Induction of annexin-1 at transcriptional and post-transcriptional level in rat brain by methylprednisolone and the 21-aminosteroid U74389F
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 pr...
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| Format: | Article |
| Language: | English |
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Wiley
1996-01-01
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| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/S0962935196000531 |
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| author | P. H. Voermans K. G. Go G. J. Ter Horst M. H. J. Ruiters E. Solito L. Parente |
| author_facet | P. H. Voermans K. G. Go G. J. Ter Horst M. H. J. Ruiters E. Solito L. Parente |
| author_sort | P. H. Voermans |
| collection | DOAJ |
| description | 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 promoter was incorporated into the cDNA using a 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 on sections of the brain. The probe was visualized by an immunoassay using an antidigoxigenin 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 indicated 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. |
| format | Article |
| id | doaj-art-dca6146a3d2646dbb7000a69730e3053 |
| institution | Kabale University |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 1996-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-dca6146a3d2646dbb7000a69730e30532025-02-03T01:26:00ZengWileyMediators of Inflammation0962-93511466-18611996-01-015537037810.1155/S0962935196000531Induction of annexin-1 at transcriptional and post-transcriptional level in rat brain by methylprednisolone and the 21-aminosteroid U74389FP. H. Voermans0K. G. Go1G. J. Ter Horst2M. H. J. Ruiters3E. Solito4L. Parente5Department of Neurosurgery, University Hospital Groningen, Hanzeplein. 1, P.O. Box 30.001, Groningen NL-9700 RB, The NetherlandsDepartment of Neurosurgery, University Hospital Groningen, Hanzeplein. 1, P.O. Box 30.001, Groningen NL-9700 RB, The NetherlandsDepartment of Biological Psychiatry, University Hospital Groningen, Hanzeplein. 1, P.O. Box 30.001, Groningen NL-9700 RB, The NetherlandsBioMedical Technology Center, University Hospital Groningen, Hanzeplein. 1, P.O. Box 30.001, Groningen NL-9700 RB, The NetherlandsInstitut Cochin de Génétique Moléculaire, Paris, FranceIstituto di Farmacología e Farmacognosía, Palermo, ItalyBrain 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 promoter was incorporated into the cDNA using a 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 on sections of the brain. The probe was visualized by an immunoassay using an antidigoxigenin 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 indicated 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.http://dx.doi.org/10.1155/S0962935196000531 |
| spellingShingle | P. H. Voermans K. G. Go G. J. Ter Horst M. H. J. Ruiters E. Solito L. Parente Induction of annexin-1 at transcriptional and post-transcriptional level in rat brain by methylprednisolone and the 21-aminosteroid U74389F Mediators of Inflammation |
| title | Induction of annexin-1 at transcriptional and post-transcriptional level in rat brain by methylprednisolone and the 21-aminosteroid U74389F |
| title_full | Induction of annexin-1 at transcriptional and post-transcriptional level in rat brain by methylprednisolone and the 21-aminosteroid U74389F |
| title_fullStr | Induction of annexin-1 at transcriptional and post-transcriptional level in rat brain by methylprednisolone and the 21-aminosteroid U74389F |
| title_full_unstemmed | Induction of annexin-1 at transcriptional and post-transcriptional level in rat brain by methylprednisolone and the 21-aminosteroid U74389F |
| title_short | Induction of annexin-1 at transcriptional and post-transcriptional level in rat brain by methylprednisolone and the 21-aminosteroid U74389F |
| title_sort | induction of annexin 1 at transcriptional and post transcriptional level in rat brain by methylprednisolone and the 21 aminosteroid u74389f |
| url | http://dx.doi.org/10.1155/S0962935196000531 |
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