Biol. Neonate 32: 33 -42 (1977)

Regulation of Cyclic Nucleotide Phosphodiesterase Activity in Rhesus Fetal Muscle Clarissa H. Beatty, Paul T. Herrington and Rose Mary Bocek Division of Perinatal Physiology, Oregon Regional Primate Research Center, Beaverton, Oreg. and Department of Biochemistry, University of Oregon Medical School, Portland, Oreg.

Key Words. Fetal rhesus muscle • Guanosine 3':5'-monophosphate • Subcellular distribution of cyclic nucleotide phosphodiesterases • Regulation of activity of cyclic nucleotide phosphodiester­ ases

Introduction

Cyclic nucleotides are involved in regulating a number of biological processes. In rapidly growing muscle, levels of cyclic nucleotides are of particular interest since both cyclic adeno­ sine 3':5'-monophosphate (cyclic AMP) and cyclic guanosine 3':5'-monophosphate (cyclic GMP) appear to influence growth and differen­ tiation of myoblasts (16, 33) and fibroblasts (19). We have previously reported that levels of

cyclic AMP and adenylate cyclase activity (4, 5) are higher in fetal than in adult rhesus muscle (Macaca mulatto). Many investigators believe that cyclic AMP and cyclic GMP function in biologic regulation as opposing influences (9, 26), although recent evidence indicates that this hypothesis is not always true (20, 21, 26). Conversion of cyclic GMP and cyclic AMP to the 5'-nucleotide is the only physiological mechanism known to terminate their action. Most tissues appear to contain at least two

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Abstract. In both fetal and adult rhesus muscle, low levels of guanosine 3':5'-monophosphate (cyclic GMP; 0.25-5.0 yM) stimulated the adenosine 3':5'-monophosphate (cyclic AMP)phosphodiesterase (PDE) at low substrate levels; the fetal enzyme was more sensitive than the adult (supernatant and particulate fractions). At high levels of substrate cyclic AMP (25-100 ¡jM ), hydrolysis was not influenced by either 10 or 20 ijM cyclic GMP but the fetal and adult series were equally inhibited by 40 /uW cyclic GMP. In the supernatant fraction cyclic AMP inhibition of cyclic GMP hydrolysis increased with increasing cyclic AMP levels. There was no difference between fetal and adult muscle preparations in percent inhibition; however, in the adult series, the inhibition was noncompetitive whereas in the fetal series the inhibition was competitive. The Kj for cyclic AMP was 40 juM for fetal and 100 juM for adult cyclic GMP-PDE. The values for cyclic GMP-PDE and both cyclic AMP-PDE enzymes were higher in the fetal series.

Bcatty/Herrington/Bocek

cyclic nucleotide phophodiesterase (PDE) ac­ tivities which differ in kinetic behavior, in relative affinities for their substrates (cyclic AMP and cyclic GMP), in molecular weight, and in differential activation and inhibition (26, 29). Although the cyclases, especially adenylate cyclase are known to be under metabolic regu­ lation, less is known about control of the PDEs. Developmental changes in tissues from rats and guinea pigs have been studied (7, 31). In the experiments reported here, we studied the kinetics of the fetal PDEs and their sensi­ tivity to the potential biological regulators cyclic GMP and cyclic AMP. We also deter­ mined the percentage of cyclic GMP-PDE and cyclic AMP-PDE in the supernatant and particu­ late fractions obtained from fetal and adult rhesus muscle homogenates by differential centrifugation. The cyclic nucleotides appear to play a central role in cell growth and metabo­ lism and it is important to know the ontogenic development of the enzymes involved in their metabolism.

Materials and Methods Rhesus fetuses (M. mulatto), at 150 days gesta­ tional age, and adult rhesus monkeys were used. (The average gestational age in our colony is 165 days.) The mothers were anesthesized intramuscularly with 1.0 mg/kg of Sernylan (l-|phenylcyclohexyl|piperdine hydrochloride) and the fetuses, obtained by cesarean section, were anesthesized with Ketaject (dl-2-|och!oropheny!|-2-|mcthylamino| cyclohexanone hydro­ chloride). Samples of muscle were obtained from the biceps-triceps groups of the upper arms, freed of fat and connective tissue, and cither frozen in a Wollenberger clamp cooled in liquid nitrogen or used immediately unfrozen. No difference was observed in the cyclic nucleotide PDE activity after freezing. Adult rhesis monkeys were anesthesized with Ketaject or Sernylan and samples of the sartorius muscle obtained at biopsy were treated similarly. There was no difference in the data obtained with the two

anesthetics. Both the biceps-triceps group and the sartorius are predominantly red muscles and give similar results for the purpose of this experiment. Preparation o f Cellular Fractions A 10% (w/v) homogenate in 0.25 M sucrose-10 mM Tris, pH 7.4, was prepared. Fresh muscle was weighed and minced in a small amount of sucrosc-Tris buffer. Frozen tissue was weighed in the frozen state, thawed in a small amount of cold buffer, and minced. Onehalf of the required volume of medium was added and the tissue was homogenized in a Polytron ST-20 for 10 sec at ’/« the maximum setting. Homogenization with a conical glass-glass system produced virtually the same percentage distribution of PDE in the 100,000? fractions. However, homogenization in the polytron was faster. After centrifugation for 5 min at 500?, and 0 °C the supernatant was decanted and the pre­ cipitate was resuspended in the remainder of the sucrose-Tris buffer and rehomogenized for 10 sec at the same setting. This second homogenate was centri­ fuged at 500? for 5 min. The supernatants were combined and centrifuged for 10 min at 650?. An aliquot of this fraction was saved for enzyme assay, and the remainder was centrifuged at 10,000? for 30 min, decanted and then centrifuged for 60 min at 100,000?. The final 100,000? supernatant was designated the soluble fraction. The particulate frac­ tions from the 10,000 and 100,000? centrifugations were washed by resuspension in 12 ml of fresh buffer and recentrifugation. The resultant pellets were re­ suspended in 1-1.5 ml sucrose-Tris buffer. All frac­ tions were quick-frozen in dry ice-acetone and stored at - 20 °C; at this temperature, the activity remained unchanged for at least 3 months. Since over 87% of the total cyclic nucleotide PDE activity of fetal and adult skeletal muscle is in the 100,000? supernatant fraction, we used this fraction to investigate the effect of cyclic nucleotides on the hydrolysis of the other nucleotide. Beavo et al. (2) also reported that most of the cyclic nucleotide PDE activity is in the superna­ tant fraction of adult rat skeletal muscle. Cyclic Nucleotide PDF A ssavs Cyclic nucleotide PDE activities w'erc determined by a modification of l.oten and Sneyd's (14) two-stage assay, previously described (3). The triated cyclic nucleotides, hydrolyzed by PDE to the 'H-5'-dcrivative were converted to JH-adcnosinc or JH-guanosine and isolated with a Bio-Rad AG1-X2 (200-400) mesh

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34

Cyclic Nucleotide Phosphodiesterases in Fetal Muscle

column for counting. The amount of hydrolyzed cyclic nucleotides were calculated from the specific activity of the substrate and expressed in terms of nanomoles of cyclic nucleotide hydrolyzed per minute per milligram of protein. The concentrations of en­ zyme preparations were such that less than 25% of the substrate was hydrolyzed. Substrate concentrations are indicated in the tables and graphs and the level of 3H-cyclic GMP was 0.07 ;uCi/assay tube (8.4 Ci/mmol) and of JH-cyclic AMP was 0.02 /iCi/assay tube (38.2 Ci/mmol). Radioactivity in the blanks was usual­ ly less than 0.5% of the total added to the assay. Protein content of the various fractions was deter­ mined by the method of Cowry et al. (15). Bovine serum albumin was used as a standard. Km and Kj values were calculated by the method of least squares. Cyclic GMP Cyclic GMP was determined by a competitive binding protein assay. Frozen tissues were homoge­ nized in 6% TCA containing 0.05 N HC1. Supernatants were extracted 5 times with 3 times their volume of diethyl ether. The aqueous phase was applied to a neutral alumina column, eluted with 0.6 M Tris pH 7.4 onto a Bio-Rad AG1-X2 (200- 400 mesh) column. Cyclic AMP and cyclic GMP were differentially eluted

35

with 0.05 and 0.5 N HC1, respectively (17). An aliquot of the cluatc was evaporated to dryness, redissolved in 50 mM Tiis buffer, pH 7.5, and assayed by a competi­ tive-binding assay (8).

Results

This paper concerns our studies of cyclic GMP-PDE as well as cyclic AMP-PDE and the distribution of activity in the subcellular frac­ tions isolated from fetal and adult muscle by differential centrifugation. Previously, we have reported that the cyclic AMP-PDE activity of crude muscle homogenates was higher in muscle from the rhesus fetus than that from the adult (3). The enzyme activity in the 650 g superna­ tant was assumed to be 100%. Over 80% of the cyclic nucleotide PDE activity was found in the 100,000g supernatant fraction and only 5% in the combined 10.000 and 100,000 g particu­ lates (table 1). Fetal muscle did not differ from adult in percent distribution of cyclic GMP-

Table I. Distribution of cyclic nucleotide PDE activities in subcellular fractions of muscle homogenates from adult and 150-day fetal rhesus monkeys 100 vM cyclic AMP

20 fcM cyclic GMP

0.75 juA/ cyclic AMP

adult

fetal

adult

fetal

adult

fetal

20.6

2.0

3.2

17.9

49.0

1.4 82 4.3

0.8 89 2

1.2 92 3.9

0.4 86 1.3

1.3 87 4.0

Activity, nmol hydrolyzed/g wet weight -m in '1 650 g supernatant 4.4 Percent activity in subcellular fractions 10,000 g particulate 1.1 100,000 g supernatant 86 100,000 g particulate 0.4

Tissues were homogenized in 0.25 M sucrose-10 mM Tris, pH 7.4 and fractionated by differential centrifuga­ tion. Reaction mixtures contained 0.02 MCi 3H-cyclic AMP (38.2 Ci/mmol), or 0.07 /aCi 3H-cyclic GMP (8.4 Ci/mmol) 40 mM Tris buffer pH 8.0, 2 nvW MgClj, 3.75 mM mercaptoethanol, and 1% bovine serum albu­ min plus enzyme preparation in a final volume of 0.2 ml. Cyclic nucleotide substrate concentrations were as indicated. Protein concentrations averaged 0.05 mg (supernatant) and 0.1 mg (particulate) in the fetal and 0.12 mg (supernatant and particulate) in the adult series.

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Fraction

36

Beatty/Herrington/Bocek

PDE activity or in high and low Km cyclic AMP-PDE activities in the cell fractions. Linear Lineweaver-Burk plots were obtained for cyclic GMP-PDE in both the supernatant and the 100,000£ particulate of rhesus fetal muscle

(fig. 1). However, nonlinear plots were obtained with cyclic AMP as a substrate and this suggests the presence of two cyclic AMP-PDE activities (a high and a low substrate affinity forms) and/or a single enzyme undergoing negative

Fig. 1 . Lineweaver-Burk plots for the hydrolysis of cyclic GMP by 100,000 g supernatant and particulate of homogenates from 150-day rhesus fetal muscle. Substrate concentrations ranged from 0.55 to 20 ¡jM.

Velocities arc expressed as nmol cyclic GMP hydro­ lyzed min-1 mg protein'*1. Conditions of assay de­ scribed in table 1.

Table II. Kinetic values for cyclic nucletode PDE in 100,000 g supernatant and pellet from homogenates of muscle from 150-day fetal and adult rhesus monkeys Cyclic GMP-PDE Km, fiM

100,000g supernatant 7 Adult 9.2 ± 1.9 Fetus 2 13 100,000g particulate Adult 3 8.5 ± 1.7 Fetus 2 27'

n

Vmax

16 -s 6 140' 2.4 ± 0.4 851

Cyclic AMP-PDE low affinity

high affinity

KnvM*/

Vmax

Km,ßM

Vmax

4 2

13 ± 1.8 231

55 ± 12 2321

4.3 ± 0.5 9.9'

35 ± 6 155'

2 2

18 43

6.9J 94

2.8 4.7

4.22 231

Values are means * SD. Substrate concentrations ranged from 0.7 to 20 nM cyclic GMP or from 0.8 to 100 *vW cyclic AMP. Velocities are in terms of nanomoles of cyclic nucleotide hydrolyzed m in '1 100 mg p ro tein''. 1 Values more than 3 times the SD for the average adult series. ! Supernatant versus pellet 7 9 times different.

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n

37

Cyclic Nucleotide Phosphodiesterases in Fetal Muscle

Table III. Cyclic GMP levels in fetal and adult rhesus muscle Cyclic GMP

Fetal 100-day 150-day Adult

19 13 6

pmol/mg nitrogen

1.18 0.51 0.192

2 fetuses in the 100-day series, 5 in the 150-day series and 5 in the adult series (duplicate analyses on duplicate tissue samples). Cyclic GMP determined by a competitive protein-binding assay.

cooperative regulation (supernatant and par­ ticulate fractions of fetal muscle). These results agree with data obtained by other workers on adult tissues in a variety of other species (1, 18, 22, 26). The apparent Km values of both the cyclic GMP-PDE and the low and high affinity cyclic AMP-PDE enzymes appeared to be higher in the fetal than in the adult series (table II). The Vmax values for the cyclic GMP-PDE were 9 times and 35 times higher in the fetal than the adult series for the supernatant and particulate fractions, respectively (table II). The Vmax values for cyclic AMP-PDE in the two fractions were also higher in the fetal series. The data on cyclic AMP-PDE agrees with earlier results on crude muscle homogenates (3). The level of cyclic GMP is higher in fetal than in adult skeletal muscle; we have previous­ ly reported that cyclic AMP is also higher in fetal muscle. Several studies have shown cyclic AMP levels to be a function of age, particularly in the rat brain (30). We know of no values for cyclic GMP in fetal tissues. The value for cyclic

Cyclic GMP

\iM

Fig. 2. Effect of cyclic GMP on the hydrolysis of cyclic AMP by cyclic AMP-PDE in the 100,000 g supernatant fraction of homogenates of 150-day fetal and adult rhesus muscle. Values are means. Perpen­ dicular lines above bars represent one standard error. Numbers inside bars represent number of experiments on 4 fetal and 3 adult monkeys. Substrate levels of 1- 5 tiM cyclic AMP gave similar results; pooled values were graphed.

GMP in adult rhesus skeletal muscle (table III) is lower than reported for adult rat skeletal muscle, 18 ± 1.5 pmol/g wet weight (25). In adult tissues it has been shown that cyclic GMP modifies the activity of cyclic AMP-PDE (12, 22, 23, 27). Since the level of cyclic nucleotides is higher in fetal than in adult muscle, we were particularly interested in the effect of each nucleotide on the hydrolysis of the opposite nucleotide in fetal muscle. There­ fore, we determined the effect of low levels of cyclic GMP (0.25 5.0 nM) on the rate of cyclic AMP hydrolysis by the supernatant fraction of muscle homogenates. In both fetal and adult muscle, low levels of cyclic GMP stimulated the cyclic AMP-PDE at low substrate levels (fig. 2);

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pmol/g wet weight

Beatty/Herrington/Bocek

Fig. 3. Inhibition by cyclic AMP of cyclic GMPPDE in the 100,000 g supernatant from homogenates of fetal and adult muscle. Substrate level of cyclic GMP, 10 idM, Conditions of assay described in table I.

Fig. 4. Inhibition by cyclic AMP of cyclic GMPPDE from the 100,000 g supernatant of homogenates from fetal and adult muscle. Velocities are expressed as nmol cyclic GMP hydrolyzed min"' mg protein"'. Conditions of assay described in table I.

the fetal enzyme was more sensitive that the adult. At high levels of substrate cyclic AMP (25—100 pA/), hydrolysis was not influenced by either 10 or 20 yM cyclic GMP but was in­ hibited by 40 pA/ cyclic GMP in both the fetal and adult series ( - 1 3 ± 2% and -1 1 ± 2%, respectively, n = 6). When enzyme preparations from the particulate fraction of adult and fetal muscle were assayed under conditions identical to those in figure 2, the results were similar to those for the supernatant fraction (data not shown). Cyclic AMP has also been reported to modi­ fy cyclic GMP-PDE activity (22, 23). In the supernatant fraction from homogenates of fetal and adult rhesus muscle, cyclic AMP inhibition of cyclic GMP hydrolysis increased with in­ creasing cyclic AMP levels (fig. 3). There was no difference between fetal and adult muscle prep­ arations in percent inhibition; however, the type of cyclic AMP inhibition differed between fetal and adult muscle (fig. 4). In the adult series cyclic AMP noncompetitively inhibited

cyclic GMP-PDE; in the fetal series, the inhibi­ tion was competitive. The Kj for cyclic AMP was 40 yM for fetal and 100 yM for adult cyclic GMP-PDE. Insulin, a hormone with growth-stimulating properties, decreases the level of cyclic AMP and increases the level of cyclic GMP in tissues such as fat cells and lymphocytes (6, 10. 11), and there is evidence that this hormone acts in some tissues by stimulating cyclic AMP-PDE (13, 14, 23). Insulin has been reported to increase low-Km cyclic AMP-PDE of frog skele­ tal muscle (32). Its action in mammalian muscle is still uncertain. Insulin had no effect on the activity of the low-Km cyclic AMP-PDE in the soluble or particulate fractions of homogenates of muscle fiber groups incubated for 45 min (data not shown). Many agents inhibit cyclic nucleotide PDEs, and the sensitivity of the various PDEs to these agents often differ widely (29). We have deter­ mined the effect of Ro20 1724 (the 3-butoxy-4-methoxy derivative of 4-(3,4-dimethoxy-

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38

Cyclic Nucleotide Phosphodiesterases in I-'etal Muscle

39

distribution in the intracellular space, although compartmentalization may well have occurred. 40 nM is above the physiological range. Inas­ much as cyclic GMP can both stimulate and Inhibitor Inhibition, %' 0.5 mM ----------------------------------------------- inhibit the hydrolysis of cyclic AMP, the cellu­ 5 *iAi cyclic GMP 10 iiM cyclic GMP lar content of cyclic AMP in both fetal and adult muscle may be controlled by cyclic-GMPfetal adult fetal adult induced changes in cyclic AMP-PDE. In several adult tissues of other species cyclic GMP has 67 60 54 Caffeine 51 been reported to be an allosteric effector of 62 Theophylline 73 66 61 56 50 R o 2 0 -1724 70 51 cyclic AMP hydrolysis (12, 22, 27, 28). Activa­ tion of cyclic AMP-PDE by low levels of cyclic 1 Control values were taken as 100%. GMP has been demonstrated and at higher concentrations each nucleotide inhibits the hydrolysis of the other. Cyclic AMP in concentrations of 1—5 0 /uM benzyl]-2-imidazolidinone), caffeine, and theo­ inhibited the hydrolysis of cyclic GMP in the phylline on the hydrolysis of cyclic GMP in fetal and adult muscle (table IV). All three supernatant fractions of both fetal and adult inhibitors had similar effects on the hydrolysis rhesus muscle. Since the intracellular concentra­ of cyclic GMP and cyclic AMP. However, cyclic tion of cyclic AMP is 0.8—1.5/uAf in rhesus GMP-PDE activity in fetal muscle was more fetal muscle and 0.4-0.8 juM in adult muscle, sensitive to all three inhibitors than that in the level of endogenous nucleotide appears to be high enough to inhibit cyclic GMP degrada­ adult muscle. tion in vivo, even in the absence of compart­ mentalization. The details are still unclear but both cyclic AMP and cyclic GMP are probably Discussion involved in regulating the degradation of the We have reported previously that cyclic AMP other nucleotide in fetal muscle. The difference between the types of inhiblevels in rhesus fetal muscle were higher than those in adult muscle (4). We now report that tion by cyclic AMP of cyclic GMP hydrolysis — the level of cyclic GMP was also higher in fetal competitive in the fetal and noncompetitive in titan in adult muscle. Furthermore, low levels the adult supernatant series - indicates a differ­ of cyclic GMP stimulated the hydrolysis of ence between the fetal and adult PDE enzymes. cyclic AMP when the concentration of cyclic Although both cyclic nucleotides are high in AMP was below 5 ¡jM. The fetal enzyme was fetal skeletal muscle, we cannot postulate that more sensitive to stimulation than the adult cyclic GMP and cyclic AMP affect growth enzyme. At a higher concentration (40/iAf) similarly since the ratio of these two nucleo­ cyclic GMP was inhibitory. The lower concen­ tides may also be important. In fetal muscle trations of cyclic GMP used in these experi­ this ratio of cyclic AMP/cyclic GMP is 16 at ments are close to the physiological range since 100 days, 30 at 150 days and 50 in the fully the intracellular level in fetal muscle was calcu­ developed adult muscle. Evidence that cyclic lated to be about 0.04 /aAf assuming uniform AMP acts to induce synthesis of cyclic AMP-

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Table IV. Effect of inhibitors of cyclic GMP-PDE from 100,000/? supernatant of fetal and adult muscle from the rhesus monkey

40

tion is necessary to establish exact kinetic values for cyclic AMP-PDE. However, maxi­ mum enzyme activities should be measured before purification since purification usually leads to loss of activity.

Acknowledgements Publication No. 909 of the Oregon Regional Pri­ mate Research Center, supported in part by grant RR-00163 from the National Institutes of Health, by Public Health Service Research Grants HD-06069 and HD-06425 from the National Institutes of Child Health and Human Development, by General Research Support Grant RR-05694 from the National Institutes of Health, and by the Muscular Dystrophy Associa­ tions of America, Inc.

References 1 Appleman, M.M. and Terasaki, W.L.: Regulation of cyclic nucleotide phosphodiesterase; in Drum­ mond, Grecngard and Robison, Advances in cyclic nucleotide research, vol. 5, pp. 153-161 (Raven Press, New York 1975). 2 Beavo, J.A.; Hardman, J.G., and Sutherland, E.W.: Hydrolysis of cyclic guanosine and adenosine 3',5' monophosphates by rat and bovine tissues. J. biol. Chem.245: 5649-5655 (1970). 3 Bocek, R.M. and Beatty, C.H.: Cyclic AMP phosphodiesterase activity in fetal and adult mus­ cle of the rhesus monkey. Devi Biol. 48: 382- 391 (1976). 4 Bocek, R.M.; Young, M.K., and Beatty, C.H.: Cyclic AMP in developing muscle of the rhesus monkey: effect of prostaglandin E,. Biol. Neonate 28: 92-105 (1976). 5 Bocek, R.M.; Young, M.K., and Beatty, C.ll.: Ef­ fect of insulin and epinephrine on the carbohy­ drate metabolism and adenylate cyclase activity of rhesus fetal muscle. Pediat. Res. 7: 787-793 (1973). 6 Craig, J.W.; Rail, T.W., and Lamer, J.: The in­ fluence of insulin and epinephrine on adenosine 3',5' phosphate and glycogen transferase in muscle. Biochim. biophys. Acta 177: 213-219 (1969).

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PDE (21, 24) may explain the high levels of both in the fetal tissues; the same relationship probably functions when cyclic GMP levels are high. The levels of the cyclic nucleotides are deter­ mined largely, if not entirely, by a balance between the activities of adenylate and guanylate cyclases, which form the compounds and of PDEs, which degrade them. Therefore, low PDE and/or high cyclase activity could explain the high level of cyclic nucleotides in fetal muscle. We reported previously that the adeny­ late cyclase activity of fetal rhesus muscle is higher than that in adult muscle (5). In our latest experiments the Vmax values for cyclic GMP-PDE and high and low affinity cycle AMP-PDE were 4 to 35 times greater in the fetal than in the adult series in both the supernatant and particulate fractions. Hence, the high levels of cyclic nucleotides found in fetal muscle cannot be explained in terms of a decrease in their degradation. While this manu­ script was in preparation, Davis and Kuo (7) reported on the ontogenetical changes in cyclic nucleotides of guinea pig lung, liver, brain and heart. The relative activity during development for both cyclic GMP-PDE and cyclic AMP-PDE were fetus > neonate > adult. Developmental alterations in cyclic nucleotide activities have also been reported in tissues of rats (31). Davis and Kuo (7) also reported that fetal guinea pig lung was richer in cyclic GMP-PDE than in cyclic AMP-PDE (soluble fraction). In rhesus muscle cyclic AMP-PDE was more active than cyclic GMP-PDE in all the fractions except the fetal particulate. However, there is no way of estimating from this data the relative activities in vivo. Assuming that the Lineweaver-Burk plots indicate the presence of two isozymes of cyclic AMP-PDE, the Vmax values for the high and low Km enzymes will be influenced by the presence of the other enzyme. Further purifica­

Beatty/Herrington/Bocek

7 Davis, C.W. and Kuo, J.F.: Ontogenetic changes in levels of phosphodiesterase for adenosine 3':5'monophosphate and guanosine 3':5'-monophosphatc in the lung, liver, brain and heart from guin­ ea pigs. Biochim. biophys. Acta 444: 554 562 (1976). 8 Dinnendahi, V.: A rapid and simple procedure for the determination of guanosine 3’,5'-monophosphate by use of the protein-binding method. Archs Pharmacol. 284: 55-61 (1974). 9 Goldberg, N.D.; Haddox, M.K.; Nicol, S.E.; Glass, D.B.; Sanford, C.H.; Kuehl, F.A., jr., and Estensen, R.: Biologic regulation through opposing influ­ ences of cyclic GMP and cyclic AMP: the Yin Yang hypothesis; in Drummond, Greengard and Robi­ son, Advances in cyclic nucleotide research, vol. 5, pp. 307—330 (Raven Press, New York 1975). 10 llliano, G.; Tell, G.; Siegel, M.I., and Cuatracasas, P.: Guanosine 3',5'-cyclic monophosphate and the action of insulin and acetyl choline. Proc. natn. Acad. Sei. 70: 24 43 - 2447 (19 7 3). 11 lsaksson, O.; Rosberg, S., and Eden, S.: Influence of insulin on cyclic AMP level in rat diaphragm; in Drummond, Greengard and Robison, Advances in cyclic nucleotide research, vol. 5, pp. 805 (Raven Press, New York 1975). 12 Klotz, U. and Stock. K.: Influence of cyclic guano­ sine 3',5’-monophosphate on the enzymatic hydro­ lysis of adenosine 3’,5’-monophosphate. Archs Pharmacol. 274: 54 - 62 (1972). 13 Kono, T.; Robinson, F.W., and Sarver, J.A.: Insulin sensitive phosphodiesterase. J. biol. Chcm. 250: 7826-7835 (1975). 14 Loten, E.G. and Sncyd, J.G.T.: An effect of insulin on adipose-tissue adenosine 3’,5'-cyclic monophos­ phate phosphodiesterase. Biochem. J. 120: 187193(1970). 15 Lowry, O.H.; Rosebrough, N.J.; Farr, A.L., and Randall, R.J.: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193: 265-275 (1951). 16 Mandel, J.-L. and Pearson, M.: Effect of insulin and cyclic nucleotides on differentiation of a myo­ blast cell line; in Drummond, Greengard and Robi­ son, Advances in cyclic nucleotide research, vol. 5, p. 832 (Raven Press, New York 1975). 17 Mao, C.C. and Guidotti, A.: Simultaneous isolation of adenosine 3',5'-cyclic monophosphate (cAMP) and guanosine 3',5'-cyclic monophosphate (cGMP)

41

in small tissue samples. Analyt. Biochem. 59: 63-68 (1974). 18 Marks, F. and Raab, I.: The second messenger sys­ tem of mouse epidermis. IV. Cyclic AMP and cy­ clic GMP phosphodiesterase. Biochim. biophys. Acta 334: 368-377 (1974). 19 Miller, Z.; Lovelace, E.; Gallo, M., and Pastan, L: Cyclic guanosine monophosphate and cellular growth. Science 190: 1213-1215 (1975). 20 Nesbitt, J.A., III.; Anderson, W.B.; Miller, Z.; Pas­ tan, I.; Russell, T.R., and Gospodarowicz, D.: Guanylate cyclase and cyclic guanosine 3',5'-monophosphatc phosphodiesterase activities and cyclic guanosine 3',5’-monophosphate levels in normal and transformed fibroblasts in culture. J. biol. Chem. 251: 2344-2352 (1976). 21 Pastan, I.; Johnson, G.S., and Anderson, W.B.: Role of cyclic nucleotides in growth control. A. Rev. Biochem. 44: 491 -522 (1975). 22 Russell, T.R.; Terasaki, W.L., and Appleman, M.M.: Separate phosphodiesterases for the hydro­ lysis of cyclic adenosine 3',5'-monophosphatc and cyclic guanosine 3’,5'-monophosphate in rat liver. J. biol. Chem. 248: 1334-1340 (1973). 23 Sakai, T.; Thompson, W.J.; Lavis, V.R., and Wil­ liams, R.H.: Cyclic nucleotide phosphodiesterase activities from isolated fat cells, correlation of subcellular distribution with effects of nucleotides and insulin. Archs Biochem. Biophys. 162: 331-339 (1974) . 24 Schwartz, J.P. and Passonneau, J.V.: Correlation between intracellular cyclic nucleotide levels and cyclic nucleotide phosphodiesterase activity in C-6 glioma and C-1300 neuroblastoma cells. Abstract. Fed. Proc. Fed. Am. Socs exp. Biol. 34: 694 (1975) . 25 Steiner, A.L.; Pagliara, A.S.; Chase, L.R., and Kipnis, D.M.: Radioimmunoassay for cyclic nucleo­ tides. J. biol. Chem. 247: 1114-1120 (1972). 26 Strada, S.J. and Pledger, W.J.: The role of cyclic nucleotides in cell growth and development: regu­ lation and characterization of cyclic nucleotide phosphodiesterases in mammalian cells; in Weiss, Cyclic nucleotides in disease, pp. 3 -3 3 (University Park Press, Baltimore 1975). 27 Terasaki, W.L. and Appleman, M.M.: The role of cyclic GMP in the regulation of cyclic AMP hydro­ lysis. Metabolism 24: 311-319 (1975). 28 Vanlmvegen, R.G. and Thompson, W.J.: Regula­ tion of particulate cyclic nucleotide phosphodies­

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Cyclic Nucleotide Phosphodiesterases in Fetal Muscle

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30

31

32

terases. Abstract. Fed. Proc. Fed. Am. Socs exp. Biol. 34: 261 (1975). Weiss, B.: Differential activation and inhibition of the multiple forms of cyclic nucleotide phosphodi­ esterase; in Drummond Greengard and Robison, Advances in cyclic nucleotide research, vol. 5, pp. 195-211 (Raven Press, New York 1975). Weiss, B. and Strada, S.J.: Adenosine 3’,5’-monophosphate during fetal and post natal development; in Boreus, Fetal pharmacology, pp. 205-235 (Raven Press, New York 1973). Williams, R.H. and Thompson, W.J.: Effect of age upon guanyl cyclase, adenyl cyclase and cyclic nucleotide phosphodiesterase in rats. Proc. Soc. exp. Biol. Med. 143: 382-387 (1973). Woo, Y.-T. and Manery, J.F.: Cyclic AMP phos­ phodiesterase activity at the external surface of in­

tact skeletal muscles and stimulation of the en­ zyme by insulin. Archs Biochem. Biophys. 154: 510-519 (1973). 33 Zalin, R. and Montague, W.: Changes in the level of 3',5'-cyclic adenosine monophosphate and the ac­ tivities of adenylate cyclase and protein kinase dur­ ing the differentation of primary cultures of chick myoblasts. Abstract; in Drummond, Greengard and Robison, Advances in cyclic nucleotide research, vol 5, p. 832 (Raven Press, New York 1975).

Clarissa H. Beatty, Oregon Regional Primate Research Center, 505 NW 185th Avenue, Beaverton, OR 97005 (USA)

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29

Beatty/Herrington/Bocek

Regulation of cyclic nucleotide phosphodiesterase activity in rhesus fetal muscle.

Biol. Neonate 32: 33 -42 (1977) Regulation of Cyclic Nucleotide Phosphodiesterase Activity in Rhesus Fetal Muscle Clarissa H. Beatty, Paul T. Herring...
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