Assessment of chromosomal integrity using a novel live-cell imaging technique in mouse embryos produced by intracytoplasmic sperm injection.

Background: Intracytoplasmic sperm injection (ICSI) is a technique in which sperm are injected directly into unfertilized oocytes, whereby offspring can be obtained even with dysfunctional sperm. Despite its advantages in human and animal reproductive technology, the low rate of resultant live offspring is perturbing. One major cause is thought to be embryonic chromosomal abnormalities.

Long-term, six-dimensional live-cell imaging for the mouse preimplantation embryo that does not affect full-term development.

Mammalian preimplantation embryonic development is achieved by tightly coordinated regulation of a great variety of temporal and spatial changes. Therefore, it would be valuable to analyze these events three-dimensionally and dynamically. We have previously developed a live-cell imaging method based on the expression of fluorescent proteins, using mRNA injection and time-lapse florescence microscopy.

Further characterization of the PW peptide family that inhibits neuron differentiation in Hydra.

From an evolutionary point of view, Hydra has one of the most primitive nervous systems among metazoans. Two different groups of peptides that affect neuron differentiation were identified in a systematic screening of peptide signaling molecules in Hydra. Within the first group of peptides, a neuropeptide, Hym-355, was previously shown to positively regulate neuron differentiation.

Tet-on inducible system combined with in ovo electroporation dissects multiple roles of genes in somitogenesis of chicken embryos.

The in ovo electroporation technique in chicken embryos has enabled investigators to uncover the functions of numerous developmental genes. In this technique, the ubiquitous promoter, CAGGS (CMV base), has often been used for overexpression experiments. However, if a given gene plays a role in multiple steps of development and if overexpression of this gene causes fatal consequences at the time of electroporation, its roles in later steps of development would be overlooked.

Efficient establishment of mouse embryonic stem cell lines from single blastomeres and polar bodies.

Recently, ES cell lines were established from single blastomeres taken from eight-cell embryos in mice and humans with success rates of 4% and 2%, respectively, which suggests that the method could be used in regenerative medicine to reduce ethical concerns over harm to embryos. However, those studies used other ES cells as supporting cells. Here, we report a simple and highly efficient method of establishing mouse ES cell lines from single blastomeres, in which single blastomeres are simply plated onto a feeder layer of mouse embryonic fibroblasts with modified ES cell medium.

Successful mouse cloning of an outbred strain by trichostatin A treatment after somatic nuclear transfer.

Although the somatic cloning technique has been used for numerous applications and basic research of reprogramming in various species, extremely low success rates have plagued this technique for a decade. Further in mice, the "clonable" strains have been limited to mainly hybrid F1 strains such as B6D2F1. Recently, we established a new efficient cloning technique using trichostatin A (TSA) which leads to a 2-5 fold increase in success rates for mouse cloning of B6D2F1 cumulus cells.

Mesenchymal-to-epithelial transition during somitic segmentation: a novel approach to studying the roles of Rho family GTPases in morphogenesis.

During early development in vertebrates, cells change their shapes dramatically both from epithelial to mesenchymal and also from mesenchymal to epithelial, enabling the body to form complex tissues and organs. Using somitogenesis as a novel model, Rho family GTPases have recently been shown to play essential and differential roles in individual cell behaviors in actual developing embryos. Levels of Cdc42 activity provide a binary switch wherein high Cdc42 levels allow the cells to remain mesenchymal, while low Cdc42 levels produce epithelialization.

Pax 2 expression in mesodermal segmentation and its relationship with EphA4 and Lunatic-fringe during chicken somitogenesis.

In the Pax gene family, which encodes DNA-binding proteins, Pax 2 has been known to play important roles in the formation of the midbrain/hindbrain boundary, eye, inner ear and kidney in vertebrates (Bioessays 19 (1997) 755). In this article, we report a segmentally regulated pattern of Pax 2 expression during chicken somitogenesis. Pax 2 mRNA is localized in the rostral end of the unsegmented presomitic mesoderm (PSM), abutting anteriorly on a prospective segmentation border.

Pax 2 expression in mesodermal segmentation and its relationship with EphA4 and Lunatic-fringe during chicken somitogenesis.

In the Pax gene family, which encodes DNA-binding proteins, Pax 2 has been known to play important roles in the formation of the midbrain/hindbrain boundary, eye, inner ear and kidney in vertebrates (Bioessays 19 (1997) 755). In this article, we report a segmentally regulated pattern of Pax 2 expression during chicken somitogenesis. Pax 2 mRNA is localized in the rostral end of the unsegmented presomitic mesoderm (PSM), abutting anteriorly on a prospective segmentation border.