Deciphering the role of importin alpha in mammalian central neurons - Hippocampal gene expression profiling in importin alpha5-deleted mice
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PROVIDER: PRJNA417195 | ENA |
REPOSITORIES: ENA
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Project description:Introduction: The mechanisms governing synaptonuclear transport, and the transcriptional pathways that are at the core of neuronal disorders are poorly understood. The importin family of nuclear import factors has pivotal roles in such pathways, due to their involvement in intracellular transport from both synapse to soma and cytoplasm to nucleus. Mammals express six different isoforms of importin α, which bind an Importin β to form transport complexes for specific cargoes. Methods: We addressed the roles of importin α isoforms in mammalian CNS by evaluation of a systematic series of importin α knockout mice. A comprehensive battery of behavioral tests was used to investigate spontaneous and novelty-induced locomotion, basal ganglia function, motor coordination, neuromuscular integration, anxiety-related behaviors and memory. Subsequent analyses using acute-brain slice electrophysiology, RNA-seq, bioinformatics, imaging and pharmacology was used in order to delineate the contribution of genes and signaling pathways involved in the identified phenotype and screen for new therapeutic approaches. Results and Discussion: During the course of comprehensive behavioral profiling of importin α mouse mutants, we identified an importin α knockout (KO) mouse with significantly reduced anxiety levels. Electrophysiological recordings in acute hippocampal slices showed a reduced short-term and presynaptic plasticity (paired-pulse facilitation deficit, lower PTP), while LTP was not affected in these mutants. We then performed transcriptional profiling (RNA-seq) from hippocampal extracts of wild type versus mutant animals after exposure to an anxiogenic drug. Computational analysis revealed differences in the expression of genes and the involvement of transcription factors. Drugs that activate LXR or the Sphingosine 1 phosphate receptor have robust anxiolytic effects, while a Sphk1 blocker reverses anxiolysis in the importin α5 knockout mouse. Thus, importin α5 influences anxiety by regulating nuclear import in neurons.
2018-11-22 | GSE106546 | GEO
Project description:To test whether nuclear importin alpha2 can regulate transcription, we sought to examine gene expression changes in cells with nuclear accumulation of importin alpha2 by performing microarray analysis. We designed an experiment in which EGFP-fused full-length importin alpha2 was transfected into HeLa cells. To exclude the possibility that exogenous, full-length importin alpha2 protein enhances nuclear transport of karyophilic proteins such as transcription factors and thereby directly influences gene expression, a mutant of importin alpha2 which is mutated in the C-terminal CAS-binding domain was also transfected, so that it is never recycled to the cytoplasm and shows complete nuclear localization. We undertook to investigate whether there were changes in gene expression common to cells expressing the full-length importin alpha2 and the C-terminal mutant (C-mutant) isoform. EGFP vs. EGFP-importin alpha2 full length, or EGFP vs EGFP-importin alpha2 CAS-binding mutant. One replicate each. EGFP-expressing cells are used as a control.
2010-11-24 | E-GEOD-25303 | biostudies-arrayexpress
Project description:RNA-sequencing of mouse adult hippocampal nestin+ progenitor cells & dcx+ progenitor cells from wild-type animals, and animals in which the gene Nfix was deleted.
2018-01-30 | GSE109817 | GEO
Project description:To test whether nuclear importin alpha2 can regulate transcription, we sought to examine gene expression changes in cells with nuclear accumulation of importin alpha2 by performing microarray analysis. We designed an experiment in which EGFP-fused full-length importin alpha2 was transfected into HeLa cells. To exclude the possibility that exogenous, full-length importin alpha2 protein enhances nuclear transport of karyophilic proteins such as transcription factors and thereby directly influences gene expression, a mutant of importin alpha2 which is mutated in the C-terminal CAS-binding domain was also transfected, so that it is never recycled to the cytoplasm and shows complete nuclear localization. We undertook to investigate whether there were changes in gene expression common to cells expressing the full-length importin alpha2 and the C-terminal mutant (C-mutant) isoform.
2010-11-24 | GSE25303 | GEO
Project description:Here we show that importin alpha (α) 3 (KPNA4) controls pain responsiveness in peripheral sensory neurons. An importin α3 knockout mouse showed reduced responsiveness to noxious heat or chemical pain, as well as greater tolerance to neuropathic pain. Similar findings were obtained upon acute knockdown of importin α3 in peripheral sensory neurons. Transcriptome analyses and immunohistochemistry indicated that importin α 3 deficient neurons were impaired in nuclear import of c-Fos. Forty days after spared nerve injury acute down-regulation of importin α3, c-FOS, Jun and the dominat negative AAV9 viruses rescue the neuropathic pain phenotype. In silico screens identified importin α3 deficiency mimicking drug leads that attenuated neuropathic pain and reduced c-Fos nuclear localization. Hence, perturbing c-Fos nuclear import by importin α3 can provide analgesia at peripheral loci in the nervous system
2020-08-17 | GSE137515 | GEO
Project description:Spermatogenesis, a fundamental process in male reproduction, is driven by an ordered series of events, which rely on the trafficking of specific proteins between nucleus and cytoplasm. The importin α family of proteins mediates movement of specific cargo proteins when bound to importin β. Importin α genes have distinct expression patterns in mouse testis, implying they may have unique roles during mammalian spermatogenesis. Here we use a loss-of-function approach to specifically determine the role of importin α7 (Kpna6) in spermatogenesis and male fertility. We show that ablation of importin α7 in male mice leads to infertility and has multiple cumulative effects on both germ cells and Sertoli cells culminating in oligozoospermia. Importin α7-deficient mice exhibit an impaired Sertoli cell function, including loss of Sertoli cells from the seminiferous epithelium and a compromised nuclear transport of the androgen receptor. Furthermore, our data demonstrate devastating defects in spermiogenesis that are accompanied by a disturbed histone-protamine-exchange in elongating spermatids, resulting in incomplete sperm maturation and a massive loss of sperms. Our work uncovers the essential role of importin α7 in spermatogenesis and hence in male fertility.
2021-09-20 | GSE160969 | GEO
Project description:RNA-sequencing of mouse adult hippocampal progenitor cells in which Nfix was deleted.
| PRJNA432080 | ENA
Project description:Importin-b1, a key member of karyopherins, is composed of 18-20 tandem repeats of the HEAT motif, each of which contains two antiparallel alpha helices linked by a turn. With the help of adaptor protein importin-α, importin-b1 recognizes cargo with NLS in the cytoplasm, and ferries the complex through the NPC into the nucleus. Several studies have shown that importin-b1 is overexpressed in some cancers, especially advanced prostate cancer, and is associated with poor prognosis and drug resistances.Here, the anti-CRPC (castration-resistant prostate cancer) screening of a natural diterpenoid library followed by target fishing led to the identification of a highly potent importin-b1 inhibitor, daphnane diterpenoid DD1(YDGY-3).To further dissect the influence of importin-b1 inhibitors on gene transcription programs, we analyzed H3K27ac chip-seq of C4-2B cells with the treatment of DD1 (YDGY-3).
2022-09-01 | GSE179316 | GEO
Project description:Importin-b1, a key member of karyopherins, is composed of 18-20 tandem repeats of the HEAT motif, each of which contains two antiparallel alpha helices linked by a turn. With the help of adaptor protein importin-α, importin-b1 recognizes cargo with NLS in the cytoplasm, and ferries the complex through the NPC into the nucleus. Several studies have shown that importin-b1 is overexpressed in some cancers, especially advanced prostate cancer, and is associated with poor prognosis and drug resistances.Here, the anti-CRPC (castration-resistant prostate cancer) screening of a natural diterpenoid library followed by target fishing led to the identification of a highly potent importin-b1 inhibitor, daphnane diterpenoid DD1.And the optimized analog is DD1-Br.To further dissect the influence of importin-b1 inhibitors on gene transcription programs, we analyzed RNA-seq transcriptomes of C4-2B cells with the treatment of DD1 (YDGY-3), DD1-Br (YDGY-3BR) or importazole (IPZ).
2022-09-01 | GSE179238 | GEO
Project description:Importin 13 is a bidirectional transport receptor that mediates both nuclear import and export of proteins. To expand the range of human importin 13 cargoes, we performed importin 13 binding experiments in combination with quantitative stable isotope labeling with amino acids in cell culture (SILAC) and mass spectrometry. Apart from known importin 13 interaction partners, such as Ubc9 and eIF1A, more than 300 potential importin 13 cargoes were identified, several of which could be validated using cellular and biochemical assays.
2018-04-20 | PXD008459 | Pride