Article
Expression profiles of cytokines and chemokines at different prenatal time-points in a rat model of open spinal dysraphism
Expressionsprofile proinflammatorischer Zytokine und Chemokine während des pränatalen Zeitverlaufes in einem spinalen Dysraphie-Modell in der Ratte
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Authors
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Gesa Cohrs
- Universitätsklinikum Schleswig-Holstein, Klinik für Neurochirurgie, Kiel, Deutschland
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Ann-Kathrin Blumenröther
- Universitätsklinikum Schleswig-Holstein, Klinik für Neurochirurgie, Kiel, Deutschland
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Jan-Philip Sürie
- Universitätsklinikum Schleswig-Holstein, Klinik für Neurochirurgie, Kiel, Deutschland
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Michael Synowitz
- Universitätsklinikum Schleswig-Holstein, Klinik für Neurochirurgie, Kiel, Deutschland
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Janka Held-Feindt
- Universitätsklinikum Schleswig-Holstein, Klinik für Neurochirurgie, Kiel, Deutschland
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Friederike Knerlich-Lukoschus
- Universitätsklinikum Schleswig-Holstein, Klinik für Neurochirurgie, Kiel, Deutschland; Asklepios Klinik Sankt Augustin, Klinik für Kinderneurochirurgie, Sankt Augustin, Deutschland
| Published: | June 26, 2020 |
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Outline
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Objective: Cellular and molecular mechanisms induced by the hypothesized "second hit", which underlies progressive functional decline of the myelomeningocele (mmc) placode, are not well understood. We previously identified key players of post-traumatic lesion cascades in human mmc tissues obtained during post-natal repair. We went back to bench and investigated these mediators in the prenatal course in standardized conditions of an mmc-animal model.
Methods: A retinoic acid mmc model was established using time-dated Sprague-Dawley rats, which were gavage-fed with all-trans retinoic acid (RA; 60 mg/kg) dissolved in olive oil at day E10. Controls received olive oil only. Fetuses were obtained at E16, E18, E22. Spinal cord tissues were screened by real-time RT-PCR for cytokines and chemokines, known to play a role in lesion cascades in the central nervous system. Proinflammatory cytokines which were found on elevated mRNA level like TNFalpha (TNFa), Interleukin-1beta (IL-1b), and their receptors were analysed further by immunohistochemistry (IHC) and double-immunofluorescence-labelling (DIF) with cellular markers. Representing neuro-restorative markers the ligand-receptor pair Erythropoeitin (Epo) and EpoR were investigated. Fetal control sc tissue was obtained at the respective time points.
Results: IL-1b showed significant induction at E22, its IL1-RI was induced at E16 and E22, in IHC staining was confined to the matrix and marginal layer. Co-staining of IL-1b and Vimentin (VIM) suggests astroglial cells as a possible source of these mediators, whereas IL1-RI co-stained with VIM and Iba1, a marker for microglia. TNFa/-R showed significant induction at E22, DIF confirmed co-staining with Iba1 for the ligand-receptor pair. CXCL12 and CXCR4 showed elevated mRNA levels in controls and mmc, as these cytokines play a crucial role in developmental processes, mRNA levels drop throughout the prenatal time course as expected. The fraktalkine CX3CL1 and its receptor CX3CR1 show significant elevation throughout the investigated time course, there is no significant induction compared to controls. Epo is induced significantly at E16, EpoR shows high levels at E22.
Conclusion: Pro-inflammatory cytokines like TNFa/-R, IL-1b/-RI exhibited a time-dependent prenatal expression in the retinoic induced mmc rat model. Epo/-R also exhibited different expression patterns compared to controls. These mediators provide potential trial targets in the development of adjuvant therapies.
