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Heterokaryons in the analysis of genes and gene regulation

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Summary

Cytological and chemical analysis of heterokaryons, the immediate product of cell fusion, offer new possibilities for studying the factors responsible for genetic regulation in eukaryotic cells. In comparison with proliferating cell hybrids the heterokaryon state offers the important advantage that a heterokaryon contains two complete genomes since chromosome loss does not occur, but since segregation and recombination are absent, heterokaryons cannot be used for gene mapping in the same way as proliferating cell hybrids. However, if two cell types carrying different genetic defects are fused the analysis can be used for studies of gene complementation.

The biological information obtained with heterokaryons has emphasized the role of the cytoplasm in the control of nuclear activity. When a G1 nucleus is brought into contact with the cytoplasm of an S phase cell the G1 nucleus is stimulated to synthesize DNA. If the nucleus is brought into a mitotic cell, the chromatin of the G1 nucleus is forced to condense into prematurely condensed chromosomes. Inactive nuclei such as the dormant chick erythrocyte nucleus will be stimulated to initiate RNA and DNA synthesis when brought into contact with an active cytoplasm by cell fusion. Specific nuclear proteins have been shown to be responsible for this process of reactivation.

Other inactive nuclei such as the nuclei of macrophages and spermatozoa have likewise been shown to be reactivated by fusion with active cells. The degree of activation in all of these cases appears to be determined by the state of the active cell. Inactive nuclei are activated to the same level as the active nucleus but seldom beyond this level.

If differentiated cells are fused with undifferentiated cells, usually the differentiated character is lost rapidly after fusion. This observation is in agreement with several studies on proliferating cell hybrids indicating some type of negative control of differentiated properties. In heterokaryons obtained by fusion of cells of a similar type of histotypic differentiation usually coexpression of the differentiated markers is observed.

Zusammenfassung

Die cytologische und chemische Analyse des Heterokaryons, des unmittelbaren Produktes der Zellfusion, erschließt neue Möglichkeiten für das Studium von Faktoren, die für die genetische Regulation in eukaryoten Zellen verantwortlich sind. Im Vergleich zu proliferierenden Zellhybriden führt der Heterokaryon-Zustand den wichtigen Vorteil mit sich, daß er zwei vollständige Genome enthält, da Chromosomen verlust nicht stattfindet. Da aber Segregation und Rekombination fehlen, können Heterokaryone nicht in der gleichen Art wie proliferierende Zellhybriden zur Genlokalisation gebraucht werden. Wenn jedoch zwei Zelltypen mit unterschiedlichem genetischen Defekt fusioniert werden, kann man die Analyse von Heterokaryonen zum Studium der Genkomplementierung anwenden.

Die biologische Information, die man durch Heterokaryone erhält, hat die Rolle des Cytoplasmas bei der Kontrolle der Kernaktivität hervorgehoben. Wenn ein G1-Kern mit dem Cytoplasma einer S-Phasen-Zelle in Kontakt gebracht wird, wird der G1-Kern zur DNA-Synthese stimuliert. Bringt man den Kern in eine mitotische Zelle, wird das Chromatin des G1-Kernes gezwungen, in vorzeitig kondensierte Chromosomen zu kondensieren. Inaktive Kerne, wie z. B. der dormante Hühnererythrocytkern, werden dazu stimuliert, die RNA- und DNA-Synthese zu beginnen, wenn sie bei der Zellfusion mit aktivem Cytoplasma in Kontakt gebracht werden. Spezifische Kernproteine, die sich im Cytoplasma von sich aktiv teilenden Zellen finden, haben sich als verantwortlich für diesen Prozeß der Reaktivierung erwiesen.

Bei anderen inaktiven Kernen, wie z. B. Makrophagen und Spermakernen, konnte vergleichsweise gezeigt werden, daß sie durch Fusion mit aktiven Zellen reaktiviert wurden. Das Ausmaß der Aktivierung in den erwähnten Fällen scheint vom Zustand der aktiven Zelle bestimmt zu werden. Inaktive Kerne werden bis zum gleichen Niveau wie der aktive Kern aktiviert, aber seltener darüber hinaus.

Wenn man differenzierte Zellen mit undifferenzierten Zellen fusioniert, geht der Differenzierungscharakter gewöhnlicherweise schnell nach der Fusion verloren. Diese Beobachtung stimmt mit mehreren Studien an proliferierenden Zellhybriden überein, wobei eine gewisse Art von negativer Kontrolle der differenzierten Eigenschaften angedeutet wird. Bei Heterokaryonen, die man durch Fusion von Zellen ähnlicher histiotypischer Differenzierung erhalten hat, beobachtet man gewöhnlich Coexpression der differenzierten Eigenschaften.

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  1. Institute of Human Genetics, University of Aarhus, Aarhus, Denmark

    Jesper Zeuthen

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Zeuthen, J. Heterokaryons in the analysis of genes and gene regulation. Hum Genet 27, 275–301 (1975). https://doi.org/10.1007/BF00278421

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