A Neural Model of How the Cortical Subplate Coordinates the Laminar Development of Orientation and Ocular Dominance Maps

Air Force Office of Scientific Research (F49620-98-1-0108, F49620-0 1-1-0397); Defense Advanced Research Projects Agency and the Office of Naval Research (N00014-95-1-0409); National Science Foundation (IIS-97-20333); Office of Naval Research (N00014-01-1-0624

Development of Enteric Neurons and Muscularis Macrophages

The enteric nervous system (ENS) is a complex interconnected network of neurons and glia in the bowel wall that regulates intestinal motility, blood flow, and epithelial function. The ENS also controls aspects of inflammatory signaling within the bowel. To perform these tasks, there are at least 20 types of enteric neuron and four types of enteric glia. Although much is known about early events in ENS development, signals governing the development of specific neuronal subtypes and communication with neighboring cell types within the bowel remain poorly understood. One fundamental hypothesis is that diverse trophic factors support distinct neuronal populations in the bowel. Based on our observations that the hepatocyte growth factor (HGF) receptor Met is expressed in all ENS precursors and in a subset of adult enteric neurons, we investigated the role of HGF and MET in the ENS. We found that mice lacking functional MET receptor in the ENS (Met cKO mice) exhibit defects in a subset of CGRP-expressing sensory neurons in the myenteric plexus. These sensory cells, known as intrinsic primary afferent neurons (IPANs), are responsible for transmitting signals such as bowel stretch and villus deformation from the bowel lumen to other myenteric neurons. Met cKO mice have altered CGRP-IR neurite patterning with a corresponding failure to trigger peristalsis in response to villus stroking, but a normal response to bowel stretch. This work suggests that MET is important for the development and function of a subset of IPANs the bowel. HGF has also been known for more than a decade to protect the bowel from injury in colitis models. It had been assumed that HGF’s protective effects were mediated by MET present on bowel epithelial cells. Using Met cKO mice that are missing MET in the ENS, but have normal MET in gut epithelium, we showed that neuronal HGF signaling contributes to the protective effects of HGF in the bowel that have been previously reported. Met cKO mice have significantly more bowel injury following treatment with dextran sodium sulfate (DSS) to induce colitis. Furthermore, these animals show reduced proliferation of epithelial cells in the context of injury. Together these findings suggest that that HGF/MET signaling is important for development and function of a subset IPANs and that these cells regulate intestinal motility and epithelial cell proliferation in response to bowel injury. Our studies of Met cKO mice highlighted how defects in neurite patterning of enteric neurons can profoundly affect bowel function. Surprisingly little is known about factors that govern appropriate formation of neuronal connections in the bowel. To identify other cues important for enteric neuron axon patterning, we used broad microarray gene expression profiling. Gene expression in embryonic day 17.5 (E17.5) ENS cells was compared to gene expression in surrounding cells to identify genes encoding cell surface receptors or adhesion molecules enriched in the ENS that have known roles in axon pathfinding. We found that the semaphorin receptor Plexin-A4 was highly enriched in the ENS and confirmed this by in situ hybridization (ISH) at E17.5. Immunohistochemistry using a commercial antibody to Plexin-A4 suggested that Plexin-A4 was found in a subset of calretinin-IR neurons. However, staining of Plexin A4 knockout (Plexin A4 KO) bowel revealed this antibody to be non-specific. Despite testing two other Plexin-A4 antibodies, we were unable to determine which enteric neurons produce Plexin A4. We analyzed the ENS of Plexin A4 knockout (Plexin A4 KO) mice but could not identify any gross defects in neurite patterning. Additionally, functional analysis of Plexin A4 KO mice did not reveal any defects in the peristaltic reflex of these animals. As is true for other parts of the nervous system, it is likely that Plexin A4 acts redundantly with Plexin A2 (also enriched in the ENS) and any effects on axon pathfinding in the ENS would only be revealed in Plexin A4/Plexin A2 double knockout mice. Remarkably, the non-specific Plexin A4 antibody we had been using also labeled a population of poorly characterized muscularis macrophages within the bowel muscularis externa, and we decided to study these cells. Bowel macrophages integrate a variety of environmental stimuli to assume either a pro-inflammatory or tissue-protective phenotype. A growing body of evidence suggests that neuronal cholinergic and noradrenergic signaling dampens the inflammatory phenotype of muscularis macrophages found in close contact with enteric neurons. Additionally, it’s been suggested that enteric neurons produce CSF1, the main survival factor for muscularis macrophages. This would imply that these macrophages would be abnormal when the ENS is missing. Surprisingly, we found that muscularis macrophage colonization of the bowel precedes colonization by enteric neurons and that the main source of CSF1 during development is non-neuronal. Furthermore, Ret knockout mice that are completely missing enteric neurons in the small bowel and colon contain normal numbers of well patterned macrophages. Additionally, these macrophages do not differ in their expression of activating cell surface markers, or in their ex-vivo response to lipopolysaccharide (LPS) stimulation. These studies help clarify the role of ENS in the homeostasis and activation of muscularis macrophages, suggesting that, at least developmentally, enteric neurons are dispensable for muscularis macrophage survival and do not alter the baseline inflammatory status of muscularis macrophages

Visual Cortex

The neurosciences have experienced tremendous and wonderful progress in many areas, and the spectrum encompassing the neurosciences is expansive. Suffice it to mention a few classical fields: electrophysiology, genetics, physics, computer sciences, and more recently, social and marketing neurosciences. Of course, this large growth resulted in the production of many books. Perhaps the visual system and the visual cortex were in the vanguard because most animals do not produce their own light and offer thus the invaluable advantage of allowing investigators to conduct experiments in full control of the stimulus. In addition, the fascinating evolution of scientific techniques, the immense productivity of recent research, and the ensuing literature make it virtually impossible to publish in a single volume all worthwhile work accomplished throughout the scientific world. The days when a single individual, as Diderot, could undertake the production of an encyclopedia are gone forever. Indeed most approaches to studying the nervous system are valid and neuroscientists produce an almost astronomical number of interesting data accompanied by extremely worthy hypotheses which in turn generate new ventures in search of brain functions. Yet, it is fully justified to make an encore and to publish a book dedicated to visual cortex and beyond. Many reasons validate a book assembling chapters written by active researchers. Each has the opportunity to bind together data and explore original ideas whose fate will not fall into the hands of uncompromising reviewers of traditional journals. This book focuses on the cerebral cortex with a large emphasis on vision. Yet it offers the reader diverse approaches employed to investigate the brain, for instance, computer simulation, cellular responses, or rivalry between various targets and goal directed actions. This volume thus covers a large spectrum of research even though it is impossible to include all topics in the extremely diverse field of neurosciences

Human metabolic adaptations and prolonged expensive neurodevelopment: A review

1.	After weaning, human hunter-gatherer juveniles receive substantial (≈3.5-7 MJ day^-1^), extended (≈15 years) and reliable (kin and nonkin food pooling) energy provision.
2.	The childhood (pediatric) and the adult human brain takes a very high share of both basal metabolic rate (BMR) (child: 50-70%; adult: ≈20%) and total energy expenditure (TEE) (child: 30-50%; adult: ≈10%).
3.	The pediatric brain for an extended period (≈4-9 years-of-age) consumes roughly 50% more energy than the adult one, and after this, continues during adolescence, at a high but declining rate. Within the brain, childhood cerebral gray matter has an even higher 1.9 to 2.2-fold increased energy consumption. 
4.	This metabolic expensiveness is due to (i) the high cost of synapse activation (74% of brain energy expenditure in humans), combined with (ii), a prolonged period of exuberance in synapse numbers (up to double the number present in adults). Cognitive development during this period associates with volumetric changes in gray matter (expansion and contraction due to metabolic related size alterations in glial cells and capillary vascularization), and in white matter (expansion due to myelination). 
5.	Amongst mammals, anatomically modern humans show an unique pattern in which very slow musculoskeletal body growth is followed by a marked adolescent size/stature spurt. This pattern of growth contrasts with nonhuman primates that have a sustained fast juvenile growth with only a minor period of puberty acceleration. The existence of slow childhood growth in humans has been shown to date back to 160,000 BP. 
6.	Human children physiologically have a limited capacity to protect the brain from plasma glucose fluctuations and other metabolic disruptions. These can arise in adults, during prolonged strenuous exercise when skeletal muscle depletes plasma glucose, and produces other metabolic disruptions upon the brain (hypoxia, hyperthermia, dehydration and hyperammonemia). These are proportional to muscle mass.
7.	Children show specific adaptations to minimize such metabolic disturbances. (i) Due to slow body growth and resulting small body size, they have limited skeletal muscle mass. (ii) They show other adaptations such as an exercise specific preference for free fatty acid metabolism. (iii) While children are generally more active than adolescents and adults, they avoid physically prolonged intense exertion. 
8.	Childhood has a close relationship to high levels of energy provision and metabolic adaptations that support prolonged synaptic neurodevelopment. 
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The dendritic engram

Accumulating evidence from a wide range of studies, including behavioral, cellular, molecular and computational findings, support a key role of dendrites in the encoding and recall of new memories. Dendrites can integrate synaptic inputs in non-linear ways, provide the substrate for local protein synthesis and facilitate the orchestration of signaling pathways that regulate local synaptic plasticity. These capabilities allow them to act as a second layer of computation within the neuron and serve as the fundamental unit of plasticity. As such, dendrites are integral parts of the memory engram, namely the physical representation of memories in the brain and are increasingly studied during learning tasks. Here, we review experimental and computational studies that support a novel, dendritic view of the memory engram that is centered on non-linear dendritic branches as elementary memory units. We highlight the potential implications of dendritic engrams for the learning and memory field and discuss future research directions

ApoE4 effects on the structural covariance brain networks topology in Mild Cognitive Impairment

The Apolipoprotein E isoform E4 (ApoE4) is consistently associated with an elevated risk of developing late-onset Alzheimer's Disease (AD). However, little is known about his potential genetic modulation on the structural covariance brain networks during prodromal stages like Mild Cognitive Impairment (MCI). The covariance phenomenon is based on the observation that regions correlating in morphometric descriptors are often part of the same brain system. In a first study, I assessed the ApoE4-related changes on the brain network topology in 256 MCI patients, using the regional cortical thickness to define the covariance network. The cross-sectional sample selected from the ADNI database was subdivided into ApoE4-positive (Carriers) and negative (non-Carriers). At the group-level, the results showed a significant decrease in characteristic path length, clustering index, local efficiency, global connectivity, modularity, and increased global efficiency for Carriers compared to non-Carriers. Overall, I found that ApoE4 in MCI shaped the topological organization of cortical thickness covariance networks. In the second project, I investigated the impact of ApoE4 on the single-subject gray matter networks in a sample of 200 MCI from the ADNI database. The patients were classified based on clinical outcome (stable MCI versus converters to AD) and ApoE4 status (Carriers versus non-Carriers). The effects of ApoE4 and disease progression on the network measures at baseline and rate of change were explored. The topological network attributes were correlated with AD biomarkers. The main findings showed that gray matter network topology is affected independently by ApoE4 and the disease progression (to AD) in late-MCI. The network measures alterations showed a more random organization in Carriers compared to non-Carriers. Finally, as additional research, I investigated whether a network-based approach combined with the graph theory is able to detect cerebrovascular reactivity (CVR) changes in MCI. Our findings suggest that this experimental approach is more sensitive to identifying subtle cerebrovascular alterations than the classical experimental designs. This study paves the way for a future investigation on the ApoE4-cerebrovascular interaction effects on the brain networks during AD progression. In summary, my thesis results provide evidence of the value of the structural covariance brain network measures to capture subtle neurodegenerative changes associated with ApoE4 in MCI. Together with other biomarkers, these variables may help predict disease progression, providing additional reliable intermediate phenotypes

Endocytic trafficking is required for neuron cell death through regulating TGF-beta signaling in \u3ci\u3eDrosophila melanogaster\u3c/i\u3e

Programmed cell death (PCD) is an essential feature during the development of the central nervous system in Drosophila as well as in mammals. During metamorphosis, a group of peptidergic neurons (vCrz) are eliminated from the larval central nervous system (CNS) via PCD within 6-7 h after puparium formation. To better understand this process, we first characterized the development of the vCrz neurons including their lineages and birth windows using the MARCM (Mosaic Analysis with a Repressible Cell Marker) assay. Further genetic and MARCM analyses showed that not only Myoglianin (Myo) and its type I receptor Baboon is required for neuron cell death, but also this death signal is extensively regulated by endocytic trafficking in Drosophila melanogaster. We found that clathrin-mediated membrane receptor internalization and subsequent endocytic events involved in Rab5-dependent early endosome and Rab11-dependent recycling endosome differentially participate in TGF-β [beta] signaling. Two early endosome-enriched proteins, SARA and Hrs, are found to act as a cytosolic retention factor of Smad2, indicating that endocytosis mediates TGF-β [beta] signaling through regulating the dissociation of Smad2 and its cytosolic retention factor

Activity-dependent refinement of the developing visual system. A comparative study across retinal ganglion cell populations and target nuclei

The formation of the mammalian visual system is a complex process that takes place in several phases and includes neurogenesis, axon guidance, axonal refinement and circuit assembly. The last stage of this process occurs after birth but before eye opening. During this period, axon terminals from retinal ganglion cells (RGCs) first extensively arborize in the different visual nuclei and then refine and establish appropriate connections. It is known that the spontaneous activity generated in the immature retina during perinatal ages plays an important role in this axonal refinement process but it is not clear to what extent such retinal activity differentially influences the refinement of the distinct populations of RGCs when they project to specific visual nuclei. To address this issue we have generated conditional mouse lines to alter spontaneous activity in different populations of RGCs and we have analyzed the projection patterns of RGCs in different visual nuclei in each of these mouse lines. Our results show that the alteration of spontaneous activity in RGCs affects axon refinement in the image-forming nuclei such as the lateral geniculate nucleus and the superior colliculus, supporting previous publications. Interestingly, we also observed that, although to a lesser extent than in the image-forming nuclei, retinal spontaneous activity correlation is important for the refinement of RGC axons in the non-image-forming nuclei such as the pretectal olive nucleus or the suprachiasmatic nucleus

Human metabolic adaptations and prolonged expensive neurodevelopment: A review

1.	After weaning, human hunter-gatherer juveniles receive substantial (≈3.5-7 MJ day^-1^), extended (≈15 years) and reliable (kin and nonkin food pooling) energy provision.
2.	The childhood (pediatric) and the adult human brain takes a very high share of both basal metabolic rate (BMR) (child: 50-70%; adult: ≈20%) and total energy expenditure (TEE) (child: 30-50%; adult: ≈10%).
3.	The pediatric brain for an extended period (≈4-9 years-of-age) consumes roughly 50% more energy than the adult one, and after this, continues during adolescence, at a high but declining rate. Within the brain, childhood cerebral gray matter has an even higher 1.9 to 2.2-fold increased energy consumption. 
4.	This metabolic expensiveness is due to (i) the high cost of synapse activation (74% of brain energy expenditure in humans), combined with (ii), a prolonged period of exuberance in synapse numbers (up to double the number present in adults). Cognitive development during this period associates with volumetric changes in gray matter (expansion and contraction due to metabolic related size alterations in glial cells and capillary vascularization), and in white matter (expansion due to myelination). 
5.	Amongst mammals, anatomically modern humans show an unique pattern in which very slow musculoskeletal body growth is followed by a marked adolescent size/stature spurt. This pattern of growth contrasts with nonhuman primates that have a sustained fast juvenile growth with only a minor period of puberty acceleration. The existence of slow childhood growth in humans has been shown to date back to 160,000 BP. 
6.	Human children physiologically have a limited capacity to protect the brain from plasma glucose fluctuations and other metabolic disruptions. These can arise in adults, during prolonged strenuous exercise when skeletal muscle depletes plasma glucose, and produces other metabolic disruptions upon the brain (hypoxia, hyperthermia, dehydration and hyperammonemia). These are proportional to muscle mass.
7.	Children show specific adaptations to minimize such metabolic disturbances. (i) Due to slow body growth and resulting small body size, they have limited skeletal muscle mass. (ii) They show other adaptations such as an exercise specific preference for free fatty acid metabolism. (iii) While children are generally more active than adolescents and adults, they avoid physically prolonged intense exertion. 
8.	Childhood has a close relationship to high levels of energy provision and metabolic adaptations that support prolonged synaptic neurodevelopment. 
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