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Biomedical subjects

G S Roth

Publications and source records attributed to G S Roth.

At least 19 recordsLinked to original sources
PubMed1995

Decreased m3-muscarinic and alpha 1-adrenergic receptor stimulation of PIP2 hydrolysis in parotid gland membranes from aged rats: defect in activation of G alpha q/11.

m3-Muscarinic cholinergic receptor (m3-AChR) and alpha 1-adrenergic receptor (alpha 1-AR) stimulation of phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis (by a PIP2-specific phospholipase C, PLC) in rat parotid gland membranes is mediated via activation of alpha subunits of the Gq/11 family of G-proteins. This study examines m3-AChR and alpha 1-AR stimulation of PIP2 hydrolysis in membranes isolated from parotid glands of old (24 months) and young (3 months) rats (old and young rat membranes). Old rat membranes exhibited reduced stimulation of PIP2 hydrolysis in response to the addition of guanosine-5'-O-(3-thiotrisphosphate) (GTP gamma S) alone or GTP gamma S plus either carbachol (m3-AChR agonist) or epinephrine (alpha 1-AR agonist). This reduction in receptor-stimulated PIP2 hydrolysis was not due to a decrease in PLC activity per se since cholate-solubilized PLC activity was similar in old and young rat membranes. Additionally, these membranes exhibited comparable, immunologically detectable, levels of PLC beta 3, G alpha q/11, and G beta. In the presence of 10 microM AlCl3 and 10 mM NaF, stimulation of PIP2 hydrolysis in both old and young rat membranes was similar. Preincubation of membranes from old rats with GTP gamma S induced a time-dependent increase in the rate of PIP2 hydrolysis and, with 20 min preincubation, the rates of hydrolysis in old and young rat membranes were not statistically different. In aggregate, these data indicate that there is a defect in the activation of G alpha q/11 in parotid gland membranes from old rats.

Adenylyl Cyclases↗
PubMed1995

Age-related impairment in striatal muscarinic cholinergic signal transduction is associated with reduced membrane bilayer width measured by small angle X-ray diffraction.

In order to determine whether age-related changes in neuronal membrane structure contribute to previously reported changes in muscarinic cholinergic signal transduction, striata from 3, 13 and 23 month old F344 male rats were examined for both carbachol-stimulated low Km GTPase activity and membrane one-dimensional electron density profile using small angle X-ray diffraction. Increasing age was associated with both a reduction in stimulated GTPase activity and a decrease in membrane bilayer width. These findings suggest the possibility that fundamental membrane structural changes may contribute to alterations in signal transduction seen with aging.

Aging↗
PubMed1995

Partial restoration of impaired alpha 1-adrenergic responsiveness in parotid cells of aged rats by S-adenosylmethionine treatment.

The age related decrease in alpha 1-adrenergic stimulated inositol 1, 4, 5 trisphosphate (IP3) production in parotid cells of aged rats can be partially restored by treatment with S-adenosylmethionine (SAM). This effect is completely blocked by S-adenosyl homocysteine (SAH) and occurs in association with an increase in the conversion of phosphatidylethanolamine to phosphatidylcholine and a decrease in membrane viscosity. In contrast, SAM treatment actually inhibits stimulated IP3 production in cells of young rats. The membrane viscosity of these cells is lower than that of those from aged rats. Although conversion of phosphatidylethanolamine to phosphatidylcholine is enhanced, no further decrease in membrane viscosity is elicited in young cell preparations. These findings suggest that age changes in the membrane environment may result in impaired alpha 1-adrenergic signal transduction and that such alterations may be at least partially reversible by SAM treatment.

Adrenergic alpha-1 Receptor Agonists↗
PubMed1995

Identification of neuronal programmed cell death in situ in the striatum of normal adult rat brain and its relationship to neuronal death during aging.

Apoptotic neurons have been identified in normal adult rat striatum by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling technique. This observation suggests that neuronal programmed cell death starts at an early stage of adult life and may contribute to the aging associated neuronal loss. In addition, the frequency of apoptotic cells was found to significantly increase in old rats, which implies that aging itself accelerates the process.

Aging↗
PubMed1995

Effects of aging and dietary restriction on activity of monkey serum in promoting fibroblast migration.

In order to determine whether serum modified cellular aging in vivo, we previously studied the effects of serum from various mammals of different ages on cell functions such as proliferation and migration, and reported that cell migration was more greatly inhibited by serum from old donors than cell proliferation [1]. Moreover, since dietary restriction has been reported to extend lifespan and slow the aging rate of some animals [2], we wondered whether sera from dietary restricted and control monkeys of various ages might exhibit reduced aging effects on cell migration. When serum from young adult (3-5 years old) monkeys was added to plain medium, the migration of human fetal skin fibroblasts was very strongly inhibited compared to FBS. Surprisingly, sera from adult (6-11 years old) and old (more than 18 years old) monkeys caused significantly less migration-inhibitory activity than serum from young adult monkeys although sera from adult and old monkeys were much more inhibitory to cell migration than FBS. Dietary restriction only caused marginal effects on serum migration-promoting activity in a few monkey groups. The inhibition of cell migration caused by monkey serum was not brought about by cytotoxic effects since monkey serum stimulated cell proliferation as well as fetal bovine serum. These results indicate that the effects of aging on monkey serum migration-promoting activity are much more pronounced than those of dietary restriction.

Aging↗
PubMed1995

Age-specific alterations in muscarinic stimulation of K(+)-evoked dopamine release from striatal slices by cholesterol and S-adenosyl-L-methionine.

The present experiments were carried out in order to test the hypothesis that age-related signal transduction (ST) deficits may occur as a result of structural changes in the membrane that are reflected partially as increased membrane microviscosity. Oxotremorine (oxo) enhancement of K(+)-evoked release of dopamine (K(+)-ERDA) was examined in superfused striatal slices from mature (6 months) and old (24 months) Wistar rats incubated (1 or 4 h, 37 degrees C) with graded concentrations of S-adenosyl-L-methionine (SAM) or cholesterol hemisuccinate (CHO) in a modified Krebs medium. Tissue was then assessed for one of the following: (a) the degree of oxo-enhanced K(+)-ERDA, (b) carbachol stimulated low Km GTPase activity, or (c) alterations in membrane microviscosity. In other experiments the tissue was incubated in CHO followed by SAM (or the reverse), and oxo-enhanced K(+)-ERDA examined. Results indicated that SAM treatment increased all the parameters in the striatal tissue from old animals, while CHO had selective, opposite effects in the striatal tissue obtained from young animals. CHO-SAM, or the reverse, produced the same pattern of results. These results suggest that ST deficits may involve age-related structural alterations in membranes that interfere with receptor-G protein coupling/uncoupling.

Aging↗
PubMed1995

Influence of age, sex, and dietary restriction on intracellular free calcium responses of CD4+ lymphocytes in rhesus monkeys (Macaca mulatta).

The influence of aging and dietary restriction on increase in intracellular free calcium ([Ca2+]i) of CD4+ lymphocytes from Macaca mulatta was examined after stimulation with anti-CD3 mAb. We used a flow cytometric assay with the dye indo-1 and either direct or reciprocal immunofluorescent staining to identify CD4+ cells. After stimulation with anti-CD3 mAb, intracellular free calcium responses were reduced in CD4+ lymphocytes from old male and female ad libitum fed monkeys compared to young and adult male or female monkeys. Old female monkeys had significantly lower [Ca2+]i than did old male monkeys. The reduced responses were in part related to a decreased percentage of responding cells. Dietary restriction of males over a four-year period did not alter [Ca2+]i response compared to ad libitum fed male monkeys. Female monkeys of all ages (which were restricted only for four months) also had similar [Ca2+]i responses to ad libitum fed controls. Our data suggest that age-related changes in [Ca2+]i responses are similar between humans and M. mulatta, and that over these intervals, no effects of caloric restrictions can be detected.

Aging↗
PubMed1995

Membrane alterations as causes of impaired signal transduction in Alzheimer's disease and aging.

Changes in cell-membrane composition in normal aging and in Alzheimer's and other age-related diseases appear to result in impaired neurotransmitter-triggered signal transduction. The impaired signal transduction seems to be related to dysfunctions in the coupling of G proteins to their receptors and effectors. Direct demonstration of altered physiochemical properties of brain tissue of patients with Alzheimer's disease has been achieved by small-angle X-ray diffraction. In this disease, thinner membranes correlate with a 30% decrease in moles of cholesterol:phospholipid. Such changes can affect directly the coupling and uncoupling properties of G proteins, and can account for signal transduction deficits. These findings offer a complementary alternative to the beta-amyloid hypothesis, and an opportunity to consider new types of therapeutic interventions.

Aging↗
PubMed1995

Dissociation of striatal GTPase and dopamine release responses to muscarinic cholinergic agonists in F344 rats: influence of age and dietary manipulation.

There is evidence that dietary lipids and age both influence neuronal membrane composition and receptor G protein-linked signal transduction, but very little information is available on the interaction between these two factors. To investigate this, we obtained striata from 2, 12, and 22-month-old male F344 rats who were fed either a high-cholesterol, high-saturated fat or low-fat diet for 1 month. The striata were assayed for muscarinic agonist-stimulated low-Km GTPase activity using 10(-3) M carbachol and 10(-5) M oxotremorine and for KCl-evoked dopamine release enhancement by 10(-5) M oxotremorine. Membrane cholesterol and phospholipid content and phospholipid class composition were also determined. Mature animals showed significant but divergent changes in GTPase activity and dopamine release for high-cholesterol and low-fat diets: GTPase activity decreased, whereas dopamine release increased in these groups. Alterations in GTPase activity but not in dopamine release were inversely correlated with the cholesterol/phospholipid molar ratio. Old control animals showed reductions in both GTPase activity and oxotremorine-enhanced dopamine release compared with young animals. Whereas none of the experimental diets affected GTPase activity in old animals, the low-fat diet produced a marked decrease in dopamine release. In contrast to mature and old groups, young rats showed no significant change in either GTPase or dopamine release, suggesting a relative "resistance" to such dietary lipid modulation. The observed dissociation in GTPase and dopamine release responses to diet may reflect differing effects of these diets on discrete membrane lipid domains that preferentially influence different signal transduction components. The substantial age-related differences in striatal membrane response to dietary lipid modulation may represent the effects of underlying age differences in membrane lipid metabolism, structure, and/or dynamics. Our findings support the work of other groups that have shown that brain membranes are susceptible to modification by exogenous lipids. They also suggest the need for a more systematic examination of the influence of age on the response to other types of dietary lipid changes.

Aging↗
PubMed1995

Energy balance in rhesus monkeys (Macaca mulatta) subjected to long-term dietary restriction.

Male rhesus monkeys of various age groups representative of the species life span were fed ad libitum amounts (controls) or 30% less food than control monkeys of comparable age and body weight. Despite significantly lowered energy intake and body weight, the amount of energy lost in the feces, and fecal energy density (concentration) were not altered in diet-restricted (DR) monkeys, compared to age- and weight-matched controls. Absolute energy expenditure (EE; 24-hr) was consistently lower in DR monkeys, but this trend was not statistically significant. Expressed as a function of metabolic mass (body weight, metabolic body size, lean mass), 24-hr EE was not different in monkeys subjected to long-term DR, compared to controls. Calculations of net energy (intake-loss), as an index of energy balance, revealed that energy expenditure generally exceeded energy intake in all juvenile and adult group monkeys. However, this discrepancy was not statistically different from zero, suggesting that most animals were in energy balance. Also, there was no difference between control and DR animals with respect to energy balance. Diet restriction induced significant reductions in the absolute amount of lean body mass; however, percent (of total weight) lean and fat mass did not differ from controls.

Animals↗
PubMed1995

Aging and food restriction alter some indices of bone metabolism in male rhesus monkeys (Macaca mulatta).

Food restriction increases life span, reduces aging rate and affects a wide variety of biological functions. In rats, food restriction delays bone growth and reduces bone density and mineral content. We report the effects of aging and long-term (> 6.0 y) food restriction on several indices of bone growth and metabolism in rhesus monkeys (Macaca mulatta). Food allotments for controls approximated free access consumption, whereas food-restricted monkeys received 30% less food on a body weight basis. Cross-sectional and longitudinal age effects on serum alkaline phosphatase paralleled those reported for humans. Food restriction induced a significant delay in the developmental decline (to adult levels) in total alkaline phosphatase and significantly suppressed serum interleukin 6 concentrations, particularly in younger monkeys. Also, food restriction slowed skeletal growth, as reflected by shorter crown-rump length, and significantly reduced total body bone mineral content, but not bone mineral density, measured by dual energy X-ray absorptiometry. Analyses of serum parathyroid hormone, calcium, phosphate and osteocalcin concentrations suggested that the effects on skeletal growth were not related to alterations in calcium and phosphate homeostasis or a primary defect in bone formation. These findings suggest that long-term food restriction delays skeletal development in male rhesus monkeys while allowing the development of a reduced but otherwise normal skeleton.

Aging↗
PubMed1995

Diet restriction in rhesus monkeys lowers fasting and glucose-stimulated glucoregulatory end points.

Male rhesus monkeys (Macaca mulatta) of different age groups representing the species life span were fed ad libitum or a 30% reduced calorie diet over a 7-yr period. During the first 2-3 yr of this longitudinal study, glucose and insulin levels were not altered by diet restriction (DR). However, reductions in fasting blood glucose became apparent in DR animals after 3-4 yr. At the end of the 6th yr of study, glycated hemoglobin was measured, and intravenous glucose tolerance tests (IVGTTs) were conducted. Maximum glucose levels reached during IVGTTs increased with age but were lower in DR animals compared with controls. Several measures of the insulin response (baseline, maximum, and integrated areas under curve) increased with age and were lower in DR monkeys. With the exception of glycated hemoglobin, which was not different in monkeys subjected to DR, these findings confirm previous studies in rodents demonstrating that DR alters glucose metabolism and may be related to the antiaging action of this intervention.

Aging↗
PubMed1995

Aging-related changes in rat striatal D2 receptor mRNA-containing neurons: a quantitative nonradioactive in situ hybridization study.

In situ hybridization of a digoxigenin-labeled oligonucleotide probe combined with computer-assisted image assessment was used to directly visualize D2 receptor mRNA-containing neurons in rat striata, and quantify age-related changes in the sizes and relative mRNA content of these neurons. It was found that: (1) numbers of D2 mRNA-containing neurons appear to decrease in striata of aged rats, (2) relative amounts of D2 receptor mRNA decrease in striatal neurons of aged rats, and (3) neurons of all sizes appear to be lost, with the greatest absolute decrease in those smaller than 90 microns 2. These data confirm the well documented age-associated loss of D2 dopamine receptors, and further suggest that both neuronal death and reduced transcription contribute to this decrement.

Aging↗
PubMed1994

Area specific alterations in muscarinic stimulated low Km GTPase activity in aging and Alzheimer's disease: implications for altered signal transduction.

Carbachol-stimulated low Km GTPase activity (an index of muscarinic receptor-G protein coupling) was examined in hippocampus, basal ganglia, orbital frontal gyrus and superior frontal gyrus obtained from mature, aged and Alzheimer's Disease (AD) groups. Results indicated that carbachol-stimulated low Km GTPase activities in basal ganglia were as follows: mature controls > aged > AD, and there was a trend toward a similar pattern of decline in the hippocampus. No differences were seen in the two cortical areas examined; however, carbachol-stimulated low Km GTPase activity was small in the mature controls. Importantly, there were significant negative correlations between disease duration and carbachol-stimulated low Km GTPase activity in all areas examined except the orbital frontal gyrus. The longer the duration of the disease the lower the carbachol-stimulated low Km GTPase activity. Results suggest that age and disease-related changes in mAChR-G protein interactions in the basal ganglia may contribute to reduced signal transduction (ST). In addition, since decreased carbachol-stimulated low Km GTPase activity has also been observed in the aged rat; thus, investigations of the factors involved in decrements in signal transduction in the aged rat may be useful for understanding these alterations in aged humans or victims of AD.

Aged↗
PubMed1994

Cellular and molecular mechanisms of impaired dopaminergic function during aging.

One important cause of impaired motor function during aging is deterioration of the dopamine system. Such motor deficits in experimental animals can be closely related to loss of striatal dopamine receptors, and similar observations have now been made in humans. Two mechanisms account for the age-related decrease in striatal dopamine receptor levels: loss of receptor-containing neurons and reduced rates of receptor synthesis. The striatal neurons affected by aging appear to reside in a kainic-acid-sensitive population. Attempts to mimic those death mechanisms which occur in vivo using cultured neurons suggest that large D2-dopamine-receptor-containing cells may be the most vulnerable. Whether dopamine itself, the endogenous neurotransmitter for the cells, may ultimately be toxic to these neurons remains to be determined. The levels of D2-receptor mRNA in the surviving neurons is reduced during aging. This decrement is apparently due to a decreased rate of mRNA biosynthesis. Future experiments must therefore focus on the regulatory elements of this gene in order to determine why its transcription is selectively affected by aging. Finally, various interventions have been shown to delay or reverse the age changes characteristic of the dopaminergic system. Both dopamine receptors and motor function have been manipulated by diet and exercise as well as 6-OH-dopamine lesions and estrogen and prolactin administration. The possibility that such treatments might eventually be utilized therapeutically has become increasingly real as our knowledge of the affected cellular and molecular processes continues to expand.

Aging↗
PubMed1994

Age-related changes in DNA synthesis stimulated by epinephrine and isoproterenol in primary cultured rat hepatocytes.

We examined epinephrine- and isoproterenol-stimulated DNA synthesis in primary cultured hepatocytes from 6-, 12-, and 24-month-old rats. Epinephrine-stimulated DNA synthesis in 6-month-old rat hepatocytes began after 20 h and reached a maximum at 50 h. Similarly, isoproterenol-stimulated DNA synthesis in 6-month-old rat hepatocytes began after 10 h and reached a maximum at 45 h. In contrast, both epinephrine- and isoproterenol-stimulated DNA synthesis in 12- and 24-month-old rat hepatocytes were reduced approximately 40-60% and 80%, respectively, as compared to that at 6 months. Both epinephrine- and isoproterenol-stimulated DNA synthesis were strongly inhibited by the beta-adrenergic antagonist, propranolol, but not by the alpha 1-adrenergic antagonist, prazosin, or the alpha 2-adrenergic antagonist, yohimbine. However, in the presence of EGF, epinephrine-stimulated DNA synthesis activity was inhibited by prazosin but not by propranolol. These results indicate that stimulated DNA synthesis in rat hepatocytes declines with age and that there are two different pathways for epinephrine-stimulated DNA synthesis in the presence or absence of EGF.

Adrenergic alpha-Antagonists↗