Award details

Quantification of gamete health by fluorescent microscopy

Reference	BB/C511105/1
Principal Investigator / Supervisor	Professor Helen Picton
Co-Investigators / Co-Supervisors	Dr Sarah Harris, Dr David Miller
Institution	University of Leeds
Department	School of Medicine
Funding type	Research
Value (£)	39,443
Status	Completed
Type	Research Grant
Start date	01/01/2005
End date	30/09/2005
Duration	9 months

Abstract

Since the application of assisted reproduction technologies (ARTs) to the commercial production of cattle and sheep, basic research into cellular and molecular aspects of gametogenesis and embryogenesis in these species has been opened up. Gametes and embryos from these animals have also proved invaluable as models for human oocyte and pre-implantation embryo development as relatively few of these precious cells are available for research. However, despite much research attention, reproductive technologies remain relatively inefficient and costly, especially in humans. One of the main reasons for the failure of ART in all species is the poor developmental potential (i.e. quality) of the gametes and the failure of embryos to implant after transfer. While the low efficiency of assisted reproduction is due in part to DNA damage in sperm and a high frequency of chromosome abnormalities in oocytes, the problem can also be attributed to a general lack of understanding of the molecular and cell biology of gametogenesis in large animals and humans. Following the development of molecular biology techniques which are sensitive at the single cell level, we have been able to isolate a number of novel genes which may be involved in the production of fertile oocytes. To confirm the functional significance of these genes it is necessary to use antisense technologies to induce gene silencing in vitro. To be effective these methods need to be supported by functional assays of gamete health. Using the latest advances in fluorescent optics and digital image processing, it should now be possible to quantify gamete health and developmental potential by non-invasive measurements of cellular function such as the distribution and activity of mitochondria and to link these measurements to studies of gene expression and chromosome composition in the same cell. The non-invasive nature of the proposed fluorescent assays will also enable us to combine measurements of gamete health with post fertilisation events such as embryo morphology, embryo cleavage and blastocyst production rates, embryo metabolism and gene expression. Thus non-invasive assays will be used dynamically to quantify the quality of individual oocytes and sperm after in vitro manipulation, but without inducing terminal damage in the cells. This research will be of direct relevance to the success of assisted reproduction as there is considerable heterogeneity both between individual gametes and between donors. The primary objectives of this proposal are therefore to purchase and set up a fluorescent imaging system for living cells which will both facilitate investigations of gene function in gametes in vitro and enable the non-invasive quantification of gamete quality by the measurement of defined biological functions within individual cells. The requested workstation is multi-tasking and can be upgraded in future to accommodate different markers of cell function so it will serve our research needs for many years to come. The ability to quantify gamete health will be invaluable in supporting our continuing investigations into: the molecular and cellular biology of gametogenesis; the impact of culture environment and assisted reproduction on gamete quality; and the quantification of sperm mitochondria and nuclear dynamics in response to external stress.

Summary

unavailable

Committee	Closed Committee - Animal Sciences (AS)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	Research Equipment Initiative 2004 (RE4) [2004]
Funding Scheme	X – not Funded via a specific Funding Scheme

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