Immunolocalization of Bex protein in the mouse brain and olfactory system

Koo, Jae Hyung; Saraswati, Manda; Margolis, Frank L.

doi:10.1002/cne.20486

Cited by 38 publications

(47 citation statements)

References 25 publications

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“…CaM (20µM, Sigma) was incubated with either recombinant mouse Bex1 protein (20µM) [12] or each of the synthetic Bex1 peptides (20µM, Biopolymer Lab, UMB) in 100mM TrisHCl (pH 7.2) and 0.1mM CaCl 2 or 2mM EGTA for 1 h at the room temperature. The bound complexes were resolved by 15% non-denaturing PAGE and visualized by staining with Coomassie Brilliant Blue.…”

Section: Gel Mobility Shift Assaymentioning

confidence: 99%

“…After centrifugation (14,000 rpm, 30 min) the extract was incubated with calmodulin-agarose in the presence of 0.1mM calcium or 2mM EGTA at 4°C for overnight. The resin was washed with 20 mM HEPES buffer to remove unbound proteins and then extracted with SDS sample buffer and analyzed by Western blot using rabbit antibody against Bex1 [12].…”

Section: Calmodulin-agarose Pull Down Assaymentioning

confidence: 99%

“…Total RNA preparation and RT-PCR were performed as described by Koo et al [12] using the primers shown in Supplementary Material Table 1.…”

Section: Rt-pcrmentioning

confidence: 99%

“…They were homogenized on ice in 10 volumes of 20 mM Hepes/NaOH buffer (pH 7.5) containing protease inhibitor cocktail (Roche, Germany). The preparation of tissue extracts and western blotting were performed as described previously [12] using various antibodies (Supplementary Material Table 2). …”

Section: Western Immunoblotting Of Tissue Extractsmentioning

confidence: 99%

“…The Bex1 gene (Brain Expressed X-linked gene) is located on the X-chromosome and encodes a protein of the same name (Bex1 protein, ~15 kDa), which is expressed in several tissues [12,13]. There are several reasons why a Bex1 and CaM interaction during skeletal muscle regeneration would be important to elucidate.…”

Section: Introductionmentioning

confidence: 99%

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Bex1 knock out mice show altered skeletal muscle regeneration

Koo

Smiley

Lovering

et al. 2007

Biochemical and Biophysical Research Communications

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Bex1 and Calmodulin (CaM) are upregulated during skeletal muscle regeneration. We confirm this finding and demonstrate the novel finding that they interact in a calcium-dependent manner. To study the role of Bex1 and its interaction with CaM in skeletal muscle regeneration, we generated Bex1 knock out (Bex1-KO) mice. These mice appeared to develop normally and are fertile, but displayed a functional deficit in exercise performance compared to wild type (WT) mice. After intramuscular injection of cardiotoxin, which causes extensive and reproducible myotrauma followed by recovery, regenerating muscles of Bex1-KO mice exhibited elevated and prolonged cell proliferation, as well as delayed cell differentiation, compared to WT mice. Thus, our results provide the first evidence that Bex1-KO mice show altered muscle regeneration, and allow us to propose that the interaction of Bex1 with Ca2+/CaM may be involved in skeletal muscle regeneration.

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Section: Gel Mobility Shift Assaymentioning

confidence: 99%

Section: Calmodulin-agarose Pull Down Assaymentioning

confidence: 99%

“…Total RNA preparation and RT-PCR were performed as described by Koo et al [12] using the primers shown in Supplementary Material Table 1.…”

Section: Rt-pcrmentioning

confidence: 99%

Section: Western Immunoblotting Of Tissue Extractsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Bex1 knock out mice show altered skeletal muscle regeneration

Koo

Smiley

Lovering

et al. 2007

Biochemical and Biophysical Research Communications

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Sodium/calcium exchanger expression in the mouse and rat olfactory systems

Pyrski

Koo

Polumuri

et al. 2007

J of Comparative Neurology

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Sodium/calcium (Na(+)/Ca(2+)) exchangers are membrane transport systems that regulate Ca(2+)-homeostasis in many eukaryotic cells. In olfactory and vomeronasal sensory neurons ligand-induced olfactory signal transduction is associated with influx and elevation of intracellular Ca(2+), [Ca(2+)](i). While much effort has been devoted to the characterization of Ca(2+)-related excitation and adaptation events of olfactory chemosensory neurons (OSNs), much less is known about mechanisms that return [Ca(2+)](i) to the resting state. To identify proteins participating in the poststimulus Ca(2+)-clearance of mouse OSNs, we analyzed the expression of three potassium (K(+))-independent (NCX1, 2, 3) and three K(+)-dependent (NCKX1, 2, 3) Na(+)/Ca(2+) exchangers. In situ hybridization showed that mRNAs of all six Na(+)/Ca(2+) exchangers coexist in neurons of the olfactory and vomeronasal systems, and that some are already detectable in the embryo. Of these, NCX1 and NCKX1 represent the most and least abundant mRNAs, respectively. Moreover, immunohistochemistry revealed that the NCX1, 2, and 3 proteins are expressed in nearly all neurons of the olfactory epithelium, the vomeronasal organ, the septal organ of Masera, and the Grueneberg ganglion. These three exchanger proteins display different expression profiles in dendrites, knobs, and plasma membranes of OSNs and in sustentacular cells. Furthermore, we show that NCX1 mRNA in rat olfactory mucosa is expressed as 8 alternative splice variants. This is the first comprehensive analysis of Na(+)/Ca(2+) exchanger expression in the mammalian olfactory system. Our results suggest that Ca(2+)-extrusion by OSNs utilizes multiple different Na(+)/Ca(2+) exchangers and that different subtypes are targeted to different subcellular compartments.

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Mouse olfactory sensory neurons express 10,000 genes

Sammeta

Bose

et al. 2007

J of Comparative Neurology

116

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Olfactory epithelial cells from olfactory marker protein-green fluorescent protein (OMP-GFP) mice were separated by fluorescence-activated cell sorting into a GFP+ sample enriched in mature olfactory sensory neurons (OSNs) and a GFP- sample enriched in all other cells. GeneChip expression profiling of these samples provided a predictive measure of expression in OSNs. Validation tests comparing the ratio of GFP+/GFP- signal intensity against expression patterns from in situ hybridization for 189 mRNAs proved statistically significant and provided probabilities of expression in OSNs scaled according to the signal intensity ratios. These probabilities predict that, among 11,596 mRNAs detected in the GFP+ sample, more than 10,000 are expressed in OSNs. Transcripts and overrepresented categories of mRNAs detected in the GFP+ sample agreed with known properties of OSNs and predict additional properties. For example, ciliogenesis and spermatogenesis were overrepresented, consistent with similarities between OSN cilia and sperm flagella. Chromatin assembly mRNAs were expressed throughout the OSN cell lineage, consistent with the hypothesis that chromatin remodeling plays a role in OSN differentiation. We detected numerous signaling proteins and receptors, such as 30 nonchemosensory G-protein-coupled receptors, including the presynaptic glutamate receptor mGlur4 and the Wnt receptor Fzd3. The largest group of mRNAs, however, was the hundreds of transcriptional regulators that presumably determine the OSN phenotype. The absence of OMP protein in OMP-GFP mice had no detectable effect on mRNA abundance. Within limits prescribed by the nature of microarray data and the in situ hybridization validation, these data should be useful in directing further experiments on OSN function.

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Immunolocalization of Bex protein in the mouse brain and olfactory system

Cited by 38 publications

References 25 publications

Bex1 knock out mice show altered skeletal muscle regeneration

Bex1 knock out mice show altered skeletal muscle regeneration

Sodium/calcium exchanger expression in the mouse and rat olfactory systems

Mouse olfactory sensory neurons express 10,000 genes

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