A microinjection procedure to introduce "paternal" mitochondria from a source other than spermatozoa into fertilized mouse eggs is described. When a mitochondrial suspension isolated from the testes or liver of Mus molossinus mice was microinjected into fertilized eggs of CD1 mice, the microinjected zygotes survived, developed normally, and offspring were produced. Mus molossinus mitochondrial DNA can be distinguished from CD1 mitochondrial DNA by Southern blot analyses using restriction enzymes such as Eco R1, Xba 1, or Spe 1. Although up to 120 viable mitochondria were injected, no exogenous mitochondrial DNA was detected in fetal samples or in the brain, liver, heart, testis, or ovary of the mature progeny. Under the experimental conditions used, similar results were obtained when mitochondria from the testes of New Zealand black mice or from testes of Syrian hamsters were microinjected into fertilized CD1 mouse eggs. Failure to detect the exogenous mitochondrial DNA under our assay conditions suggests that microinjected mitochondria from testis or liver did not selectively replicate during embryonic development. The "foreign" mitochondria appear to have the same fate during early embryogenesis as the mitochondria of the spermatozoon.