[Studies of the correlation between fragile sites and giant panda's breeding problem].

An unique culture system has been established which can effectively indicate fragile sites induced by BrdU on giant panda chromosomes. After we cultured the lymphocytes of giant panda for 96 hours and treated it with low concentration of BrdU (10 micrograms/ml) for only a short time (4 hours), the areas of high-frequency-expressed fragile sites were successfully located on the centromeres of No. 2 and No. 12 chromosomes. After statistic analysis, we found that the frequency of fragile sites on No. 2 and No. 12 chromosomes are notably different among pandas. What is more important, the former is relative to giant pandas' breeding conditions and correlation coefficiency (r equals to -0.772). The result indicates that high frequency of fragile sites on No. 2 chromosome is unfavorable to giant panda's breeding and survival of filial generation.

[Studies on late replication bands of giant panda (Ailuropoda melanoleuca) chromosomes].

Using the cultured giant panda (Ailuropoda melanoleuca) lymphocytes as experimental material, we carried out the terminal marking on the chromosomes which were in replication by adding BrdU (a final concentration: 10 micrograms/ml) about four hours before harvesting the cells. The chromosomes marked by BrdU were proceeded by staining with acridine orange solution (0.05%), irradiated by ultraviolet and counter-stained by Giemsa, we obtained clear chromosomes replication patterns. According to the different replication bands, every chromosome's characteristics in late replication behavior could be identified. In the two X chromosomes of female individual, one X chromosome is obviously much later than the other one. Especially in the large area near centromere on the long arm of late replicating X chromosome. In the male individual, there is also a large area on the long arm of chromosome Y which replicates very late, but the end of long arm of chromosome Y replicates much earlier.

In vitro maturation of follicular oocytes of the Giant Panda (Ailuropoda melanoleuca): a case report.

The Giant Panda is an endangered species that would benefit from biotechnological assistance in reproduction. However, because there are only a few of these animals left in the world, scientists hesitate to use them for research procedures. We were fortunate to obtain ovaries from a Giant Panda that died of hepatic cirrhosis during the nonbreeding season. Oocytes were harvested within 4 h of death by dissecting the ovarian cortex in physiological saline and collecting the cumulus-oocyte complexes from the fluid, and then were classified into large (> 125 microns) and small (100 to 124 microns) follicular oocytes and placed in TCM199 supplemented with FSH (10 micrograms/mL) and LH (20 micrograms/mL). After culture for 22 h at 37 degrees C in air with 5% CO2, response was evaluated by growth of oocytes and presence of the first polar body. Of the 26 large follicular oocytes that were harvested, 12 were considered suitable for IVM, and 14 were degenerated, had a broken zona pellucida or had lost some cytoplasm. Of the 12 cultured oocytes, all grew to a mean diameter of 141.1(SD = +/- 6.7, n = 12), and 4 released the first polar body. None of the small follicular oocytes showed growth or other signs of maturation. We conclude from our preliminary results that it is possible to obtain functional Giant Panda oocytes from ovaries obtained post mortem during the nonbreeding season.