We propose to investigate nucleo-cytoplasmic interactions which take place during normal development and to determine the mechanism by which these interactions are controlled. The role of various components of the mammalian zygote (male genome, female genome, egg cytoplasm) in development will be analyzed using recently developed nuclear transfer techniques. Our first major objective will be to determine the reasons for developmental failure of biparental androgenones (embryos with two male pronuclei) and biparental gynogenones (two female pronuclei). Development of chimeric embryos produced by aggregation of gynogenetic or androgenetic and normal embryos will be monitored in order to determine whether embryo- or cell-lethality is ultimately responsible for death of isoparental embryos. Haploid nuclei obtained from successive stages of male gametogenesis will be transferred into zygotes from which male nuclei were removed. These experiments will address the timing of appearance of functional differences between the male and the female genome. The second major objective will be to analyze development of enucleated zygotes into which nuclei from older developmental stages are transferred. We will determine if repeated nuclear transfer will improve the development of such embryos. The use of cytoplasm from older embryos (4- and 8-cell stage) as the nuclear recipient will be explored. Using 2D gel analysis we will monitor the changes in genome expression following nuclear transfer. Nuclear morphology, changes in nuclear proteins (lamins) and the structure of the mitotic apparatus in these nuclear transfer embryos will be investigated.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD017720-07
Application #
3314739
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1983-08-01
Project End
1991-07-31
Budget Start
1989-08-01
Budget End
1990-07-31
Support Year
7
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Wistar Institute
Department
Type
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Latham, K E; Garrels, J I; Solter, D (1993) Two-dimensional gel analysis of protein synthesis. Methods Enzymol 225:473-89
Latham, K E; Solter, D (1993) Transplantation of nuclei to oocytes and embryos. Methods Enzymol 225:719-32
Latham, K E; Beddington, R S; Solter, D et al. (1993) Quantitative analysis of protein synthesis in mouse embryos. II: Differentiation of endoderm, mesoderm, and ectoderm. Mol Reprod Dev 35:140-50
Lamb, B T; Satyamoorthy, K; Solter, D et al. (1992) A DNA element that regulates expression of an endogenous retrovirus during F9 cell differentiation is E1A dependent. Mol Cell Biol 12:4824-33
Latham, K E; Solter, D; Schultz, R M (1992) Acquisition of a transcriptionally permissive state during the 1-cell stage of mouse embryogenesis. Dev Biol 149:457-62
Latham, K E; Garrels, J I; Chang, C et al. (1992) Analysis of embryonic mouse development: construction of a high-resolution, two-dimensional gel protein database. Appl Theor Electrophor 2:163-70
Latham, K E; Solter, D (1991) Effect of egg composition on the developmental capacity of androgenetic mouse embryos. Development 113:561-8
Lamb, B T; Satyamoorthy, K; Li, L et al. (1991) CpG methylation of an endogenous retroviral enhancer inhibits transcription factor binding and activity. Gene Expr 1:185-96
Latham, K E; Solter, D; Schultz, R M (1991) Activation of a two-cell stage-specific gene following transfer of heterologous nuclei into enucleated mouse embryos. Mol Reprod Dev 30:182-6
Latham, K E; Garrels, J I; Chang, C et al. (1991) Quantitative analysis of protein synthesis in mouse embryos. I. Extensive reprogramming at the one- and two-cell stages. Development 112:921-32

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