title: Expression of annexin and annexin-mRNA in rat brain under influence of steroid drugs.
abstract: Brain tissue of rats pretreated with methylprednisolone or with the 21-aminosteroid U74389F, and that of untreated control rats, was assessed for the expression of Annexin-1 (Anx-1) and the transcription of its mRNA. For this purpose Anx-1 cDNA was amplified and simultaneously a T7-RNA-polymerase promotor was incorporated into the cDNA using Polymerase Chain Reaction (PCR). Then digoxigenin-11-UTP was incorporated into the transcribed cRNA with T7-RNA-polymerase. With this probe in situ hybridization was carried out in sections of the brain. The probe was visualized by an immunoassay using an anti-digoxigenin antibody conjugate. Anx-1 protein was assessed by means of immunohistochemistry using a polyclonal antibody. The various brain areas of the control animals showed an appreciable amount of Anx-1 at mRNA or protein level; on the other hand, the animals which had been pretreated with either steroid, showed a more intense Anx-1 mRNA signal than the controls in many areas. In the pretreated animals Anx-1 immunostaining was unchanged in cortex, basal ganglia, amygdala and septum, but more intense in hippocampus, hypothalamus and thalamus. In ependyma, choroid plexus, meninges, and vascular walls there was no Anx-1 mRNA transcription detectable. An opposite profile was shown by the Anx-1 immunoreactivity, the protein was present in control animals as well as the steroid-pretreated animals, suggesting that here the protein was either from systemic origin, or has diffused from adjacent structures. The results indicate that Anx-1 mRNA transcription is upregulated by either steroid, and that in the untreated animals there is a resting level of Anx-1 mRNA transcription, presumably reflecting physiological influences on Anx-1 expression.
authors.affiliation: Department of Neurosurgery, University Hospital Groningen, The Netherlands.
authors.forename: P H
authors.initials: PH
authors.lastname: Voermans
authors.forename: K G
authors.initials: KG
authors.lastname: Go
authors.forename: G J
authors.initials: GJ
authors.lastname: ter Horst
authors.forename: M H
authors.initials: MH
authors.lastname: Ruiters
authors.forename: E
authors.initials: E
authors.lastname: Solito
authors.forename: L
authors.initials: L
authors.lastname: Parente
mesh_terms: D000818:Animals; D017305:Annexin A1 / Q000096:biosynthesis / Q000235:genetics*; D001921:Brain / Q000187:drug effects* / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008775:Methylprednisolone / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D011281:Pregnatrienes / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats
chemical_list: D017305:Annexin A1; D011281:Pregnatrienes; D012333:RNA, Messenger; C074965:U 74389F; D008775:Methylprednisolone