DOCSLIB.ORG

A New Model for Vertebrate Mineralization Via Stabilized

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
A New Model for Vertebrate Mineralization Via Stabilized bioRxiv preprint doi: https://doi.org/10.1101/2020.04.08.031989; this version posted April 10, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-ND 4.0 International license. A new model for vertebrate mineralization via stabilized amorphous calcium carbonate for avian eggshell formation Lilian Stapane1, Nathalie Le Roy1, Jacky Ezagal1, Alejandro B. Rodriguez-Navarro2, Valérie Labas3, Lucie Combes-Soia3, Maxwell T. Hincke4, Joël Gautron1* From 1BOA, INRAE, Université de Tours, 37380 Nouzilly, France ; 2Departmento de Mineralogia y Petrologia, Universidad de Granada, 18071 Granada, Spain ; 3UMR PRC, INRAE 85, CNRS 7247, Université de Tours, IFCE, 37380 Nouzilly, France ; 4Department of Innovation in Medical Education, and Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Canada K1H8M5 Running title: New model via stabilized ACC for avian eggshell formation * To whom correspondence should be addressed: BOA, INRAE, Université de Tours, 37380 Nouzilly, France; [email protected]; Tel. +33 2 47 42 75 40. Keywords: Biomineralization, Avian eggshell, extracellular vesicle, ACC transport, matrix proteins. ABSTRACT mineralization of otoconia, which are calcium Amorphous calcium carbonate (ACC) carbonate biomineralized structures present in is an unstable mineral phase, which is all vertebrates and necessary for balance and progressively transformed into aragonite or sensing linear acceleration. calcite in biomineralization of marine invertebrate shells or avian eggshells, respectively. We have previously proposed a Biomineralization is an extremely model of vesicular transport to provide widespread process by which living organisms stabilized ACC in chicken uterine fluid where produce minerals; over 60 different biominerals mineralization takes place. Herein, we report have been identified, including calcium further experimental evidence for this model. carbonates and phosphates (1). The formation We confirmed the presence of extracellular of biominerals requires high local vesicles (EVs) that contain ACC in uterine fluid concentrations of calcium to form the using transmission electron microscopy and crystalline polymorph. Recently, the transient elemental analysis. We also demonstrate high amorphous mineral phase of calcium levels of expression of vesicular markers in the phosphates and carbonates has been recognized oviduct segments where eggshell is formed. as a soluble and highly reactive source to Moreover, proteomics and facilitate the formation of the complex shapes immunofluorescence confirmed the presence of of observed crystalline biomineral structures. major vesicular, mineralization-specific and The importance of amorphous calcium eggshell matrix proteins in the uterus and in carbonate (ACC) as a transient mineral phase purified EVs. We propose a comprehensive role was reported in the calcification of calcium for EVs in eggshell mineralization, in which carbonate (CaCO3) biominerals (calcite, annexins transfer calcium into vesicles and aragonite) (2-5). ACC is a metastable carbonic anhydrase 4 catalyzes the formation of polymorph of CaCO3, and can provide high - HCO3 , for accumulation of ACC in vesicles. concentrations of ions for rapid physiological We hypothesize that ACC is stabilized by biomineralization (6). The high solubility of this ovalbumin and/or lysozyme or additional transitory phase requires stabilization by vesicle proteins identified in this study. Finally, regulatory molecules (2,5,7-11). Extracellular EDIL3 and MFGE8 are proposed to serve as vesicles (EVs) were reported as candidates to guidance molecules to target EVs to the play a role in the stabilization of ACC (5,9-11). mineralization site. We therefore report for the Experimental evidence for EVs in ACC- first time experimental evidence for the mediated calcification in CaCO3 biominerals is components of vesicular transport to supply ACC in vertebrate biomineralization. These results could give insight to understand the 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.04.08.031989; this version posted April 10, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-ND 4.0 International license. New model via stabilized ACC for avian eggshell formation only available for invertebrate structures such as that EDIL3 and MFGE8 bind to EVs budding sea urchin spines (calcite) and molluscan shells from uterine cells into the uterine fluid, in order (2,12,13). to guide vesicular transport of stabilized ACC Amongst vertebrates, the avian class for delivery to the mineralizing site and (Aves) appeared around 91 million years ago moreover prevent non-specific precipitation and is divided into Paleognathae (ancient birds (29). such as ostrich, rhea and emu) and Neognathae In order to test this hypothesis, we used (modern birds such as chicken, turkey or zebra microscopic techniques to investigate finch) (14). All birds produce eggs with hard exocytosis activity at the plasma membrane of shells composed of calcitic calcium carbonate, uterine cells that could be a source of EVs in the which provides a biomineral barrier that allows uterine fluid. We also used elemental analysis to appropriate development of the embryo within verify the presence of ACC inside these EVs an autonomous chamber (15). In addition to and we quantified gene expression of a variety defense against physical aggression, the of validated EV components in the oviduct eggshell protects the egg contents against segments and other tissues. Finally, we purified microbial contamination, regulates water and EVs from UF and demonstrated the presence of gaseous exchange, and provides calcium for key vesicular proteins in these vesicles. This embryonic bone calcification (16,17). The experimental study is the first to demonstrate chicken eggshell is a widely utilized vesicular transport of ACC in vertebrates, experimental model. Eggshell formation occurs which we propose supports the rapid eggshell in the uterine segment of the hen oviduct and is biomineralization process in birds. one of the fastest processes of vertebrate biomineralization (18-20). Approximately 6 g Results Transmission electron microscopy of uterine of CaCO3 are deposited in a rapid period with a mineralization rate of 0.32 g/h (21). During this epithelial cells and uterine fluid extracellular process, the uterine cells secrete The presence of vesicles in the tissues organic and mineral eggshell precursors into the and milieu involved in shell mineralization was uterine fluid (UF) where mineralization takes investigated by transmission electron place (22-24). Both mineral and organic microscopy (TEM). Ultra-thin negatively precursors interact to produce the specific stained sections of uterus were examined by eggshell texture and its resulting mechanical TEM to investigate exocytosis activity adjacent properties (19,25). This process requires to the luminal site of mineralization (Fig. 1). delivery of large amounts of calcium and Uterine ciliated cells possess numerous carbonates at the site of eggshell calcification, vacuolar and vesicular structures as well as which are continuously supplied from the blood dense and light granules (Fig. 1A). At higher across the uterine epithelium (26). The active magnification (Fig. 1B-D) we observed vesicles transepithelial transfer of calcium and in the cell cytoplasm, their accumulation at the carbonates is well described and constitutes the apical plasma membrane, and their budding to current model for eggshell calcification (23,27). generate EVs in the adjacent luminal uterine In chicken eggshell, the important role fluid (Fig. 1C-D). Figure 1E and F, show EVs of ACC during mineralization has been in the uterine fluid, with vesicle diameters in the described (28). ACC aggregates are first 100-400 nm range. Vesicle membranes massively deposited on the shell membranes appeared electron-dense compared to their and transform into calcite during initial internal regions. This density is due to organic calcification of the mammillary knobs (organic material comprising the membranes cores where the primary nucleation occurs). (glycoproteins, proteins, lipids…), which was ACC is also required at the mineralization edge stained by uranyl acetate. However, some during the overall eggshell calcification internal regions also appeared electron-dense. process, in order to provide material for the These TEM observations demonstrated the growth of columnar calcite crystals that presence of vesicles in the luminal uterine fluid constitute the palisade layer (28). In a recent adjacent to the apical region of uterine cells. study, we used bioinformatics tools, gene The uterine epithelium contains ciliated and expression and protein quantification to explore non-ciliated cells, and the same results were the role of EDIL3 and MFGE8 in chicken observed in proximity to non-ciliated cells (data eggshell biomineralization. We hypothesized not shown). UF was also subjected to TEM 2 bioRxiv preprint doi: https://doi.org/10.1101/2020.04.08.031989; this version posted April 10, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It
Load more

Recommended publications
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • CHML Promotes Liver Cancer Metastasis by Facilitating Rab14 Recycle
    ARTICLE https://doi.org/10.1038/s41467-019-10364-0 OPEN CHML promotes liver cancer metastasis by facilitating Rab14 recycle Tian-Wei Chen1,2,3, Fen-Fen Yin1,2, Yan-Mei Yuan1,2, Dong-Xian Guan1, Erbin Zhang1,2, Feng-Kun Zhang1,2, Hao Jiang1,2, Ning Ma1,2, Jing-Jing Wang1, Qian-Zhi Ni1,2, Lin Qiu1, Jing Feng3, Xue-Li Zhang3, Ying Bao4, Kang Wang5, Shu-Qun Cheng5, Xiao-Fan Wang6, Xiang Wang7, Jing-Jing Li1,2 & Dong Xie1,2,8,9 Metastasis-associated recurrence is the major cause of poor prognosis in hepatocellular 1234567890():,; carcinoma (HCC), however, the underlying mechanisms remain largely elusive. In this study, we report that expression of choroideremia-like (CHML) is increased in HCC, associated with poor survival, early recurrence and more satellite nodules in HCC patients. CHML promotes migration, invasion and metastasis of HCC cells, in a Rab14-dependent manner. Mechanism study reveals that CHML facilitates constant recycling of Rab14 by escorting Rab14 to the membrane. Furthermore, we identify several metastasis regulators as cargoes carried by Rab14-positive vesicles, including Mucin13 and CD44, which may contribute to metastasis- promoting effects of CHML. Altogether, our data establish CHML as a potential promoter of HCC metastasis, and the CHML-Rab14 axis may be a promising therapeutic target for HCC. 1 CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Xuhui district 200031, China. 2 University of Chinese Academy of Sciences, Chinese Academy of Sciences, Xuhui district 200031, China. 3 Department of General Surgery, Fengxian Hospital Affiliated to Southern Medical University, 6600 Nanfeng Road, Shanghai 201499, China.
    [Show full text]
  • Rabbit Anti-RABEP1 Antibody-SL19721R
    SunLong Biotech Co.,LTD Tel: 0086-571- 56623320 Fax:0086-571- 56623318 E-mail:[email protected] www.sunlongbiotech.com Rabbit Anti-RABEP1 antibody SL19721R Product Name: RABEP1 Chinese Name: RABEP1蛋白抗体 Neurocrescin; Rab GTPase binding effector protein 1; RAB5EP; Rabaptin 4; Rabaptin Alias: 5; Rabaptin 5alpha; RABPT5; RABPT5A; Renal carcinoma antigen NY REN 17; Renal carcinoma antigen NYREN17. Organism Species: Rabbit Clonality: Polyclonal React Species: Human,Mouse,Rat, ELISA=1:500-1000IHC-P=1:400-800IHC-F=1:400-800ICC=1:100-500IF=1:100- 500（Paraffin sections need antigen repair） Applications: not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user. Molecular weight: 99kDa Cellular localization: The cell membrane Form: Lyophilized or Liquid Concentration: 1mg/ml immunogen: KLH conjugated synthetic peptide derived from human RABEP1:501-600/862 Lsotype: IgGwww.sunlongbiotech.com Purification: affinity purified by Protein A Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year Storage: when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C. PubMed: PubMed RABEP1 is a Rab effector protein acting as linker between gamma-adaptin, RAB4A and RAB5A. It is involved in endocytic membrane fusion and membrane trafficking of Product Detail: recycling endosomes. Stimulates RABGEF1 mediated nucleotide exchange on RAB5A.
    [Show full text]
  • Expression Pattern of the PRDX2, RAB1A, RAB1B, RAB5A and RAB25 Genes in Normal and Cancer Cervical Tissues
    INTERNATIONAL JOURNAL OF ONCOLOGY 46: 107-112, 2015 Expression pattern of the PRDX2, RAB1A, RAB1B, RAB5A and RAB25 genes in normal and cancer cervical tissues ANDREJ NIKOSHKOV1, KRISTINA BROLIDEN2, SANAZ ATTARHA1,3, VITALI SVIATOHA3, ANN-CATHRIN HELLSTRÖM4, MIRIAM MINTS1 and SONIA ANDERSSON1 1Department of Women's and Children's Health, Division of Obstetrics and Gynecology, Karolinska Institute, Karolinska University Hospital Solna, 171 76 Stockholm; 2Department of Medicine Solna, Unit of Infectious Diseases, Center for Molecular Medicine, Karolinska Institute, Karolinska University Hospital, 171 76 Stockholm; 3Department of Oncology-Pathology, Karolinska Institute, 171 76 Stockholm; 4Department of Gynecological Oncology, Radiumhemmet, Karolinska University Hospital, 171 76 Stockholm, Sweden Received July 11, 2014; Accepted August 28, 2014 DOI: 10.3892/ijo.2014.2724 Abstract. Cervical cancer is the second most prevalent RAB1B transcript occurs in normal cervical tissue and in malignancy among women worldwide, and additional preinvasive cervical lesions; not in invasive cervical tumors. objective diagnostic markers for this disease are needed. Given the link between cancer development and alternative Introduction splicing, we aimed to analyze the splicing patterns of the PRDX2, RAB1A, RAB1B, RAB5A and RAB25 genes, which Alternative splicing is a process in which either different are associated with different cancers, in normal cervical combinations of exons or parts of exons are employed to tissue, preinvasive cervical lesions and invasive cervical generate new transcripts through the use of alternative-splicing tumors, to identify new objective diagnostic markers. Biopsies sites. More than 90% of human intron-containing genes of normal cervical tissue, preinvasive cervical lesions and undergo alternative splicing, which allows a single transcript invasive cervical tumors, were subjected to rapid amplification to encode a number of RNAs that produce various protein of cDNA 3' ends (3' RACE) RT‑PCR.
    [Show full text]
  • Rab5 and Rab4 Regulate Axon Elongation in Thexenopus Visual System
    The Journal of Neuroscience, January 8, 2014 • 34(2):373–391 • 373 Cellular/Molecular Rab5 and Rab4 Regulate Axon Elongation in the Xenopus Visual System Julien Falk, Filip A. Konopacki, Krishna H. Zivraj, and Christine E. Holt Department of Physiology, Development, and Neuroscience, Anatomy Building, University of Cambridge, Cambridge CB2 3DY, United Kingdom The elongation rate of axons is tightly regulated during development. Recycling of the plasma membrane is known to regulate axon extension; however, the specific molecules involved in recycling within the growth cone have not been fully characterized. Here, we investigated whether the small GTPases Rab4 and Rab5 involved in short-loop recycling regulate the extension of Xenopus retinal axons. We report that, in growth cones, Rab5 and Rab4 proteins localize to endosomes, which accumulate markers that are constitutively recycled. Fluorescence recovery after photo-bleaching experiments showed that Rab5 and Rab4 are recruited to endosomes in the growth cone, suggesting that they control recycling locally. Dynamic image analysis revealed that Rab4-positive carriers can bud off from Rab5 endosomes and move to the periphery of the growth cone, suggesting that both Rab5 and Rab4 contribute to recycling within the growth cone. Inhibition of Rab4 function with dominant-negative Rab4 or Rab4 morpholino and constitutive activation of Rab5 decreases the elongation of retinal axons in vitro and in vivo, but, unexpectedly, does not disrupt axon pathfinding. Thus, Rab5- and Rab4-mediated control of endosome trafficking appears to be crucial for axon growth. Collectively, our results suggest that recycling from Rab5-positive endosomes via Rab4 occurs within the growth cone and thereby supports axon elongation.
    [Show full text]
  • Rab5 (RAB5A) (NM 004162) Human Tagged ORF Clone – RC203873L4
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for RC203873L4 Rab5 (RAB5A) (NM_004162) Human Tagged ORF Clone Product data: Product Type: Expression Plasmids Product Name: Rab5 (RAB5A) (NM_004162) Human Tagged ORF Clone Tag: mGFP Symbol: RAB5A Synonyms: RAB5 Vector: pLenti-C-mGFP-P2A-Puro (PS100093) E. coli Selection: Chloramphenicol (34 ug/mL) Cell Selection: Puromycin ORF Nucleotide The ORF insert of this clone is exactly the same as(RC203873). Sequence: Restriction Sites: SgfI-MluI Cloning Scheme: ACCN: NM_004162 ORF Size: 645 bp This product is to be used for laboratory only. Not for diagnostic or therapeutic use. View online » ©2021 OriGene Technologies, Inc., 9620 Medical Center Drive, Ste 200, Rockville, MD 20850, US 1 / 2 Rab5 (RAB5A) (NM_004162) Human Tagged ORF Clone – RC203873L4 OTI Disclaimer: The molecular sequence of this clone aligns with the gene accession number as a point of reference only. However, individual transcript sequences of the same gene can differ through naturally occurring variations (e.g. polymorphisms), each with its own valid existence. This clone is substantially in agreement with the reference, but a complete review of all prevailing variants is recommended prior to use. More info OTI Annotation: This clone was engineered to express the complete ORF with an expression tag. Expression varies depending on the nature of the gene. RefSeq: NM_004162.3, NP_004153.2 RefSeq Size: 2548 bp RefSeq ORF: 648 bp Locus ID: 5868 UniProt ID: P20339, A0A024R2K1 Domains: ras, RAN, RAS, RHO, RAB Protein Families: Druggable Genome Protein Pathways: Amyotrophic lateral sclerosis (ALS), Endocytosis MW: 23.7 kDa Gene Summary: The small GTPases Rab are key regulators of intracellular membrane trafficking, from the formation of transport vesicles to their fusion with membranes.
    [Show full text]
  • Systematic Elucidation of Neuron-Astrocyte Interaction in Models of Amyotrophic Lateral Sclerosis Using Multi-Modal Integrated Bioinformatics Workflow
    ARTICLE https://doi.org/10.1038/s41467-020-19177-y OPEN Systematic elucidation of neuron-astrocyte interaction in models of amyotrophic lateral sclerosis using multi-modal integrated bioinformatics workflow Vartika Mishra et al.# 1234567890():,; Cell-to-cell communications are critical determinants of pathophysiological phenotypes, but methodologies for their systematic elucidation are lacking. Herein, we propose an approach for the Systematic Elucidation and Assessment of Regulatory Cell-to-cell Interaction Net- works (SEARCHIN) to identify ligand-mediated interactions between distinct cellular com- partments. To test this approach, we selected a model of amyotrophic lateral sclerosis (ALS), in which astrocytes expressing mutant superoxide dismutase-1 (mutSOD1) kill wild-type motor neurons (MNs) by an unknown mechanism. Our integrative analysis that combines proteomics and regulatory network analysis infers the interaction between astrocyte-released amyloid precursor protein (APP) and death receptor-6 (DR6) on MNs as the top predicted ligand-receptor pair. The inferred deleterious role of APP and DR6 is confirmed in vitro in models of ALS. Moreover, the DR6 knockdown in MNs of transgenic mutSOD1 mice attenuates the ALS-like phenotype. Our results support the usefulness of integrative, systems biology approach to gain insights into complex neurobiological disease processes as in ALS and posit that the proposed methodology is not restricted to this biological context and could be used in a variety of other non-cell-autonomous communication
    [Show full text]
  • Proteomic Profiling and Genome-Wide Mapping of O-Glcnac Chromatin
    ARTICLE https://doi.org/10.1038/s41467-020-19579-y OPEN Proteomic profiling and genome-wide mapping of O-GlcNAc chromatin-associated proteins reveal an O-GlcNAc-regulated genotoxic stress response Yubo Liu 1,3, Qiushi Chen 2,3, Nana Zhang1, Keren Zhang2, Tongyi Dou1, Yu Cao1, Yimin Liu1, Kun Li1, ✉ ✉ Xinya Hao1, Xueqin Xie1, Wenli Li1, Yan Ren 2 & Jianing Zhang 1 fi 1234567890():,; O-GlcNAc modi cation plays critical roles in regulating the stress response program and cellular homeostasis. However, systematic and multi-omics studies on the O-GlcNAc regu- lated mechanism have been limited. Here, comprehensive data are obtained by a chemical reporter-based method to survey O-GlcNAc function in human breast cancer cells stimulated with the genotoxic agent adriamycin. We identify 875 genotoxic stress-induced O-GlcNAc chromatin-associated proteins (OCPs), including 88 O-GlcNAc chromatin-associated tran- scription factors and cofactors (OCTFs), subsequently map their genomic loci, and construct a comprehensive transcriptional reprogramming network. Notably, genotoxicity-induced O- GlcNAc enhances the genome-wide interactions of OCPs with chromatin. The dynamic binding switch of hundreds of OCPs from enhancers to promoters is identified as a crucial feature in the specific transcriptional activation of genes involved in the adaptation of cancer cells to genotoxic stress. The OCTF nuclear factor erythroid 2-related factor-1 (NRF1) is found to be a key response regulator in O-GlcNAc-modulated cellular homeostasis. These results provide a valuable clue suggesting that OCPs act as stress sensors by regulating the expression of various genes to protect cancer cells from genotoxic stress.
    [Show full text]
  • Small Rab Gtpases in Intracellular Vesicle Trafficking:The Case Of
    International Journal of Molecular Sciences Review Small Rab GTPases in Intracellular Vesicle Trafficking: The Case of Rab3A/Raphillin-3A Complex in the Kidney Olga Martinez-Arroyo 1,† , Estela Selma-Soriano 2,†, Ana Ortega 1,* , Raquel Cortes 1,‡ and Josep Redon 1,3,*,‡ 1 Cardiometabolic and Renal Risk Research Group, INCLIVA Biomedical Research Institute, 46010 Valencia, Spain; [email protected] (O.M.-A.); [email protected] (R.C.) 2 Physiopathology of Cellular and Organic Oxidative Stress Group, University of Valencia, 46100 Valencia, Spain; [email protected] 3 CIBERObn, Carlos III Institute, 28029 Madrid, Spain * Correspondence: [email protected] (A.O.); [email protected] (J.R.); Tel.: +34-658-909-676 (A.O. & J.R.) † These authors contributed equally as first authors to this work. ‡ These authors contributed equally as senior authors to this work. Abstract: Small Rab GTPases, the largest group of small monomeric GTPases, regulate vesicle traf- ficking in cells, which are integral to many cellular processes. Their role in neurological diseases, such as cancer and inflammation have been extensively studied, but their implication in kidney disease has not been researched in depth. Rab3a and its effector Rabphillin-3A (Rph3A) expression have been demonstrated to be present in the podocytes of normal kidneys of mice rats and humans, around vesicles contained in the foot processes, and they are overexpressed in diseases with proteinuria. In addition, the Rab3A knockout mice model induced profound cytoskeletal changes in podocytes of high glucose fed animals. Likewise, RphA interference in the Drosophila model produced structural Citation: Martinez-Arroyo, O.; and functional damage in nephrocytes with reduction in filtration capacities and nephrocyte number.
    [Show full text]
  • Rab Proteins and the Compartmentalization of the Endosomal System
    Downloaded from http://cshperspectives.cshlp.org/ on September 26, 2021 - Published by Cold Spring Harbor Laboratory Press Rab Proteins and the Compartmentalization of the Endosomal System Angela Wandinger-Ness1 and Marino Zerial2 1Department of Pathology MSC08 4640, University of New Mexico HSC, Albuquerque, New Mexico 87131 2Max Planck Institute of Molecular and Cell Biology and Genetics, 01307 Dresden, Germany Correspondence: [email protected]; [email protected] Of the approximately 70 human Rab GTPases, nearly three-quarters are involved in endo- cytic trafficking. Significant plasticity in endosomal membrane transport pathways is closely coupled to receptor signaling and Rab GTPase-regulated scaffolds. Here we review current literature pertaining to endocytic Rab GTPaselocalizations, functions, and coordination with regulatory proteins and effectors. The roles of Rab GTPasesin (1) compartmentalization of the endocytic pathway into early, recycling, late, and lysosomal routes; (2) coordination of individual transport steps from vesicle budding to fusion; (3) effector interactomes; and (4) integration of GTPase and signaling cascades are discussed. he general working principle of Rab coded by interacting proteins (Wittinghofer TGTPases is that they contribute to the struc- et al. 1993). Guanine-nucleotide exchange fac- tural and functional identity of intracellular or- tors (GEFs) and GTPase-activating proteins ganelles. These functions rely on the versatile (GAPs) catalyze the exchange and hydrolysis GTP/GDP cycle for the assembly of multipro- reactions and, therefore, act as regulators of tein machineries on the cytoplasmic surface of the GTP–GDP cycle (Yoshimura et al. 2010; intracellular membranes. Rab GTPase protein Wu et al. 2011; Barr 2013; Guo et al. 2013; Kot- assemblies are spatially and temporally regulat- ting and Gerwert 2013).
    [Show full text]
  • KCNQ1 Antibody (N-Term) Affinity Purified Rabbit Polyclonal Antibody (Pab) Catalog # Ap12143a
    10320 Camino Santa Fe, Suite G San Diego, CA 92121 Tel: 858.875.1900 Fax: 858.622.0609 KCNQ1 Antibody (N-term) Affinity Purified Rabbit Polyclonal Antibody (Pab) Catalog # AP12143a Specification KCNQ1 Antibody (N-term) - Product Information Application WB,E Primary Accession P51787 Other Accession NP_000209.2, NP_861463.1 Reactivity Human Host Rabbit Clonality Polyclonal Isotype Rabbit Ig Calculated MW 74699 Antigen Region 4-33 KCNQ1 Antibody (N-term) - Additional Information KCNQ1 Antibody (N-term) (Cat. #AP12143a) western blot analysis in MDA-MB435 cell line Gene ID 3784 lysates (35ug/lane).This demonstrates the Other Names KCNQ1 antibody detected the KCNQ1 protein Potassium voltage-gated channel subfamily (arrow). KQT member 1, IKs producing slow voltage-gated potassium channel subunit alpha KvLQT1, KQT-like 1, Voltage-gated KCNQ1 Antibody (N-term) - Background potassium channel subunit Kv71, KCNQ1, KCNA8, KCNA9, KVLQT1 This gene encodes a voltage-gated potassium channel Target/Specificity required for the repolarization phase of the This KCNQ1 antibody is generated from cardiac action rabbits immunized with a KLH conjugated potential. The gene product can form synthetic peptide between 4-33 amino acids heteromultimers with two other from the N-terminal region of human potassium channel proteins, KCNE1 and KCNQ1. KCNE3. Mutations in this gene are associated with hereditary long QT Dilution syndrome (also known as WB~~1:1000 Romano-Ward syndrome), Jervell and Lange-Nielsen syndrome and Format familial atrial fibrillation. The gene is located in Purified polyclonal antibody supplied in PBS a region of with 0.09% (W/V) sodium azide. This chromosome 11 that contains a number of antibody is purified through a protein A contiguous genes, which are column, followed by peptide affinity abnormally imprinted in cancer and the purification.
    [Show full text]
  • 5. Rab Proteins
    Rabaptin5 is recruited to endosomes by Rab4a and Rabex5 to regulate endosome maturation Inauguraldissertation zur Erlangung der Würde eines Doktors der Philosophie vorgelegt der Philosophisch-Naturwissenschaftlichen Faktultät der Universität Basel von Simone Kälin aus Einsiedeln (SZ) Basel, 2014 Genehmigt von der Philosophisch-Naturwissenschaftlichen Fakultät auf Antrag von Prof. Dr. Martin Spiess Prof. Dr. Kurt Ballmer Basel, den 18. Februar 2014 Prof. Dr. Jörg Schibler Dekan 3 Acknowledgments I would like to express my sincerest thanks to the following people: Martin Spiess for giving me the opportunity to work on this project and for his continuous support. My thesis committee members, Kurt Ballmer and Markus Rüegg, for your time, helpful dis- cussions and advice. David Hirschmann for data contributions and experimental advice Nicole Beuret, who had always an open ear for technical questions and for always offering a helping hand. All the past and present members of the Spiess group for a great working atmosphere, an- swering questions, giving input and relaxing coffee and lunch breaks: Cristina Baschong, Julia Birk, Dominik Buser, Erhan Demirci, Franziska Hasler, Sonja Huser, Tina Junne, Lucyna Kocik, Deyan Mihov and Barry Shortt. Andrijana Kriz and Philipp Berger for introducing me to the MultiLabel system Aurélien Rizk for helping me with the quantitation of the endosome size 4 Summary Membrane trafficking between organelles is fundamental to the existence of eukaryotic cells. A multitude of proteins is involved in membrane trafficking, acting as building blocks for transport carriers, regulators of transport, and targeting and fusion factors. One important group of regulators are the Rab GTPases. They serve as multifaceted organizers of almost all membrane trafficking related processes in eukaryotic cells.
    [Show full text]