Method for quantitative in situ hybridization histochemistry and image analysis applied for Homer1a gene expression in rat brain.
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.
Ambesi-Impiombato
Alberto
A
Laboratory 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.
D'Urso
Giordano
G
Muscettola
Giovanni
G
de Bartolomeis
Andrea
A
D000465:Algorithms; D000704:Analysis of Variance; D000818:Animals; D001345:Autoradiography / Q000379:methods; D002352:Carrier Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D000076122:Disks Large Homolog 4 Protein; D018492:Dopamine Antagonists / Q000494:pharmacology; D001330:Electronic Data Processing; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D006220:Haloperidol / Q000494:pharmacology; D006651:Histocytochemistry / Q000379:methods; D000072233:Homer Scaffolding Proteins; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D017403:In Situ Hybridization / Q000379:methods*; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009479:Neuropeptides / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D016548:Prosencephalon / Q000187:drug effects / Q000378:metabolism*; D015347:RNA Probes / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D013462:Sulfur Radioisotopes / Q000493:pharmacokinetics
D002352:Carrier Proteins; D000076122:Disks Large Homolog 4 Protein; C501449:Dlg4 protein, rat; D018492:Dopamine Antagonists; D000072233:Homer Scaffolding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D015347:RNA Probes; D012333:RNA, Messenger; D013462:Sulfur Radioisotopes; C033652:postsynaptic density proteins; D006220:Haloperidol