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Biomedical subjects

C Y Lu

Publications and source records attributed to C Y Lu.

At least 181 records · Page 10Linked to original sources

Beta-adrenergic stimulation evokes a rapid, Ca2+-dependent stimulation of endocytosis, hexose and amino acid transport associated with increased Ca2+ fluxes in mouse kidney cortex.

The beta-adrenergic agonist 1-isoproterenol evokes an acute (less than 5 min) stimulation of endocytosis, hexose transport and amino acid transport, measured by the temperature-sensitive uptake of HRP, 3H-DG and 14C-AIB, in mouse kidney cortex slices. This stimulation is concentration dependent and is maximal at 10(-8)-10(-7) M isoproterenol. Peroxidase cytochemistry showed that the hormonal increase in HRP uptake is confined to proximal tubules. The rapid membrane response is abolished in a calcium-free medium and by the beta-adrenergic antagonist propranolol, indicating Ca2+- and beta-adrenoreceptor-dependence. Isoproterenol (1 microM) rapidly (less than 30 sec) stimulates the influx and efflux of 45Ca in cortex slices. Isoproterenol also decreased mitochondrial 45Ca and increased soluble 45Ca. These results indicate that beta-adrenergic stimulation of membrane transport functions involves an increased influx of extracellular calcium and a mobilization of intracellular (mitochondrial) calcium. An increase in cytosolic Ca2+ concentration appears to be the regulatory signal for these membrane transport processes.

Amino Acids↗

Androgenic stimulation of endocytosis, amino acid and hexose transport in mouse kidney cortex involves increased calcium fluxes.

Testosterone was previously shown to induce an early (less than 1 min) receptor-dependent stimulation of endocytosis, hexose and amino acid transport in mouse kidney cortex (Koenig, H., Goldstone, A. and Lu, C.Y. (1982) Biochem. Biophys. Res. Commun. 104, 165-172). Testosterone (10(-8) M) has now been found to stimulate rapidly (less than 30 s) the influx and efflux of 45Ca2+ in cortex slices. Testosterone also decreased mitochondrial 45Ca and augmented soluble 45Ca, indicating a mobilization of intracellular calcium. Incubation of cortex slices in calcium-free medium without or with 2.5 mM EGTA decreased basal endocytosis, hexose and amino acid transport and blocked the hormonal response. 100 microM verapamil blocked the hormonal response without affecting basal transport. The calcium ionophore A23187 rapidly stimulated endocytosis, hexose and amino acid transport. These data indicate that androgenic stimulation of membrane transport functions involves an increased influx of extracellular calcium and a mobilization of intracellular calcium. Increased cytosolic Ca2+ is probably the regulatory signal for these transport processes.

Amino Acids↗

Beta-adrenergic stimulation of Ca2+ fluxes, endocytosis, hexose transport, and amino acid transport in mouse kidney cortex is mediated by polyamine synthesis.

We recently found that the beta-adrenergic agonist 1-isoproterenol evokes a rapid (less than 5 min) Ca2+- and receptor-dependent stimulation of endocytosis, hexose transport, and amino acid transport in mouse renal cortex involving proximal tubule cells. This response is associated with increased Ca2+ fluxes and a mobilization of mitochondrial calcium, suggesting that stimulus-response (stimulus-"transport") coupling is mediated by cytosolic Ca2+. We show here that 1 microM isoproterenol evokes a rapid (less than 60 sec) transient increase in the activity of ornithine decarboxylase followed by an early (less than 2 min) sustained increase in putrescine, spermidine, and spermine concentrations in mouse kidney cortex slices in vitro. Small doses of isoproterenol (down to 24 nmol/kg) elicited a rapid (less than 2 min) increase in polyamines in vivo. The ornithine decarboxylase inhibitor alpha-difluoromethylornithine (5 mM) suppressed the testosterone-induced increase in polyamine levels and rates of endocytosis, hexose transport, and amino acid transport, measured by horseradish peroxidase, [14C]aminoisobutyric acid, and deoxy[3H]glucose uptake. alpha-Difluoromethylornithine also blocked the isoproterenol-induced increase in 45Ca influx and efflux and 45Ca redistribution; 0.5 mM putrescine nullified alpha-difluoromethylornithine inhibition and restored the increment in polyamines, 45Ca fluxes, endocytosis, hexose transport, and amino acid transport. These data implicate polyamine synthesis in isoproterenol stimulation of Ca2+ fluxes and membrane transport processes and support a model for signal transduction and stimulus-response coupling in which ornithine decarboxylase activation and polyamine synthesis play a pivotal role in regulating Ca2+ fluxes. In this model the polyamines generate local Ca2+ signals by stimulating Ca2+ influx or mobilizing intracellular calcium (or both) through a cation exchange reaction.

Amino Acids↗

Ontogeny of murine macrophages: functions related to antigen presentation.

Macrophage function in neonates was dissected into four components: antigen uptake and catabolism, cytotoxicity, antigen presentation, and the production of the lymphostimulatory molecule interleukin-1 (also called thymocyte mitogenic protein or lymphocyte-activating factor). The uptake and catabolism of 125I-labeled Listeria monocytogenes was equivalent in macrophages from adult and neonatal mice. However, interactions between macrophages from neonates, heat-killed Listeria organisms, and immune T lymphocytes were impaired, and no cytocidal macrophages capable of killing tumor cells were generated. Previous studies with cells from adult mice had established that the development of cytocidal macrophages required Ia-bearing, antigen-presenting macrophages and histocompatibility at I-A between macrophages and T cells. To circumvent this requirement for antigen-presenting macrophages, an assay was used in which lymphokine was added directly to the macrophages from neonates. Strong cytocidal activity resulted. Thus, our studies confirmed that macrophages from neonates present antigen poorly but can acquire cytocidal function provided that the need for antigen-presenting function is bypassed. Similar conclusions were reached for the secretion of interleukin-1. Macrophages from neonates spontaneously secreted as much mediator as macrophages from adults, and the secretion was increased after the ingestion of heat-killed Listeria organisms or endotoxin. However, the marked increase in interleukin-1 production that follows antigen-macrophage-lymphocyte interaction was best seen in macrophages from adults. Macrophages from neonates could be activated to ingest C3b-coated sheep erythrocytes.

Animals↗

Testosterone-mediated sexual dimorphism of the rodent heart. Ventricular lysosomes, mitochondria, and cell growth are modulated by androgens.

The ventricular myocardium was studied in A/J mice and in Sprague-Dawley rats. In male mice, the ventricles were slightly larger and the specific activities of the lysosomal hydrolases, beta-glucuronidase, hexosaminidase, beta-galactosidase, and arylsulphatase, and the inner mitochondrial enzyme cytochrome c oxidase were substantially higher than in female mice. Orchiectomy abolished this sex difference. Testosterone administration induced myocardial hypertrophy and accretion of RNA and protein without altering the DNA, and substantial increases in the activities of the lysosomal hydrolases and cytochrome c oxidase. However, the mitochondrial membrane enzyme monoamine oxidase was unaffected by sex, orchiectomy, and testosterone administration. Heart lysosomes from male mice showed a smaller structure-linked latency of the lysosomal enzymes and a greater fragility of the lysosomal membrane to osmotic and mechanical stress than those from female mice. This sex difference was also abolished by orchiectomy and restored by testosterone replacement. Similar sex differences were observed in the rat with respect to heart size, acid hydrolase activities, and lysosomal enzyme latency and membrane stability. These findings indicate that endogenous androgens regulate myocardial cell growth, the activity of enzymes associated with lysosomes and the inner mitochondrial membrane, and some physiochemical properties of lysosomes.

Animals↗

Control of macrophage Ia expression in neonatal mice--role of a splenic suppressor cell.

The control of macrophage expression of I region-associated antigens (Ia) in neonatal mice was studied by comparing responses of neonatal and adult mice to immune vs nonimmune stimuli. Adults generated peritoneal exudates rich in Ia-bearing macrophages in response to i.p. injection of live Listeria monocytogenes, Listeria-immune T cells, and heat-killed Listeria, or a soluble mediator termed macrophage Ia-recruiting factor (MIRF). Neonates failed to respond to these stimuli. In contrast, both neonates and adults generated Ia-negative peritoneal exudates when stimulated with thioglycollate. A neonatal spleen cell that blocked the response of adults both to immune T cells and heat-killed Listeria and to MIRF was identified and characterized. Some of the suppressor cells appeared to be early precursors of the phagocytic lineage that develop into mature monocyte-macrophages. Suppression was apparently mediated by metabolites of arachidonic acid since indomethacin and aspirin in vivo blocked the effect. Similar suppressor activity was found in adult bone marrow and in adult resident peritoneal exudate cells. Thus, the phagocytic line autoregulates its surface expression of Ia in both neonatal and adult mice. This mechanism becomes particularly pointed during early development and could contribute to the lack of immunity during ontogeny.

Animals↗

Glucocorticoids and aspirin inhibit and cyproterone acetate enhances the androgenic response in mouse kidney.

Glucocorticoids and aspirin antagonize the androgenic response in mouse kidney, but not in ventral prostate or seminal vesicles. These agents impeded the testosterone-mediated increase in kidney weight, cytochrome c oxidase, and lysosomal hydrolases and urinary excretion of lysosomal hydrolases and proteins. They also attenuated the testosterone-induced decrease in enzyme latency and membrane stability of kidney lysosomes. In contrast, the antiandrogen cyproterone acetate is weakly androgenic in kidney and potentiates testosterone-induced lysosomal enzymuria and proteinuria (synandrogenic effect).

Animals↗

Androgens regulate cell growth, lysosomal hydrolases and mitochondrial cytochrome c oxidase in mouse aorta.

The aorta in male mice shows higher activities of several lysosomal hydrolases and of cytochrome c oxidase, an inner mitochondrial membrane enzyme, than in female mice. Orchiectomy abolishes this sex difference, whereas testosterone administration induces an accretion of RNA and protein and elevated activities of lysosomal hydrolases and cytochrome c oxidase. However, the outer mitochondrial membrane enzyme monoamine oxidase is unaffected by sex, orchiectomy or testosterone. Thus, androgens regulate cell growth and enzymes associated with lysosomes and the inner mitochondrial membrane.

Androgens↗

Ia-bearing macrophages in athymic mice: antigen presentation and regulation.

Ia-bearing macrophages from spleens and peritoneal cavities of athymic (nu/nu) and euthymic (nu/+) were compared. Macrophages from both strains of mice had equal antigen-presenting ability. The basal numbers of unstimulated Ia-bearing peritoneal macrophages were equal, but only euthymic mice recruited large numbers of Ia-bearing macrophages after Listeria infection. The i.p. injections of a "macrophage Ia-recruiting factor" induced exudates rich in Ia-bearing macrophages in both athymic and euthymic mice. These data suggest 2 levels of control of Ia-bearing macrophages: a basal T cell-independent level and a stimulated level dependent on mature T cells.

Animals↗

Androgens regulate mitochondrial cytochrome c oxidase and lysosomal hydrolases in mouse skeletal muscle.

The gastrocnemius, a fast-twitch white muscle, and the soleus, a slow-twitch red muscle, were studied in A/J mice. The specific activities of the lysosomal hydrolases, beta-D-glucuronidase, hexosaminidase, beta-D-galactosidase and arylsulphatase, the inner-mitochondrial-membrane enzyme cytochrome c oxidase, and the outer-mitochondrial-membrane enzyme monoamine oxidase, were greater in the soleus than in the gastrocnemius. The specific activities of the lysosomal hydrolases and cytochrome c oxidase in the gastrocnemius and soleus were substantially higher in male mice than in female mice. Orchiectomy abolished this sex difference. Testosterone increased the activities of the lysosomal hydrolases and cytochrome c oxidase and coincidentally induced muscle hypertrophy and an accretion of protein and RNA, but total DNA remained constant. Monoamine oxidase was unaffected by sex, orchiectomy and testosterone. These findings indicate that endogenous androgens regulate the activity of enzymes associated with lysosomes and the inner mitochondrial membrane, as well as muscle fibre growth in mouse skeletal muscle.

Animals↗

Testosterone-mediated sexual dimorphism of mitochondria and lysosomes in mouse kidney proximal tubules.

In kidney proximal tubules of male mice the mitochondria are larger and more electron-lucent, autophagic vacuoles and lysosomes (predominantly myeloid bodies) more numerous and voluminous, and exocytosed intraluminal myeloid bodies more common than in females. Males also have higher kidney activities of mitochondrial cytochrome c oxidase and lysosomal hydrolases, and excrete larger quantities of hydrolases and protein in the urine. Orchiectomy evokes the feminine pattern whereas testosterone administration induces the male pattern. Endogenous testosterone modulates mitochondrial structure and function and enhances the activity of the lysosomal-vacuolar system in proximal tubule cells.

Animals↗

During ontogeny, Ia-bearing accessory cells are found early in the thymus but late in the spleen.

The ontogeny of Ia-bearing accessory cells was studied in mice. Ia-bearing adherent cells from the thymus, consisting predominantly of macrophages, were found from birth. These adherent cells were able to present antigen, as measured by their ability to induce immune T-cell proliferation. In contrast, Ia-bearing adherent cells from the spleen were not found until the second week of life, and their antigen-presentation function was not present until later. The differential ontogeny of Ia-bearing accessory cells at these sites may be important in both development of immune competence and the restriction of autoimmunity.

Animals↗

Lysosomal enzyme secretion in rat ventral prostate. Secretagogue action of testosterone and dibutyryl cyclic AMP.

Rat ventral prostate slices, when incubated in vitro, secrete protein, lysosomal enzymes and newly phosphorylated components in a time- and temperature-dependent manner. Testosterone and dibutyryl cyclic AMP immediately stimulate secretion and 32Pi incorporation. Acid phosphatase and beta-N-acetylhexosaminidase are secreted as acidic isoenzymes presumably contained within primary lysosomes.

Acid Phosphatase↗

Testosterone and 6-N,2'-O-dibutyryladenosine 3':5'-cyclic monophosphate stimulate protein and lysosomal enzyme secretion in rat seminal vesicle.

Rat seminal-vesicle secretion was studied in vitro in a slice-incubation system. Seminal-vesicle slices were preincubated with 32Pi for 15 min, rinsed, and incubated in an isotope-free 'chase' medium for up to 4h. Gland slices spontaneously discharged protein, three lysosomal hydrolases and trichloroacetic acid-insoluble 32P into the medium in a time- and temperature-dependent manner. Testosterone (10 muM) and dibutyryl cyclic AMP (1 mM) stimulated the discharge of protein, acid hydrolases and trichloroacetic acid-insoluble 32P, and also stimulated the incorporation of 32Pi into trichloroacetic acid-insoluble components. The acid phosphatase and beta-N-acetylhexosaminidase isoenzymes were separated by isoelectric focusing. These hydrolases were secreted into the medium as acidic isoenzymes, presumably contained within primary lysosomes, whereas they occurred largely as less acidic and basic isoenzymes in the glandular tissue.

Acid Phosphatase↗