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  • Stem cell-derived neural stem/progenitor cell supporting factor is an autocrine/paracrine survival factor for adult neural stem/progenitor cells. 12832409

    Recent evidence suggests that adult neural stem/progenitor cells (ANSCs) secrete autocrine/paracrine factors and that these intrinsic factors are involved in the maintenance of adult neurogenesis. We identified a novel secretory molecule, stem cell-derived neural stem/progenitor cell supporting factor (SDNSF), from adult hippocampal neural stem/progenitor cells by using the signal sequence trap method. The expression of SDNSF in adult central nervous system was localized to hippocampus including dentate gyrus, where the neurogenesis persists throughout life. In induced neurogenesis status seen in ischemically treated hippocampus, the expression of SDNSF was up-regulated. As functional aspects, SDNSF protein provided a dose-dependent survival effect for ANSC following basic fibroblast growth factor 2 (FGF-2) withdrawal. ANSCs treated by SDNSF also retain self-renewal potential and multipotency in the absence of FGF-2. However, SDNSF did not have mitogenic activity, nor was it a cofactor that promoted the mitogenic effects of FGF-2. These data suggested an important role of SDNSF as an autocrine/paracrine factor in maintaining stem cell potential and lifelong neurogenesis in adult central nervous system.
    Document Type:
    Reference
    Product Catalog Number:
    MAB4324

  • Splicing regulator SC35 is essential for genomic stability and cell proliferation during mammalian organogenesis. 17526736

    The members of the SR family of splicing regulators were initially characterized for their critical roles in constitutive and regulated splicing. They are implicated in different aspects of gene expression processes, including transcription, RNA stability, mRNA transport, and translational control. While knockout studies have demonstrated their essential functions during animal development, the pathway(s) leading to a specific cellular phenotype remains poorly understood. We report here that the SR protein SC35 controls cell proliferation during pituitary gland development but is completely dispensable in terminal differentiated mature cardiomyocytes in mice. We show that loss of SC35 in mouse embryonic fibroblasts induces G2/M cell cycle arrest and genomic instability, resulting at least in part from p53 hyperphosphorylation and hyperacetylation. While p53 hyperphosphorylation appears related to ATM activation, its hyperacetylation has been attributed to the increased expression of the acetyltransferase gene p300 and the aberrant splicing of the deacetylase gene SirT1. These findings reveal the involvement of SC35 in specific pathways in regulating cell proliferation and genomic stability during mammalian organogenesis and suggest its potential function in tumorigenesis.
    Document Type:
    Reference
    Product Catalog Number:
    07-131
    Product Catalog Name:
    Anti-Sirt1(Sir2) Antibody

  • Immunohistochemical characterization of chicken pituitary cells containing the vasotocin VT2 receptor. 18548280

    Research in mammals has demonstrated a variety of regulatory effects of vasopressin and oxytocin on endocrine functions of the anterior pituitary gland. Less evidence is available regarding the hypophysiotropic action of arginine vasotocin (AVT) comprising vasopressic and oxytocic activities in birds. Some hypophysiotropic effects of AVT may result from its interactions with brain circuits controlling pituitary functions, whereas others are caused by its direct affect on pituitary cells. Use of an antiserum to the vasotocin receptor VT2 (VT2R) has revealed numerous immunoreactive cells in the anterior pituitary gland of the chicken. The objective of the present study has been to identify endocrine phenotypes of chicken pituitary cells containing VT2R by means of immunohistochemical labeling. VT2R immunoreactivity has been found in all cells immunoreactive for adrenocorticotropin and alpha-melanotropin. Approximately 10% of labeled lactotropes are also immunoreactive for VT2R and lie around the anatomical boundary dividing the cephalic and caudal lobes. In both corticotropes/melanotropes and lactotropes, immunoreactive VT2R is present in a narrow layer outlining cell bodies. Immunoreactive VT2R is not found in gonadotropes, thyrotropes, or somatotropes. These results provide evidence for the important role of VT2Rs in mediating effects of AVT on endocrine secretion from corticotropes and, partially, from lactotropes.
    Document Type:
    Reference
    Product Catalog Number:
    AB5087
    Product Catalog Name:
    Anti-Melanocyte Stimulating Hormone α Antibody

  • Coordination of lipid droplet-associated proteins during the transition period of Holstein dairy cows. 21426973

    Dairy cows often experience negative energy balance with the onset of lactation, and severe or prolonged negative energy balance can contribute to declines in overall fitness. Energy stores, in the form of adipose tissue triacylglycerides, are mobilized during times of energy deficit, and recent research has implicated several proteins associated with the lipid droplet as lipolytic regulators. The objective of this study was to determine if these novel proteins associated with lipolytic regulation are altered with the changing metabolic demands of lactation. Weekly blood samples were collected from 26 Holstein cows from 21 d before expected parturition through 28 d postpartum, and again at 150 d postpartum. Serum nonesterified fatty acids, glycerol, and β-hydroxybutyrate were measured. Energy balance was calculated from daily feed intake and milk yield, weekly body weight, and monthly milk component measurements. Adipose tissue biopsies were taken 21 d before expected parturition (-21 d) and at 5, 21, and 150 d postpartum. Semiquantitative Western blotting was used to measure abundance of hormone-sensitive lipase (HSL), phosphorylated HSL, perilipin, phosphorylated perilipin (PPLIN), adipose triglyceride lipase (ATGL), and comparative gene identity-58 (CGI-58). Abundance of ATGL was less at 5 and 21 d in milk (DIM) compared with -21 and 150 DIM, even though cows were in negative energy balance and experiencing increased rates of lipolysis in early lactation. In contrast, phosphorylated HSL and PPLIN increased with increasing lipolysis immediately after parturition. Additionally, PPLIN was negatively correlated with milk yield at 5, 21, and 150 d postpartum, and negatively correlated with feed intake and energy balance at 21 d postpartum. This result is consistent with the hypothesis that phosphorylation of perilipin is responsive to signals for increased triaclyglyceride mobilization. Finally, a consistent negative correlation between abundance of perilipin and CGI-58 proteins was observed throughout the transition period. These results confirm that novel lipolytic proteins in adipose tissue are regulated at the level of protein abundance and phosphorylation during the periparturient period and into mid lactation.Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
    Document Type:
    Reference
    Product Catalog Number:
    AB10200

  • Coordination of cell proliferation and anterior-posterior axis establishment in the mouse embryo. 21427142

    During development, the growth of the embryo must be coupled to its patterning to ensure correct and timely morphogenesis. In the mouse embryo, migration of the anterior visceral endoderm (AVE) to the prospective anterior establishes the anterior-posterior (A-P) axis. By analysing the distribution of cells in S phase, M phase and G2 from the time just prior to the migration of the AVE until 18 hours after its movement, we show that there is no evidence for differential proliferation along the A-P axis of the mouse embryo. Rather, we have identified that as AVE movements are being initiated, the epiblast proliferates at a much higher rate than the visceral endoderm. We show that these high levels of proliferation in the epiblast are dependent on Nodal signalling and are required for A-P establishment, as blocking cell division in the epiblast inhibits AVE migration. Interestingly, inhibition of migration by blocking proliferation can be rescued by Dkk1. This suggests that the high levels of epiblast proliferation function to move the prospective AVE away from signals that are inhibitory to its migration. The finding that initiation of AVE movements requires a certain level of proliferation in the epiblast provides a mechanism whereby A-P axis development is coordinated with embryonic growth.
    Document Type:
    Reference
    Product Catalog Number:
    07-145
    Product Catalog Name:
    Anti-phospho-Histone H3 (Ser28) Antibody

  • P2 receptor-mediated signaling in spherical bushy cells of the mammalian cochlear nucleus. 19571200

    Purinoreceptors of the P2 family contribute strongly to signaling in the cochlea, but little is known about the effects of purinergic neurotransmission in the central auditory system. Here we examine P2 receptor-mediated signaling in the large spherical bushy cells (SBCs) of Mongolian gerbils around the onset of acoustically evoked signal processing (P9-P14). Brief adenosine 5'-O-(3-thiotriphosphate) (ATPgammaS) application evoked inward current, membrane depolarization, and somatic Ca2+ signals. Moreover, ATPgammaS changed the SBCs firing pattern from phasic to tonic, when the application was synchronized with depolarizing current injection. This bursting discharge activity was dependent on [Ca2+]i and Ca2+-dependent protein kinase (PKC) activity and is presumably caused by modulation of low-threshold K+ conductance. Activation of P2Y1 receptors could not evoke these changes per se, thus it was concluded that the involvement of P2X receptors seems to be necessary. Ca2+ imaging data showed that both P2X and P2Y1 receptors mediate Ca2+ signals in SBCs where P2Y1 receptors most likely activate the PLC-IP3 (inositol trisphosphate) pathway and release Ca2+ from internal stores. Immunohistochemical staining confirmed the expression of P2X2 and P2Y1 receptor proteins in SBCs, providing additional evidence for the involvement of both receptors in signal transduction in these neurons. Purinergic signaling might modulate excitability of SBCs and thereby contribute to regulation of synaptic strength. Functionally, the increase in firing rate mediated by P2 receptors could reduce temporal precision of the postsynaptic firing, e.g., phase locking, which has an immediate effect on signal processing related to sound localization. This might provide a mechanism for adaptation to the ambient acoustic environment.
    Document Type:
    Reference
    Product Catalog Number:
    Multiple
    Product Catalog Name:
    Multiple