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Effects Of Microtubule Stabilizing And Recovery Agents On Vitrified Bovine Oocytes, Emily Dzekunskas
Effects Of Microtubule Stabilizing And Recovery Agents On Vitrified Bovine Oocytes, Emily Dzekunskas
LSU Master's Theses
Oocyte vitrification, while beneficial for research and species conservation, has limited success due to cryoinjury to the meiotic spindle. This study aimed to evaluate treatments to improve meiotic spindle recovery after vitrification by treating bovine oocytes with either a microtubule stabilizing agent, or a microtubule recovery agent. In the first two experiments, Taxol and epothilone B were used to treat bovine oocytes before vitrification. Both compounds have microtubule stabilizing properties and are known antimitotic compounds commonly used in cancer treatment. Oocytes were incubated with one of the microtubule stabilizing agents for 15 minutes before vitrification at one of three concentrations …
Sperm Decondensation And Male Pronuclear Formation In Bovine Intracytoplasmic Sperm Injection, Lauren Gatenby
Sperm Decondensation And Male Pronuclear Formation In Bovine Intracytoplasmic Sperm Injection, Lauren Gatenby
LSU Master's Theses
This study assessed the effects both DTT and progesterone have on bovine spermatozoa to induce decondensation or the acrosome reaction in order to facilitate male pronuclear formation after ICSI. Sperm prepared by swim-up in a 5 mM concentration of DTT displayed time dependent morphological changes resulting in decondensation over a 6-hour period. An estimated 90% of treated sperm displayed early changes in morphology after the second hour of incubation. Sperm displaying partial decondensation of the nucleus was estimated as 42% by the 3rd hour and increased to 62% by the 5th hour of incubation. Fully decondensed sperm or …
Regulation Of Oocyte Meiotic Resumption Using Camp Modulators In Bovine In Vitro Maturation, Sarah Farmer
Regulation Of Oocyte Meiotic Resumption Using Camp Modulators In Bovine In Vitro Maturation, Sarah Farmer
LSU Master's Theses
In vitro maturation (IVM) is a reproductive technique critical to in vitro embryo production (IVP) in commercial livestock industries, research, and human infertility treatment. Currently, IVP has low efficiency due to an inadequate IVM system in which premature meiotic resumption results in low oocyte viability. Meiotic arrest is regulated primarily by 3’,5’-cyclic adenosine monophosphate (cAMP), and the most successful methods of improving IVM utilize cAMP modulators to maintain high intra-oocyte cAMP, delaying the onset of maturation. This thesis includes experiments comparing standard bovine IVM to a novel extended IVM method similar to the procedure described by Albuz and colleagues (Albuz …
Effect Of Culture Conditions On Gene Expression In Manipulated Bovine Embryos, Megan Nicole Purpera
Effect Of Culture Conditions On Gene Expression In Manipulated Bovine Embryos, Megan Nicole Purpera
LSU Master's Theses
Numerous studies have reported aberrant gene expression levels attributed to suboptimal in vitro culture conditions presented to embryos. Since the culture environment is a common aspect of both in vitro production (IVP) and nuclear transfer (NT), research focusing on the in vitro culture system will have the potential to improve both techniques. This study investigated the effects of different culture systems and protein sources on the developmental competence of IVP embryos measured by cleavage and blastocyst rates, cell number, and relative abundance of oct-4, nanog, connexin 43, and GLUT-1 transcripts when compared to in vivo embryos. Experiment 1 compared IVP …
Cryopreservation Of Bovine And Caprine Oocytes By Vitrification, Sabrina Marie Luster
Cryopreservation Of Bovine And Caprine Oocytes By Vitrification, Sabrina Marie Luster
LSU Master's Theses
Cryopreservation of animal oocytes will permit germplasm of valuable or unique females to be preserved for extended times. The objective of this research was to derive a procedure to cryopreserve bovine oocytes by vitrification to be used as recipients for somatic cell nuclear transfer (SCNT). Caprine oocytes vitrified by the same procedure were assayed by cytological examination of microtubules. In the first two of three experiments, bovine oocytes matured in vitro were vitrified in a mixture of ethylene glycol (EG), dimethylsulfoxide (Me2SO) and trehalose, and then subjected to in vitro fertilization (IVF) or SCNT. For vitrification, oocytes were first exposed …
The Effect Of Holding Bovine Oocytes In Follicular Fluid On Subsequent Fertilization And Embryonic Development, Angela Marie Klumpp
The Effect Of Holding Bovine Oocytes In Follicular Fluid On Subsequent Fertilization And Embryonic Development, Angela Marie Klumpp
LSU Master's Theses
The objective of Experiment 1 was to determine the effect of bovine follicular fluid (bFF) on nuclear maturation. Treatment A (Control) oocytes were stained with Hoechst-33342 immediately after aspiration from follicles, whereas, oocytes in Treatment B were held in bFF for 12 hours at 38¢ªC and then stained to determine nuclear status. No significant difference was detected between treatment groups. Results indicate that bFF inhibits resumption of meiosis. The objective of Experiment 2 was to determine the effect of bFF on embryonic development. Oocytes in Treatment A (Control) were placed into in vitro maturation (IVM) for 22 hours followed by …