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P J Scarpace

Publications and source records attributed to P J Scarpace.

At least 19 recordsLinked to original sources
PubMed1992

Beta-adrenergic receptor activity of cerebral microvessels in experimental diabetes mellitus.

The effect of diabetes mellitus on beta-adrenergic receptor number (B(max)), receptor-cyclase coupling and adenylate cyclase (AC) activity was determined in cerebral microvessels isolated from control and streptozotocin induced diabetic rats after 5 weeks of induction of diabetes. Scatchard analysis of [125I]iodocyanopindolol (ICYP) binding indicated that the B(max) (fmol/mg) in diabetic rat cerebral microvessels (63.8 +/- 4.8) (mean +/- S.E.M.) was not significantly different from the B(max) in control rats (56.5 +/- 6.9). Isoproterenol competition of [125I]ICYP binding sites indicated that the percentage of beta-receptors expressing high affinity binding was 53.9 +/- 0.45% in control rats and 47.5 +/- 2.3% in diabetic rats. The total isoproterenol-stimulated AC activity (pmol cAMP/mg) in diabetic rats (76.7 +/- 6.1) was significantly lower than that in control rats (118.4 +/- 11.2) (P less than 0.01). However, the net isoproterenol-stimulated AC activity (i.e. total minus GTP-stimulated AC activity) was not altered in diabetes. The net sodium fluoride (NaF) stimulated AC activity in diabetic rats (109.5 +/- 11.4) was significantly lower than the control rats (154.3 +/- 16.3) (P less than 0.05). It is concluded that diabetes mellitus in rats is associated with reduced post receptor activation of adenylate cyclase in cerebral microvessels while the beta-adrenergic receptor density, affinity and receptor-cyclase coupling are not significantly altered.

Adenylyl Cyclases↗
PubMed1992

Beta-adrenergic responsiveness in cultured aorta smooth muscle cells. Effects of subculture and aging.

beta-Adrenoceptor-mediated vasorelaxation is diminished in vessels from a variety of aged species including humans. This phenomenon was studied for the first time in cultured aorta smooth muscle cells (ASMC) from young (4- to 6-month) and old (24- to 26-month) F-344 rats. Cyclic AMP (cAMP) accumulation was assessed following isoproterenol and forskolin stimulations in primary cultures and after 1-4 passages of aorta smooth muscle cells. Isoproterenol and forskolin increased cAMP accumulation 6- and 10-fold, respectively, in primary cultures from young rats. Isoproterenol stimulation was reduced markedly in passaged cells. Forskolin stimulation was unaffected, indicating passage-related phenotypic changes in receptor-mediated stimulation, but not in post-receptor adenylate cyclase activation. The response to isoproterenol was diminished in old animals, but that to forskolin was unaltered. Thus, cultured ASMC from F-344 rats are highly responsive to beta-adrenoceptor stimulation and demonstrate age-related changes, but undergo phenotypic modulation during passage.

Adenylyl Cyclases↗
PubMed1992

A biochemical analysis of parotid and submandibular salivary gland function with age after simultaneous stimulation with pilocarpine and isoproterenol in female NIA Fischer 344 rats.

This analysis of physiological, biochemical and molecular changes related to aging was made in 3-, 12- and 24-month-old rats. The salivary gland weight/body weight ratio and the structural membrane proteins did not change with age for either gland, but a significant age-related decline in DNA synthesis for both glands was detected, unrelated to the hormonal responsiveness at the level of the plasma membrane. There was a marked increase in the concentration of soluble proteins in adolescent parotid gland and, for the two older age groups, in submandibular gland. The saliva flow rate was different when expressed as volume per time, as volume per time and g glandular wet weight, and/or kg body weight. The concentration of secreted proteins was not affected by age in either gland. The total amount of proteins secreted over 30 min revealed no age-related perturbation for the parotid gland, but showed a significant age-related increase in submandibular saliva. Sodium dodecyl sulphate-polyacrylamide gel analysis revealed changes in the protein bands between 39 and 50 kDa in the Coomassie blue-stained gels from 12-month-old animals. Amylase showed an initial increase (12 months), followed by a marked decline in its activity in parotid saliva. The glandular supernatant had low residual cellular amylase activity after stimulation. Therefore, secretory impairment with age after pilocarpine-isoproterenol stimulation was excluded. Analysis of total RNA showed a pronounced decrease of amylase mRNA in the parotid gland between 12 and 24 months of age. No amylase mRNA was expressed in any of the submandibular samples. For epidermal growth factor, total saliva showed a decrease with age. It seemed that the submandibular gland followed the same picture with age as the parotid gland, with a specific decline in the biosynthesis of single secretory proteins.

Aging↗
PubMed1992

Desipramine desensitizes beta-adrenergic signal transduction in rat salivary glands.

The consequences of the tricyclic antidepressant drugs include sedation, orthostatic hypotension and salivary dysfunction. It was reported that administration of desipramine resulted in a decrease in the concentration of secreted salivary protein. Tricyclic antidepressants may indirectly alter salivary function as a result of their action on the central nervous system to decrease adrenergic neural transmission or, alternately, may act directly on salivary glands to modulate beta-adrenergic signal transduction. To investigate the latter possibility, the effects of administration of desipramine (DMI) for 28 days was assessed on beta-adrenergic and post-receptor signal transduction in the parotid and submandibular glands of the rat and the reversibility of this treatment following a washout period of 15 days. Both glands demonstrated desensitization of the isoproterenol-stimulated activity, and in the parotid gland treatment with DMI decreased the post-receptor signal transduction as well. The washout period of 15 days completely reversed the desensitization in the parotid gland and partially reversed the effects in the submandibular gland. These data suggest that desipramine-induced attenuation of beta-adrenergic signal transduction is not limited to the brain and that these direct effects on salivary glands may explain the salivary dysfunction observed after administration of desipramine.

Adenylyl Cyclases↗
PubMed1992

Influence of exercise and age on myocardial beta-adrenergic receptor properties.

The bradycardia following physical training may be mediated by an alteration in beta-adrenergic receptor number or agonist affinity. We characterized the interaction between age and exercise on myocardial beta-adrenergic receptor number and agonist affinity in 4- and 24-month-old F344 rats to test the hypothesis that the effects of training should be blunted in older rats. beta-adrenergic receptor density was unchanged with age or training. The total number of receptors per heart increased with age due to increased ventricle weight. With training, in the senescent rats the total number of receptors decreased due to a reduced amount of homogenate protein recovered from the ventricle, the significance of which is unknown. The receptor agonist dissociation constant for isoproterenol was determined in both the absence and presence of beta,gamma-imidoguanosine 5'-triphosphate [Gpp(NH)p] and did not change with age or training. Neither training nor age influenced beta-adrenergic receptor characteristics, suggesting that training bradycardia is not mediated by an alteration in beta-adrenergic receptors.

Aging↗
PubMed1992

The association of E. coli peritonitis with an impaired and delayed fever response in senescent rats.

Infection is one of the leading causes of death in elderly humans, and the importance of the early diagnosis of severe infection is undisputed. In the elderly a delay in diagnosis is often due to a reduced or absent fever. To understand more fully the pathogenesis of fever in senescence, we assessed the febrile response to E. coli peritonitis in 3-, 12-, and 24-month-old rats. Baseline temperatures were unchanged with age. Following infection with 1 x 10(8) CFU E. coli, a fever was evident in 2.8 h in the young, 3.9 h in the 12-month-old rats, and delayed until 5.8 h in the senescent rats. The magnitude of the fever was quantitatively less in the older rats compared with the two younger age groups throughout the time course of the fever. Because beta-adrenergic-mediated thermogenesis in brown adipose tissue has been implicated in the genesis of fever, we also assessed adenylate cyclase activity in this tissue. There was a progressive age-related decrease in both receptor- and postreceptor-stimulated adenylate cyclase activity. Our findings indicate there is both a delay in the onset of the fever and a reduced febrile response in the senescent rats following E. coli infection.

Adenylyl Cyclases↗
PubMed1992

Thermogenesis and mitochondrial GDP binding with age in response to the novel agonist CGP-12177A.

The ability to regulate body temperature diminishes with age in both humans and rodents. To investigate whether attenuation of sympathetically activated thermogenesis in brown adipose tissue (BAT) may account for the loss of thermoregulation with age, we assessed O2 consumption and body temperature in response to norepinephrine and the specific BAT beta-adrenergic agonist CGP-12177A in 6-, 18-, and 24-mo-old rats. In addition, the effects of this agonist on interscapular BAT mitochondrial GDP binding in young and senescent rats were determined. CGP-12177A rapidly induced an elevation in O2 consumption, which peaked at 25 min, followed by a decline over 4 h. The peak increase in O2 consumption over baseline and the cumulative 4-h response were decreased with age [P less than 0.02, analysis of variance (ANOVA)]. CGP-12177A induced an increase in body temperature that paralleled but appropriately lagged behind the increase in O2 consumption and that was decreased with age (P less than 0.02, ANOVA). The norepinephrine-induced increase in O2 consumption was also reduced with age but was not paralleled by a change in body temperature and was associated with a four- to fivefold increase in physical activity. In young rats CGP-12177A increased the number of available BAT mitochondrial GDP binding sites at 20 and 60 min post-injection, but in senescent rats GCP-12177A was unable to increase GDP binding. These data indicate that CGP-12177A is a novel agonist for BAT thermogenesis. With age there is a reduced capacity for thermogenesis that involves a failure to increase GDP binding, either due to a diminished amount of uncoupling protein with age or a failure to unmask reserve GDP binding sites.

Adipose Tissue, Brown↗
PubMed1992

Impaired febrile response with age: role of thermogenesis in brown adipose tissue.

We demonstrated previously that in Escherichia coli-infected rats, the heat necessary for the febrile response is a result of thermogenesis in brown adipose tissue (BAT). To investigate whether senescent rats have an impaired febrile response to infection and whether such an impairment is a result of attenuated sympathetically activated thermogenesis in BAT, we assessed body temperature and the increase in mitochondrial guanosine 5'-diphosphate (GDP) binding sites in interscapular BAT in response to E. coli administration in young and senescent male F-344 rats. There was a significant delay of 2 hr in the onset of fever in the older animals. In addition, in senescent rats, the peak fever (1.0 +/- 0.1 delta degrees C vs 2.2 +/- 0.1) and the cumulative fever (383 +/- 43 delta degrees C.min vs 775 +/- 69) were significantly less than in the young rats (P less than 0.005). Baseline levels of GDP binding were the same in young and old rats. In young rats, during the rising phase of the fever, E. coli infection resulted in a 50% increase in the density of GDP binding sites in BAT mitochondria. In contrast, there was no increase in GDP binding in the older rats following infection. The failure to increase GDP binding may be a result of a reduced ability to unmask reserve GDP binding sites. Alternatively, there may be fewer total GDP binding sites (masked and unmasked) in senescent rats and these sites may already be unmasked. Collectively, these data suggest that the impaired febrile response with age is due to reduced thermogenesis in BAT.

Adipose Tissue, Brown↗
PubMed1992

Beta-adrenergic receptors and salivary gland secretion during aging.

Beta-adrenergic signal transduction is primarily responsible for the control of the protein secretions by salivary cells. To examine the relationship between beta-adrenergic signal transduction and beta-adrenergic agonist-stimulated salivary secretion, we simultaneously assessed beta-adrenergic receptor number and pilocarpine-isoproterenol-stimulated salivary flow and secreted proteins in parotid and submandibular glands from 3-, 12- and 24-month-old female NNIA F-344 rats. There were no age-related changes in the density of beta-adrenergic receptors in the parotid gland or in the submandibular gland. In the parotid gland there was a significant increase in saliva flow rate in the oldest age group and no changes in the amount of total proteins secreted over 30 min. However, when normalized to gland weight, flow rate was unchanged and the amount of total secreted proteins decreased with age. In the submandibular gland there were age-related increases in both absolute volume and total secreted protein, but when normalized to gland weight there were no longer changes with age. Changes in flow rate were paralleled by reciprocal changes in protein secretory function such that changes in the salivary protein concentrations for the most part were unchanged with age for both the parotid and the submandibular gland. These parameters were compared to our previous data on adenylate cyclase activity, and collectively, these data suggest that in the submandibular gland salivary secretory function does not correlate with changes in beta-adrenergic receptor density or isoproterenol-stimulated adenylate cyclase activity.

Aging↗
PubMed1991

Beta-adrenergic receptor activity of cerebral microvessels is reduced in aged rats.

The effect of age on beta- (beta) adrenergic receptor number (Bmax) and adenylate cyclase (AC) activity was determined in microvessels isolated from male F-344 rats at 3, 18, and 24 months of age. Scatchard analysis of [125I]iodocyanopindolol (ICYP) binding indicated reduced Bmax (fmol/mg) of microvessels isolated from 24 month old rats (27.2 +/- 4.9) compared with 3 month old (50.4 +/- 5.2) and 18 month old rats (p less than 0.01) (61.4 +/- 7.6). The basal AC activity (pmol cAMP/mg) in 24 month old rats (32.0 +/- 6.7) and in 18 month old rats (30.4 +/- 2.1) were significantly reduced compared to the basal activity in the young (50.1 +/- 4.2). The net isoproterenol or NaF stimulated AC activity in 24 month old rats (zero and 15.6 +/- 8.5 respectively) was also reduced compared to young rats (10.1 +/- 3.9 and 166.0 +/- 21.2 respectively). It is concluded that aging is associated with reduced isoproterenol stimulated AC activity of cerebral microvessels. This reduction is the product of reduced beta-adrenergic receptor number and reduced activity of AC in aged rat cerebral microvessels.

Adenylyl Cyclases↗
PubMed1991

Beta-adrenergic signal transduction in aging parotid and submandibular salivary glands.

This study assessed agonist- and post-receptor-stimulated adenylate cyclase (AC) activity in parotid and submandibular salivary glands from female F-344 rats of 3, 12, and 24 months of age. Isoproterenol-stimulated dose-response activation of adenylate cyclase was unchanged between 3 and 12 months but decreased at 24 months (p less than .05). Forskolin-stimulated AC activity, representing catalytic unit activity, was decreased at 24 months in the parotid (p less than .05) and at 12 months (p less than .05) and 24 months (p less than .01) in the submandibular gland. Beta-adrenergic signal transduction in salivary glands stimulates the secretion of salivary proteins that have important functions in the maintenance of oral health.

Adenylyl Cyclases↗
PubMed1991

Escherichia coli peritonitis activates thermogenesis in brown adipose tissue: relationship to fever.

Fever is a complex and important nonspecific, host defense mechanism against infection. The generation of the heat necessary to increase body temperature may involve thermogenesis in brown adipose tissue. To investigate whether the febrile response to Escherichia coli peritonitis involves thermogenesis in brown adipose tissue, we assessed whole rat oxygen consumption and brown adipose tissue mitochondrial guanosine 5'-diphosphate binding. Non-lethal doses of E. coli, 1 x 10(6) to 1 x 10(8) colony forming units, induced a fever for greater than 8 h. In contrast, a dose of 1 x 10(9) colony forming units resulted in a progressive hypothermia culminating in death. A 48% increase in oxygen consumption (p less than 0.05) in E. coli-infected rats occurred almost immediately, preceded the development of the fever, and was sustained throughout the fever. There was a highly significant correlation (r = 0.736, p less than 0.01) between oxygen consumption and body temperature for both control and infected animals. Guanosine 5'-diphosphate binding assessed by multi-point Scatchard analysis of [3H]guanosine 5'-diphosphate binding to isolated mitochondria was increased by 45.4 +/- 7.3% at 1.75 h and by 31.9 +/- 9.0% at 3.5 h (p less than 0.05). The greater increase was during the rising phase of the fever. Unexpectedly, a lethal dose of 5 x 10(9) colony forming units of E. coli also increased guanosine 5'-diphosphate binding sites by 54.4 +/- 14.2% (p less than 0.05) despite a hypothermia of -1.71 +/- 0.29 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗
PubMed1991

Adenylate cyclase agonist properties of CGP-12177A in brown fat: evidence for atypical beta-adrenergic receptors.

Thermogenesis in brown adipose tissue (BAT) is stimulated by catecholamine activation of adenylate cyclase through the beta-adrenergic receptor. Recently it was reported that the beta-adrenergic antagonist CGP-12177A stimulates oxygen consumption in BAT. To investigate the mechanism of action of CGP-12177A in BAT, we assessed the inhibitory and stimulatory affects of CGP-12177A on the adenylate cyclase system in myocardial and BAT membranes from rats. CGP-1277A inhibited isoproterenol-stimulated adenylate cyclase activity in a dose-dependent manner, with an inhibitory constant (Ki) of 1.94 +/- 0.18 microM in BAT and 0.49 +/- 0.11 microM in the heart. However, in the absence of isoproterenol, CGP-12177A stimulated adenylate cyclase in BAT with two components of activation, and half-maximal stimulation occurred at 1 microM and 1.5 mM. In contrast, CGP-12177A did not stimulate adenylate cyclase activity in heart membranes. Propranolol inhibited the isoproterenol-stimulated activity with a potency that was one log less in BAT compared with heart. Propranolol fully blocked the high-affinity component but only weakly blocked the low-affinity component of CGP-12177A-stimulated activity in BAT. Pindolol was also less potent in BAT but inhibited the CGP-12177A-stimulated activity in a manner similar to the inhibition of the isoproterenol-stimulated activity, suggesting the CGP-12177A activation was beta-receptor mediated. Binding curves of [125I]iodocyanopindolol ([125I]ICYP) in competition with CGP-12177A demonstrated a shift to lower affinity in the presence of beta,gamma-imidoguanosine 5'-triphosphate, indicating that CGP-12177A has agonist properties with respect to the [125I]ICYP binding site.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclases↗
PubMed1991

Beta-adrenergic function in aging. Basic mechanisms and clinical implications.

Catecholamines have an important endocrine and neuroendocrine role in mediating a variety of autonomic functions. One consequence of normal aging, in particular in the cardiovascular system, is a decline in beta-adrenergic function associated with an alteration in responsiveness to beta-adrenergic therapy. The intrinsic ability for muscle contractility or relaxation is maintained with age and there appears to be an alteration in the process linking the receptor with the contractile or relaxation mechanisms. In rats, beta-adrenergic receptor density decreases with age in adipose tissues and most brain areas, is unchanged in lymphocytes, heart and lung, and increases in the liver. In humans, there are no receptor changes with age in either lymphocytes or brain. In contrast, the number of high-affinity receptors (or coupled receptors) decreases with age in most tissues. In addition, there is a decrease in membrane adenylate cyclase activity or cellular production of cyclic adenosine monophosphate (adenosine 3',5'-cyclic phosphate; cAMP). Plasma noradrenaline (norepinephrine) concentration increases with age. The reduced receptor number in some tissues (down-regulation), the reduced high-affinity receptors and the reduced hormone-stimulated adenylate cyclase activity with age suggests receptor desensitisation to increased plasma noradrenaline concentration. The inability of older animals to desensitise to beta-adrenergic agonists further supports this hypothesis. However, there is an additional post-receptor reduction in catalytic unit activity with age independent of desensitisation. Medications directed at the beta-adrenergic system are commonly used in the elderly. Many of the data on the impact of age on clinical responses are conflicting or unavailable. Concomitant disease, functional status, nutritional state and polypharmacy may play an even greater role than age. However, the available data can be used to guide the selection of therapy, anticipate side effects, and predict potential interactions with other medications and diseases.

Aged↗
PubMed1991

Agonist-induced desensitization of liver alpha 1-receptors; implications for assessment of PI hydrolysis.

Alpha 1-adrenergic stimulation of phosphatidyl inositide (PI) hydrolysis was measured in liver slices and in cultured hepatocytes prelabeled with 3H-myoinositol. Incubation of cultured hepatocytes for 15 min with l-epinephrine stimulated the release of 3H-inositol phosphates with an EC50 of 0.28 +/- 0.19 microM and Emax representing 2.88 +/- 0.39% of incorporated radiolabel. Pretreatment of cultures with epinephrine resulted in reductions both in the maximum response and in the sensitivity to epinephrine. In liver slices, epinephrine elicited a similar maximum response (3.38 +/- 0.32%), but the sensitivity to epinephrine (EC50 = 2.27 +/- 1.27 microM) was considerably lower than for cultured cells. Prior treatment of liver slices with 200 microM l-epinephrine had no effect either on the maximum response to l-epinephrine or on the sensitivity to epinephrine. The affinity of alpha 1-adrenergic receptors for agonists was reduced 2.11 fold during incubation of liver slices for 60 min at 37 degrees C in the absence of added agonist (as is required for 3H-myoinositol labeling). Agonist affinity was not further reduced when 200 microM l-epinephrine was included in the incubation. It was concluded that alpha 1-adrenergic stimulation of PI hydrolysis in liver undergoes agonist-induced desensitization. It was also concluded that a limitation exists in the methodology for measuring PI hydrolysis in tissue slices prelabeled with 3H-myoinositol, in that this method assesses the activity of receptors in a desensitized state.

Animals↗
PubMed1990

Forskolin activation of adenylate cyclase in rat myocardium with age: effects of guanine nucleotide analogs.

Beta-adrenergic and post-receptor activation of adenylate cyclase decreases with age in the rat myocardium. Forskolin-stimulated adenylate cyclase activity decreases with age to the same extent as NaF or isoproterenol stimulation, suggesting that a loss of catalytic unit activity accounts for the loss of activity with age. However, recent evidence suggests that there are both a guanine nucleotide-dependent and independent component of forskolin activation. We assessed the possible role of each of these components with age by measuring forskolin dose-response stimulation of adenylate cyclase activity in the presence and absence of guanosine 5'-O-(2-thiodiphosphate) (GDP-beta S) and beta,gamma-imidoguanine 5'-triphosphate (Gpp(NH)p) in myocardial membranes from F-344 rats of 3, 12, and 24 months of age. Maximal forskolin stimulation of adenylate cyclase was least in the 24- (121 +/- 21 pmol cAMP/min/mg) as compared with either the 12- (212 +/- 29) or 3- (190 +/- 19) month-old rats. The presence of GDP-beta S had no effect on either the EC50 or maximum activity, and the age-related decline persisted. Gpp(NH)p plus isoproterenol enhanced forskolin activation but the effect was additive and not synergistic. There was no effect on the EC50, and the maximum activity was least in the 24-month-old rats. These data reconfirm the loss of catalytic unit activity with age and indicate that the reduced activity with age is independent of the presence or absence of activated G-protein.

Adenylyl Cyclases↗
PubMed1990

Alpha 1-adrenergic stimulation of inositide hydrolysis in liver of senescent rats.

Hydrolysis of phosphatidyl inositides (PI) was measured in response to alpha 1-adrenergic agonist in slices of liver prepared from F-344 female rats aged 6-, 15- and 25 months. Tissue was labeled with [3H]inositol and release of inositol phosphates was measured following a 60-min incubation with a range of concentrations of l-epinephrine. The response to epinephrine was completely blocked by prazosin, indicating the involvement of alpha 1-adrenergic receptors. The maximum response to epinephrine represented release of 13-15% of radioactivity incorporated into the pool of cellular lipids and was unchanged with age. The concentration of epinephrine required for half-maximal stimulation was 1-1.5 microM and was also unchanged with age. These data suggest that, although senescence is accompanied by alterations with age in the liver alpha 1-receptor, alpha 1-stimulation of PI hydrolysis is unchanged.

Aging↗