Luminance Information Is Required for the Accurate Estimation of Contrast in Rapidly Changing Visual Contexts.

Summary Visual perception scales with changes in the visual stimulus, or contrast, irrespective of background illumination. However, visual perception is challenged when adaptation is not fast enough to deal with sudden declines in overall illumination, for example, when gaze follows a moving object from bright sunlight into a shaded area. Here, we show that the visual system of the fly employs a solution by propagating a corrective luminance-sensitive signal. We use in vivo 2-photon imaging and behavioral analyses to demonstrate that distinct OFF-pathway inputs encode contrast and luminance. Predictions of contrast-sensitive neuronal responses show that contrast information alone cannot ex…

TAF-ChIP: An ultra-low input approach for genome wide chromatin immunoprecipitation assay

Chromatin immunoprecipitation (ChIP) followed by next generation sequencing is an invaluable and powerful technique to understand transcriptional regulation. However, ChIP is currently limited by the requirement of large amount of starting material. This renders studying rare cell populations very challenging, or even impossible. Here, we present a tagmentation-assisted fragmentation ChIP (TAF-ChIP) and sequencing method to generate high-quality datasets from low cell numbers. The method relies on Tn5 transposon activity to fragment the chromatin that is immunoprecipitated, thus circumventing the need for sonication or MNAse digestion to fragment. Furthermore, Tn5 adds the sequencing adapto…

TAF-ChIP: an ultra-low input approach for genome-wide chromatin immunoprecipitation assay

The authors present a novel method for obtaining chromatin profiles from low cell numbers without prior nuclei isolation. The method is successfully implemented in generating epigenetic profile from 100 cells with high signal-to-noise ratio.

An optimal population code for global motion estimation in local direction-selective cells

AbstractNervous systems allocate computational resources to match stimulus statistics. However, the physical information that needs to be processed depends on the animal’s own behavior. For example, visual motion patterns induced by self-motion provide essential information for navigation. How behavioral constraints affect neural processing is not known. Here we show that, at the population level, local direction-selective T4/T5 neurons in Drosophila represent optic flow fields generated by self-motion, reminiscent to a population code in retinal ganglion cells in vertebrates. Whereas in vertebrates four different cell types encode different optic flow fields, the four uniformly tuned T4/T5…

The physiological basis for the computation of direction selectivity in theDrosophilaOFF pathway

AbstractInDrosophila, direction-selective neurons implement a mechanism of motion computation similar to cortical neurons, using contrast-opponent receptive fields with ON and OFF subunits. It is not clear how the presynaptic circuitry of direction-selective neurons in the OFF pathway supports this computation, because all major inputs are OFF-rectified neurons. Here, we reveal the biological substrate for motion computation in the OFF pathway. Three interneurons, Tm2, Tm9 and CT1, also provide information about ON stimuli to the OFF direction-selective neuron T5 across its receptive field, supporting a contrast-opponent receptive field organization. Consistent with its prominent role in mo…

Author response: ON selectivity in the Drosophila visual system is a multisynaptic process involving both glutamatergic and GABAergic inhibition

First-order visual interneurons distribute distinct contrast and luminance information across ON and OFF pathways to achieve stable behavior

The accurate processing of contrast is the basis for all visually guided behaviors. Visual scenes with rapidly changing illumination challenge contrast computation because photoreceptor adaptation is not fast enough to compensate for such changes. Yet, human perception of contrast is stable even when the visual environment is quickly changing, suggesting rapid post receptor luminance gain control. Similarly, in the fruit fly Drosophila, such gain control leads to luminance invariant behavior for moving OFF stimuli. Here, we show that behavioral responses to moving ON stimuli also utilize a luminance gain, and that ON-motion guided behavior depends on inputs from three first-order interneuro…

m6A RNA methylation regulates promoter proximal pausing of RNA Polymerase II

AbstractRNA Polymerase II (RNAP II) pausing is essential to precisely control gene expression and is critical for development of metazoans. Here, we show that the m6A RNA modification regulates promoter-proximal RNAP II pausing. The m6A methyltransferase complex (MTC), with the nuclear reader Ythdc1, are recruited to gene promoters. Depleting the m6A MTC leads to a decrease in RNAP II pause release and in Ser2P occupancy on the gene body, and affects nascent RNA transcription. Tethering Mettl3 to a heterologous gene promoter is sufficient to increase RNAP II pause release, an effect that relies on its m6A catalytic domain. Collectively, our data reveal an important link between RNAP II paus…

(A,B) In vivo GCaMP6f signals recorded in layers M1, M5 and M9/10 of Mi1 (A) and Tm3 (B) neurons, before (blue, green) and after (gray, red) application of 0 (sham), 1, 5 or 100 µM PTX. (C,D) Bar plot showing the quantification of the ON step in (A,B). Sample sizes: sham, n = 5 (89); 1 µM, n = 5 (68); 5 µM, n = 5 (64); 100 µM, n = 5 (89). All traces show mean ± SEM. All sample sizes are given as number of flies (number of cells). *: p<0.05, **: p<0.01, ***: p<0.001, tested with a one-way ANOVA and a post-hoc unpaired t-test with Bonferroni-Holm correction for multiple comparisons.

Sensory systems sequentially extract increasingly complex features. ON and OFF pathways, for example, encode increases or decreases of a stimulus from a common input. This ON/OFF pathway split is thought to occur at individual synaptic connections through a sign-inverting synapse in one of the pathways. Here, we show that ON selectivity is a multisynaptic process in the Drosophila visual system. A pharmacogenetics approach demonstrates that both glutamatergic inhibition through GluClα and GABAergic inhibition through Rdl mediate ON responses. Although neurons postsynaptic to the glutamatergic ON pathway input L1 lose all responses in GluClα mutants, they are resistant to a cell-type-specifi…

NECAB2 participates in an endosomal pathway of mitochondrial stress response at striatal synapses

Synaptic signaling depends on ATP generated by mitochondria. Due to extensive connectivity, the striatum is especially vulnerable to mitochondrial dysfunction and thus requires efficient mitochondrial quality control. We found that the neuronal calcium-binding protein NECAB2 ensures synaptic function in the striatum by increasing mitochondrial efficiency. NECAB2 associates with early endosomes and mitochondria at striatal synapses. Loss of NECAB2 dysregulates proteins of the endosomal ESCRT machinery and oxidative phosphorylation. Mitochondria from NECAB2-deficient mice are more abundant but less efficient. These mitochondria exhibit increased respiration and superoxide production but produ…

The physiological basis for contrast opponency in motion computation in Drosophila

This dataset contains traces (dF/F0) from in vivo two-photon calcium imaging from Tm1, tm2, Tm4, Tm9, CT1, and T5 neurons from responses to ONOFF fullfield flashes, ON and OFF bars, and moving sinewaves.

First-order visual interneurons distribute distinct contrast and luminance information across ON and OFF pathways to achieve stable behavior

Source data of the paper Ketkar, G&uuml;r, Molina-Obando et al. 2022, eLife. We analyzed the behavioral contribution and physiological response properties of first order interneurons L1, L2 and L3 in the Drosophila melanogaster visual system. Data are sorted by figures and comprise either behavioral measurements of flies walking on an air-cushioned ball while being shown visual stimuli, or in vivo two photon microscopy recordings of L1-L3 calcium responses. Please find all relevant information to use the data in the README file. The code to analyze the data, either written in Matlab or Python, is found at https://github.com/silieslab/Ketkar-Gur-MolinaObando-etal2022