Kindled Seizure‐induced c‐fos and Prodynorphin mRNA Expressions are Unrelated in the Rat Brain

Abstract

Levels of mRNA for c-fos and prodynorphin were studied by in situ hybridization in adjacent coronal sections taken from kindled rats 30-60 min after the last seizure. Within this time frame, expression of both genes was induced in multiple brain areas. Anatomical colocalization of the induced gene expressions was found in the hippocampus. Induction of c-fos in the dentate gyrus was bilateral and symmetrical in a subgroup of rats, ipsilateral in another subgroup and absent in a third subgroup. However, no relative increase was observed in the ipsilateral compared with the contralateral prodynorphin expression in the dentate gyrus when c-fos expression was induced ipsilaterally only. These observations suggest that, at variance with other experimental situations, Fos is not involved in the mechanisms of kindled seizure-induced activation of prodynorphin transcription in the rat forebrain.

Functional antagonism between nociceptin/orphanin FQ (N/OFQ) and corticotropin-releasing factor (CRF) in the rat brain: evidence for involvement of the bed nucleus of the stria terminalis

“…In situ hybridization was performed as previously described (Simonato et al 1996;Rodi et al 2002). The in situ hybridization mixture contained 50% deionized formamide, 2× sodium Tris EDTA (STE), 5× Denhart's solution, 100 mg/ml single-stranded DNA, 100 μg/ml transfer RNA, 0.05% sodium pyrophosphate, and 60 ng/ml [ 33 P]-riboprobe.…”

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“…In situ hybridization was performed as described previously (Simonato et al, 1996). Briefly, sections were incubated overnight at 52°C with 40 l of hybridization mixture (50% deionized formamide, 2 sodiumTris-EDTA, 5ϫ Denhart's solution, 100 g/ml single-stranded DNA, 100 g/ml tRNA, 0.05% sodium pyrophosphate, and 60 ng/ml [ 33 P]-riboprobe).…”

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“…42 The in situ hybridization mixture contained 50% deionized formamide, 0.6 m NaCl, 2 mm EDTA, 20 mm Tris (2 × NTE), 5 × Denhardt's solution, 100 g/ml ssDNA, 100 g/ml tRNA, 0.05% sodium pyrophosphate and 60 ng/ml 33 P-riboprobe. Sections were incubated overnight at 52°C with 40 l hybridization mixture.…”