Antisense-mediated isoform switching of steroid receptor coactivator-1 in the central nucleus of the amygdala of the mouse brain
| العنوان: | Antisense-mediated isoform switching of steroid receptor coactivator-1 in the central nucleus of the amygdala of the mouse brain |
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| المؤلفون: | Ioannis Zalachoras, Annemieke Aartsma-Rus, Nicole A. Datson, Willeke M. C. van Roon-Mom, Suzanne R. de Kreij, Lisa T C M van Weert, Yves Aubert, Onno C. Meijer, Gwendolynn Grootaers |
| المصدر: | BMC Neuroscience BMC Neuroscience, 14, 5 BMC Neuroscience, Vol 14, Iss 1, p 5 (2013) BMC Neuroscience, 14, pp. 5 BMC Neuroscience, 14 |
| سنة النشر: | 2013 |
| مصطلحات موضوعية: | Male, Time Factors, DCN MP - Plasticity and memory, RNA Splicing, Laser Capture Microdissection, Glucocorticoid receptor, Biology, lcsh:RC321-571, Oligodeoxyribonucleotides, Antisense, Exon, Cellular and Molecular Neuroscience, Mice, Nuclear Receptor Coactivator 1, Glial Fibrillary Acidic Protein, medicine, Animals, Protein Isoforms, Antisense oligonucleotide, Receptor, lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry, General Neuroscience, Central nucleus of the amygdala, Methodology Article, lcsh:QP351-495, Brain, Amygdala, Molecular biology, Exon skipping, Cell biology, Nuclear receptor coactivator 1, Mice, Inbred C57BL, Cell nucleus, medicine.anatomical_structure, lcsh:Neurophysiology and neuropsychology, Nuclear receptor, Steroid receptor coactivator-1, Phosphopyruvate Hydratase, Leukocyte Common Antigens |
| الوصف: | Contains fulltext : 125235.pdf (Publisher’s version ) (Open Access) BACKGROUND: Antisense oligonucleotide (AON)-mediated exon skipping is a powerful tool to manipulate gene expression. In the present study we investigated the potential of exon skipping by local injection in the central nucleus of the amygdala (CeA) of the mouse brain. As proof of principle we targeted the splicing of steroid receptor coactivator-1 (SRC-1), a protein involved in nuclear receptor function. This nuclear receptor coregulator exists in two splice variants (SRC-1a and SRC-1e) which display differential distribution and opposing activities in the brain, and whose mRNAs differ in a single SRC-1e specific exon. METHODS: For proof of principle of feasibility, we used immunofluorescent stainings to study uptake by different cell types, translocation to the nucleus and potential immunostimulatory effects at different time points after a local injection in the CeA of the mouse brain of a control AON targeting human dystrophin with no targets in the murine brain. To evaluate efficacy we designed an AON targeting the SRC-1e-specific exon and with qPCR analysis we measured the expression ratio of the two splice variants. RESULTS: We found that AONs were taken up by corticotropin releasing hormone expressing neurons and other cells in the CeA, and translocated into the cell nucleus. Immune responses after AON injection were comparable to those after sterile saline injection. A successful shift of the naturally occurring SRC-1a:SRC-1e expression ratio in favor of SRC-1a was observed, without changes in total SRC-1 expression. CONCLUSIONS: We provide a proof of concept for local neuropharmacological use of exon skipping by manipulating the expression ratio of the two splice variants of SRC-1, which may be used to study nuclear receptor function in specific brain circuits. We established that exon skipping after local injection in the brain is a versatile and useful tool for the manipulation of splice variants for numerous genes that are relevant for brain function. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 1471-2202 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::222d3e3f8573861bf5fe371ac0b51f89 https://hdl.handle.net/1887/98603 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....222d3e3f8573861bf5fe371ac0b51f89 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14712202 |
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