Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About MCB
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
Molecular and Cellular Biology
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About MCB
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
Articles

Targeted Deletion of the Gene Encoding the La Autoantigen (Sjögren's Syndrome Antigen B) in B Cells or the Frontal Brain Causes Extensive Tissue Loss

Sergei Gaidamakov, Olga A. Maximova, Hyongi Chon, Nathan H. Blewett, Hongsheng Wang, Amanda K. Crawford, Amanda Day, Natalie Tulchin, Robert J. Crouch, Herbert C. Morse III, Robert D. Blitzer, Richard J. Maraia
Sergei Gaidamakov
aIntramural Research Programs of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Olga A. Maximova
bLaboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hyongi Chon
aIntramural Research Programs of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nathan H. Blewett
aIntramural Research Programs of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hongsheng Wang
cLaboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Amanda K. Crawford
aIntramural Research Programs of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Amanda Day
aIntramural Research Programs of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Natalie Tulchin
dDepartment of Pathology, Mount Sinai School of Medicine, New York, New York, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Robert J. Crouch
aIntramural Research Programs of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Herbert C. Morse III
cLaboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Robert D. Blitzer
eDepartment of Pharmacology and Systems Therapeutics, Mount Sinai School of Medicine, New York, New York, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Richard J. Maraia
aIntramural Research Programs of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA
fCommissioned Corps, U.S. Public Health Service, Washington, DC, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/MCB.01010-13
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • FIG 1
    • Open in new tab
    • Download powerpoint
    FIG 1

    A conditional La allele. (A) Schematic representation of La (Ssb) alleles. (B) Tissue-specific detection of αCaMKII Cre-mediated rearrangement in a 12-week-old LaF/− CreCaMKII mouse. FB, frontal brain; HB, hindbrain; Hrt, heart; Liv, liver; Spl, spleen; cn, control, in which no DNA was added to the PCR mixture; Lane M, molecular size markers. Lengths of the expected PCR products are indicated on the right.

  • FIG 2
    • Open in new tab
    • Download powerpoint
    FIG 2

    Reduction in La protein levels and decreased frontal brain mass following targeted deletion of La. (A) Immunohistochemistry of the hippocampal CA1 regions of 11-week-old αCamKIICre La-deleted and control mice, as indicated. (Upper row) Immunohistochemistry with anti-mLa antibody (35); (middle row) H&E stained section; (bottom row) antitubulin antibody-stained section. (B) αCamKIICre La-deleted and control mice brains as indicated. Double-sided arrows superimposed on the hindbrains (HB) are the same size; the FB (front brain) is also indicated. (C) Comparison of average brain wet weights of 10 αCamKIICre La-deleted and 10 control mice. Error bars indicate standard deviations. P = 1.2 × 10−5. All of these samples were obtained without prior perfusion-fixation.

  • FIG 3
    • Open in new tab
    • Download powerpoint
    FIG 3

    La deletion in αCamKIICre animals causes developmental postnatal loss of frontal brain mass. (A) Development of αCamKIICre La-deleted and sibling control brains at 16 and 32 postnatal weeks. Double-sided arrows on the hindbrain (HB) images are the same size; FB, front brain. (B) Immunohistochemistry of neocortex stained with anti-mLa antibody and H&E counterstaining of 16-week-old mouse cortices. (C) Plot of brain mass versus age of αCamKIICre La-deleted and control mice. For mice aged 13 weeks and older, P values ranged from 0.004 (13 weeks) to 0.0001 (17 and 25 weeks). All of the samples were obtained by dissection after perfusion-fixation.

  • FIG 4
    • Open in new tab
    • Download powerpoint
    FIG 4

    Progressive neurodegeneration in the cerebral cortex and hippocampus of La-deleted mice. Representative sections of the forebrains from the La-deficient (LaF/− CreCaMKII) and control littermates (LaF/−) at 12 weeks (A and B), 16 weeks (C and D), and 65 weeks (E and F). Areas of the frontal cortex and hippocampus (CA1 cells) that are outlined in the overview images (H&E, left panels) are shown at a higher magnification in the adjacent, corresponding Nissl-stained sections. Note there was some decrease in cortical thickness and a more pronounced decrease in neuronal density in the hippocampus (CA1) of 16-week-old La-deleted mice than in control littermates. Cortical atrophy and neuron loss in the hippocampus (CA1) were especially pronounced in 65-week-old La-deleted mice compared to controls (compare panels F and E). Bars in the Nissl insets (cortex and hippocampus), 100 μm. (G to J) High-magnification immunohistochemistry images of representative regions of the somatosensory cortex (G and H) and retrosplenial cortex (I and J), stained with anti-mLa antibody and H&E counterstaining, from 16-week-old control brains (upper panels) and matched littermate La knockout mutant brains (lower panels). (K and L) High-magnification immunohistochemistry results for the hippocampal regions of 12-week-old control (K) and matched littermate La knockout mutant (L) mouse brains stained with anti-mLa antibody and H&E counterstaining. The upper panels show low magnification images, which were further magnified for the middle and lower panels, including the circular insets, as annotated.

  • FIG 5
    • Open in new tab
    • Download powerpoint
    FIG 5

    La deletion by Mb1Cre causes reduced spleen size, agammaglobulenemia, and absence of cells of the B lineage. (A) Photographs of freshly isolated spleens from typical control and La-deleted 17-week-old mice. The mean ± standard deviation spleen weights from eight control and eight La-deleted mice are indicated under the images. (B) Absence of serum IgG in La-deleted mice. Lane 1, control mouse serum; lane 2, from La-deleted mouse serum. (C to E) Developmental arrest of La-depleted B lineage cells. BM (C), spleen (D), and PerC cells (E) were stained with the indicated Abs and analyzed by flow cytometry using the indicated gating parameters. The numbers are frequencies of cells falling in each gate. Data are representative of multiple mice with similar results. (C) BM IgM+ B220lo cells (top panels) were gated to identify pro-B cells (CD43+ B220+) and pre-B cells (CD43− B220+) (lower panels). (D) Spleen cells were stained to identify B cells (IgM+ B220+) and non-B cells (IgM− B220−) (top panels). (E) PerC cells were examined using the gating parameters shown.

  • FIG 6
    • Open in new tab
    • Download powerpoint
    FIG 6

    Northern blot analysis revealed altered pre-tRNA metabolism in La mutant forebrains. (A to C) A blot containing total RNA isolated from the frontal cortex was probed, autoradiographed, stripped, and reprobed with probes to detect different intermediates in the maturation pathway for pre-tRNATyr, as indicated. The cartoons to the left of panels A and B are schematic representations of the species detected; rectangles reflect exons, lines reflect the intron or 3′ trailer, and the thick line reflects the position of the probe. (C) The same blot, probed for U12 snRNA to serve as a loading control. (D) Graph of the ratio of lower versus upper band intensities from panel A and a similar probing of another matched sample set. Error bars reflect results of two independent experiments from different sample sets. (E) The same blot, probed for spliced mature tRNATyr.

Tables

  • Figures
  • TABLE 1

    Alleles evaluated in this study

    Allele descriptionDesignation
    La knockoutLa−
    La wild typeLa+
    La flip (before Cre)LaF
    La floxed (after Cre)Lax
    Mb1 CreMb1Cre
    Mb1 wild typeMb1+
    αCaMKII CreCreCaMKII
PreviousNext
Back to top
Download PDF
Citation Tools
Targeted Deletion of the Gene Encoding the La Autoantigen (Sjögren's Syndrome Antigen B) in B Cells or the Frontal Brain Causes Extensive Tissue Loss
Sergei Gaidamakov, Olga A. Maximova, Hyongi Chon, Nathan H. Blewett, Hongsheng Wang, Amanda K. Crawford, Amanda Day, Natalie Tulchin, Robert J. Crouch, Herbert C. Morse III, Robert D. Blitzer, Richard J. Maraia
Molecular and Cellular Biology Dec 2013, 34 (1) 123-131; DOI: 10.1128/MCB.01010-13

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this Molecular and Cellular Biology article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Targeted Deletion of the Gene Encoding the La Autoantigen (Sjögren's Syndrome Antigen B) in B Cells or the Frontal Brain Causes Extensive Tissue Loss
(Your Name) has forwarded a page to you from Molecular and Cellular Biology
(Your Name) thought you would be interested in this article in Molecular and Cellular Biology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Targeted Deletion of the Gene Encoding the La Autoantigen (Sjögren's Syndrome Antigen B) in B Cells or the Frontal Brain Causes Extensive Tissue Loss
Sergei Gaidamakov, Olga A. Maximova, Hyongi Chon, Nathan H. Blewett, Hongsheng Wang, Amanda K. Crawford, Amanda Day, Natalie Tulchin, Robert J. Crouch, Herbert C. Morse III, Robert D. Blitzer, Richard J. Maraia
Molecular and Cellular Biology Dec 2013, 34 (1) 123-131; DOI: 10.1128/MCB.01010-13
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • INTRODUCTION
    • MATERIALS AND METHODS
    • RESULTS
    • DISCUSSION
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

Related Articles

Cited By...

About

  • About MCB
  • Editor in Chief
  • Editorial Board
  • Policies
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Article Types
  • Ethics
  • Contact Us

Follow #MCBJournal

@ASMicrobiology

       

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 0270-7306; Online ISSN: 1098-5549