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Comparative analysis of Drd1+ Medium Spiny Neurons and Drd2+ Medium Spiny Neurons

ABSTRACT: The cellular heterogeneity of the brain confounds efforts to elucidate the biological properties of distinct neuronal populations. Using Bacterial Artificial Chromosome (BAC) transgenic mice which express EGFP-tagged ribosomal protein L10a in defined cell populations, we have developed a methodology to affinity purify polysomal mRNAs from genetically defined cell populations in the brain. The utility of this approach is illustrated by the comparative analysis of four types of neurons, revealing hundreds of genes that distinguish these four cell populations. We find that even two morphologically indistinguishable, intermixed subclasses of medium spiny neuron display vastly different translational profiles and present examples of the physiological significance of such differences. This genetically targeted Translating Ribosome Affinity Purification (TRAP) methodology is a generalizable method useful for the identification of molecular changes in any genetically defined cell type in response to genetic alterations, disease, or pharmacological perturbations. Keywords: Cell Type Comparison

ORGANISM(S): Mus musculus

PROVIDER: GSE13384 | GEO | 2008/11/14

SECONDARY ACCESSION(S): PRJNA113951

REPOSITORIES: GEO

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Comparative analysis of Drd1+ Medium Spiny Neurons, Drd2+ Medium Spiny Neurons, cocaine treatment

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Project description:The cellular heterogeneity of the brain confounds efforts to elucidate the biological properties of distinct neuronal populations. Using Bacterial Artificial Chromosome (BAC) transgenic mice which express EGFP-tagged ribosomal protein L10a in defined cell populations, we have developed a methodology to affinity purify polysomal mRNAs from genetically defined cell populations in the brain. The utility of this approach is illustrated by the comparative analysis of four types of neurons, revealing hundreds of genes that distinguish these four cell populations. We find that even two morphologically indistinguishable, intermixed subclasses of medium spiny neuron display vastly different translational profiles and present examples of the physiological significance of such differences. This genetically targeted Translating Ribosome Affinity Purification (TRAP) methodology is a generalizable method useful for the identification of molecular changes in any genetically defined cell type in response to genetic alterations, disease, or pharmacological perturbations. Keywords: Cell Type Comparison For each cell population, D1 and D2, three independent TRAP replicates were collected, and total RNA from the immunoprecipitates were amplified and hybridized. Data were normalized with the GC-RMA algorithm, and expression values on each chip were normalized to that chip’s 50th percentile. Data were then converted to log2 scale. We recommend that only genes where more than one sample has a normalized intensity larger than 16 (4 in log2 scale) should be kept in the analysis.

2008-11-14 | E-GEOD-13384 | biostudies-arrayexpress

Comparative analysis of Drd1+ Medium Spiny Neurons, Drd2+ Medium Spiny Neurons, cocaine treatment

Project description:The cellular heterogeneity of the brain confounds efforts to elucidate the biological properties of distinct neuronal populations. Using Bacterial Artificial Chromosome (BAC) transgenic mice which express EGFP-tagged ribosomal protein L10a in defined cell populations, we have developed a methodology to affinity purify polysomal mRNAs from genetically defined cell populations in the brain. The utility of this approach is illustrated by the comparative analysis of four types of neurons, revealing hundreds of genes that distinguish these four cell populations. We find that even two morphologically indistinguishable, intermixed subclasses of medium spiny neuron display vastly different translational profiles and present examples of the physiological significance of such differences. This genetically targeted Translating Ribosome Affinity Purification (TRAP) methodology is a generalizable method useful for the identification of molecular changes in any genetically defined cell type in response to genetic alterations, disease, or pharmacological perturbations. Keywords: Cell Type Comparison For each cell type and treatment (cell = D1 or D2, treatment = acute or chronic, saline or cocaine) three independent TRAP replicates were collected, and total RNA from the immunoprecipitates were amplified and hybridized. Data were normalized with the GC-RMA algorithm, and expression values on each chip were normalized to that chip’s 50th percentile. Data were then converted to log2 scale. We recommend that only genes where more than one sample has a normalized intensity larger than 16 (4 in log2 scale) should be kept in the analysis.

2008-11-14 | E-GEOD-13386 | biostudies-arrayexpress

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