Go to JCI Insight
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
  • Clinical Research and Public Health
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Gastroenterology
    • Immunology
    • Metabolism
    • Nephrology
    • Neuroscience
    • Oncology
    • Pulmonology
    • Vascular biology
    • All ...
  • Videos
    • Conversations with Giants in Medicine
    • Video Abstracts
  • Reviews
    • View all reviews ...
    • Complement Biology and Therapeutics (May 2025)
    • Evolving insights into MASLD and MASH pathogenesis and treatment (Apr 2025)
    • Microbiome in Health and Disease (Feb 2025)
    • Substance Use Disorders (Oct 2024)
    • Clonal Hematopoiesis (Oct 2024)
    • Sex Differences in Medicine (Sep 2024)
    • Vascular Malformations (Apr 2024)
    • View all review series ...
  • Viewpoint
  • Collections
    • In-Press Preview
    • Clinical Research and Public Health
    • Research Letters
    • Letters to the Editor
    • Editorials
    • Commentaries
    • Editor's notes
    • Reviews
    • Viewpoints
    • 100th anniversary
    • Top read articles

  • Current issue
  • Past issues
  • Specialties
  • Reviews
  • Review series
  • Conversations with Giants in Medicine
  • Video Abstracts
  • In-Press Preview
  • Clinical Research and Public Health
  • Research Letters
  • Letters to the Editor
  • Editorials
  • Commentaries
  • Editor's notes
  • Reviews
  • Viewpoints
  • 100th anniversary
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
Single-cell Tsc1 knockout during corticogenesis generates tuber-like lesions and reduces seizure threshold in mice
David M. Feliciano, … , Jean-Claude Platel, Angélique Bordey
David M. Feliciano, … , Jean-Claude Platel, Angélique Bordey
Published March 14, 2011
Citation Information: J Clin Invest. 2011;121(4):1596-1607. https://doi.org/10.1172/JCI44909.
View: Text | PDF
Technical Advance Neuroscience Article has an altmetric score of 3

Single-cell Tsc1 knockout during corticogenesis generates tuber-like lesions and reduces seizure threshold in mice

  • Text
  • PDF
Abstract

Tuberous sclerosis complex (TSC) is an autosomal dominant disorder characterized by mutations in Tsc1 or Tsc2 that lead to mammalian target of rapamycin (mTOR) hyperactivity. Patients with TSC suffer from intractable seizures resulting from cortical malformations known as tubers, but research into how these tubers form has been limited because of the lack of an animal model. To address this limitation, we used in utero electroporation to knock out Tsc1 in selected neuronal populations in mice heterozygous for a mutant Tsc1 allele that eliminates the Tsc1 gene product at a precise developmental time point. Knockout of Tsc1 in single cells led to increased mTOR activity and soma size in the affected neurons. The mice exhibited white matter heterotopic nodules and discrete cortical tuber-like lesions containing cytomegalic and multinucleated neurons with abnormal dendritic trees resembling giant cells. Cortical tubers in the mutant mice did not exhibit signs of gliosis. Furthermore, phospho-S6 immunoreactivity was not upregulated in Tsc1-null astrocytes despite a lower seizure threshold. Collectively, these data suggest that a double-hit strategy to eliminate Tsc1 in discrete neuronal populations generates TSC-associated cortical lesions, providing a model to uncover the mechanisms of lesion formation and cortical hyperexcitability. In addition, the absence of glial reactivity argues against a contribution of astrocytes to lesion-associated hyperexcitability.

Authors

David M. Feliciano, Tiffany Su, Jean Lopez, Jean-Claude Platel, Angélique Bordey

×

Figure 6

Lack of astrogliosis in the tuber-like lesions.

Options: View larger image (or click on image) Download as PowerPoint
Lack of astrogliosis in the tuber-like lesions.
(A–F) Photograph of mRFP...
(A–F) Photograph of mRFP+ cells (red) and GFAP immunostaining (green) in the ipsilateral (A, B, D, and E) and contralateral (C and F) in P28 and P15 Tsc1fl/mut mice electroporated at E15 (A–C) and E16 (D–F), respectively. (G–I) Photograph of mRFP+ cells (red) and GS (green) immunostaining in the ipsilateral (G and H) and contralateral (I) in P28 Tsc1fl/mut mice electroporated at E15. (J and K) Photograph of mRFP fluorescence (red, J), GS (blue, K), and GFP fluorescence (green) in a P15 CAG-GFP × Tsc1fl/mut mouse electroporated with pCAG-mRFP and pCAG-Cre:GFP at E16. Inset in J shows PCR gels from genomic DNA obtained from E16-electroporated cortical tissue containing electroporated astrocytes but no electroporated neurons, microdissected from slices obtained from a P28 Tsc1fl/fl mouse in which no mRFP+ neurons were visible. (L and M) Higher-magnification photographs of GS staining and GFP fluorescence from the boxed region in K. Arrows indicate GFP+ astrocytes (i.e., GS+), and arrowheads indicate GFP– astrocytes that exhibit the same soma size. (N) Photograph of GFP fluorescence (green) and pS6 immunostaining (blue) in a CAG-GFP × Tsc1fl/mut mouse electroporated at E16. (O and P) Higher-magnification photographs of pS6 staining in GFP+ cells from the boxed region in N. Arrows point to GFP+ astrocytes that are pS6 negative. (Q) Quantification of Tsc1fl/mut and Tsc1fl/WT astrocytic soma size, as outlined by GS staining. Scale bars: 300 μm (A–F), 140 μm (G–I), 70 μm (J, K, and N), 20 μm (L, M, O, and P).

Copyright © 2025 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

Sign up for email alerts

Referenced in 1 Wikipedia pages
121 readers on Mendeley
See more details