Pertussis toxin-catalyzed ADP-ribosylation of a G protein in mouse oocytes, eggs, and preimplantation embryos: Developmental changes and possible functional roles
- Univ. of Pennsylvania, Philadelphia (USA)
G proteins, which in many somatic cells serve as mediators of signal transduction, were identified in preimplantation mouse embryos by their capacity to undergo pertussis toxin-catalyzed ADP-ribosylation. Two pertussis toxin (PT) substrates with Mr = 38,000 and 39,000 (alpha 38 and alpha 39) are present in approximately equal amounts. Relative to the amount in freshly isolated germinal vesicle (GV)-intact oocytes, the amount of PT-catalyzed ADP-ribosylation of alpha 38-39 falls during oocyte maturation, rises between the one- and two-cell stages, falls by the eight-cell and morula stages, and increases again by the blastocyst stage. The decrease in PT-catalyzed ADP-ribosylation of alpha 38-39 that occurs during oocyte maturation, however, does not require germinal vesicle breakdown (GVBD), since inhibiting GVBD with 3-isobutyl-1-methyl xanthine (IBMX) does not prevent the decrease in the extent of PT-catalyzed ADP-ribosylation. A biologically active phorbol diester (12-O-tetradecanoyl phorbol 13-acetate), but not an inactive one (4 alpha-phorbol 12,13-didecanoate, 4 alpha-PDD), totally inhibits the increase in PT-catalyzed ADP-ribosylation of alpha 38-39 that occurs between the one- and two-cell stage; TPA inhibits cleavage, but not transcriptional activation, which occurs in the two-cell embryo. In contrast, cytochalasin D, genistein, or aphidicolin, each of which inhibits cleavage of one-cell embryos, or alpha-amanitin or H8, each of which inhibits transcriptional activation but not cleavage of one-cell embryos, have little or inhibitory effects on the increase in PT-catalyzed ADP-ribosylation of alpha 38-39. Results of immunoblotting experiments using an antibody that is highly specific for alpha il-3 reveal the presence of a cross-reactive species of Mr = 38,000 (alpha 38) in the GV-intact oocyte, metaphase II-arrested egg, and one-, two-cell embryos.
- OSTI ID:
- 7011533
- Journal Information:
- Developmental Biology; (USA), Vol. 139:2; ISSN 0012-1606
- Country of Publication:
- United States
- Language:
- English
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OOCYTES
CELL DIFFERENTIATION
PHORBOL ESTERS
BIOLOGICAL EFFECTS
PROTEINS
METABOLISM
ADP
CELL MEMBRANES
CLEAVAGE
ELECTROPHORESIS
INHIBITION
METHIONINE
MICE
MOLECULAR WEIGHT
NAD
ONTOGENESIS
SULFUR ISOTOPES
TOXINS
TRACER TECHNIQUES
TRANSCRIPTION
AMINO ACIDS
ANIMALS
ANTIGENS
CARBOXYLIC ACIDS
CARCINOGENS
CELL CONSTITUENTS
COENZYMES
CRYSTAL STRUCTURE
DRUGS
ESTERS
GERM CELLS
ISOTOPE APPLICATIONS
ISOTOPES
LIPOTROPIC FACTORS
MAMMALS
MATERIALS
MEMBRANES
MICROSTRUCTURE
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
RODENTS
TOXIC MATERIALS
VERTEBRATES
560300* - Chemicals Metabolism & Toxicology
550501 - Metabolism- Tracer Techniques