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Biology subjects

Cermak, N.

Publications and source records attributed to Cermak, N..

6 recordsLinked to original sources

GraFT: Graph Filtered Temporal Dictionary Learning for Functional Neural Imaging

Optical imaging of calcium signals in the brain has enabled researchers to observe the activity of hundreds-to-thousands of individual neurons simultaneously. Current methods predominantly focus on matrix factorization and aim at detecting neurons in the imaged field-of-view, and then inferring the corresponding time-traces. The explicit locality constraints on the cell shapes additionally limits the applicability to optical imaging at different scales (i.e., dendritic or widefield data). Here we present a new method that frames the problem of isolating independent fluorescing components as a dictionary learning problem. Specifically, we focus on the time-traces, which are the main quantity used in scientific discovery, and learn the dictionary of time traces with the spatial maps acting as the presence coefficients encoding which pixels the time traces are active in. Furthermore, we present a novel graph filtering model which redefines connectivity between pixels in terms of their shared temporal activity, rather than spatial proximity. This model greatly eases the ability of our method to handle data with complex non-local spatial structure, such as dendritic imaging. We demonstrate important properties of our method, such as robustness to initialization, implicitly inferring number of neurons and simultaneously detecting different neuronal types, on both synthetic data and real data examples. Specifically, we demonstrate applications of our method to calcium imaging both at the dendritic, somatic, and widefield scales.

neuroscience↗

Whole-organism behavioral profiling reveals a role for dopamine in state-dependent motor program coupling in C. elegans

Animal behaviors are commonly organized into long-lasting states that coordinately impact the generation of diverse motor outputs such as feeding, locomotion, and grooming. However, the neural mechanisms that coordinate these diverse motor programs remain poorly understood. Here, we examine how the distinct motor programs of the nematode C. elegans are coupled together across behavioral states. We describe a new imaging platform that permits automated, simultaneous quantification of each of the main C. elegans motor programs over hours or days. Analysis of these whole-organism behavioral profiles shows that the motor programs coordinately change as animals switch behavioral states. Utilizing genetics, optogenetics, and calcium imaging, we identify a new role for dopamine in coupling locomotion and egg-laying together across states. These results provide new insights into how the diverse motor programs throughout an organism are coordinated and suggest that neuromodulators like dopamine can couple motor circuits together in a state-dependent manner.

neuroscience↗

Stimjim: open source hardware for precise electrical stimulation

Electrical stimulation is a simple and powerful tool to perturb and evoke neuronal activity in order to understand the function of neurons and neural circuits. Despite this, devices that can provide precise current or voltage stimulation are expensive and closed-source. Here, we introduce Stimjim, a capable and inexpensive ($200 USD) open-source instrument for electrical stimulation that combines both function generation and electrical isolation. Stimjim provides microsecond temporal resolution with microampere or millivolt scale precision on two electrically isolated output channels. We demonstrate Stimjims utility both in vitro by precisely stimulating brain slices, and in vivo by training mice to perform intracranial self-stimulation (ICSS) for brain stimulation reward. During ICSS, Stimjim enables the experimenter to smoothly tune the strength of reward-seeking behavior by varying either the output frequency or amplitude. We envision Stimjim will enable new kinds of experiments due to its open-source and scalable nature.

neuroscience↗

Modular assembly of polysaccharide-degrading microbial communities in the ocean

Many complex biological systems such as metabolic networks can be divided into functional and organizational subunits, called modules, which provide the flexibility to assemble novel multi-functional hierarchies by a mix and match of simpler components. Here we show that polysaccharide-degrading microbial communities in the ocean can also assemble in a modular fashion. Using synthetic particles made of a variety of polysaccharides commonly found in the ocean, we showed that the particle colonization dynamics of natural bacterioplankton assemblages can be understood as the aggregation of species modules of two main types: a first module type made of narrow niche-range primary degraders, whose dynamics are controlled by particle polysaccharide composition, and a second module type containing broad niche-range, substrate-independent taxa whose dynamics are controlled by interspecific interactions, in particular cross-feeding via organic acids, amino acids and other metabolic byproducts. As a consequence of this modular logic, communities can be predicted to assemble by a sum of substrate-specific primary degrader modules, one for each complex polysaccharide in the particle, connected to a single broad-niche range consumer module. We validate this model by showing that a linear combination of the communities on single-polysaccharide particles accurately predicts community composition on mixed-polysaccharide particles. Our results suggest thus that the assembly of heterotrophic communities that degrade complex organic materials follow simple design principles that can be exploited to engineer heterotrophic microbiomes.

microbiology↗

Linking single-cell measurements of mass, growth rate, and gene expression

We introduce a microfluidic platform that enables single-cell mass and growth rate measurements upstream of single-cell RNA-sequencing (scRNA-seq) to generate paired single-cell biophysical and transcriptional data sets. Biophysical measurements are collected with a serial suspended microchannel resonator platform (sSMR) that utilizes automated fluidic state switching to load individual cells at fixed intervals, achieving a throughput of 120 cells per hour. Each single-cell is subsequently captured downstream for linked molecular analysis using an automated collection system. From linked measurements of a murine leukemia (L1210) and pro-B cell line (FL5.12), we identify gene expression signatures that correlate significantly with cell mass and growth rate. In particular, we find that both cell lines display a cell-cycle signature that correlates with cell mass, with early and late cell-cycle signatures significantly enriched amongst genes with negative and positive correlations with mass, respectively. FL5.12 cells also show a significant correlation between single-cell growth efficiency and a G1-S transition signature, providing additional transcriptional evidence for a phenomenon previously observed through biophysical measurements alone. Importantly, the throughput and speed of our platform allows for the characterization of phenotypes in dynamic cellular systems. As a proof-of-principle, we apply our system to characterize activated murine CD8+ T cells and uncover two unique features of CD8+ T cells as they become proliferative in response to activation: i) the level of coordination between cell cycle gene expression and cell mass increases, and ii) translation-related gene expression increases and shows a correlation with single-cell growth efficiency. Overall, our approach provides a new means of characterizing the transcriptional mechanisms of normal and dysfunctional cellular mass and growth rate regulation across a range of biological contexts.

bioengineering↗

Rapid, inexpensive measurement of synthetic bacterial community composition by Sanger sequencing

Simple synthetic bacterial communities are powerful tools for studying microbial ecology and evolution, as they enable rapid iteration between controlled laboratory experiments and theoretical modeling. However, their utility is hampered by the lack of fast, inexpensive, and accurate methods for quantifying bacterial community composition. For instance, while next-generation amplicon sequencing can be very accurate, high costs (>$30 per sample) and turnaround times (>1 month) limit the nature and pace of experiments. Here, we introduce a new approach for quantifying composition in synthetic bacterial communities based on Sanger sequencing. First, for a given community, we PCR-amplify a universal marker gene (here, the 16S rRNA gene), which yields a mixture of amplicons. Second, we sequence this amplicon mixture in a single Sanger sequencing reaction, which produces a \"mixed\" electropherogram with contributions from each community member. We also sequence each community members marker gene individually to generate \"individual\" electropherograms. Third, we fit the mixed electropherogram as a linear combination of time-warped individual electropherograms, thereby allowing us to estimate the fractional amplicon abundance of each strain within the community. Importantly, our approach accounts for retention-time variability in electrophoretic signals, which is crucial for accurate compositional estimates. Using synthetic communities of marine bacterial isolates, we show that this approach yields accurate and reproducible abundance estimates for two-, four-, and seven-strain bacterial communities. Furthermore, this approach can provide results within one day and costs ~$5 USD per sample. We envision this approach will enable new insights in microbial ecology by increasing the number of samples that can be analyzed and enabling faster iteration between experiments and theory. We have implemented our method in a free and open-source R package called CASEU (\"Compositional Analysis by Sanger Electropherogram Unmixing\"), available at https://bitbucket.org/DattaManoshi/caseu.

bioinformatics↗