International Mammalian Genome Society

The 14th International Mouse Genome Conference (2000)

H15. Mouse Primordial Germ Cells are Highly Sensitive to ENU Mutagenesis

T. Shibuya, H. Sui and T. Hara
Hatano Research Institute, Food and Drug Safety Center, Hadano, Kanagawa, JAPAN

Primordial germ cells (PGC) are the ancestor cells of both male and female gamete in sexually reproducing animals. If a mutation is induced in a PGC, all germ cells derived from it possess the same mutation (cluster mutations).

Using a mouse specific locus test, we have found that N-ethyl-N-nitrosourea (ENU) induces recessive mutations at a relatively high rate in male mice PGC at 8.5, 10.5 and 13.5 days of development (G8.5, G10.5 and G13.5, respectively) with those in G10.5 the highest, followed by G8.5 and then G13.5. The induction of mutations was dose-dependent in these three developmental stages of PGC.

Cluster mutations were common in G8.5 and G10.5 PGC at the higher doses of ENU, and that was the cause of the apparent high mutation rate in those stages. The number of PGC in G8.5 and G10.5 are very small, and mouse PGC proliferate rapidly at those stages with a doubling time of about 16 hr. Thus, treatment with ENU would give rise to a clonal expansion of mutated PGC.

In general, mouse saturation mutagenesis experiments with ENU are carried out in spermatogonia. We will discuss the advantage and disadvantage of using PGC instead.

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