@phdthesis{12531, abstract = {All visual experiences of the vertebrates begin with light being converted into electrical signals by the eye retina. Retinal ganglion cells (RGCs) are the neurons of the innermost layer of the mammal retina, and they transmit visual information to the rest of the brain. It has been shown that RGCs vary in their morphology and genetic profiles, moreover they can be unambiguously grouped into subtypes that share the same morphological and/or molecular properties. However, in terms of RGCs function, it remains unclear how many distinct types there are and what response properties their typology relies on. Even given the recent studies that successfully classified RGCs in a patch of the retina [1] and in scotopic conditions [2], the question remains whether the found subtypes persist across the entire retina. In this work, using a novel imaging method, we show that, when sampled from a large portion of the retina, RGCs can not be clearly divided into functional subtypes. We found that in photopic conditions, which implies more prominent natural scene statistic differences across the visual field, response properties can be exhibited by cells differently depending on their location in the retina, which leads to formation of a gradient of features rather than distinct classes. This finding suggests that RGCs follow a global organization across the visual field of the animal, adapting each RGC subtype to the requirements imposed by the natural scene statistics.}, author = {Kirillova, Kseniia}, issn = {2791-4585}, pages = {46}, publisher = {Institute of Science and Technology Austria}, title = {{Panoramic functional gradients across the mouse retina}}, doi = {10.15479/at:ista:12531}, year = {2023}, } @article{12226, abstract = {Background: Biases of DNA repair can shape the nucleotide landscape of genomes at evolutionary timescales. The molecular mechanisms of those biases are still poorly understood because it is difficult to isolate the contributions of DNA repair from those of DNA damage. Results: Here, we develop a genome-wide assay whereby the same DNA lesion is repaired in different genomic contexts. We insert thousands of barcoded transposons carrying a reporter of DNA mismatch repair in the genome of mouse embryonic stem cells. Upon inducing a double-strand break between tandem repeats, a mismatch is generated if the break is repaired through single-strand annealing. The resolution of the mismatch showed a 60–80% bias in favor of the strand with the longest 3′ flap. The location of the lesion in the genome and the type of mismatch had little influence on the bias. Instead, we observe a complete reversal of the bias when the longest 3′ flap is moved to the opposite strand by changing the position of the double-strand break in the reporter. Conclusions: These results suggest that the processing of the double-strand break has a major influence on the repair of mismatches during single-strand annealing.}, author = {Pokusaeva, Victoria and Diez, Aránzazu Rosado and Espinar, Lorena and Pérez, Albert Torelló and Filion, Guillaume J.}, issn = {1474-760X}, journal = {Genome Biology}, publisher = {Springer Nature}, title = {{Strand asymmetry influences mismatch resolution during single-strand annealing}}, doi = {10.1186/s13059-022-02665-3}, volume = {23}, year = {2022}, } @article{12288, abstract = {To understand the function of neuronal circuits, it is crucial to disentangle the connectivity patterns within the network. However, most tools currently used to explore connectivity have low throughput, low selectivity, or limited accessibility. Here, we report the development of an improved packaging system for the production of the highly neurotropic RVdGenvA-CVS-N2c rabies viral vectors, yielding titers orders of magnitude higher with no background contamination, at a fraction of the production time, while preserving the efficiency of transsynaptic labeling. Along with the production pipeline, we developed suites of ‘starter’ AAV and bicistronic RVdG-CVS-N2c vectors, enabling retrograde labeling from a wide range of neuronal populations, tailored for diverse experimental requirements. We demonstrate the power and flexibility of the new system by uncovering hidden local and distal inhibitory connections in the mouse hippocampal formation and by imaging the functional properties of a cortical microcircuit across weeks. Our novel production pipeline provides a convenient approach to generate new rabies vectors, while our toolkit flexibly and efficiently expands the current capacity to label, manipulate and image the neuronal activity of interconnected neuronal circuits in vitro and in vivo.}, author = {Sumser, Anton L and Jösch, Maximilian A and Jonas, Peter M and Ben Simon, Yoav}, issn = {2050-084X}, journal = {eLife}, keywords = {General Immunology and Microbiology, General Biochemistry, Genetics and Molecular Biology, General Medicine, General Neuroscience}, publisher = {eLife Sciences Publications}, title = {{Fast, high-throughput production of improved rabies viral vectors for specific, efficient and versatile transsynaptic retrograde labeling}}, doi = {10.7554/elife.79848}, volume = {11}, year = {2022}, } @article{9955, abstract = {Neurons can change their classical neurotransmitters during ontogeny, sometimes going through stages of dual release. Here, we explored the development of the neurotransmitter identity of neurons of the avian nucleus isthmi parvocellularis (Ipc), whose axon terminals are retinotopically arranged in the optic tectum (TeO) and exert a focal gating effect upon the ascending transmission of retinal inputs. Although cholinergic and glutamatergic markers are both found in Ipc neurons and terminals of adult pigeons and chicks, the mRNA expression of the vesicular acetylcholine transporter, VAChT, is weak or absent. To explore how the Ipc neurotransmitter identity is established during ontogeny, we analyzed the expression of mRNAs coding for cholinergic (ChAT, VAChT, and CHT) and glutamatergic (VGluT2 and VGluT3) markers in chick embryos at different developmental stages. We found that between E12 and E18, Ipc neurons expressed all cholinergic mRNAs and also VGluT2 mRNA; however, from E16 through posthatch stages, VAChT mRNA expression was specifically diminished. Our ex vivo deposits of tracer crystals and intracellular filling experiments revealed that Ipc axons exhibit a mature paintbrush morphology late in development, experiencing marked morphological transformations during the period of presumptive dual vesicular transmitter release. Additionally, although ChAT protein immunoassays increasingly label the growing Ipc axon, this labeling was consistently restricted to sparse portions of the terminal branches. Combined, these results suggest that the synthesis of glutamate and acetylcholine, and their vesicular release, is complexly linked to the developmental processes of branching, growing and remodeling of these unique axons.}, author = {Reyes‐Pinto, Rosana and Ferrán, José L. and Vega Zuniga, Tomas A and González‐Cabrera, Cristian and Luksch, Harald and Mpodozis, Jorge and Puelles, Luis and Marín, Gonzalo J.}, issn = {1096-9861}, journal = {Journal of Comparative Neurology}, number = {2}, pages = {553--573}, publisher = {Wiley}, title = {{Change in the neurochemical signature and morphological development of the parvocellular isthmic projection to the avian tectum}}, doi = {10.1002/cne.25229}, volume = {530}, year = {2022}, } @article{7551, abstract = {Novelty facilitates formation of memories. The detection of novelty and storage of contextual memories are both mediated by the hippocampus, yet the mechanisms that link these two functions remain to be defined. Dentate granule cells (GCs) of the dorsal hippocampus fire upon novelty exposure forming engrams of contextual memory. However, their key excitatory inputs from the entorhinal cortex are not responsive to novelty and are insufficient to make dorsal GCs fire reliably. Here we uncover a powerful glutamatergic pathway to dorsal GCs from ventral hippocampal mossy cells (MCs) that relays novelty, and is necessary and sufficient for driving dorsal GCs activation. Furthermore, manipulation of ventral MCs activity bidirectionally regulates novelty-induced contextual memory acquisition. Our results show that ventral MCs activity controls memory formation through an intra-hippocampal interaction mechanism gated by novelty.}, author = {Fredes Tolorza, Felipe A and Silva Sifuentes, Maria A and Koppensteiner, Peter and Kobayashi, Kenta and Jösch, Maximilian A and Shigemoto, Ryuichi}, journal = {Current Biology}, number = {1}, pages = {P25--38.E5}, publisher = {Elsevier}, title = {{Ventro-dorsal hippocampal pathway gates novelty-induced contextual memory formation}}, doi = {10.1016/j.cub.2020.09.074}, volume = {31}, year = {2021}, }