Beta-defensin overexpression induces progressive muscle degeneration in mice.

Defensins comprise a family of cationic antimicrobial peptides characterized by conserved cysteine residues. They are produced in various organs including skeletal muscle and are identified as key elements in the host defense system as potent effectors. At the same time, defensins have potential roles in the regulation of inflammation and, furthermore, can exert cytotoxic effects on several mammalian cells. Here, we developed transgenic mice overexpressing mouse beta-defensin-6 to explore the pathophysiological roles of the defensin family as a novel mediator of inflammatory tissue injury. Unexpectedly, the transgenic mice showed short lifespan, poor growth, and progressive myofiber degeneration with functional muscle impairment, predominant centronucleated myofibers, and elevated serum creatine kinase activity, as seen in human muscular dystrophy. Furthermore, some of the transgenic myofibers showed IkappaBalpha accumulation, which would be related to the myofiber apoptosis of limb-girdle muscular dystrophy type 2A. The present findings may unravel a concealed linkage between the innate immune system and the pathophysiology of degenerative diseases.

Long-term cryopreservation of mouse sperm.

The objective was to determine if mouse sperm can maintain their fertilizing ability after being frozen for >10 y and whether the offspring derived from these sperm had normal fertilizing ability and phenotype. We cryopreserved sperm from six strains of mice (C57BL/6J, DBA/2N, BALB/cA, C3H/HeJ, B6D2F1 and B6C3F1) in a solution containing 18% (w/v) raffinose and 3% (w/v) skim milk, and preserved them in liquid nitrogen for >10 y. To assess the normality and fertilizing ability of these sperms, they were thawed and used for in vitro fertilization of oocytes of the same strains. Fertilization rates for C57BL/6J, DBA/2N, BALB/cA, C3H/HeJ, B6D2F1 and B6C3F1 were 66.4, 92.3, 72.8, 32.9, 60.3 and 53.7%, respectively. Furthermore, 38.3, 15.0, 43.3, 26.1, 38.3 and 16.7% of the embryos transferred to pseudopregnant females developed and produced live offspring that had normal phenotype and fertility.

Ovarian follicular development stimulated by leuprorelin acetate plus human menopausal gonadotropin in chimpanzees.

We attempted ovarian stimulation using gonadotropins in 14 chimpanzees. Subjects were given a single administration of leuprorelin acetate, followed by repeated administration of human menopausal gonadotropin (hMG) for 16-21 days. During the dosing period, the ovarian follicle diameter and count were measured by transvaginal ultrasonography. The hormone administration induced the development of multiple follicles, and multiple oocytes were subsequently retrieved. However, the follicle count was decreased, suggesting atresia, in some subjects. Statistically, the final follicle diameter was dependent on the dosing duration and the hMG dose in the late stage, while the maximum follicle count during hMG administration was dependent on age and the hMG dose in the early stage. Five subjects showed mild ovarian hyperstimulation syndrome (OHSS)-like symptoms with a high serum estradiol (E2) concentration. These results suggest that leuprorelin acetate plus hMG administration successfully stimulates the development of multiple ovarian follicles for oocyte retrieval and that the serum E2 concentration is predictive of OHSS-like symptoms in chimpanzees.

Cytoplasmic dysmorphisms in metaphase II chimpanzee oocytes.

Prior to fertilization by intracytoplasmic sperm injection (ICSI), cytoplasmic organization was evaluated in metaphase II chimpanzee oocytes obtained from stimulated ovaries. The findings demonstrate a high frequency of anomalies that are remarkably similar to the types of cytoplasmic dysmorphisms reported for human oocytes used in IVF. Similar to the human, the occurrence of these anomalies was oocyte- and animal-specific and associated with reduced competence as indicated by embryo development in vitro to the blastocyst stage.