Abstract
Embryos from the cross C57BL/6JHan x T which are heterozygous among other for 4 recessive coat-color genes were treated in utero by injection of a mutagen into the peritoneal cavity of the mother animal. If this treatment did lead in a pigment precursor cell to an alteration of the wild type allele of one of the four genes under study or to its loss, a brownish or light to dark gray color spot in the adult black coat could be seen. White midventral spots which also occur are predominantly the result of pigment cell killing, while the nonwhite spots with perhaps the exception of the white-gray should be the result of either gene mutations or recombinational processes. Of the numerical and structural chromosome aberrations produced only the few come into consideration which are able to pass the filter of several mitoses.
The following substances in the given concentrations were effective in this test system: 0.40 mmole/kg Ethyl methanesulfonate (EMS), 0.25 mmole/kg N-Methyl-N′-nitro-N-nitrosoguanidine (MNNG), 0.02 mmole/kg 2-[Bis-(2-chlo-roethyl) amino]-tetrahydro-2H-1,3,2-oxazaphosphorine 2-oxide (Endoxan®), 0.1 mmole/kg Dimethylnitrosamine (DMNA), 1 mmole/kg Trichloroethylene, 0.1 mmole/kg 4-Nitroquinoline 1-oxide (4-NQO), 0.006 mmole/kg Mitomycin C, 2 mmole/kg 2-Aminopurine (2-AP), 0.3 mmole/kg Acridine orange (AO), and 0.03 mmole/kg Hycanthone methanesulfonate. In addition, dose-effect relationships with Hycanthone were determined. At doses up to 0.2 mmole/kg Diethylnitrosamine (DENA) did not induce measurable amounts of color spots.
All these results clearly show that the test system described here is very sensitive to mutagens of all different types of action. Thus, it can be expected that the mammalian spot test will become a suitable procedure for routine testing of environmental mutagens.
Zusammenfassung
Embryonen der Kreuzung C57BL/6JHan x T, u. a. heterozygot für 4 rezessive Fellfarbmutationen, wurden durch Injektion eines Mutagens in die Peritonealhöhle des Muttertieres im Uterus behandelt. Wurde hierdurch in irgendeiner Pigmentzelle das Wildtypallel eines der vier Fellfarbgene geändert oder fiel es ganz aus, so war — nach mehreren Teilungsschritten der betroffenen Zelle — auf dem schwarzen Fell des erwachsenen Tieres ein Fleck in entweder bräunlicher oder hell-bis dunkelgrauer Farbe zu erkennen. Weiße Flecken an der Mittellinie des Bauches, welche ebenfalls in Erscheinung traten, sollten hauptsächlich die Folge einer Abtötung von Pigmentzellen sein, nichtweiße Fellflecken — mit Ausnahme vielleicht der weißgrauen — dagegen die Folge von Genmutationen oder Rekombinationsprozessen. Von numerischen und strukturellen Chromosomenaberrationen sollten nur die wenigen in Frage kommen, welche das Filter mehrerer Mitosen durchlaufen können.
Mit folgenden Substanzen wurden in nachstehenden Konzentrationen positive Ergebnisse erzielt: 0,40 mmol/kg Äthylmethansulfonat (EMS), 0,25 mmol/kg N-Methyl-N′-nitro-N-nitrosoguanidin (MNNG), 0,02 mmol/kg 2-[Bis-(2-chloräthyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorin 2-oxid (Endoxan®), 0,1 mmol/kg Dimethylnitrosamin (DMNA), 1 mmol/kg Trichloräthylen, 0,1 mmol/kg 4-Nitrochinolin-N-oxid (4-NQO), 0,006 mmol/kg Mitomycin C, 2 mmol/kg 2-Aminopurin (2-AP), 0,3 mmol/kg Acridinorange (AO) und 0,03 mmol/kg Hycanthonmethansulfonat. Mit Hycanthon wurde darüberhinaus eine Dosis/Effekt-Kurve aufgestellt. Diäthylnitrosamin (DENA) zeigte bis hin zu einer Konzentration von 0,2 mmol/kg keine Wirksamkeit.
Diese Ergebnisse zeigen deutlich, daß das beschriebene Testsystem nicht nur sehr empfindlich ist, sondern auch auf völlig verschiedenartig wirkende Mutagene anspricht. Man kann deshalb erwarten, daß sich der Fellfleckentest (Spottest) als Routinetestsystem für Umweltchemikalien bewähren wird.
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Fahrig, R. The mammalian spot test (Fellfleckentest) with mice. Arch. Toxicol. 38, 87–98 (1977). https://doi.org/10.1007/BF00293666
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DOI: https://doi.org/10.1007/BF00293666