title: Efficient production of intersubspecific hybrid mice and embryonic stem cells by intracytoplasmic sperm injection.
abstract: Recently, mice and embryonic stem (ES) cells with allelic polymorphisms have been used extensively in the field of genetics and developmental biology. In this study, we examined whether intersubspecific hybrid mice and ES cells with these genotypes can be efficiently produced by intracytoplasmic sperm injection (ICSI). Frozen-thawed spermatozoa from wild-derived strains, JF1 (Mus musculus molossinus), MSM (M. m. molossinus), HMI (M. m. castaneus), and SWN (M. m. spp.), were directly injected into mature oocytes from laboratory mice ([C57BL/6 x DBA2]F1; M. m. domesticus). The in vitro and in vivo developmental capacity of F1 embryos was not significantly different among the groups (P > 0.05), and term offspring were efficiently obtained in all groups (27%-34% of transferred embryos). However, the mean body and placental weights of the offspring differed significantly with genotype (P < 5 x 10(-10)), with the HMI hybrid greatest in both body and placental weights. In an application study using these F1 offspring, we analyzed their mitochondrial DNA using intersubspecific polymorphisms and found the consistent disappearance of sperm mitochondrial DNA in the F1 progeny. In a second series of experiments, we generated F1 blastocysts by injecting MSM spermatozoa into C57BL/6 oocytes and used them to generate hybrid ES cell lines. The ES cell lines were established at a high efficiency (9 lines from 20 blastocysts) and their allelic polymorphisms were confirmed. Thus, ICSI using cryopreserved spermatozoa allows the efficient and immediate production of a number of F1 hybrid mice and ES cell lines, which can be used for polymorphic analysis of mouse genetics.
authors.affiliation: RIKEN Bioresource Center, Tsukuba, Ibaraki, Japan.
authors.forename: Akie
authors.initials: A
authors.lastname: Shinmen
authors.forename: Arata
authors.initials: A
authors.lastname: Honda
authors.forename: Mika
authors.initials: M
authors.lastname: Ohkawa
authors.forename: Michiko
authors.initials: M
authors.lastname: Hirose
authors.forename: Narumi
authors.initials: N
authors.lastname: Ogonuki
authors.forename: Misako
authors.initials: M
authors.lastname: Yuzuriha
authors.forename: Hiromi
authors.initials: H
authors.lastname: Miki
authors.forename: Keiji
authors.initials: K
authors.lastname: Mochida
authors.forename: Kimiko
authors.initials: K
authors.lastname: Inoue
authors.forename: Kuniya
authors.initials: K
authors.lastname: Abe
authors.forename: Masao
authors.initials: M
authors.lastname: Ito
authors.forename: Atsuo
authors.initials: A
authors.lastname: Ogura
mesh_terms: D000818:Animals; D002460:Cell Line; D002678:Chimera; D015925:Cryopreservation; D004272:DNA, Mitochondrial / Q000235:genetics; D047108:Embryonic Development; D053595:Embryonic Stem Cells; D004628:Embryonic Structures; D005260:Female; D005838:Genotype; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains / Q000033:anatomy & histology / Q000235:genetics; D010641:Phenotype; D011110:Polymorphism, Genetic; D012662:Semen Preservation; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa / Q000166:cytology / Q000502:physiology
chemical_list: D004272:DNA, Mitochondrial