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Biomedical subjects

J A Pachter

Publications and source records attributed to J A Pachter.

24 records · Page 2Linked to original sources

Dopamine has no effect on thyrotropin-releasing hormone mobilization of calcium from intracellular stores in rat anterior pituitary cells.

To investigate whether dopaminergic reduction of PRL secretion is caused by an inhibition of release of Ca2+ from intracellular stores, we used a perifusion system to monitor simultaneously changes in intracellular Ca2+ concentrations ([Ca2+]c) and changes in PRL secretion from rat anterior pituitary cells, and from enriched populations of lactotrophs. We eliminated influx of extracellular Ca2+ by using medium with no added Ca2+ and 0.2 mM EGTA, conditions which abolished the increase in [Ca2+]c caused by 56 mM KCl. In this low Ca2+-containing medium, 100 nM TRH induced a burst of [Ca2+]c; the magnitude of the peak was the same whether the cells were perifused with low Ca2+-containing medium for 2 or 20 min before adding TRH, indicating the source of intracellular Ca2+ was stable under these conditions. After 2 min in this medium, TRH was still able to stimulate almost as much PRL release as in medium containing 1.8 mM Ca2+, and 1 microM dopamine inhibited this release, but did not affect the magnitude of the TRH-induced increase in [Ca2+]c. Preincubation for 5 min with dopamine did not affect the ability of a 30-sec incubation with TRH to stimulate the accumulation of inositol phosphate, inositol bisphosphate, and inositol trisphosphate, either in medium containing 1.8 mM Ca2+, or in low Ca2+-containing medium. Preincubation with dopamine for 5 min had no effect on TRH-induced mobilization of intracellular calcium. A source of Ca2+ is needed to refill internal Ca2+-stores discharged by TRH, and as dopamine lowered [Ca2+]c which might fill these stores, we tested to see if dopamine prevented.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Interactions between vasoactive intestinal peptide and dopamine in the rabbit retina: stimulation of a common adenylate cyclase.

Although 3,4-dihydroxyphenylethylamine (dopamine, DA) and vasoactive intestinal peptide (VIP) have been reported to stimulate adenylate cyclase activity in the rabbit retina, possible interactions between VIP-sensitive and DA-sensitive adenylate cyclase systems have not been previously investigated. To elucidate the interactions between these two putative transmitter-stimulated cyclase systems, the effects of VIP, DA, and VIP + DA on the conversion of [alpha-32P]ATP to [32P]cyclic AMP in rabbit retinal homogenates were measured. VIP stimulated adenylate cyclase activity in a biphasic manner, suggesting that two classes of VIP receptors may be involved in the induction of cyclic AMP formation. DA was less potent than VIP, and stimulated cyclase activity with a monophasic dose-response curve. When assayed together, these stimulations were partially nonadditive, implying the existence of a common adenylate cyclase pool that may be stimulated by both putative neurotransmitters. The dopaminergic antagonist (+)-butaclamol completely blocked dopaminergic stimulation, but had no significant effect on VIP-induced stimulation, indicating that VIP interacts with specific VIP receptor sites, which are distinct from the dopaminergic receptor sites. Furthermore, the specific D-2 dopaminergic receptor agonist LY141865 demonstrated no inhibitory effect on adenylate cyclase activity, suggesting that the interaction between the VIP- and DA-sensitive adenylate cyclase systems does not result from a D-2 receptor-mediated cyclase inhibition in the rabbit retina. Finally, at maximally effective concentrations, DA and VIP were less potent than fluoride or forskolin in the stimulation of cyclic AMP formation, suggesting that adenylate cyclase pools that are not sensitive to DA and VIP may also be present in this retina.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclases↗

Viscometric and fluorometric studies of deoxyribonucleic acid interactions of several new anthracyclines.

Interactions involved in the binding of the anthracycline analogues adriamycin, carminomycin, pyrromycin, musettamycin, marcellomycin, and aclacinomycin to calf thymus DNA and covalently closed circular PM-2 DNA have been studied. Fluorescence quenching experiments revealed that denaturation of calf thymus DNA and increasing ionic strength each resulted in a marked decrease in the DNA binding affinities of all of the anthracyclines studied. These results suggest that intercalative and electrostatic interactions are both important in the DNA binding of these analogues. Viscometric studies indicated that under high ionic strength conditions which negated electrostatic effects, all of the anthracyclines induced an unwinding--rewinding process of the closed superhelical PM-2 DNA typically observed for DNA intercalators. Relative to the 26 degrees unwinding angle of ethidium bromide, anthracyclines with a daunomycinone-like aglycon induced an unwinding angle of approximately 13 degrees. This differed slightly from the unwinding angles of 10.3 degrees-11.1 degrees which were induced by anthracyclines with a pyrromycinone-like aglycon. Increases in the length of the glycosidic side chain did not elicit significant differences in PM-2 DNA unwinding ability, implying a lack of effect of glycosidic side chain length on the anthracycline intercalation process. The unwinding angles also showed little sensitivity to decreases in ionic strength, suggesting that the fraction of bound anthracycline molecules which are in a nonintercalated state is similar to the fraction of bound ethidium bromide molecules which are in a nonintercalated state.

Antibiotics, Antineoplastic↗

Deoxyribonucleic acid binding studies on several new anthracycline antitumor antibiotics. Sequence preference and structure--activity relationships of marcellomycin and its analogues as compared to adriamycin.

The deoxyribonucleic acid (DNA) binding characteristics of adriamycin and several new anthracycline glycosides, including marcellomycin, aclacinomycin, rudolfomycin, musettamycin, and pyrromycin, have been studied. The fluorescence spectra were determined for all six anthracyclines, and the fluorescence quenching effects caused by interactions with the natural DNAs poly(dAdT)--poly(dAdT) and poly(dGdC) were characterized. Binding parameters were determined by Scatchard analyses of results obtained by spectrofluorometric titrations of anthracyclines with DNA. Consistent with earlier structure--activity relationship studies of nucleic acid synthesis inhibitory effects, the results demonstrate a correlation between the length of the glycosidic side chain and DNA binding affinity. In addition, the sugar residue 2-deoxyfucose appears to confer greater DNA binding ability than do the sugars rednosamine and cinerulose when present in the terminal position of the glycosidic side chain, also in agreement with earlier studies. The sequence preference of anthracycline--DNA interaction has been examined by using DNAs of varying GC content, including the naturally occurring calf thymus DNA (43% GC), Clostridium perfringens DNA (28% GC), and Micrococcus luteus DNA (72% GC) and the synthetic double-stranded copolymers poly(dGdC)--poly(dGdC) and poly(dAdT)--POLY(DAdT). The results demonstrate that although adriamycin shows an absolute requirement for GC sequences for DNA binding, marcellomycin and its analogues showed no such sequence requirement. Furthermore, an AT preference for DNA binding was demonstrated with marcellomycin and its analogues.

Aclarubicin↗