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The thin-layer isoelectric focusing of lactate dehydrogenase isoenzymes in rabbit lens parts and in intraocular tissues

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Summary

Lactate dehydrogenase (LDH) isoenzymes of rabbit lens and other intraocular tissues are separated by thin-layer isoelectric focusing and localized as discrete groups of multiple bands with defined isoelectric points after staining by the tetrazolium method.

In the rabbit lens parts, the predominant isoenzymes are LDH-4 and LDH-5. The bands show microheterogeneity, are composed of 2–4 subcomponents, and the patterns show a distribution of the liver type. The activity of the LDH-4 decreases and that of LDH-5 increases in an order given by the equator, anterior and posterior cortex, and nucleus. LDH-3 remains almost constant in all lens parts. LDH-4 is composed of two subcomponents, one of which, the most cathodic with higher isoelectric point, is almost absent in the lens nucleus.

Of the LDH localized in the rabbit intraocular tissues, only the retina shows a pattern of five isoenzymes also of the liver type. In all intraocular tissues LDH-3, -4, and -5 are very prominent, show also microheterogeneity of their isoenzyme bands, and are each composed of 4–6 subcomponents. LDH-1 and-2 show only one isofocused component.

Species specificity is shown of the LDH isoenzymes in the rabbit, mouse, dog, and calf lens.

Zusammenfassung

Mit dem Verfahren der Isoelektrofokussierung auf Dünnschichtplatten werden die Isoenzyme der Laktatdehydrogenase aus Linse, Hornhaut, Iris, Netzhaut und Aderhaut getrennt. Nach Anfärbung mit der Tetrazoliummethode können verschiedene LDH-Isoenzyme, die aus mehreren Einzelbanden bestehen, aufgrund der definierten isoelektrischen Punkte unterschieden werden.

In den einzelnen Linsenbereichen sind LDH-4 und LDH-5 die vorherrschenden Isoenzymformen. Die Banden zeigen Mikroheterogenität, bestehen meist aus 2–4 Unterkomponenten und entsprechen im Isoenzymmuster dem Lebertyp. Die Aktivität der LDH-4 vermindert sich in der Reihenfolge: Linsenäquator, vordere und hintere Linsenrinde und Linsenkern; die Aktivität der LDH-5 steigt im Gegensatz dazu an, LDH-3 ist in allen Linsenteilen gleich. Im Linsenkern ist die eine der beiden Unterkomponenten der LDH-4, die am stärksten kathodische Form mit höherem isoelektrischen Punkt, kaum vorhanden. Von den anderen untersuchten Augengeweben zeigt lediglich die Netzhaut ein Isoenzymmuster mit 5 Formen vom Lebertyp. In Hornhaut, Iris und Aderhaut sind die LDH-3, LDH-4 und LDH-5 prominent. Die einzelnen Isoenzymbanden weisen ebenfalls Mikroheterogenität auf, sie bestehen aus 4–6 Unterkomponenten. LDH-1 und LDH-2 treten immer nur als eine einzige Bande auf.

Die Verteilung der LDH Isoenzyme in den Linsen von Kaninchen, Maus, Hund und Kalb zeigt artspezifische Unterschiede.

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Authors and Affiliations

  1. Division of Biochemistry of the Eye, Clinical Institute for Experimental Ophthalmology, University of Bonn, D-5300, Bonn-Venusberg, Federal Republic of Germany

    J. Bours, H. Neuhaus & O. Hockwin

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  1. J. Bours
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  2. H. Neuhaus
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  3. O. Hockwin
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Research supported by the “Deutsche Forschungs-Gemeinschaft” (No. 249/8: Altersveränderungen von Linsen)

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Bours, J., Neuhaus, H. & Hockwin, O. The thin-layer isoelectric focusing of lactate dehydrogenase isoenzymes in rabbit lens parts and in intraocular tissues. Albrecht von Graefes Arch. Klin. Ophthalmol. 203, 9–19 (1977). https://doi.org/10.1007/BF00410043

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