Antisense-mediated isoform switching of steroid receptor coactivator-1 in the central nucleus of the amygdala of the mouse brain

BMC Neuroscience

Onno C Meijer²  Annemieke Aartsma-Rus⁴  Willeke MC van Roon-Mom⁴  Nicole A Datson⁴  Suzanne R de Kreij³  Yves Aubert³  Lisa TCM van Weert³  Gwendolynn Grootaers³  Ioannis Zalachoras¹ 

[1] Postzone C4-R, P.O. Box 9600, Leiden, 2300 RC, The Netherlands;Department of Endocrinology, Leiden University Medical Center, Albinusdreef 2, Building 1, Leiden, 2333 ZA, The Netherlands;Division of Medical Pharmacology, Leiden/Amsterdam Center for Drug Research, Leiden University/Leiden University Medical Center, Leiden, The Netherlands;Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands

关键词: Brain;    Glucocorticoid receptor;    Exon skipping;    Antisense oligonucleotide;    Steroid receptor coactivator-1;   

Others  :  1140611
DOI  :  10.1186/1471-2202-14-5

 received in 2012-09-07, accepted in 2012-12-26,  发布年份 2013

【 摘 要 】

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.

【 授权许可】

   
2013 Zalachoras et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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