By Gilbert S. Greenwald (auth.), Cornelia P. Channing, John M. Marsh, William A. Sadler (eds.)
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Extra resources for Ovarian Follicular and Corpus Luteum Function
Although the highest concentration of d-cAMP promoted a 52% decline in LHstimulated AC activity, the cyclase still retained a 5-fold responsiveness to stimulation by LH. These results indicate that d-cAMP, in concentrations which have previously been shown to promote 32 MARY HUNZICKER-DUNN ET AL. I NaF BSA Additions to 2 hour Incubation Fig. 4. ) on AC activity. Follicles from estrous rabbits were incubated for 2 h at 37 C in KRB-glucose containing BSA, 10 ~g/ml LH, or d-cAMP in the concentrations indicated on the figure.
L.... ·,,······ •. 2 10 Mn in 2nd Sto~ of Incubation (mM) Fig. 6. Effect of Mn 2+ on the AC activity of porcine follicular membranes. Graafian follicle membranes (18 ~g protein; see legend to Fig. 5 for details and ref. 14) were incubated in 3 stages at 30 C as diagrammed on the figure. 67 times those desired in the third stage of the incubation). (b) In the second stage of the incubation (10 min), Mn 2+ (in concentrations indicated on the figure) was added to the first incubation mixture in a volume of 10 ~l.
Percent desensitization is calculated as the decline in LH-stimulated AC activity measured with LH present in all 3 stages of the incubation vs. that with LH present only in the AC assay (stage 3). Zero percent reversal of desensitization corresponds to values obtained in the absence of added phosphatase (BSA control; 21% desensitization). Complete reversal of LH-induced desensitization was obtained using a phosphatase-enriched preparation (containing 15 ~g protein and exhibiting 110 pmoles/min/mg protein specific activity) obtained from calf thymus nuclei (not shown).