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Molekulare Netzwerke von Hypoxie und neuronaler Apoptose in der Cochlea

Molecular networks of hypoxia and neuronal apoptosis in the cochlea

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Zusammenfassung

Hintergrund

In organotypischen Kulturen zeigt die Region des Modiolus (MOD) von neugeborenen Ratten eine 4‑fach höhere Rate des Zelltods als die Region des Corti-Organs (OC). Die unterschiedliche Vulnerabilität geht mit einer differenziellen Expression zahlreicher Gene einher.

Methodik

Organotypische Kulturen von OC und MOD von 3–5 Tage alten Ratten wurden einer norm- bzw. hypoxischen (pO2: 10–20 mmHg; 5 h) Atmosphäre ausgesetzt. 24 h nach Anlegen der Kultur wurde die Zelltodrate bestimmt und die Expression mittels c‑DNA-Microarray untersucht. Mithilfe der DAVID-Datenbank wurden aus einer Liste von 60 Genen mit veränderter Expression biologische Prozesse entsprechend der Gene-Ontology-Datenbank (GO) zugeordnet. Molekulare Netzwerke wurden mithilfe der Datenbanken STRING und ConsensusPathDB erstellt.

Ergebnisse

Das Netzwerk der GO-Annotationen „Hypoxie“, „Entzündung“ und „mechanischer Stimulus“ deutet auf das Vorliegen von 2 Gen-Clustern, einem Cluster mit proinflammatorischen Genen (Ccl3, Cxcl2, Cxcr4, Ccl20) und einem Cluster mit hypoxieassoziierten Genen (c-Jun, Hif1a und Vegfa). Das Netzwerk der GO-Annotationen „positive und negative neuronale Apoptose“ lässt vermuten, dass die unterschiedliche Expression der Gene c-Jun, Ngfr und Casp3 von entscheidender Bedeutung für die Regulation des programmierten Zelltods von neuronalen Zellen des OC und MOD ist.

Schlussfolgerung

Während c‑JUN als ein wichtiger Modulator des Gleichgewichts zwischen Zelltod und Überleben wirkt, scheinen die Assoziationen von NGFR und CASP3 bedeutsam für die Einleitung des Zelltods zu sein. Die Auswertung und Anwendung von Erkenntnissen aus biostatistischen Datenbanken sind ein wichtiges Mittel für das Verständnis der Funktion von einzelnen Genen und Gen-Clustern in medizinisch relevanten biologischen Prozessen.

Abstract

Background

In organotypic cultures, the modiolus (MOD) region of newborn rats shows a fourfold higher rate of cell death than the organ of Corti (OC). The differing vulnerability of OC and MOD cells is related to differential expression of numerous genes (DEG).

Materials and methods

Organotypic cultures of OC and MOD of 3–5-day-old rats were exposed to a normoxic or a hypoxic (pO2 10–20 mmHg; 5 h) atmosphere. Cell death rate and gene expression as detected by c‑DNA microarray analysis were determined 24 h after the culture was created. Genes with modified expression (n = 60) were analyzed for biological processes according to the DAVID Gene Ontology Database (GO). Molecular networks were created using the STRING and ConsensusPathDB databases.

Results

The network of the GO annotations “hypoxia”, “inflammation”, and “mechanical stimulus” indicates the existence of two gene clusters: a cluster with pro-inflammatory genes (Ccl3, Cxcl2, Cxcr4, Ccl20) and a cluster with hypoxia-associated genes (e.g., c-Jun, Hif1a, and Vegfa). The network of the GO annotations “positive and negative regulation of neuron apoptotic process” suggests that the differential expression of c-Jun, Ngfr, and Casp3 is important for regulation of programmed cell death in neuronal cells of the OC and MOD.

Conclusion

While c‑JUN acts as an important modulator of the balance between cell death and survival, the associations of NGFR and CASP3 seem to be significant for the initiation of cell death. The evaluation and application of findings from biostatistical databases is important for understanding the function of individual genes and gene clusters in medically relevant biological processes.

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Authors and Affiliations

  1. Tinnituszentrum, Charité – Universitätsmedizin Berlin, Campus Charité Mitte, Charitéplatz 1, 10117, Berlin, Deutschland

    J. Gross &  B. Mazurek

  2. Klinik für Hals‑, Nasen‑, Ohrenheilkunde, Charité – Universitätsmedizin Berlin, Campus Virchow-Klinikum, Berlin, Deutschland

    H. Olze

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Correspondence to J. Gross.

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J. Gross, H. Olze und B. Mazurek geben an, dass kein Interessenkonflikt besteht.

Dieser Beitrag beinhaltet keine von den Autoren durchgeführten Studien an Menschen oder Tieren.

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Gross, J., Olze, H. & Mazurek, B. Molekulare Netzwerke von Hypoxie und neuronaler Apoptose in der Cochlea. HNO 66, 677–685 (2018). https://doi.org/10.1007/s00106-018-0539-9

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