Quantitative proteomic analysis for relative protein abundance between DRM and non-raft fraction.

<p>Quantitative proteomic analysis for relative protein abundance between DRM and non-raft fraction.</p

Characterization of the functions and proteomes associated with membrane rafts in chicken sperm

<div><p>Cellular membranes are heterogeneous, and this has a great impact on cellular function. Despite the central role of membrane functions in multiple cellular processes in sperm, their molecular mechanisms are poorly understood. Membrane rafts are specific membrane domains enriched in cholesterol, ganglioside G_M1, and functional proteins, and they are involved in the regulation of a variety of cellular functions. Studies of the functional characterization of membrane rafts in mammalian sperm have demonstrated roles in sperm-egg binding and the acrosomal reaction. Recently, our biochemical and cell biological studies showed that membrane rafts are present and might play functional roles in chicken sperm. In this study, we isolated membrane rafts from chicken sperm as a detergent-resistant membranes (DRM) floating on a density gradient in the presence of 1% Triton X-100, and characterized the function and proteomes associated with these domains. Biochemical comparison of the DRM between fresh and cryopreserved sperm demonstrated that cryopreservation induces cholesterol loss specifically from membrane rafts, indicating the functional connection with reduced post-thaw fertility in chicken sperm. Furthermore, using an avidin-biotin system, we found that sperm DRM is highly enriched in a 60 KDa single protein able to bind to the inner perivitelline layer. To identify possible roles of membrane rafts, quantitative proteomics, combined with a stable isotope dimethyl labeling approach, identified 82 proteins exclusively or relatively more associated with membrane rafts. Our results demonstrate the functional distinctions between membrane domains and provide compelling evidence that membrane rafts are involved in various cellular pathways inherent to chicken sperm.</p></div

Binding affinity of the low density DRM to the IPVL.

<p>The DRM (fraction 2 and 3) and non-raft (fraction 8) proteins were biotinylated and then co-incubated with the IPVL. After washing away unbound proteins, the IPVL was treated with FITC-conjugated streptavidin (A). The binding affinity of the DRM and non-raft proteins were measured by quantification of fluorescence intensity using LAS AF software, as described (B). IPVL binding proteins of the low density DRM and non-raft were purified using the NeutrAvidin-biotin system and were processed for immunoblotting. Data were expressed as mean ± SEM (n = 3). The different letters denote significant difference (<i>P</i> < 0.05). Bar = 20 μm.</p

The distribution of lipid contents in the sucrose gradient of DRM isolated from fresh and cryopreserved sperm.

<p>Sperm DRM were separated into fractions based on their relative buoyancies. Numbers denote fractions from top (1) to bottom (8) of the tube, with fraction 1 representing the lowest density. Quantification of cholesterol (A) and G_M1 (B) was performed in the 8 fractions as described. Data are expressed as mean ± SEM (n = 3–6). The different letters denote significant differences between the fractions of the same set (<i>P</i> < 0.05). Asterisks denote significant differences between fresh and cryopreserved sperm (<i>P</i> < 0.05). Fractions of sperm DRM were processed for SDS-PAGE and subjected to immunoblotting for presence of β-tubulin (C). Fresh and cryopreserved sperm were labeled with Filipin III (D) or CTB-Alexa 488 (E). Images were acquired with a same exposure time between fresh and cryopreserved sperm (n = 3). Bar = 10 μm.</p

DRM-specific proteins.

<p>DRM-specific proteins.</p

Calcium Signaling Pathway Is Associated with the Long-Term Clinical Response to Selective Serotonin Reuptake Inhibitors (SSRI) and SSRI with Antipsychotics in Patients with Obsessive-Compulsive Disorder

<div><p>Background</p><p>Selective serotonin reuptake inhibitors (SSRI) are established first-line pharmacological treatments for obsessive-compulsive disorder (OCD), while antipsychotics are used as an augmentation strategy for SSRI in OCD patients who have either no response or a partial response to SSRI treatment. The goal of the present study was to identify genetic variants and pathways that are associated with the long-term clinical response of OCD patients to SSRI or SSRI with antipsychotics.</p><p>Methods</p><p>We first performed a genome-wide association study of 96 OCD patients to examine genetic variants contributing to the response to SSRI or SSRI with antipsychotics. Subsequently, we conducted pathway-based analyses by using Improved Gene Set Enrichment Analysis for Genome-wide Association Study (i-GSEA4GWAS) to examine the combined effects of genetic variants on the clinical response in OCD.</p><p>Results</p><p>While we failed to detect specific genetic variants associated with clinical responses to SSRI or to SSRI with an atypical antipsychotic at genome-wide levels of significance, we identified 8 enriched pathways for the SSRI treatment response and 5 enriched pathways for the treatment response to SSRI with an antipsychotic medication. Notably, the calcium signaling pathway was identified in both treatment responses.</p><p>Conclusions</p><p>Our results provide novel insight into the molecular mechanisms underlying the variability in clinical response to SSRI and SSRI with antipsychotics in OCD patients.</p></div

Pathway-based analysis for the results of GWAS for the treatment response in OCD.

<p>Pathway-based analysis for the results of GWAS for the treatment response in OCD.</p

Association results of GWAS for the treatment response in OCD.

<p>Association results of GWAS for the treatment response in OCD.</p

Treatment response group and clinical symptoms of patients with obsessive-compulsive disorder.

<p>Treatment response group and clinical symptoms of patients with obsessive-compulsive disorder.</p

How ‘Circumpolar’ is Ainu Music? Musical and Genetic Perspectives on the History of the Japanese Archipelago

<div><p>Understanding the cultural and genetic origins of the Ainu of northern Japan has important implications for understanding the history of the Japanese archipelago. Ethnomusicologists have tended to emphasise connections between Ainu music and a ‘circumpolar’ culture area. However, the ‘dual structure’ model from physical anthropology describes the Ainu as descendants of the first inhabitants of Japan with minimal circumpolar influence. To examine Ainu musical diversity empirically from a comparative perspective, we analysed 680 traditional songs from two Ainu and 33 surrounding East Asian and circumpolar populations. The Ainu repertoire contained a majority (∼50%) of unique stylistic song-types and lower frequencies of types shared with circumpolar (∼40%) and East Asian (∼10%) populations. These frequencies were similar to the corresponding frequencies of mitochondrial DNA types within the Ainu gene pool (∼50%, ∼30% and ∼20%, respectively), consistent with an emerging ‘triple structure’ model of Japanese archipelago history.</p></div