Life Sciences Commons^™

Controlling Feeding Behavior By Chemical Or Gene-Directed Targeting In The Brain: What’S So Spatial About Our Methods?, Arshad Khan

Arshad M. Khan, Ph.D.

Intracranial chemical injection (ICI) methods have been used to identify the locations in the brain where feeding behavior can be controlled acutely. Scientists conducting ICI studies often document their injection site locations, thereby leaving kernels of valuable location data for others to use to further characterize feeding control circuits. Unfortunately, this rich dataset has not yet been formally contextualized with other published neuroanatomical data. In particular, axonal tracing studies have delineated several neural circuits originating in the same areas where ICI injection feeding-control sites have been documented, but it remains unclear whether these circuits participate in feeding control. However, comparing …

Prolactin And Fmri Response To Skf38393 In The Baboon, Brad D. Miller, Lauren A. Marks, Jonathan M. Koller, Blake J. Newman, G Larry Bretthorst, Kevin J. Black

Kevin J. Black, MD

Background: This study’s goal was to provide dose-response data for a dopamine agonist in the baboon using standard methods (replicate measurements at each dose, across a range of doses), as a standard against which to subsequently validate a novel pharmacological MRI (phMRI) method. Dependent variables were functional MRI (fMRI) data from brain regions selected a priori, and systemic prolactin release. Necessary first steps included estimating the magnitude and time course of prolactin response to anesthesia alone and to various doses of agonist. These first steps (“time course studies”) were performed with three agonists, and the results were used to select …

Inwardly Rectifying Potassium Channels In Drosophila, Zhuo Luan, Hong-Sheng Li

zhuo luan

Inwardly rectifying potassium channels (Kir) are a special subset of potassium selective ion channels which pass potassium more easily into rather than out of the cell. These channels mediate a variety of cellular functions, including control of membrane resting potential, maintenance of potassium homeostasis and regulation of cellular metabolism. Given the existence of fifteen Kir genes in mammals, current genetic studies using mutant animals that lack a single channel may have missed many important physiological functions of these channels due to gene redundancy. This issue can be circumvented by using a simple model organism like Drosophila, whose genome encodes only …

Alcohol-Induced Il-1beta In The Brain Is Mediated By Nlrp3/Asc Inflammasome Activation That Amplifies Neuroinflammation, Dora Lippai, Shashi Bala, Jan Petrasek, Timea Csak, Ivan Levin, Evelyn Kurt-Jones, Gyongyi Szabo

Gyongyi Szabo

Alcohol-induced neuroinflammation is mediated by proinflammatory cytokines, including IL-1beta. IL-1beta production requires caspase-1 activation by inflammasomes-multiprotein complexes that are assembled in response to danger signals. We hypothesized that alcohol-induced inflammasome activation contributes to increased IL-1beta in the brain. WT and TLR4-, NLRP3-, and ASC-deficient (KO) mice received an ethanol-containing or isocaloric control diet for 5 weeks, and some received the rIL-1ra, anakinra, or saline treatment. Inflammasome activation, proinflammatory cytokines, endotoxin, and HMGB1 were measured in the cerebellum. Expression of inflammasome components (NLRP1, NLRP3, ASC) and proinflammatory cytokines (TNF-alpha, MCP-1) was increased in brains of alcohol-fed compared with control mice. Increased …

Genetics In Non-Genetic Model Systems, Carlos Lois, James Groves

Carlos Lois

The past few decades have seen the field of genetic engineering evolve at a rapid pace, with neuroscientists now equipped with a wide range of tools for the manipulation of an animal's genome in order to study brain function. However, the number of species to which these technologies have been applied, namely the fruit fly, C. elegans, zebrafish and mouse, remains relatively few. This review will discuss the variety of approaches to genetic modification that have been developed in such traditional 'genetic systems', and highlight the progress that has been made to translate these technologies to alternative species such as …

Wandering Neuronal Migration In The Postnatal Vertebrate Forebrain, Benjamin Scott, Timothy Gardner, Ni Ji, Michale Fee, Carlos Lois

Carlos Lois

Most non-mammalian vertebrate species add new neurons to existing brain circuits throughout life, a process thought to be essential for tissue maintenance, repair, and learning. How these new neurons migrate through the mature brain and which cues trigger their integration within a functioning circuit is not known. To address these questions, we used two-photon microscopy to image the addition of genetically labeled newly generated neurons into the brain of juvenile zebra finches. Time-lapse in vivo imaging revealed that the majority of migratory new neurons exhibited a multipolar morphology and moved in a nonlinear manner for hundreds of micrometers. Young neurons …

Genetic Labeling Of Neuronal Subsets Through Enhancer Trapping In Mice, Wolfgang Kelsch, Alberto Stolfi, Carlos Lois

Carlos Lois

The ability to label, visualize, and manipulate subsets of neurons is critical for elucidating the structure and function of individual cell types in the brain. Enhancer trapping has proved extremely useful for the genetic manipulation of selective cell types in Drosophila. We have developed an enhancer trap strategy in mammals by generating transgenic mice with lentiviral vectors carrying single-copy enhancer-detector probes encoding either the marker gene lacZ or Cre recombinase. This transgenic strategy allowed us to genetically identify a wide variety of neuronal subpopulations in distinct brain regions. Enhancer detection by lentiviral transgenesis could thus provide a complementary method for …

Similarity Of Visual Selectivity Among Clonally Related Neurons In Visual Cortex, Gen Ohtsuki, Megumi Nishiyama, Takashi Yoshida, Tomonari Murakami, Mark Histed, Carlos Lois, Kenichi Ohki

Carlos Lois

Neurons in rodent visual cortex are organized in a salt-and-pepper fashion for orientation selectivity, but it is still unknown how this functional architecture develops. A recent study reported that the progeny of single cortical progenitor cells are preferentially connected in the postnatal cortex. If these neurons acquire similar selectivity through their connections, a salt-and-pepper organization may be generated, because neurons derived from different progenitors are intermingled in rodents. Here we investigated whether clonally related cells have similar preferred orientation by using a transgenic mouse, which labels all the progeny of single cortical progenitor cells. We found that preferred orientations of …

Increasing Heterogeneity In The Organization Of Synaptic Inputs Of Mature Olfactory Bulb Neurons Generated In Newborn Rats, Wolfgang Kelsch, Shuyin Sim, Carlos Lois

Carlos Lois

New neurons are added into the mammalian olfactory bulb throughout life, but it remains unknown whether the properties of new neurons generated in newborn animals differ from those added during adulthood. We compared the densities of glutamatergic synapses of granule cells (GCs) generated in newborn and adult rats over extended periods of time. We observed that, whereas adult-born GCs maintained stable cell-to-cell variability of synaptic densities soon after they integrated into the circuit, cell-to-cell variability of synaptic densities of neonatal-born GCs increased months after their integration. We also investigated whether the synaptic reorganization induced by sensory deprivation occurred differently in …

Coincident Generation Of Pyramidal Neurons And Protoplasmic Astrocytes In Neocortical Columns, Sanjay Magavi, Drew Friedmann, Garrett Banks, Alberto Stolfi, Carlos Lois

Carlos Lois

Astrocytes, one of the most common cell types in the brain, are essential for processes ranging from neural development through potassium homeostasis to synaptic plasticity. Surprisingly, the developmental origins of astrocytes in the neocortex are still controversial. To investigate the patterns of astrocyte development in the neocortex we examined cortical development in a transgenic mouse in which a random, sparse subset of neural progenitors undergoes CRE/lox recombination, permanently labeling their progeny. We demonstrate that neural progenitors in neocortex generate discrete columnar structures that contain both projection neurons and protoplasmic astrocytes. Ninety-five percent of developmental cortical columns labeled in our system …

Optogenetic Manipulation Of Neural Activity In Freely Moving Caenorhabditis Elegans, Andrew Leifer, Christopher Fang-Yen, Marc Gershow, Mark Alkema, Aravinthan Samuel

Christopher Fang-Yen

We present an optogenetic illumination system capable of real-time light delivery with high spatial resolution to specified targets in freely moving Caenorhabditis elegans. A tracking microscope records the motion of an unrestrained worm expressing channelrhodopsin-2 or halorhodopsin in specific cell types. Image processing software analyzes the worm's position in each video frame, rapidly estimates the locations of targeted cells and instructs a digital micromirror device to illuminate targeted cells with laser light of the appropriate wavelengths to stimulate or inhibit activity. Because each cell in an unrestrained worm is a rapidly moving target, our system operates at high speed ( …

Human-Specific Histone Methylation Signatures At Transcription Start Sites In Prefrontal Neurons, Hennady Shulha, Jessica Crisci, Denis Reshetov, Jogender Tushir, Iris Cheung, Rahul Bharadwaj, Hsin-Jung Chou, Isaac Houston, Cyril Peter, Amanda Mitchell, Wei-Dong Yao, Richard Myers, Jiang-Fan Chen, Todd Preuss, Evgeny Rogaev, Jeffrey Jensen, Zhiping Weng, Schahram Akbarian

Hsin-Jung Chou

Cognitive abilities and disorders unique to humans are thought to result from adaptively driven changes in brain transcriptomes, but little is known about the role of cis-regulatory changes affecting transcription start sites (TSS). Here, we mapped in human, chimpanzee, and macaque prefrontal cortex the genome-wide distribution of histone H3 trimethylated at lysine 4 (H3K4me3), an epigenetic mark sharply regulated at TSS, and identified 471 sequences with human-specific enrichment or depletion. Among these were 33 loci selectively methylated in neuronal but not non-neuronal chromatin from children and adults, including TSS at DPP10 (2q14.1), CNTN4 and CHL1 (3p26.3), and other neuropsychiatric susceptibility …

Dopamine Transporter Endocytic Trafficking In Striatal Dopaminergic Neurons: Differential Dependence On Dynamin And The Actin Cytoskeleton., Haley Melikian

Haley Melikian

Dopaminergic signaling profoundly impacts rewarding behaviors, movement, and executive function. The presynaptic dopamine (DA) transporter (DAT) recaptures released DA, thereby limiting synaptic DA availability and maintaining dopaminergic tone. DAT constitutively internalizes and PKC activation rapidly accelerates DAT endocytosis, resulting in DAT surface loss. Longstanding evidence supports PKC-stimulated DAT trafficking in heterologous expression studies. However, PKC-stimulated DAT internalization is not readily observed in cultured dopaminergic neurons. Moreover, conflicting reports implicate both classic and nonclassic endocytic mechanisms mediating DAT trafficking. Prior DAT trafficking studies relied primarily upon chronic gene disruption and dominant-negative protein expression, or were performed in cell lines and cultured …

Does Experience In Talking Facilitate Speech Repetition?, Linda Shuster, Donna Moore, Gang Chen, Dennis Ruscello, William Wonderlin

Linda Shuster

No abstract provided.

Pleiotropic And Isoform-Specific Functions For Pitx2 In Superior Colliculus And Hypothalamic Neuronal Development, Mindy Waite

Mindy Waite