Digital reconstruction, quantitative morphometric analysis, and membrane properties of bipolar cells in the rat retina.

A basic principle of neuroscience is that structure reflects function. This has led to numerous attempts to characterize the complete morphology of types of neurons throughout the central nervous system. The ability to acquire and analyze complete neuronal morphologies has advanced with continuous technological developments for over 150 years, with progressive refinements and increased understanding of the precise anatomical details of different types of neurons. Bipolar cells of the mammalian retina are short-range projections neurons that link the outer and inner retina. Their dendrites contact and receive input from the terminals of the light-sensing photoreceptors in the outer plexiform layer and their axons descend through the inner nuclear and inner plexiform layers to stratify at different levels of the inner plexiform layer. The stratification level of the axon terminals of different types of bipolar cells in the inner plexiform layer determines their synaptic connectivity and is an important basis for the morphological classification of these cells. Between 10 and 15 different types of cone bipolar cells have been identified in different species and they can be divided into ON-cone bipolar cells (that depolarize to the onset of light) and OFF-cone bipolar cells (that depolarize to the offset of light). Different types of cone bipolar cells are thought to be responsible for coding and transmitting different features of our visual environment and generating parallel channels that uniquely filter and transform the inputs from the photoreceptors. There is a lack of detailed morphological data for bipolar cells, especially for the rat, where biophysical mechanisms have been most extensively studied. The work presented in this thesis provides the groundwork for the future goal of developing morphologically realistic compartmental models for cone and rod bipolar cells. First, the contribution of gap junctions to the membrane properties, specifically input resistance, of bipolar cells was investigated. Gap junctions are ubiquitous within the retina, but it remains to be determined whether the strength of coupling between specific cell types is sufficiently strong for the cells to be functionally coupled via electrical synapses. There are gap junctions between the same class of bipolar cells, and this appears to be a common circuit motif in the vertebrate retina. Surprisingly, our results suggested that the gap junctions between OFF-cone bipolar cells do not support consequential electrical coupling. This provides an important first step both to elucidate the potential roles for these gap junctions, and also for the development of compartmental models for cone bipolar cells. Second, from image stacks acquired from multiphoton excitation microscopy, quantitative morphological reconstructions and detailed morphological analysis were performed with fluorescent dye-filled cone and rod bipolar cells. Compared to previous descriptions, the extent and complexity of branching of the axon terminals was surprisingly high. By precisely quantifying the level of stratification of the axon terminals in the inner plexiform layer, we have generated a reference system for the reliable classification of individual cells in future studies that are focused on correlating physiological and morphological properties. The workflow that we have implemented can be readily extended to the development of morphologically realistic compartmental models for these neurons.Doktorgradsavhandlin

Digital reconstruction and quantitative morphometric analysis of bipolar cells in live rat retinal slices

Bipolar cells convey signals from photoreceptors in the outer retina to amacrine and ganglion cells in the inner retina. In mammals, there are typically 10–15 types of cone bipolar cells and one type of rod bipolar cell. Different types of cone bipolar cells are thought to code and transmit different features of a complex visual stimulus, thereby generating parallel channels that uniquely filter and transform the photoreceptor outputs. Differential synaptic connectivity and expression of ligand- and voltage-gated ion channels are thought to be important mechanisms for processing and filtering visual signals. Whereas the biophysical basis for such mechanisms has been investigated more extensively in rat retina, there is a lack of quantitative morphological data necessary for advancing the structure–function correlation in this species, as recent connectomics investigations have focused on mouse retina. Here, we performed whole-cell recordings from cone and rod bipolar cells in rat retinal slices, filled the cells with fluorescent dyes, and acquired image stacks by multiphoton excitation microscopy. Following deconvolution, we performed digital reconstruction and morphometric analysis of 25 cone and 14 rod bipolar cells. Compared to previous descriptions, the extent and complexity of branching of the axon terminal was surprisingly high. By precisely quantifying the level of stratification of the axon terminals in the inner plexiform layer, we have generated a reference system for reliable classification of individual cells in future studies focused on correlating physiological and morphological properties. The implemented workflow can be extended to the development of morphologically realistic compartmental models for these neurons.publishedVersio

Digital reconstruction, quantitative morphometric analysis, and membrane properties of bipolar cells in the rat retina.

A basic principle of neuroscience is that structure reflects function. This has led to numerous attempts to characterize the complete morphology of types of neurons throughout the central nervous system. The ability to acquire and analyze complete neuronal morphologies has advanced with continuous technological developments for over 150 years, with progressive refinements and increased understanding of the precise anatomical details of different types of neurons. Bipolar cells of the mammalian retina are short-range projections neurons that link the outer and inner retina. Their dendrites contact and receive input from the terminals of the light-sensing photoreceptors in the outer plexiform layer and their axons descend through the inner nuclear and inner plexiform layers to stratify at different levels of the inner plexiform layer. The stratification level of the axon terminals of different types of bipolar cells in the inner plexiform layer determines their synaptic connectivity and is an important basis for the morphological classification of these cells. Between 10 and 15 different types of cone bipolar cells have been identified in different species and they can be divided into ON-cone bipolar cells (that depolarize to the onset of light) and OFF-cone bipolar cells (that depolarize to the offset of light). Different types of cone bipolar cells are thought to be responsible for coding and transmitting different features of our visual environment and generating parallel channels that uniquely filter and transform the inputs from the photoreceptors. There is a lack of detailed morphological data for bipolar cells, especially for the rat, where biophysical mechanisms have been most extensively studied. The work presented in this thesis provides the groundwork for the future goal of developing morphologically realistic compartmental models for cone and rod bipolar cells. First, the contribution of gap junctions to the membrane properties, specifically input resistance, of bipolar cells was investigated. Gap junctions are ubiquitous within the retina, but it remains to be determined whether the strength of coupling between specific cell types is sufficiently strong for the cells to be functionally coupled via electrical synapses. There are gap junctions between the same class of bipolar cells, and this appears to be a common circuit motif in the vertebrate retina. Surprisingly, our results suggested that the gap junctions between OFF-cone bipolar cells do not support consequential electrical coupling. This provides an important first step both to elucidate the potential roles for these gap junctions, and also for the development of compartmental models for cone bipolar cells. Second, from image stacks acquired from multiphoton excitation microscopy, quantitative morphological reconstructions and detailed morphological analysis were performed with fluorescent dye-filled cone and rod bipolar cells. Compared to previous descriptions, the extent and complexity of branching of the axon terminals was surprisingly high. By precisely quantifying the level of stratification of the axon terminals in the inner plexiform layer, we have generated a reference system for the reliable classification of individual cells in future studies that are focused on correlating physiological and morphological properties. The workflow that we have implemented can be readily extended to the development of morphologically realistic compartmental models for these neurons

Vignette detection and reconstruction of composed ornaments with a strengthened autoencoder

A strengthened autoencoder formed by placing an object detector upstream of a decoder is here developed in the context of the model-helped human analysis of composed ornaments from a dictionary of vignettes. The detection part is in charge to detect regions of interest containing some vignette features, and the decoding part to ensure vignette reconstruction with a relative quality depending on feature match. Images of ornaments without typographical composition are generated in order to properly assess the performance of each of the two parts

Differential contribution of gap junctions to the membrane properties of ON- and OFF-bipolar cells of the rat retina

Gap junctions are ubiquitous within the retina, but in general, it remains to be determined whether gap junction coupling between specific cell types is sufficiently strong to mediate functionally relevant coupling via electrical synapses. From ultrastructural, tracer coupling and immunolabeling studies, there is clear evidence for gap junctions between cone bipolar cells, but it is not known if these gap junctions function as electrical synapses. Here, using whole-cell voltage-clamp recording in rat (male and female) retinal slices, we investigated whether the gap junctions of bipolar cells make a measurable contribution to the membrane properties of these cells. We measured the input resistance (RN) of bipolar cells before and after applying meclofenamic acid (MFA) to block gap junctions. In the presence of MFA, RN of ON-cone bipolar cells displayed a clear increase, paralleled by block of the electrical coupling between these cells and AII amacrine cells in recordings of coupled cell pairs. For OFF-cone and rod bipolar cells, RN did not increase in the presence of MFA. The results for rod bipolar cells are consistent with the lack of gap junctions in these cells. However, for OFF-cone bipolar cells, our results suggest that the morphologically identified gap junctions between these cells do not support a junctional conductance that is sufficient to mediate effective electrical coupling. Instead, these junctions might play a role in chemical and/or metabolic coupling between subcellular compartments

Digital reconstruction and quantitative morphometric analysis of bipolar cells in live rat retinal slices

Bipolar cells convey signals from photoreceptors in the outer retina to amacrine and ganglion cells in the inner retina. In mammals, there are typically 10–15 types of cone bipolar cells and one type of rod bipolar cell. Different types of cone bipolar cells are thought to code and transmit different features of a complex visual stimulus, thereby generating parallel channels that uniquely filter and transform the photoreceptor outputs. Differential synaptic connectivity and expression of ligand- and voltage-gated ion channels are thought to be important mechanisms for processing and filtering visual signals. Whereas the biophysical basis for such mechanisms has been investigated more extensively in rat retina, there is a lack of quantitative morphological data necessary for advancing the structure–function correlation in this species, as recent connectomics investigations have focused on mouse retina. Here, we performed whole-cell recordings from cone and rod bipolar cells in rat retinal slices, filled the cells with fluorescent dyes, and acquired image stacks by multiphoton excitation microscopy. Following deconvolution, we performed digital reconstruction and morphometric analysis of 25 cone and 14 rod bipolar cells. Compared to previous descriptions, the extent and complexity of branching of the axon terminal was surprisingly high. By precisely quantifying the level of stratification of the axon terminals in the inner plexiform layer, we have generated a reference system for reliable classification of individual cells in future studies focused on correlating physiological and morphological properties. The implemented workflow can be extended to the development of morphologically realistic compartmental models for these neurons

First-line single-agent pembrolizumab for PD-L1-positive (tumor proportion score ≥ 50%) advanced non-small cell lung cancer in the real world: impact in brain metastasis: a national French multicentric cohort (ESCKEYP GFPC study)

International audienceFew real-world data are available in patients with advanced metastatic non-small cell lung cancer (NSCLC) treated with first-line immunotherapy, particularly in those with brain metastases at treatment initiation

First-line single-agent pembrolizumab for PD-L1-positive (tumor proportion score ≥ 50%) advanced non-small cell lung cancer in the real world: impact in brain metastasis: a national French multicentric cohort (ESCKEYP GFPC study)

International audienceFew real-world data are available in patients with advanced metastatic non-small cell lung cancer (NSCLC) treated with first-line immunotherapy, particularly in those with brain metastases at treatment initiation

"Zähmung der Bellona" oder Ökonomie der Gewalt? Überlegungen zur Kultur des Krieges im Ancien régime

International audienc