Search PubMed⌕ Search

Biomedical subjects

R A Wallace

Publications and source records attributed to R A Wallace.

At least 55 records · Page 3Linked to original sources
PubMed1988

Interaction of permanently uncharged dopamine analogs with the D-2 dopaminergic receptor.

The purpose of this study was to determine if structural analogs of dopamine in which the side chain nitrogen has been replaced by a permanently uncharged monomethylsulfide, monomethylselenide or sulfoxide group are capable of binding to the striatal D-2 dopamine receptor and acting as agonists at this receptor. All the permanently uncharged dopamine analogs were found to bind to the D-2 dopamine receptor as evidenced by their abilities to inhibit significantly [3H]spiperone binding to striatal homogenates. However, the inhibition of [3H]spiperone binding by the uncharged dopamine analogs was incomplete and was almost abolished by the addition of NaCl (125 mM) to the incubation medium or by the addition of dopamine or quinpirole at a concentration that that saturates the high-affinity state of the D-2 dopamine receptor. These effects of NaCl, dopamine and quinpirole suggest that the uncharged dopamine analogs bind primarily to the high-affinity state of the D-2 dopamine receptor. Whether the uncharged monomethylsulfide and sulfoxide analogs could function as dopamine agonists at the striatal D-2 dopamine receptor was assessed by determining the abilities of these compounds to inhibit the K+-evoked release of [3H]acetylcholine from striatal slices. Both the monomethylsulfide and sulfoxide analogs inhibited the K+-evoked release of [3H]acetylcholine, but this inhibitory effect does not appear to be due to the activation of the D-2 dopamine receptor since it was not reversed by the selective D-2 dopamine antagonist, sulpiride. Additionally, the uncharged monomethylsulfide and sulfoxide dopamine analogs were found to antagonize the ability of apomorphine to inhibit the K+-evoked release of [3H]acetylcholine, but this antagonistic effect does not appear to be due to the reversible blockade of the D-2 dopamine receptor since it was not reduced by increasing the concentration of apomorphine. Therefore, while the permanently uncharged analogs of dopamine appear to bind to the high-affinity state of the D-2 dopamine receptor, they are not dopamine agonists or antagonists at the striatal D-2 dopamine receptor involved in regulating the release of acetylcholine. These results suggest that a positive charge may be a requirement for the activation of the striatal D-2 dopamine receptor.

Acetylcholine↗
PubMed1988

New microspot micromanipulator for carbon dioxide laser surgery in otolaryngology. Early clinical results.

A new micromanipulator with microspot capability has been developed for carbon dioxide laser surgery in otolaryngology. The instrument features new infrared optics, which provide smaller laser spot sizes than those achievable with conventional micromanipulators (300 micron using a 400-mm operating microscope lens). The conventional red helium-neon aiming laser is replaced with a nonlaser fiberoptic image, and a power defocus control on the joystick manipulator eliminates external defocus controls. Less surrounding tissue trauma occurs with this new instrument by using a higher power density with average power settings of 1 to 2 W for cutting and ablation of tissue. Eight patients with benign laryngeal disease and one patient with dysplastic changes of the vocal cord were treated successfully without complications.

Animals↗
PubMed1987

Interaction of permanently charged analogs of dopamine with the D-2 dopaminergic receptor.

Dopamine can exist in both charged and uncharged forms at physiological pH. At present it is unclear which of these forms is responsible for dopaminergic agonist activity. The purpose of this study was to determine whether permanently charged structural analogs of dopamine containing either a nitrogen, sulfur, or selenium atom in the side chain can bind to and activate the D-2 dopamine receptor. Binding to and activation of the D-2 dopamine receptor were measured by determining the abilities of the permanently charged dopamine analogs to inhibit [3H]spiperone binding to striatal homogenates and to inhibit K+-stimulated [3H]acetylcholine release from striatal slices respectively. The quaternary ammonium, dimethylsulfonium and dimethylselenonium analogs of dopamine were all found to inhibit [3H]spiperone binding to the same extent and in a manner qualitatively similar to the parent amines, dopamine and dimethyldopamine. Thus, [3H]spiperone inhibition curves for dopamine, dimethyldopamine and the permanently charged dopamine analogs were generally shallow and fit best to a two-site binding model as indicated by computer-assisted analyses. The addition of 125 mM NaCl to the incubation medium resulted in a significant decrease in the proportion of high affinity binding sites for both the permanently charged analogs and the parent amines. Similarly, the permanently charged dopamine analogs were found to maximally inhibit the K+-stimulated release of [3H]acetylcholine to the same extent as dopamine and dimethyldopamine. However, the permanently charged analogs were less potent in inhibiting both [3H]spiperone binding and K+-stimulated [3H]acetylcholine release than dopamine and dimethyldopamine. These results show that dopamine analogs possessing a permanent positive charge in the side chain can bind to and activate the D-2 dopamine receptor. The lower potencies of the permanently charged analogs in binding to and activation of the D-2 dopamine receptor suggest that, while the ability of a compound to exist in an uncharged form is not a requirement, both charged and uncharged forms of the agonist molecule appear to play a role in D-2 dopamine agonist activity.

Acetylcholine↗
PubMed1987

Fundulus heteroclitus gonadotropin(s). I. Homologous bioassay using oocyte maturation and steroid production by isolated ovarian follicles.

Isolated ovarian follicles from several species were cultured to develop an in vitro bioassay system for Fundulus heteroclitus gonadotropin. An extract of F. heteroclitus pituitaries, when tested in heterologous systems using follicles from Rana pipiens. Xenopus laevis, and Carassius auratus, was ineffective in provoking either germinal vesicle breakdown or steroid production. In a homologous system using F. heteroclitus follicles, F. heteroclitus pituitary extract was capable of inducing both germinal vesicle breakdown and steroid production in a dose-dependent fashion. Testosterone, estradiol-17 beta, and 17 alpha-hydroxy,20 beta-dihydroprogesterone were detected in both the culture media and the follicle extracts after F. heteroclitus pituitary extract stimulation. The steroidogenic responses resulting from the pituitary extract stimulation were dependent on the size and stage of follicular development. Only large vitellogenic follicles (1.2-1.4 mm diameter) were able to produce 17 alpha-hydroxy,20 beta-dihydroprogesterone and testosterone. Small vitellogenic follicles (less than 1.2 mm) were unresponsive to stimulation by F. heteroclitus pituitary extracts as scored by either germinal vesicle breakdown or production of 17 alpha-hydroxy, 20 beta-dihydroprogesterone and testosterone. However, estradiol-17 beta production was detected in follicles of a much wider size range: Follicles as small as 0.8 mm diameter were responsive to F. heteroclitus pituitary extract stimulation and produced a large quantity of estradiol-17 beta. There was a marked seasonal sensitivity of F. heteroclitus follicles to pituitary extract stimulation in vitro. Follicles obtained from fish outside of the breeding season (January) were less responsive to stimulation by pituitary extract or steroid. The same preparation of pituitary extract was capable of provoking germinal vesicle breakdown in follicles obtained in May. Pituitary extracts prepared during October through January were also less potent than those prepared during the breeding season (February through September). We conclude that F. heteroclitus gonadotropin(s) shows a noticeable species specificity and that F. heteroclitus follicles exhibit both a season- and a size-dependent responsiveness to gonadotropin(s). Hence, with a judicious use of the appropriate types of F. heteroclitus ovarian follicles, we have been able to demonstrate that in vitro oocyte maturation and steroid production are sensitive, homologous bioassays for F. heteroclitus gonadotropin(s).

Animals↗
PubMed1987

Dopamine agonists: effects of charged and uncharged analogues of dopamine.

Dopamine, at physiological pH, may exist as either an uncharged amine or a charged ammonium species. In order to gain insight as to which species is better suited for interaction with the dopamine receptor, we have synthesized dopamine analogues in which the nitrogen atom is replaced with a neutral methyl sulfide, a neutral methyl selenide, a charged dimethylsulfonium iodide, and a charged dimethylselenonium iodide. These analogues were tested for their ability to inhibit the K+-stimulated release of [3H]acetylcholine from striatal slices. At 30 microM concentration, the charged sulfonium and selenonium salts possessed significant agonist activity while the corresponding neutral species were inactive, suggesting that a charged species is optimal for dopamine agonist activity. In addition, the methyl sulfide was converted into the corresponding sulfoxide and sulfone; however, neither of these oxidation products possessed significant activity as dopaminergic agonists.

Acetylcholine↗
PubMed1987

Charged analogues of chlorpromazine as dopamine antagonists.

Chlorpromazine (1, CPZ) is a potent dopamine antagonist that has been used widely as an antipsychotic agent. Since dopaminergic antagonists, like dopaminergic agonists, exist in solution as the charged and uncharged molecular species, it is not clear which form of the amine is most important for interaction with the dopamine receptor. Previous work from our laboratory has indicated that a variety of permanently charged species could replace the amine/ammonium moiety of dopamine and retain dopamine agonist activity. This paper describes the synthesis and dopamine antagonist activity of both the trimethylammonium iodide and the dimethylsulfonium iodide analogues of chlorpromazine. The permanently uncharged methyl sulfide analogue was also synthesized; however, due to its lack of aqueous solubility, its pharmacological activity could not be evaluated. Binding of both the dimethylsulfonium iodide and the trimethylammonium iodide analogues to D-2 dopamine receptors of rat striatal tissue was observed. The observed relative order of binding was CPZ greater than CPZ sulfonium analogue greater than CPZ ammonium analogue. These compounds had a similar order of activity in antagonizing the apomorphine-induced inhibition of potassium-induced release of [3H]acetylcholine from mouse striatal slices.

Acetylcholine↗
PubMed1986

Chromatographic resolution of chicken phosvitin. Multiple macromolecular species in a classic vitellogenin-derived phosphoprotein.

Chicken phosvitin was prepared from egg yolk by a variety of published methods, including a modification of our own original procedure. Yolk granules and all phosvitin preparations have been previously found to contain major phosphoproteins at Mr 40,000 and 33,000 and minor satellite components when electrophoresed on polyacrylamide gradient gels and stained with Stains-all. However, only our current preparation contained three additional phosphoproteins (Mr 18,000, 15,000 and 13,000) that are also present in yolk granules. Our current phosvitin preparation also appeared to have additional components when compared with other preparations by size-exclusion and anion-exchange chromatography. Particularly complex but entirely reproducible patterns were obtained by hydrophobic-interaction chromatography. However, a cross-referencing of fractions eluted by size-exclusion chromatography to the other procedures employed, including gel electrophoresis, reinforced the notion that unfractionated chicken phosvitin contains at least five major components, designated B, C, E1, E2 and F for the Mr 40,000, 33,000, 15,000, 18,000, and 13,000 phosphoproteins, respectively. Stoichiometric considerations lead us to suggest that vitellogenin I gives rise to phosvitins C and F, vitellogenin II gives rise to phosvitin B, and vitellogenin III gives rise to either phosvitin E1 or E2, but not both. Thus, a fourth, as yet undetected, vitellogenin may exist for the chicken.

Animals↗
PubMed1986

Isolation of phosvitin: retention of small molecular weight species and staining characteristics on electrophoretic gels.

Phosvitin was prepared from chicken yolk by a variety of published methods, including a new procedure modified from our own original method. Yolk granules and all phosvitin preparations were found to contain a cluster of phosphoproteins ranging in size from Mr 28,000 to 43,000 when electrophoresed on polyacrylamide gradient gels and stained with Stains-all. However, only the current preparation contained three additional phosphoproteins (Mr 13,000, 15,000, and 18,000) that are also present in yolk granules. None of these components could be demonstrated with Coomassie blue.

Animals↗
PubMed1986

Oocyte maturation in the mummichog (Fundulus heteroclitus): effects of steroids on germinal vesicle breakdown of intact follicles in vitro.

The effects of several steroids on the maturation of follicle-enclosed oocytes of the mummichog Fundulus heteroclitus in vitro were examined. At a relatively high concentration (1.0 microgram/ml), a number of different steroids, including pregnenolone, 17 alpha-hydroxypregnenolone, corticosterone, cortisol, 11-deoxycorticosterone, 11-deoxycortisol, androstenedione, testosterone, progesterone, 17 alpha-hydroxyprogesterone, 20 beta-dihydroprogesterone, and 17 alpha-hydroxy-20 beta-dihydroprogesterone, were able to induce germinal vesicle breakdown (GVBD) in prematuration oocytes. Cholesterol, 17 beta-estradiol, 11-ketotestosterone, and 11 beta-hydroxytestosterone were totally ineffective. In general, 11-oxysteroids tended to be less effective than their 11-deoxysteroid counterparts. Two 20 beta-dihydroprogestins--17 alpha-hydroxy-20 beta-dihydroprogesterone and 20 beta-dihydroprogesterone--were the most potent maturation-inducing steroids, initiating 50% GVBD at 1 ng/ml in follicles obtained from ovaries containing mature or maturing follicles in vivo, or at 2.5-4.0 ng/ml in follicles from ovaries lacking mature or maturing oocytes in vivo. These results are consistent with several previous studies involving salmonids and various other teleosts, and suggest a possible physiological role for a 20 beta-dihydroprogestin in the resumption of meiotic maturation in F. heteroclitus.

20-alpha-Dihydroprogesterone↗
PubMed1986

Changes in teleost yolk proteins during oocyte maturation: correlation of yolk proteolysis with oocyte hydration.

Yolk proteins of prematuration occytes and postmaturation eggs were compared by SDS gel electrophoresis in several teleosts, including freshwater species that produce demersal eggs, estuarine and marine species with demersal eggs, and marine species with pelagic eggs. In certain teleosts distinct changes in yolk protein banding patterns during oocyte maturation are suggestive of extensive secondary proteolysis of yolk proteins at this time; proteolysis is most pronounced in marine fishes with pelagic eggs. In many teleosts the oocyte swells by hydration during maturation; this hydration is also most pronounced in marine fishes with pelagic eggs. The extent of yolk proteolysis is well correlated with the extent of oocyte hydration during maturation.

Animals↗
PubMed1985

Phosvitins in Fundulus oocytes and eggs. Preliminary chromatographic and electrophoretic analyses together with biological considerations.

Vitellogenin serves as the plasma precursor for the yolk proteins, lipovitellin and phosvitin, in nonmammalian vertebrates. 32P-Vitellogenin was isolated from the plasma of the teleost, Fundulus heteroclitus, and was used both to label phosvitin in the ovary and to indicate the phosvitin region in preparative chromatographs of ovarian extracts on DEAE-cellulose. Crude [32P]phosvitin could be resolved further into two labeled components with shallow gradients on DEAE-cellulose and into eight labeled components by electrophoresis on 12% polyacrylamide gels. Only the two largest electrophoretically resolved components could be correlated with Coomassie Blue staining bands, but several of the smaller components could be indicated with the cationic carbocyanine dye, Stains-all. Stains-all-dyed components were also generally indicated as multiple bands. The ovary of a reproductively active female contains vitellogenic oocytes, postvitellogenic oocytes undergoing maturation prior to ovulation, and ovulated eggs. Examination of various types of follicles and eggs on polyacrylamide gels revealed that during maturation, the largest phosvitin components formed during vitellogenesis either disappear or diminish, while smaller phosvitin components appear. The transformation of phosvitin components can also be achieved in vitro by incubating prematurational follicles in a saline medium containing deoxycorticosterone. These preliminary results demonstrate that a complex array of phosvitin-like components are present within a single ovary of F. heteroclitus. We also postulate that one reason for the anomalous yolk proteins generally found thus far in teleost eggs is that some of the proteins derived from vitellogenin during vitellogenesis undergo further proteolysis during oocyte maturation.

Animals↗
PubMed1985

Major protein changes during vitellogenesis and maturation of Fundulus oocytes.

The protein content of various size follicles was measured in Fundulus heteroclitus and indicated four phases of increase relative to follicle volume: Phases I (previtellogenic; estimated to be less than 0.01 mg/mm3), II (vitellogenic; 0.20 mg/mm3), III (early maturation; 0.03 mg/mm3), and IV (late maturation; 0 mg/mm3). A pronounced and rapid size increase occurs during maturation due to hydration, but protein uptake, which was also documented cytologically, contributes to about 16% of the volume increment during early maturation. Protein incorporation appears to stop abruptly at the time of germinal vesicle breakdown, most likely reflecting an altered physiological state of the oocyte. SDS-polacrylamide gel electrophoretic patterns of various size follicles indicated that five major protein bands (molecular weights = 122, 103, 45, 26, and 20 k) accumulate during vitellogenesis and presumably are proteolytically derived from a 200-kDa vitellogenin precursor. During maturation, the 122- and 45-kDa proteins disappear and several new, lower molecular weight bands appear. Proteolysis of specific yolk proteins may thus help generate part of the osmotic pressure gradient required for water uptake during oocyte maturation.

Animals↗
PubMed1984

Effect of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) on monoamine neurotransmitters in mouse brain & heart.

The effect of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was studied on dopamine (DA), norepinephrine (NE), serotonin (5HT) and gamma-aminobutyric acid (GABA) neurons in mouse brain and on NE neurons of mouse heart. MPTP (45 mg/kg) was administered s.c. to mice twice daily for 2 consecutive days. This dosage regimen produced a decrease in the forebrain concentrations of DA and NE at 7 and 20 days after injection. In contrast, the forebrain concentrations of 5HT and GABA were not significantly decreased at either time. MPTP administration also produced a marked decrease in the uptake of 3H-DA into striatal slices and 3H-NE into cerebral cortical slices. In contrast, the uptake of 3H-NE into hypothalamic slices and the uptake of 3H-5HT into slices from several brain regions were not altered. MPTP initially reduced the concentration of NE in the heart, but unlike the persistent decreases in the forebrain concentrations of NE and DA, the NE concentration in the heart returned to control levels at approximately 20 days after MPTP administration. These results, showing that MPTP can produce a long lasting and selective decrease in the forebrain concentrations of NE and DA and in the uptake of radioactive DA and NE into brain slices, suggest that MPTP can cause the destruction of catecholamine neurons in mouse brain. In contrast, MPTP administration does not appear to produce long term changes in either 5HT or GABA neurons.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗
PubMed1983

The oocyte as an endocytic cell.

Oocytes of Xenopus laevis grow primarily by sequestering vitellogenin (VTG) selectively from the maternal bloodstream. Morphological observations have demonstrated that an endocytic system is responsible for VTG uptake. Binding studies indicate the presence of 2-28 X 10(10) surface VTG receptors per oocyte. These are continuously internalized into endosomes whether or not they are occupied by VTG, and other macromolecules may become trapped in the process. VTG-containing endosomes give rise to dense transitional yolk bodies; these fuse with yolk platelets only after the cleavage of vitellogenin. In the absence of VTG, endosomes appear to fuse directly with yolk platelets. From these observations it is postulated that receptor occupancy can act as a transmembrane signal which directs the postendocytic compartmentation of proteins. Yolk platelet proteins do not undergo subsequent turnover, whereas adventitiously incorporated protein is gradually lost from the oocyte by a dual mechanism which may involve both lysosomal proteolysis and secretion from the oocyte as a consequence of membrane recycling. Although these observations may not apply to all growing oocytes, the X. laevis oocyte nevertheless appears to be a particularly attractive experimental system for studies of endocytic compartmentation and membrane receptor recycling.

Animals↗
PubMed1983

Oogenesis in Fundulus heteroclitus. III. Vitellogenesis.

Several physiological parameters associated with oocyte growth (vitellogenesis) in the killifish, Fundulus heteroclitus, are defined. Hepatocytes from females and estrogen-treated males are highly synthetic cells, whereas those from males are relatively quiescent. A 2000,000-molecular weight phosphoprotein (vitellogenin) has been identified by sodium dodecyl sulfate (SDS)-gel electrophoresis in the plasma of females and estrogen-treated males and isolated chromatographically using a potassium phosphate gradient system on DEAE-cellulose. To inhibit proteolysis, the nontoxic inhibitor, aprotinin, was injected into fish prior to bleeding. The structure of vitellogenic follicles and the process of oocyte growth are described. Once follicles reach a diameter of 0.5 mm, their oocytes incorporate exogenous materials by micropinocytosis and sequestered material is immediately translocated to yolk spheres in the oocyte cortex. This process has been followed by using the tracer horseradish peroxidase. Exogenous proteins appear to leave the perifollicular capillaries via an intercellular route and pass through intercellular channels within the follicular epithelium and patent pore canals of the vitelline envelope before reaching the oocyte surface. [32P]Vitellogenin isolated from F. heteroclitus, [3H]vitellogenin isolated from X. laevis, and the dye trypan blue have been used to determine which sized follicles are vitellogenic (i.e., incorporate exogenous materials into growing oocytes). Morphological and autoradiographic studies are presented to confirm that sequestered macromolecules are incorporated into yolk spheres in the growing oocyte. Follicles appear to grow from 0.5 to 1.3 mm in diameter due to vitellogenesis and subsequently undergo an additional increase in size, which is primarily due to hydration, during oocyte maturation (Wallace and Selman, '78).

Animals↗