The Experts below are selected from a list of 342 Experts worldwide ranked by ideXlab platform
Jose L Lanciego - One of the best experts on this subject based on the ideXlab platform.
-
Neuroanatomical Tract-Tracing techniques that did go viral
Brain Structure and Function, 2020Co-Authors: Jose L Lanciego, Floris G WouterloodAbstract:Neuroanatomical Tracing methods remain fundamental for elucidating the complexity of brain circuits. During the past decades, the technical arsenal at our disposal has been greatly enriched, with a steady supply of fresh arrivals. This paper provides a landscape view of classical and modern tools for Tract-Tracing purposes. Focus is placed on methods that have gone viral, i.e., became most widespread used and fully reliable. To keep an historical perspective, we start by reviewing one-dimensional, standalone transport-Tracing tools; these including today’s two most favorite anterograde Neuroanatomical tracers such as Phaseolus vulgaris -leucoagglutinin and biotinylated dextran amine. Next, emphasis is placed on several classical tools widely used for retrograde Neuroanatomical Tracing purposes, where Fluoro-Gold in our opinion represents the best example. Furthermore, it is worth noting that multi-dimensional paradigms can be designed by combining different tracers or by applying a given tracer together with detecting one or more neurochemical substances, as illustrated here with several examples. Finally, it is without any doubt that we are currently witnessing the unstoppable and spectacular rise of modern molecular-genetic techniques based on the use of modified viruses as delivery vehicles for genetic material, therefore, pushing the Tract-Tracing field forward into a new era. In summary, here, we aim to provide neuroscientists with the advice and background required when facing a choice on which Neuroanatomical tracer—or combination thereof—might be best suited for addressing a given experimental design.
-
chapter 17 classic and contemporary neural Tract Tracing techniques
Diffusion MRI (Second Edition)#R##N#From Quantitative Measurement to In vivo Neuroanatomy, 2014Co-Authors: Robert J Morecraft, Gabriella Ugolini, Jose L Lanciego, Floris G Wouterlood, Deepak N. PandyaAbstract:Investigating the organization of nerve pathways is of fundamental interest in neurology and the neurosciences. In this chapter we draw attention to landmark events that have contributed to the scientific evolution of neuron Tract Tracing and cover in detail contemporary methods employed in experimental animals to trace nerve pathways. The basic anatomical Tract-Tracing methods, including the cellular mechanisms underlying Tract tracer uptake and axonal transport, are described. Specific techniques and experimental applications involving tracers transported retrogradely, anterogradely, and transneuronally are reviewed. Finally, examples of combined Neuroanatomical Tract-Tracing applications are discussed and illustrated.
-
classic and contemporary neural Tract Tracing techniques
Diffusion MRI: from quantitative measurement to in-vivo neuroanatom, 2009Co-Authors: Robert J Morecraft, Gabriella Ugolini, Jose L Lanciego, Floris G Wouterlood, Deepak N. PandyaAbstract:AbsTract Investigating the organization of nerve pathways is of fundamental interest in neurology and the neurosciences. In this chapter we draw attention to landmark events that have contributed to the scientific evolution of neuron Tract Tracing and cover in detail contemporary methods employed in experimental animals to trace nerve pathways. The basic anatomical Tract-Tracing methods, including the cellular mechanisms underlying Tract tracer uptake and axonal transport, are described. Specific techniques and experimental applications involving tracers transported retrogradely, anterogradely, and transneuronally are reviewed. Finally, examples of combined Neuroanatomical Tract-Tracing applications are discussed and illustrated.
-
high resolution Neuroanatomical Tract Tracing for the analysis of striatal microcircuits
Brain Research, 2008Co-Authors: Pascal Salin, Maria Castle, Philippe Kachidian, Pedro Barrosochinea, Iciar P Lopez, Alberto J Rico, Lydia Kerkerianle Goff, Patrice Coulon, Jose L LanciegoAbstract:Although currently available retrograde tracers are useful tools for identifying striatal projection neurons, transported tracers often remained restricted within the neuronal somata and the thickest, main dendrites. Indeed, thin dendrites located far away from the cell soma as well as post-synaptic elements such as dendritic spines cannot be labeled unless performing intracellular injections. In this regard, the subsequent use of anterograde tracers for the labeling of striatal afferents often failed to unequivocally elucidate whether a given afferent makes true contacts with striatal projections neurons. Here we show that such a technical constraint can now be circumvented by retrograde Tracing using rabies virus (RV). Immunofluorescence detection with a monoclonal antibody directed against the viral phosphoprotein resulted in a consistent Golgi-like labeling of striatal projection neurons, allowing clear visualization of small-size elements such as thin dendrites as well as dendritic spines. The combination of this retrograde Tracing together with dual anterograde Tracing of cortical and thalamic afferents has proven to be a useful tool for ascertaining striatal microcircuits. Indeed, by taking advantage of the trans-synaptic spread of RV, different subpopulations of local-circuit neurons modulating striatal efferent neurons can also be identified. At the striatal level, structures displaying labeling were visualized under the confocal laser-scanning microscope at high resolution. Once acquired, confocal stacks of images were firstly deconvoluted and then processed through 3D-volume rendering in order to unequivocally identify true contacts between pre-synaptic elements (axon terminals from cortical or thalamic sources) and post-synaptic elements (projection neurons and/or interneurons labeled with RV).
-
functional Neuroanatomical Tract Tracing analysis of changes in gene expression of brain circuits of interest
Brain Research, 2006Co-Authors: Monica Perezmanso, Maria S Aymerich, Pedro Barrosochinea, Jose L LanciegoAbstract:Neuroanatomical Tracing when considered as an isolated method produces relatively straightforward answers. Although single-, double- or even triple-Tracing paradigms produce valuable data on the organization of brain circuits, the final outcome often is too simplistic since it is not possible to elucidate the activity of these circuits. In this regard, emerging technologies contribute with additional information about the status of neuronal circuits. The laser-guided capture microdissection microscope (LCM) allows the accurate dissection of small brain areas under the microscope that could be further analyzed for gene expression or proteomics. In order to elucidate the gene expression of a given circuit of interest, we have developed a combination of methods comprising (i) fluorescent non-radioactive in situ hybridization for the detection of vGLUT2 mRNA expression combined with retrograde Tracing with Fluoro-Gold (FG; analysis performed under the confocal microscope) and (ii) laser-guided capture microdissection of brain areas containing neurons retrogradely labeled with FG followed by the measurement of changes in mRNA levels encoding for vGLUT2 by real-time PCR. Our goal was to detect changes in gene expression of the thalamostriatal pathway in unilaterally 6-OHDA lesioned rats. Taking advantage of this procedure, we found a three-fold increase in vGLUT2 mRNA expression within thalamic neurons projecting to the dopamine-depleted striatum when compared with the activity of the thalamic neurons innervating the control striatum.
Floris G Wouterlood - One of the best experts on this subject based on the ideXlab platform.
-
Neuroanatomical Tract-Tracing techniques that did go viral
Brain Structure and Function, 2020Co-Authors: Jose L Lanciego, Floris G WouterloodAbstract:Neuroanatomical Tracing methods remain fundamental for elucidating the complexity of brain circuits. During the past decades, the technical arsenal at our disposal has been greatly enriched, with a steady supply of fresh arrivals. This paper provides a landscape view of classical and modern tools for Tract-Tracing purposes. Focus is placed on methods that have gone viral, i.e., became most widespread used and fully reliable. To keep an historical perspective, we start by reviewing one-dimensional, standalone transport-Tracing tools; these including today’s two most favorite anterograde Neuroanatomical tracers such as Phaseolus vulgaris -leucoagglutinin and biotinylated dextran amine. Next, emphasis is placed on several classical tools widely used for retrograde Neuroanatomical Tracing purposes, where Fluoro-Gold in our opinion represents the best example. Furthermore, it is worth noting that multi-dimensional paradigms can be designed by combining different tracers or by applying a given tracer together with detecting one or more neurochemical substances, as illustrated here with several examples. Finally, it is without any doubt that we are currently witnessing the unstoppable and spectacular rise of modern molecular-genetic techniques based on the use of modified viruses as delivery vehicles for genetic material, therefore, pushing the Tract-Tracing field forward into a new era. In summary, here, we aim to provide neuroscientists with the advice and background required when facing a choice on which Neuroanatomical tracer—or combination thereof—might be best suited for addressing a given experimental design.
-
chapter 17 classic and contemporary neural Tract Tracing techniques
Diffusion MRI (Second Edition)#R##N#From Quantitative Measurement to In vivo Neuroanatomy, 2014Co-Authors: Robert J Morecraft, Gabriella Ugolini, Jose L Lanciego, Floris G Wouterlood, Deepak N. PandyaAbstract:Investigating the organization of nerve pathways is of fundamental interest in neurology and the neurosciences. In this chapter we draw attention to landmark events that have contributed to the scientific evolution of neuron Tract Tracing and cover in detail contemporary methods employed in experimental animals to trace nerve pathways. The basic anatomical Tract-Tracing methods, including the cellular mechanisms underlying Tract tracer uptake and axonal transport, are described. Specific techniques and experimental applications involving tracers transported retrogradely, anterogradely, and transneuronally are reviewed. Finally, examples of combined Neuroanatomical Tract-Tracing applications are discussed and illustrated.
-
classic and contemporary neural Tract Tracing techniques
Diffusion MRI: from quantitative measurement to in-vivo neuroanatom, 2009Co-Authors: Robert J Morecraft, Gabriella Ugolini, Jose L Lanciego, Floris G Wouterlood, Deepak N. PandyaAbstract:AbsTract Investigating the organization of nerve pathways is of fundamental interest in neurology and the neurosciences. In this chapter we draw attention to landmark events that have contributed to the scientific evolution of neuron Tract Tracing and cover in detail contemporary methods employed in experimental animals to trace nerve pathways. The basic anatomical Tract-Tracing methods, including the cellular mechanisms underlying Tract tracer uptake and axonal transport, are described. Specific techniques and experimental applications involving tracers transported retrogradely, anterogradely, and transneuronally are reviewed. Finally, examples of combined Neuroanatomical Tract-Tracing applications are discussed and illustrated.
-
Neuroanatomical Tract Tracing 3 molecules neurons and systems
2006Co-Authors: Lennart Heimer, Floris G Wouterlood, Laszlo Zaborszky, Jose L LanciegoAbstract:Short Retrospection.- Preembedding Immunoelectron Microscopy: Applications for Studies of the Nervous System.- Postembedding Immunogold Cytochemistry of Membrane Molecules and Amino Acid Transmitters in the Central Nervous System.- Cell and Tissue Microdissection in Combination with Genomic and Proteomic Applications.- Molecules and Membrane Activity: Single-Cell RT-PCR and Patch-Clamp Recording from Central Neurons.- Merging Structure and Function: Combination of In Vivo Extracellular and Intracellular Electrophysiological Recordings with Neuroanatomical Techniques.- Juxtacellular Labeling of Individual Neurons In Vivo: From Electrophysiology to Synaptology.- Nonradioactive In Situ Hybridization in Combination with Tract-Tracing.- Viral Tracers for the Analysis of Neural Circuits.- Dextran Amines: Versatile Tools for Anterograde and Retrograde Studies of Nervous System Connectivity.- Multiple Neuroanatomical Tract-Tracing: Approaches for Multiple Tract-Tracing.- Tract-Tracing in Developing Systems and in Postmortem Human Material Using Carbocyanine Dyes.- Combined Fluorescence Methods to Determine Synapses in the Light Microscope: Multilabel Confocal Laser Scanning Microscopy.- Advances in Understanding Cortical Function Through Combined Voltage-Sensitive Dye Imaging, Whole-Cell Recordings, and Analysis of Cellular Morphology.- From Dendrites to Networks: Optically Probing the Living Brain Slice and Using Principal Component Analysis to Characterize Neuronal Morphology.- Stereology of Neural Connections: An Overview.- Three-Dimensional Computerized Reconstruction from Serial Sections: Cell Populations, Regions, and Whole Brain.- Atlases of the Human Brain: Tools for Functional Neuroimaging.- Neuron and Network Modeling.- Functional Connectivity of the Brain: Reconstruction from Static and Dynamic Data.
-
multiple Neuroanatomical Tract Tracing approaches for multiple Tract Tracing
2006Co-Authors: Jose L Lanciego, Floris G WouterloodAbstract:Experimental Neuroanatomical Tracing techniques are fundamental to the study of the structure of the central nervous system. In the last few decades, many new methods for axonal Tracing and cell labeling have been introduced. Neuroanatomical Tracing applied as an isolated method produces relatively straightforward answers, for instance, whether there is connectivity from compartment Y in nucleus A to layer X in area B. However, questions that deal with the intrinsic complexity of brain circuits require the application of multiple-Tracing paradigms in which two or even three different tracers are combined in single histological sections. With such paradigms we can handle questions like “are the fibers arriving in layer X of area B in contact with neurons that project to compartment Z in nucleus C,” “do these projection neurons receive as well innervation from area W,” and “what is the neurochemical signature of these connectivity-identified neurons?”We illustrate this approach with examples from our studies on pallidonigral connectivity in association with nigrostriatal efferent neurons.
Philip T. Hicks - One of the best experts on this subject based on the ideXlab platform.
-
A Comparative Neuroanatomical Study of the Red Nucleus of the Cat, Macaque and Human
2013Co-Authors: Satoru Onodera, Philip T. HicksAbstract:Background: The human red nucleus (Nr) is comparatively less well-studied than that of cats or monkeys. Given the functional importance of reticular and midbrain structures in control of movement and locomotion as well as from an evolutionary perspective, we investigated the nature and extent of any differences in Nr projections to the olivary complex in quadrupedal and bipedal species. Using Neuroanatomical Tract-Tracing techniques we developed a ‘‘neural sheet’’ hypothesis allowing us to propose how rubro-olivary relations differ among the three species. Methods and Findings: Wheat germ agglutinin-horseradish peroxidase staining supports findings that the cat’s nucleus accessories medialis of Bechtrew (NB) projects mainly to the lateral bend of the principal olive. We clarified boundaries among nucleus of Darkschewitsch (ND), NB and parvicellular red nucleus (pNr) of the cat’s neural sheet. The macaque’s NDmedial accessory olivary projection is rostro-caudally organized and the dorsomedial and ventrolateral parts of the macaque’s pNr may project to the principal olive’s rostral and caudal dorsal lamella; in cat it projects as well to pNr. Myelinand Nissl-stained sections show that a well-developed dorsomedial part of the human Nr consists of densely packed cells, deriving small myelinated fibers that continue into the medial central tegmental Tract. Conclusions: Based on these findings we suggest there are distinct bipedal-quadrupedal differences for Nr projections to the olivary complex. We propose the Nr of cats and monkeys comprise the ND, NB and pNr in a zonal sheet-like structure
-
a comparative Neuroanatomical study of the red nucleus of the cat macaque and human
PLOS ONE, 2009Co-Authors: Satoru Onodera, Philip T. HicksAbstract:Background The human red nucleus (Nr) is comparatively less well-studied than that of cats or monkeys. Given the functional importance of reticular and midbrain structures in control of movement and locomotion as well as from an evolutionary perspective, we investigated the nature and extent of any differences in Nr projections to the olivary complex in quadrupedal and bipedal species. Using Neuroanatomical Tract-Tracing techniques we developed a “neural sheet” hypothesis allowing us to propose how rubro-olivary relations differ among the three species. Methods and Findings Wheat germ agglutinin-horseradish peroxidase staining supports findings that the cat's nucleus accessories medialis of Bechtrew (NB) projects mainly to the lateral bend of the principal olive. We clarified boundaries among nucleus of Darkschewitsch (ND), NB and parvicellular red nucleus (pNr) of the cat's neural sheet. The macaque's ND-medial accessory olivary projection is rostro-caudally organized and the dorsomedial and ventrolateral parts of the macaque's pNr may project to the principal olive's rostral and caudal dorsal lamella; in cat it projects as well to pNr. Myelin- and Nissl-stained sections show that a well-developed dorsomedial part of the human Nr consists of densely packed cells, deriving small myelinated fibers that continue into the medial central tegmental Tract. Conclusions Based on these findings we suggest there are distinct bipedal-quadrupedal differences for Nr projections to the olivary complex. We propose the Nr of cats and monkeys comprise the ND, NB and pNr in a zonal sheet-like structure, retaining clear nuclear boundaries and an isolated, well-developed mNr. The human NB may be distinguished from its more specialised ND (ND lies alongside a well-developed pNr) in the human central gray. Phylogenetically, the NB may have been translocated into a roll-shaped Nr in the reticular formation, the dorsomedial portion of which might correspond to the cat's and monkey's NB.
Pedro Barrosochinea - One of the best experts on this subject based on the ideXlab platform.
-
high resolution Neuroanatomical Tract Tracing for the analysis of striatal microcircuits
Brain Research, 2008Co-Authors: Pascal Salin, Maria Castle, Philippe Kachidian, Pedro Barrosochinea, Iciar P Lopez, Alberto J Rico, Lydia Kerkerianle Goff, Patrice Coulon, Jose L LanciegoAbstract:Although currently available retrograde tracers are useful tools for identifying striatal projection neurons, transported tracers often remained restricted within the neuronal somata and the thickest, main dendrites. Indeed, thin dendrites located far away from the cell soma as well as post-synaptic elements such as dendritic spines cannot be labeled unless performing intracellular injections. In this regard, the subsequent use of anterograde tracers for the labeling of striatal afferents often failed to unequivocally elucidate whether a given afferent makes true contacts with striatal projections neurons. Here we show that such a technical constraint can now be circumvented by retrograde Tracing using rabies virus (RV). Immunofluorescence detection with a monoclonal antibody directed against the viral phosphoprotein resulted in a consistent Golgi-like labeling of striatal projection neurons, allowing clear visualization of small-size elements such as thin dendrites as well as dendritic spines. The combination of this retrograde Tracing together with dual anterograde Tracing of cortical and thalamic afferents has proven to be a useful tool for ascertaining striatal microcircuits. Indeed, by taking advantage of the trans-synaptic spread of RV, different subpopulations of local-circuit neurons modulating striatal efferent neurons can also be identified. At the striatal level, structures displaying labeling were visualized under the confocal laser-scanning microscope at high resolution. Once acquired, confocal stacks of images were firstly deconvoluted and then processed through 3D-volume rendering in order to unequivocally identify true contacts between pre-synaptic elements (axon terminals from cortical or thalamic sources) and post-synaptic elements (projection neurons and/or interneurons labeled with RV).
-
functional Neuroanatomical Tract Tracing analysis of changes in gene expression of brain circuits of interest
Brain Research, 2006Co-Authors: Monica Perezmanso, Maria S Aymerich, Pedro Barrosochinea, Jose L LanciegoAbstract:Neuroanatomical Tracing when considered as an isolated method produces relatively straightforward answers. Although single-, double- or even triple-Tracing paradigms produce valuable data on the organization of brain circuits, the final outcome often is too simplistic since it is not possible to elucidate the activity of these circuits. In this regard, emerging technologies contribute with additional information about the status of neuronal circuits. The laser-guided capture microdissection microscope (LCM) allows the accurate dissection of small brain areas under the microscope that could be further analyzed for gene expression or proteomics. In order to elucidate the gene expression of a given circuit of interest, we have developed a combination of methods comprising (i) fluorescent non-radioactive in situ hybridization for the detection of vGLUT2 mRNA expression combined with retrograde Tracing with Fluoro-Gold (FG; analysis performed under the confocal microscope) and (ii) laser-guided capture microdissection of brain areas containing neurons retrogradely labeled with FG followed by the measurement of changes in mRNA levels encoding for vGLUT2 by real-time PCR. Our goal was to detect changes in gene expression of the thalamostriatal pathway in unilaterally 6-OHDA lesioned rats. Taking advantage of this procedure, we found a three-fold increase in vGLUT2 mRNA expression within thalamic neurons projecting to the dopamine-depleted striatum when compared with the activity of the thalamic neurons innervating the control striatum.
Maria S Aymerich - One of the best experts on this subject based on the ideXlab platform.
-
functional Neuroanatomical Tract Tracing analysis of changes in gene expression of brain circuits of interest
Brain Research, 2006Co-Authors: Monica Perezmanso, Maria S Aymerich, Pedro Barrosochinea, Jose L LanciegoAbstract:Neuroanatomical Tracing when considered as an isolated method produces relatively straightforward answers. Although single-, double- or even triple-Tracing paradigms produce valuable data on the organization of brain circuits, the final outcome often is too simplistic since it is not possible to elucidate the activity of these circuits. In this regard, emerging technologies contribute with additional information about the status of neuronal circuits. The laser-guided capture microdissection microscope (LCM) allows the accurate dissection of small brain areas under the microscope that could be further analyzed for gene expression or proteomics. In order to elucidate the gene expression of a given circuit of interest, we have developed a combination of methods comprising (i) fluorescent non-radioactive in situ hybridization for the detection of vGLUT2 mRNA expression combined with retrograde Tracing with Fluoro-Gold (FG; analysis performed under the confocal microscope) and (ii) laser-guided capture microdissection of brain areas containing neurons retrogradely labeled with FG followed by the measurement of changes in mRNA levels encoding for vGLUT2 by real-time PCR. Our goal was to detect changes in gene expression of the thalamostriatal pathway in unilaterally 6-OHDA lesioned rats. Taking advantage of this procedure, we found a three-fold increase in vGLUT2 mRNA expression within thalamic neurons projecting to the dopamine-depleted striatum when compared with the activity of the thalamic neurons innervating the control striatum.
-
thalamic innervation of striatal and subthalamic neurons projecting to the rat entopeduncular nucleus
European Journal of Neuroscience, 2004Co-Authors: Jose L Lanciego, Maria Castle, Nancy Gonzalo, Carlos Sanchezescobar, Maria S Aymerich, Jose A ObesoAbstract:The present study analyses the anatomical arrangement of the projections linking the Wistar rat parafascicular thalamic nucleus (PF) and basal ganglia structures, such as the striatum and the subthalamic nucleus (STN), by using Neuroanatomical Tract-Tracing techniques. Both the thalamostriatal and the striato-entopeduncular projections were topographically organized, and several areas of overlap between identified circuits were noticed, sustaining the existence of up to three separated channels within the Nauta-Mehler loop. Thalamic afferents arising from dorsolateral PF territories are in register with striatofugal neurons located in dorsolateral striatal areas, which in turn project to dorsolateral regions of the entopeduncular nucleus (ENT). Medial ENT regions are innervated by striatal neurons located within medial striatal territories, these neurons being the target for thalamic afferents coming from medial PF areas. Finally, afferents from neurons located in ventrolateral PF areas approached striatal neurons in ventral and lateral striatal territories, which in turn project towards ventral and lateral ENT regions. Efferent STN neurons projecting to ENT were found to be the apparent postsynaptic target for thalamo-subthalamic axons. The thalamo-subthalamic projection was also topographically organized. Medial, central and lateral STN territories are innervated by thalamic neurons located within medial, ventrolateral and dorsolateral PF areas, respectively. Thus, each individual PF subregion projects in a segregated fashion to specific parts of the striato-entopeduncular and subthalamo-entopeduncular systems. These circuits enabled the caudal intralaminar nuclei to modulate basal ganglia output.