The Prevention of Aging in Mammals INTRODUCTION/BACKGROUND RESULTS CONCLUSION

Females produce a gamete called an oocyte or egg just as males produce sperm. The formation of these gametes is a process of meiosis and then fertilization between these two can occur. The resulting zygote will undergo mitosis and development until it is a fully grown adult. Mitosis is part of the cell cycle where a cell is replicated and produces an identical copy of itself (Campbell 2011). Following mitosis a cell enters a large portion of the cell cycle called interphase. Interphase is comprised of the three major sub-phases G1, S, and G2. At the G1 checkpoint the cell can then be navigated to the G0 phase, which basically means the cell goes into a non-dividing dormancy state. Throughout the cell cycle there are fluctuations in the abundance of cell cycle control molecules and these molecules are primarily two types of proteins: cyclins and protein kinases. In order to be active, a kinase must be attached to a cyclin. These kinases are therefore called cyclin-dependent kinases (Cdks). Maturation-promoting factor (MPF) triggers a cells passage into mitosis after the G2 checkpoint (Campbell 2011). The level of MPF being too low would make it difficult for a cell to grow during interphase and would also slow the process of aging. A female is able to produce eggs during a certain period in her lifetime and the extension of a females reproductive period could show signs of longer life in general and younger, healthier cells. The Abbott group has figured out that the best time to work with an egg in vitro would be as soon as possible to maintain the same qualities as in vivo (Abbott et al. 1998). Since we are examining mammals in general and not simply humans, it worthy to note that the results presented in this project do not only focus on humans. In fact, it has been researched in the past by the Chi group that the optimal nutrient requirements for mouse ova may not be exactly the same for humans, stating that specific amino acids and some fatty acids may be of greater importance, as one example (Chi et al. 1988). It would be a great advancement to give these

oocytes or cells in general a longer time to mature, or reset their certain features to a youthful age. Michael Fossel has explored the potential of inserting a gene for the protein component of telomerase into aging cells in humans and has found some interesting results. This includes the lengthening of telomeres to that of a youthful telomere and even more signs of reversed aging throughout the cell body (Fossel 1998). Recently, there have been studies to reset cellular chronological age and give cells an opportunity to age at a slower rate under modified conditions. More specifically, there has been one study to examine the cumulus-oocyte complexes (COCs) and cumulus-denuded oocytes (DOs) of mice, which were freshly ovulating. These samples were placed in a Chatot-ZiomekBavister (CZB) medium. These experimenters were attempting to connect pyruvate with a property of slowing the aging process of these cells (Liu et al. 2009). This is in direct relation to our topic as we are examining the possibilities available to slow aging or even prevent aging.

Freshly ovulated mouse oocytes were cultured with a variety of pyruvate concentrations in the CZB medium that was utilized. By the end of the culturing period, each culture was observed for MPF activity. Each culture was also treated with ethanol to measure the activation rates. The COC cultures containing a small level of pyruvate exhibited both an increased rate of activation and a decreased level of MPF. These amounts of pyruvate giving these results consistently were of 0.27 and 2.5 mM concentration. The larger concentrations (5, 10, and 20 mM) of pyruvate did not exhibit a considerable difference from that of the controls. When the DO cultures were observed, it was found that they were very similar to the controls in activation rates and MPF activity. In order to study the effects of the pyruvate on aging oocytes further, fresh mouse oocytes were cultured with and without the pyruvate for 6 and 12 hours followed by in vitro insemination or were subjected to a chymotrypsin digestion assay of zona pellucida (ZP). Supplementation of pyruvate to the medium increased fertilization rates while decreased the chymotrypsin digestion time of ZP in aged oocytes. When an aging-accelerating agent was present in the medium, it was found that the pyruvate counteracted this agent in attempt to keep the cell younger and healthier.

Abbott and her colleagues found something similar to Liu in that an in vitro culture stopped cell cycle events, which were in association with the second meiotic reduction completion. These oocytes were less susceptible to parthenogenetic activation in comparison to oocytes that resided in the oviduct. Abbott found also that due to an increase in age the activity in MPF levels decrease. This helped Liu and associates to draw their conclusions on the effect pyruvate had on MPF levels on oocytes. It is our understanding that from these results that the implementation of the pyruvate into the oocytes are improving fertilizability and giving them a supplemental life, or a longer developmental capacity. The pyruvate in general acts as a sort of aging-decelerating agent. This is because it was shown to counteract a medium in which the oocytes were to grow at a more rapid rate. Also, the pyruvate was shown to consistently give positive results for increasing fertilization ratio. Two more very important manifestations of the experimentation completed by Na Liu and associates were that the MPF activity decreased and premature exocytosis or cortical granules was blocked in aged oocytes with supplementation of pyruvate. Michael Fossels study with telomeres in senescent human cells leads us to believe that in the future we will be able to implement his technique to cure diseases caused by age. He has basically reset the chronological ages of some cells and it might be possible to one day give human-kind a longer average life span and a greater maximum life span, but will not act as an elixir-of-life type of solution.