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Method for quantitative in situ hybridization histochemistry and image analysis applied for Homer1a gene expression
Alberto Ambesi-Impiombato1, Giordano D'Urso, Giovanni Muscettola
1Laboratory of Molecular Psychiatry and Psychopharmacotherapy, Section of Psychiatry, Department of Neuroscience and Behavioral Science University School of Medicine "Federico II", Edificio 18, Via Pansini 5, 80131 Naples, Italy.
Abstract:
Here we describe the detailed method for quantitative in situ hybridization histochemistry adopted in our previously published short communication on differential gene expression of the postsynaptic density proteins Homer and PSD-95 in rat forebrain after acute treatment with antipsychotic drugs [de Bartolomeis et al., Mol. Brain Res. 98 (2002) 124-129]. Specific 35S radiolabeled oligodeoxyribonucleotides were used to hybridize rat forebrain sections and data analysis was carried out on digitalized images acquired by means of a CCD camera. Special emphasis has been posed on data preprocessing options applied to a dataset obtained using a transparency scanner as an alternative image capturing method.