by area under the receiver operating characteristic curve (AUC-ROC). Combining the winning models with the knowledge already gained from Tox21 screening data are expected to improve the community’s ability to prioritize novel chemicals with respect to potential human health concern.

It has been previously reported that N-terminus of mutant huntingtin (product of the 1st exon) is sufficient to cause a Huntington’s disease (HD) pathological phenotype. In view of recent data suggesting that improper regulation of store-operated calcium (SOC) channels is involved in neurodegenerative processes, we investigated influence of expression of the mutant huntingtin N-terminal fragment (Htt138Q-1exon) on SOC entry (SOCE) in mouse neuroblastoma cells (Neuro-2a) and in primary culture of medium spiny neurons (MSNs) isolated from mice. The results show that SOCE in these cells is enhanced upon lentiviral expression of the Htt138Q-1exon. Moreover we demonstrated that RNAi-mediated knockdown of TRPC1, Orai1 or STIM1 proteins leads to dramatic reduction of abnormal SOCE in both Neuro-2a and MSNs, expressing Htt138Q-1exon. Thus we concluded that abnormal SOCE in these cells is maintained by both TRPC1- and Orai1-containing channels and required STIM1 for its activation. Furthermore, EVP4593 compound previously tested as a potential anti-HD drug in a Drosophila screening system has proved to be capable of reducing SOCE to the normal level in MSNs expressing the Htt138Q-1exon.

To any model of brain function, the variability of neuronal spike firing is a problem that needs to be taken into account. Whereas the synaptic integration can be described in terms of the original Hodgkin-Huxley (H-H) formulations of conductance-based electrical signaling, the transformation of the resulting membrane potential into patterns of spike output is subjected to stochasticity that may not be captured with standard single neuron H-H models. The dynamics of the spike output is dependent on the normal background synaptic noise present in vivo, but the neuronal spike firing variability in vivo is not well studied. In the present study, we made long-term whole cell patch clamp recordings of stationary spike firing states across a range of membrane potentials from a variety of subcortical neurons in the non-anesthetized, decerebrated state in vivo. Based on the data, we formulated a simple, phenomenological model of the properties of the spike generation in each neuron that accurately captured the stationary spike firing statistics across all membrane potentials. The model consists of a parametric relationship between the mean and standard deviation of the inter-spike intervals, where the parameter is linearly related to the injected current over the membrane. This enabled it to generate accurate approximations of spike firing also under inhomogeneous conditions with input that varies over time. The parameters describing the spike firing statistics for different neuron types overlapped extensively, suggesting that the spike generation had similar properties across neurons.

Spinocerebellar systems are likely to be crucial for cerebellar hallmark functions such as coordination. However, in terms of cerebellar functional analyses, these are perhaps among the least explored systems. The aim of the present study is to achieve activation of a single component of the spinocerebellar systems and to explore to what extent it can influence the spike output of granule cells, Golgi cells, molecular layer interneurons (stellate and basket cells) and Purkinje cells. For this purpose, we took advantage of a unique arrangement discovered in neuroanatomical studies, in which the spinal border cell component of the ventral spinocerebellar system was found to be the only the only spinocerebellar tract which ascends in the contralateral lateral funiculus and have terminations in sublobulus C1 of the paramedian lobule in the posterior cerebellum. Using electrical stimulation of this tract, we find a subset of the cerebellar cortical neurons in this region to be moderately or powerfully activated. For example, some of our granule cells displayed high intensity responses whereas the majority of the granule cells displayed no response at all. The finding that more than half of the Purkinje cells were activated by stimulation of the spinal border cell tract indicated that this system is capable of directly influencing cerebellar cortical output. The implications of these findings for the view of the integrative functions of the cerebellar cortex are discussed.