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  • 1
    ISSN: 1432-2048
    Keywords: Acetyl CoA ; Chloroplast ; Hordeum ; Metabolic pathway (phosphoglycerate to acetyl CoA) ; 3-Phosphoglycerate ; Phosphoglycerate mutase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Earlier studies on the synthesis of C3-derived amino acids, plastidic isoprenoids and fatty acids from CO2 by isolated chloroplasts in the light indicate the presence of a complete, but low-capacity, chloroplast (chlp) 3-phosphoglycerate → acetyl-CoA pathway which is predominantely active in immature (developing) chloroplasts (A. Heintze et al., 1990, Plant Physiol. 93, 1121–1127). In this paper, we demonstrate the activity of the enzymes involved i.e. chlp phosphoglycerate mutase, chlp enolase, chlp pyruvate kinase and chlp pyruvate-dehydrogenase complex (PDC), in the stroma of purified barley (Hordeum sativum L.) chloroplasts of different developmental stages. The chlp phosphoglycerate mutase was partially purified for the first time. The activities of the enzymes of this chlp pathway (except PDC) were about a magnitude lower than those of the cytosolic enzymes. The chlp PDC of barley was more active than that of spinach. The apparent K m values of the enzymes of this pathway were about 100 μM or lower except for the chlp phosphoglycerate mutase which had a K m of 1.6–1.8 mM for 3-phospho-d-glycerate. Interestingly, no appreciable change in the activity of these enzymes was observed during maturation of the chloroplasts. In contrast, the activity of the reversible NADP+-glyceraldehyde 3-phosphate dehydrogenase increased about five times (from 140 to 590 nkat per g leaf dry weight). The following hypothesis is put forward to explain the regulation of carbon metabolism during chloroplast development: 3-phospho-d-glycerate is withdrawn from a common pool by the actions of 3-phosphoglycerate kinase and NADP+-glyceraldehyde-3-phosphate dehydrogenase, the activity of which increases considerably during maturation of chloroplasts. This leads to an insufficient supply of 3-phospho-glycerate for the chlp phosphoglycerate mutase, which has a low affinity for its substrate.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 3 (1980), S. 343-349 
    ISSN: 0148-7280
    Keywords: Sperm ; capacitation ; mouse strain differences ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Intial in vivo studies were performed to observe the proportion of eggs fertillized at specific intervals after natural mating and ovulation in our research mouse colony. Proestrous females of the C57BL/10Wt, SJL/Wt inbred strains and the F1 hybrid cross (B10 × SJL or reciprocals) were paired in the after-noon with males of their respective strain and examined for vaginal plugs at the midpoint of the dark period (2400 hours). Oviducts were periodically collected from mated females, and ovulation was first observed at 4, 5.2, and 3 hours after 2400 hours in the B10, SJL, and F1 hyrid, respectively. The clutch of eggs from each ovulating female, was placed in culture, and cleavage oviduct lavage verifying female mating was placed in culture, and cleavage was used as the criterion for fertilizaition. Fifty percent of the eggs were fertilized 2.2, 5.0, and 2.5 hours after ovulation in B10, SJL, and F1 hybrid females, respectively. Because twice the legth of time was required to fertilize a similar proportion of eggs from the SJL strain as the F1 hybrid, these two strains were used for determining their rate of fertilization under more fully controlled conditions in vitro. Forty-nine percent of F1 hybrid eggs were fertilized after 4 hours incubation with SJL epididymal sperm, whereas 53% fo SJL and 56% of F1 hybrid eggs were fertilized after only 2 hours incubation with F1 hybrid epididymal sperm. Thus, using sperm from these two mouse strains, the amount of time required to fertilize approximately 50% of the eggs within a clutch both in vivo and vitro was very similar. These observations demonstrte teh validity of using this in vitro system for fertilization studies and confirm that the temporal events in sperm capacitation and egg penetration are dependent on the genotype of the sperm. Similarities in fertilization rates at specific times after ovulation or insemination in vitro imply that the initiationof sperm capacitation in vivo occurs near the time of ovulation and several hours after mating. We tentatively suggest that follicular fluid may be required for completion of mouse sperm capacitaiton in vivo.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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