title: prosc-affy-rat-185420

status: Public on Jan 13 2006

city: Phoenix

institute: Translational Genomics

summary:
The goal of the research proposed herein is to identify (i) gene products that allow identification of specific precursor populations in vivo and (ii) genetic programs that regulate the generation of lineaege-restricted precursor (LRP) cells from neuroepithelial stem cells (NSC).  As these cells play a major role in development and tissue repair, this effort will enhance both basic and clinically relevant research. Note: Funding for this micro-array analysis was granted through NINDS as supplement 3R01NS042820-02S1.
Our ability to generate essentially unlimited numbers of purified rodent NSCs, thier restricted decendants leading the generation of macroglia - and also to control the generation of each of these cell types from its immediate ancestor - provides us with a unique system for studying the multi-step nature of differentiation. Using defined conditions, samples will be isolated from each type of purified cell population. Expression profiles will be compared to reveal cell type specific markers and regulatory factors.
The generation of differentiated cell types from neuroepithelial stem cells appears to require progression through a sequence of lineaege-restricted intermediate cell stages.  In the case of at least one of the pathways involved in oliigodendrocyte generation, it now appears that all of the stages of this sequence have been defined and all of the intermediate cells have been isolated (as detailed in the Noble, M. et al. (2004)  Dev. Biol. 265, 33-52).  Studies in vitro, and consequent to cell transplantation, have enabled us to gain an increasingly detailed understanding of the differentiation potential of these cells. The microarray analysis proposed herein will provide the specific markers that will enable the LRP cells to be unambiguously identified in vivo, thereby setting the stage to develop a detailed mapping of the role of these cells during development.
RNA will be isolated from purified cell populations.  All populations will initially be derived from spinal cord.  Preparation of purified populations of each of these cell types has been described in multiple of our previous publications (Mayer, M. et al. (1994) Development 120, 142-153; Mayer-Pröschel, M. et al. (1997) Neuron 19, 773-785; Rao, M. and Mayer-Pröschel, M. (1997) Dev. Biology 188, 48-63). Total RNA from purified cell cultures will be extracted in a two step process. RNA will first be extracted by dounce homogenization in phenol/guanidine isothiocyonate solution (Trizol, Invitrogen, California) followed by isopropanol precipitation. Contaminating traces of DNA and protein will then be removed by silica-based affinity chromatography and Dnase treatment (NucleoSpin RNA Purification system, Clontech, California). Depending on the cell type, 5x105 cells will typically yield 5-15µg of high quality RNA (OD260/280=2.1 at pH7.5). In order to determine purity and molecular weight distribution, samples will be routinely tested on a Agilent Bioanalyzer. cDNA will be generated from total RNA and then be submitted to one round of amplification and labeling using the messageAmp system (Ambion). Probes are then hybridized to the Affymerix Rat 230 (A/B or 2.0) GeneChip expression array. Array data will be processed using the Micro Array Suite 5.0 (MAS, Affymetrix). Cel files containing normalized, relative expression data for all genes tested will be used for downstream statistical analysis.
Keywords: other

first: Christoph

last: Proschel


title: The transcriptome of the hypothalamoneurohypophyseal system in euhydrated and dehydrated rats

status: Public on Jan 31 2006

city: Bristol

first: DAVID

last: MURPHY

state: AVON

address: Whitson Street

department: Medicine and Dentistry

country: United Kingdom

institute: University of Bristol

summary: This SuperSeries is composed of the SubSeries listed below.

overall_design: Refer to individual Series


title: A Bioinformatic Analysis of Arginine-Sensitive Regulation of rat Hepatic Gene Expression

status: Public on Dec 31 2005

city: Providence

first: Xianne

last: Leong

state: RI

department: Medical Oncology

country: USA

institute: Rhode Island Hospital; Brown University

summary:
Goals of the Study:
1. Assess the scope of arginine-responsive hepatic gene expression using in vitro rat models.
2. Compare normal and tumorigenic cells
3.  Identify potentially novel genes and pathways that may be subject to amino acid (arginine) regulation

Background:  We previously reported that mRNA levels of the tumor associated glycoprotein amino acid transporter TA1/LAT1/ CD98 light chain arginine increase in normal hepatic cells under low arginine conditions while levels are constitutive and high in hepatic tumor cells.  This suggested LAT1 amino acid response was associated with the normal hepatic phenotype and lost in carcinogenesis and may impact cell growth and survival in the tumor microenvironment.  We sought to investigate how many and what types of genes are responsive to a change in arginine levels over 18 hrs using an in vitro model system.

Experimental design:
Differential gene expression was determined by microarrays using samples from triplicates of normal and transformed cells subjected to 18 hour arginine-deprivation compared to controls
Keywords = arginine
Keywords = hepatocyte
Keywords = cancer
Keywords = rat
Keywords = amino acid
Keywords = gene regulation
Keywords = microarray
Keywords: repeat sample

first: Hwei Xian

first: Anne

last: Lesieur-Brooks

first: Carl

last: Simkevich

first: Bonnie

last: Lau

first: Celine

last: Fugere

first: Nancy

last: Thompson


title: Expression profile between mast cells from diabetic prone and diabetic resistant rat strains

status: Public on Jun 09 2006

city: Milwaukee

first: Lisa

middle: Ann

last: Meyer

state: WI

address: 8701 Watertown Plank Rd

department: Pediatrics

institute: Medical College of Wisconsin

summary:
 Abstract
   Human type 1 diabetes mellitus (T1DM) arises through autoimmune destruction of pancreatic β cells and is modeled in many respects by the lymphopenic and spontaneously diabetic BioBreeding (BB) DRlyp/lyp rat. Previously, pre-onset expression profiling of whole DRlyp/lyp pancreatic lymph nodes (PLN) revealed innate immune activity, specifically that of mast cells and eosinophils. Furthermore, we observed that pancreatic islets of DRlyp/lyp rats as well as those of diabetes-inducible BB DR+/+ rats potentially recruit innate cells through eotaxin expression. Here we determine that lifelong eotaxin expression begins before 40 days of life and localized specifically to β cells.  In this report, we find PLN mast cells more abundant in DRlyp/lyp compared to related BB DR+/+ rats (2.1 +/-0.9% versus 0.9+/-0.4% of total cells, p<0.0001). DRlyp/lyp PLN mast cell gene expression profiling revealed an activated population and included significant overrepresentation of transcripts for mast cell protease 1, cationic trypsinogen, carboxypeptidase A, IL-5, and phospholipase Cγ. In the DR+/+ rat, which develops T1DM upon depletion of TREG cells, mast cells displayed gene expression consistent with the negative regulation of degranulation, including significant overrepresentation of transcripts encoding tyrosine phosphatase SHP-1, lipid phosphatase SHIP, and E3 ubiquitin ligase c-Cbl. To recapitulate negative mast cell regulation observed in the DR+/+, we treated DRlyp/lyp rats with the mast cell “stabilizer” cromolyn which significantly (p<0.05) delayed T1DM onset. These findings are consistent with a growing body of evidence in humans and animal models where a role for mast cells in the initiation and progression of autoimmune disease is emerging.
Keywords: Expression profile between disease and control

overall_design: Rat mast cells were isolated from freshly harvested, female d65 DR+/+ and DRlyp/lyp PLN (n=8 female animals per group) and total RNA was extracted using TRIzol Reagent (Invitrogen Life Technologies, Carlsbad CA). The GeneChip® Rat Genome 230 2.0 array was selected for this study and possesses more than 31,000 probes sets, representing more than 17,734 unique UniGenes.  Purified RNA (~50ng) was amplified using the Two-Cycle cDNA Synthesis Kit (Affymetrix, 900432) and cRNA was synthesized, labeled, fragmented, and hybridized to the Rat Genome 230 2.0 array in accordance to standard Affymetrix protocols (Affymetrix, Santa Clara, CA). Each RNA pool was analyzed in duplicate from independent RNA amplifications (4 arrays total). After hybridization, arrays were washed, stained with PE-conjugated streptavidin (Molecular Probes, Eugene, OR), and scanned


title: Comprehensive description of the transcriptome of hypothalamo-neurohypophyseal system in euhydrated and dehydrated rat

status: Public on Dec 01 2005

summary:
The hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei  are important integrative structures that regulate co-ordinated  responses to perturbations in cardiovascular homeostasis. Through  descending projections from parvocellular neurons to the brainstem,  the PVN acts as an autonomic 'premotor nucleus', regulating reflex  changes in sympathetic nerve activity that are involved in blood  pressure and blood volume regulation. Endocrine responses are  mediated through axonal projections from SON and PVN magnocellular  neurons (MCNs) to the capillaries of the posterior pituitary neural  lobe. In response to dehydration, a massive release of the  antidiuretic peptide hormone vasopressin (VP) into the circulation is  accompanied by a dramatic functional remodelling of the HNS. We have  used microarrays to comprehensively catalogue the genes expressed in  the PVN, the SON and the neurointermediate lobe (NIL) of the  pituitary gland. Further, we have identified transcripts that are  regulated as a consequence of dehydration, as well as RNAs that are  enriched in either the PVN or the SON. We suggest that these  differentially expressed genes represent candidate regulators and  effectors of HNS activity and remodelling.
Keywords: Dehydration

overall_design: In total, 10 Affymetrix Genechip Rat Genome 230 2.0 were used. The experiment compared hypothalamic supraoptic nucleus from 3 day dehydrated and control male Sprague Dawley rats (10-12 weeks). For each chip, tissue from 5 animals was pooled prior to extraction of total RNA and in total 5 chips were used for each condition using  independently prepared RNA samples from separate groups of animals.

first: JULIAN

last: PATON

first: CHARLES

middle: C

last: HINDMARCH

first: SONG

last: YAO

first: GEMMA

last: BEIGHTON


title: Expression QTL (eQTL) mapping in the anterior pituitary gland using DES-treated COPxACI F2 rats.

status: Public on Mar 31 2006

city: Madison

first: James

last: Shull

address: 1400 University Avenue

department: McArdle Laboratory for Cancer Research

institute: University of Wisconsin-Madison

summary:
Anterior pituitary glands were isolated from 21 week old male rats treated for 12 weeks with the synthetic estrogen diethylstilbestrol (DES). 43 COPxACI F2 animals were selected based on pituitary weight phenotypic extremes: 22 highest and 21 lowest pituitary weights in our F2 population. Expression profiles were determined using Affymetrix Rat Genome 230 v. 2.0 arrays. In conjunction with genomewide genotypes available for these 43 animals, linkage analysis was done using mRNA abundance of each transcript on the array as a separate quantitative phenotype. These arrays provide a resource for expression QTL (eQTL) mapping on a genomewide level in the pituitary gland.
Keywords: Estrogen Response

overall_design: 43 samples from anterior pituitary gland were analyzed. COPxACI F2 male animals were treated with DES for 12 weeks, starting at 9 weeks of age. Pituitary glands were harvested and weighed at sacrifice (21 weeks of age). The 43 samples were selected for pituitary weight phenotypic extremes: 22 highest and 21 lowest pituitary weights in our F2 population. Genomewide genotypes are available for these animals and in conjunction with the array data, genomewide expression QTL (eQTL) mapping was done.

first: Karen

middle: E

last: Deffenbacher

middle: D


title: sabba-affy-rat-168529

first: Winnie

last: Liang

state: AZ

address: 445 N. Fifth Street

summary:
Stress is a major risk factor for many cardiovascular and neuropsyhological disorders, and involved changes in gene expression
Determine target genes affected by single or repeated immboilization stress.
Stress triggers alterations in gene expresssion of many proteins, especially transcription factors. The target genes of repeated stress partially overlap, but differ from those altered by single stress.
Sprague Dawley male rats were exposed to single 2 hr immobilization stress or 2 hrs immbolization stress each for six consecultive days. Controls for single stress were untreated, and controls for repeated stress were handled for six days.
Keywords: dose response

first: Esther

last: Sabban


title: Microarray analysis of MEHP gavaged PND28 male rats

status: Public on Apr 01 2006

city: Research Triangle Park

first: Kamin

last: Johnson

state: NC

address: 6 Davis Drive

department: Biological Sciences

institute: CIIT Centers for Health Research

summary:
In rodent models, phthalate exposure alters both the fetal and pubertal testis, but the resulting histopathological changes are divergent.  This suggests that the underlying molecular and cellular phthalate mechanism may be age-dependent.  Using genome-wide expression profiling of acutely-exposed rats, the initial molecular response in pubertal rat testis following in vivo phthalate exposure was determined. For this study, postnatal day 28 rats were exposed to a single dose of 1 g/kg mono-(2-ethyl)hexyl phthalate (MEHP) and assayed at 1, 2, 3, 6, and 12 hrs thereafter using Affymetrix Rat Genome 230 2.0 Arrays.
Keywords: Time course, single dose

overall_design: At each timepoint, testes from 3 treated and 3 control (corn oil gavage) were analzyed.

middle: J

first: Stephanie

middle: A

last: Lahousse

first: Duncan

middle: G

last: Wallace

first: Linda

last: Pluta


title: Sabban-5R01NS028869-11

status: Public on Jul 06 2005

summary:
Stress leads to important adaptive physiological responses including release of catecholamines from the adrenal medulla. Repeated or prolonged stress increases the propensity for many disorders including cardiovascular disease and neuropsychiatric disorders. The susceptibility to stress is determined by a number of factors including prior experience with stress. It has been found that while there is adaptation to prolonged cold stress; it leads to sensitization when cold stressed animals are exposed to a different (novel) stressor. Our study aims to determine the mechanism of this adaptation and sensitization.
We will determine the gene expression patterns in rat adrenal medulla in response to several times of cold stress. We will also determine the changes in gene expression with single immobilization stress in adrenal medulla of rat with and without preexposure to cold stress to investigate the mechanism of sensitization in cold adapted animals.
We hypothesize that during the course of cold stress there are alterations in gene expression of several neurotransmitter related genes and transcription factors. In cold adapted animals, there is increased susceptibility to stress due to sustained changes in expression of several transcription factors, such as FRA-2 which lead to an exaggerated response to the novel stressor.
Rats were exposed to cold stress (4oC) for 1, 7 or 28 days. A parallel group was kept at room temperature (unstressed). Unstressed or rats pre-exposed to 28 days cold were immobilized for 2 hrs and killed by decapitation. The adrenal medullae, separately from right and left side, have been dissected and individually frozen at -70oC. Our recent immunoblots with the tissue from one side revealed that there is increase in tyrosine hydroxylase and c-fos with the short term cold stress and in dopamine beta hydroxylase and Fra-2 even with the prolonged cold stress. The response of several transcription factors to immobilization stress was enhanced in the cold adapted animals. RNA will be prepared with RNAqueous-Micro Isolation kit from Ambion (alternatively we can ship the tissues). We will pool samples from 4 rats per treatment to control for variability.
Keywords: dose response

overall_design: as above

first: E


title: Comprehensive description of the transcriptome of neurointermediate lobe in euhydrated and dehydrated rat

first: DAVID

last: MURPHY

summary:
The hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei  are important integrative structures that regulate co-ordinated  responses to perturbations in cardiovascular homeostasis. Through  descending projections from parvocellular neurons to the brainstem,  the PVN acts as an autonomic 'premotor nucleus', regulating reflex  changes in sympathetic nerve activity that are involved in blood  pressure and blood volume regulation. Endocrine responses are  mediated through axonal projections from SON and PVN magnocellular  neurons (MCNs) to the capillaries of the posterior pituitary neural  lobe. In response to dehydration, a massive release of the  antidiuretic peptide hormone vasopressin (VP) into the circulation is  accompanied by a dramatic functional remodelling of the HNS. We have  used microarrays to comprehensively catalogue the genes expressed in  the PVN, the SON and the neurointermediate lobe (NIL) of the  pituitary gland. Further, we have identified transcripts that are  regulated as a consequence of dehydration, as well as RNAs that are  enriched in either the PVN or the SON. We suggest that these  differentially expressed genes represent candidate regulators and  effectors of HNS activity and remodelling.
Keywords: DEHYDRATED - NIL


title: Comprehensive description of the transcriptome of PVN in euhydrated and dehydrated rat

summary:
The hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei  are important integrative structures that regulate co-ordinated  responses to perturbations in cardiovascular homeostasis. Through  descending projections from parvocellular neurons to the brainstem,  the PVN acts as an autonomic 'premotor nucleus', regulating reflex  changes in sympathetic nerve activity that are involved in blood  pressure and blood volume regulation. Endocrine responses are  mediated through axonal projections from SON and PVN magnocellular  neurons (MCNs) to the capillaries of the posterior pituitary neural  lobe. In response to dehydration, a massive release of the  antidiuretic peptide hormone vasopressin (VP) into the circulation is  accompanied by a dramatic functional remodelling of the HNS. We have  used microarrays to comprehensively catalogue the genes expressed in  the PVN, the SON and the neurointermediate lobe (NIL) of the  pituitary gland. Further, we have identified transcripts that are  regulated as a consequence of dehydration, as well as RNAs that are  enriched in either the PVN or the SON. We suggest that these  differentially expressed genes represent candidate regulators and  effectors of HNS activity and remodelling.
Keywords: DEHYDRATION, PVN


title: Effect of DES-treated Ept congenic rat lines on gene expression in the anterior pituitary gland.

summary:
Anterior pituitary glands were isolated from 21 week old male rats treated for 12 weeks with the synthetic estrogen diethylstilbestrol (DES). Three biological replicates were prepared for each of 4 congenic lines: Ept1, Ept2, Ept6, and Ept9. Congenic rat lines were constructed by introgressing COP alleles onto an ACI background within a given Ept locus. Expression profiles were determined using Affymetrix Rat Genome 230 v. 2.0 arrays. Comparison of congenic rats with the ACI and COP parental strains (see series GSE4028) will allow identification of genes whose expression is influenced by an estrogen-induced pituitary tumor (Ept) QTL.
Keywords: Estrogen Response

overall_design: 12 samples from anterior pituitary gland were analyzed. For each of 4 congenic rat lines (Ept1, Ept2, Ept6, Ept9) there were 3 biological replicates of DES-treated male animals. Congenic animals were constructed by introgressing COP alleles onto an ACI background. This was done within the linkage interval for a given Ept locus. These congenic samples can be compared to the ACI and COP parental strains (see GSE4028) to determine expression differences due to a particular chromosomal region, or QTL.


title: d'mel-affy-rat-168311

summary:
Neurological diseases disrupt the quality of the lives of patients and often lead to their death prematurely.  A common feature of most neurological diseases is the degeneration of neurons, which results from an inappropriate activation of apoptosis. Drugs that inhibit neuronal apoptosis could thus be candidates for therapeutic intervention in neurodegenerative disorders.  We have identified (and recently reported) a chemical called GW5074 that inhibits apoptosis in a variety of cell culture paradigms of neuronal apoptosis.  Additionally, we have found that GW5074 reduces neurodegeneration and improves behavioral outcome in a mouse model of Huntington's disease.  Although GW5074 is a c-Raf inhibitor, we know very little about the molecular mechanisms underlying its neuroprotective action.  Identifying genes that are regulated by GW5074 in neurons will shed insight into this issue.
We believe that neuroprotection by GW5074 involves the regulation of several genes.  Some of these genes are likely to be induced whereas the expression of other genes might be inhibited.  The specific aim is to identify genes that are differentially expressed in neurons treated with GW5074.
We believe that neuroprotection by GW5074 involves the regulation of several genes.  Some of these genes are likely to be induced whereas the expression of other genes might be inhibited.
Cultures of cerebellar granule neurons undergo apoptosis when switched from medium containing levated levels of potassium (high K+ or HK) to medium containing low potassium (LK).  Although cell death begins at about 18 h, we have found that the by 6 h after LK treatment these neurons are irreversibly committed to cell death.  We will treat cerebellar granule neuron cultures with LK medium (which induces them to undergo apoptosis) or with GW5074 (1 uM).  We will extract RNA at two time-points after treatment: 3 h and 6 h. Analysis at the two different time-points will show us whether changes in expression of specific genes is transient or sustained or whether the changes occurs early or relatively late in the process.  Neuronal cultures will be prepared from 1 week old Wistar rats.  The cultures will be maintained in culture for 1 week before treatment.  Following treatment the cells will be lysed and total RNA isolated.  The RNA will be stored at -80oC and shipped to the microarray facility for analysis.  The experiment will be done in triplicate.  Thus, for each time-point (3 or 6h treatment) we will be hope to provide 3 sets of samples (each set coming from a different culture preparation and containing lysates from cells treated with LK or GW5074).  Having samples from 3 independent cultures will mitigate any expression differences resulting from subtle variations in culture quality or in the preparation or quality of RNA.
Keywords: dose response

first: S

middle: R

last: D'Mello


title: dingl-affy-rat-50847

summary:
Epilepsy is a major neurological disorder that affects approximately 1% of the population. The processes that lead to the development of epilepsy (epileptogenesis) are largely unknown. Levetiracetam is a novel antiepileptic drug (AED) that in the kindling model inhibits epileptogenesis in addition to being effective in controlling established epilepsy. The mechanisms of action of levetiracetam as an AED and an antiepileptogenic drug are unknown. By identifying the effect of chronic levetiracetam therapy on gene expression in the brain we hope to be able to identify genes that are involved in epileptogenesis. By comparing the gene expression profiles of levetiracetam and phenytoin treatments, we hope to be able to distinguish between genes that are important for the antiepileptic (anti-seizure) effect and genes that are important for the antiepileptogenic effect of levetiracetam. Phenytoin is a well-established AED; its mechanism of action involves inhibition of sodium channels. In contrast to levetiracetam, available data suggest that phenytoin in certain situations may enhance rather than inhibit the development of epilepsy.
We will identify changes in gene expression in the hippocampus, frontal cortex, and brain stem caused by chronic treatment (3 months) with levetiracetam or phenytoin. We will search for differences in the gene expression profiles after chronic treatment with the two drugs, to explain the different clinical effects of the two drugs.
We hypothesize that chronic treatment with levetiracetam leads to changes in the expression of genes that are involved in both the antiepileptogenetic and antiepileptic effect of the drug, whereas chronic treatment with phenytoin affects genes that may be related to pro-epileptogenetic as well as antiepileptic effects of that drug. We further hypothesize that i) regional differences in the effects of levetiracetam and phenytoin may reflect important functional differences in the actions of these two drugs, and ii) there may be different mechanisms involved in their anti-seizure effect.
Male Wistar rats receive levetiracetam (150 mg/kg x 2), phenytoin (75 mg/kg x 2) or control solution (N=7 in each group) through a gastric tube twice daily for 90 days. Experimental animals and controls are housed together in the same cages in groups of three, facilitating paired comparisons among the three groups in animals that have experienced the same environment throughout the 3 month experimental period. Four hrs after the last dose animals are anesthetized, decapitated, and total RNA is extracted from brain stem, frontal cortex, and hippocampus. RNA will be sent to the TGEN facility for hybridization to the rat U230A chips. The number of chips needed is (2 drugs + 1 control) * (3 brain regions) * (7 animals per group) = 63 chips. We will initially select those genes for analysis that are called “present” by the Affymetrix software in 4 or more animals for at least one of the 6 experimental groups. Then t-tests with Benjamini & Hochberg correction for protection against multiple comparisons will be used to identify genes that are differentially expressed at a false discovery rate of .05 among any of the six groups. We will be primarily interested in the following expression profiles, analyzed as paired comparisons by the Affymetrix software:

1. phenytoin vs control in hippocampus
2. phenytoin vs control in brain stem
3. levetiracetam vs control in hippocampus
4. levetiracetam vs control in brain stem

We will then compare the profiles of both drugs in each brain region (e.g., phenytoin vs levetiracetam in hippocampus) to select genes that are drug-specific. We will also take a Venn diagram approach to identify genes that are drug-specific across both brain regions tested.
Keywords: other

first: Ray

last: Dingledine


title: Fat-type specific effects of high fat diets

status: Public on Oct 28 2005

city: Regensburg

first: Roland

last: Buettner

address: Franz-Josef-Strauss-Allee 11

department: Internal Medicine I

country: Germany

institute: University of Regensburg

summary:
High fat diet (HF) rodent models have contributed significantly to the dissection of the pathophysiology of the insulin resistance syndrome, but their phenotype varies distinctly between different studies. Here, we have analyzed gene expression patterns in livers of animals fed with different HF with varying fatty acid compositions.
Keywords: diet-gene interaction

overall_design: Male Wistar rats were fed with high fat diets (42 energy%, fat sources: lard, olive oil; coconut fat; cod liver oil). Weight, food intake, whole body insulin tolerance and plasma parameters of glucose and lipid metabolism were measured during a 12 week diet course. Liver histologies and hepatic gene expression profiles using AffymetrixR gene chips were obtained.

first: Klaus

last: Parhofer

first: Matthias

last: Woenckhaus

first: Christian

last: Wrede

first: Leoni

last: Kunz-Schughart

first: Jurgen

last: Scholmerich

first: Leo

last: Bollheimer


title: Ventral hippocampal lesion, tetrodotoxin disruption of the ventral hippocampus, and chronic administration of neuroleptics (colan-affy-rat-89421)

first: Winnie

summary:
In pevious research we have shown that the disruption of the normal development of the ventral hippocampus in rodents leads to cellular abnormalities in the frontal cortex and behavioral deficits related to schizophenia (Neurotox Res. 2002, 4(5-6):469-475). We propose the use of gene expression analysis to investigate the molecular underpinnings of these processes which may shed light on the molecular processes relevant to human schizophrenia. In addition, we seek to characterize expression differences induced by chronic administration of antipsychotic medications, which may give insight into the molecular processes involved in ameliorating psychotic symptoms.
Using both surgical and drug interventions, we aim to examine experimentally induced expression differences in the rodent  brain that are relevant to human neuropsychiatric disorders.
Disruption of the normal development of the vental hippocampus or chronic neuroleptic administration,  will result in gene expression changes in the frontal cortex of rats. Elucidation of the molecular cascades underlying these treatments will shed light on both the pathoetiology (from the lesion experiments) and theurapeutic processes (from the antipsychotic treatment experiments) involved in human schizophrenia.
We have developed 3 groups of samples:
1] Ventral hippocampal lesion in neonatal rats (treatment and controls, total N=22)
2] Tetrodotoxin disruption of the ventral hippocampus (treatment and controls, total N=20)
3] Chronic administration of neuroleptics (multiple drugs, multiple doses, and controls, total N=63)
Grand Total N=105

first: Carlo

last: Colantuoni


title: H_Alessio_05: sedentary and active lifestyles

status: Public on Jun 24 2005

city: Oxford

first: Natalie

last: Schweitzer

state: OH

institute: Miami University

summary:
The interaction of genetics and environment play a major role in health and disease. Therefore, modeling experiments to investigate gene-environment interaction in a laboratory setting would provide the opportunity to gain mechanistic insight under controlled conditions. In laboratory research it is unclear if the gene-environment interaction of a sedentary lifestyle predisposes laboratory animals to hypokinetic diseases.
Keywords: repeat sample

overall_design: We are seeking to compare gene expression and physiological markers of health and wellness in animals with: i. a sedentary lifestyle in a standard cage (CSS1-4); ii. access to twice-weekly physical activity in a large box outside a standard cage (CSA1-4); and iii. access to regular exercise in cages with running wheels (CSR1-4).

first: Helaine

middle: M

last: Alessio

middle: B

first: Ann

last: Hagerman


title: Ogle-5P01NS037520-05

last: Liang

summary:
Prolonged stress has adverse consequences for neurons in the hippocampus (HIPP). The amygdala (AMY) is a brain region that is similar to HIPP across many measures, suggesting that chronic stress might modulate AMY and HIPP function in similar ways. However, studies addressing this issue have produced surprising results. For example, a regimen of chronic stress shown to produce atrophy in HIPP neurons caused dendritic branching in AMY neurons. These and other data have revealed that excessive stress induces fundamentally opposing processes in the AMY and HIPP, such that AMY function is facilitated by levels of stress that produce deleterious effects in HIPP. The mechanisms that contribute to the opposing responses of these brain regions to chronic stress are unknown. Such knowledge may suggest novel directions for pharmacological interventions to protect the HIPP from stress-mediated damage.
We will determine gene expression patterns in the hippocampus and amygdala under basal and stressful conditions, with a particular interest in those genes that are differentially regulated across the two regions.
We hypothesize that genes that are oppositely regulated in the amygdala and hippocampus after stress mediate the opposing functional consequences of chronic stress in these two brain regions.  Moreover, genes which are differentially expressed in these regions under basal conditions may underlie the distinct responses of these regions to stress.
Rats will receive either 30sec of handling (Control group; n=15) or 3hr of immobilization stress (Stress group; n=15) each day for 14 consecutive days.  This stress paradigm has previously been shown to produce opposing effects on dendritic morphology and region-dependent behaviors for  the hippocampus versus amygdala.  Twenty-four hours after the last stress or handling session, the rats will be overanaesthetized, and the brain will be removed and placed in ice-cold buffer.  Tissue will be rapidly dissected from the basolateral complex of the amygdala and the CA3 region of the hippocampus of both hemispheres, flash frozen in liquid nitrogen, and stored at -80C until further processing.  The tissue from each brain region will be pooled across groups, yielding four samples (Control-Hippocampus, Control-Amygdala, Stress-Hippocampus, and Stress-Amygdala).  Total RNA will be extracted using standard methods, and sent to the centers.  Each sample will be run in triplicate, utilizing a total of 12 Affymetrix Rat Genome U34A gene chips.  A triplicate array assay of pooled samples has previously been shown to both substantially increase the sensitivity of detecting genes of interest and reduce the variability associated with individual microarrays.  In addition, the large number of rats used in each group will reduce the variability in gene expression associated with individual responses to chronic stress, as well as variability due to slight anatomical differences in the tissue extracted from each rat.
Keywords: dose response

first: W

last: Ogle


title: p38 MAPK overexpression in heart in vivo.

status: Public on Dec 19 2006

city: University of OULU

first: Jaana

last: Rysä

address: PO BOX 5000

department: Pharmacology and toxicology

country: Finland

institute: University of Oulu

summary:
Mitogen-activated protein kinases (MAPKs) regulate cardiomyocyte growth and apoptosis in response to extracellular stimulation, but the downstream effectors that mediate their pathophysiological effects remain poorly understood. We determined the targets and role of p38 MAPK in the heart in vivo by using local adenovirus-mediated gene transfer of constitutively active upstream kinase mitogen-activated protein kinase kinase 3b (MKK3bE) and wild-type p38α in rats. DNA microarray analysis of animals with cardiac-specific overexpression of p38 MAPK revealed that 264 genes were upregulated more than 2-fold including multiple genes controlling cell division, cell signaling, inflammation, adhesion and transcription. Several previously unknown p38 target genes were found. Using gel mobility shift assays we identified several cardiac transcription factors that were directly activated by p38 MAPK. Finally, we determined the functional significance of the altered cardiac gene expression profile by histological analysis and echocardiographic measurements, which indicated that p38 MAPK overexpression induced gene expression results in cell proliferation, myocardial inflammation and fibrosis. In conclusion, we defined the novel target genes and transcription factors as well as the functional effects of p38 MAPK in the heart. Expression profiling of p38 MAPK overexpression identified cell cycle regulatory and inflammatory genes critical for pathological processes in the adult heart.
Keywords: Gene transfer

overall_design: Left ventricular ene expression profiles three days after MKK3bE + WT p38α gene transfer were compared with those of Lac Z –treated animals by screening Affymetrix Rat Expression Set 230_2.0 Arrays (there are 5 samples in both group).

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