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M Treiman

Publications and source records attributed to M Treiman.

30 records · Page 2Linked to original sources
PubMed1988

Low-affinity uptake of [3H]choline by rat neurointermediate lobe in vitro.

Uptake of [3H]choline by rat neurointermediate lobes in vitro was investigated. The rate of uptake showed saturation with concentration of [3H]choline above 120 microM. Lowering of the incubation temperature from 32 to 4 degrees C, or increasing the concentration of K+ in the incubation medium from 5 to 100 mM, diminished the rate of uptake by 83.8% (SEM 6.9%, n = 3) or 43.1% (SEM 13.5%, n = 3), respectively. Following preloading with [3H]choline, a slow efflux (1% of the content every 20 min) of [3H]radioactivity was observed from the perifused glands. This efflux was enhanced 10-fold by increasing the concentration of K+ in the perifusion medium to 100 mM. Neither the uptake of [3H]choline, nor the subsequent basal or potassium-enhanced efflux of [3H]radioactivity were affected by reducing the concentration of Na+ from 125 to 19 mM, or by including 10 microM hemicholinium-3 in the medium during preincubation and perifusion. Replacing Ca2+ by 0.5 mM EGTA during perifusion resulted in a minor decrease (28%, SEM 7.3%, n = 7) in the potassium-enhanced [3H]radioactivity efflux. This decrease occurred only in one of the two high-potassium periods during the perifusion. In conclusion, uptake of [3H]choline by rat neurointermediate lobes was due to a low-affinity, saturable mechanism, with the efflux of [3H]radioactivity most likely representing the depolarization-facilitated outflow of [3H]choline. Autoradiography of the tissue sections showed this uptake to be localized to both neuronal and glial elements of the neural lobe, in contrast to sparse labelling of pars intermedia.

Animals↗
PubMed1985

D-1 and D-2 dopaminergic receptors regulate protein phosphorylation in the rat neurohypophysis.

Dopamine stimulates the phosphorylation of the neuron-specific synaptic vesicle proteins Synapsin I, Protein IIIa and Protein IIIb in the posterior pituitary gland of the rat [Tsou and Greengard (1982) Proc. natn. Acad. Sci. U.S.A. 79, 6075-6079]. This effect has been characterized in the present investigation. The stimulatory effect of dopamine was mimicked by the selective D-1 receptor agonist SKF 38393 and was competitively and potently inhibited by the selective D-1 receptor antagonist SKF 83509 as well as by the mixed D-1/D-2 antagonist fluphenazine. Conversely, the effect of dopamine was attenuated by a D-2 receptor agonist (LY 141865) and potentiated by a D-2 receptor antagonist (sulpiride). Norepinephrine also stimulated phosphorylation of the synaptic vesicle proteins, apparently through activation of the D-1 receptor. D-1 and D-2 dopaminergic receptors may play a role in the regulation of hormone secretion from the neurohypophysis. Evidence exists that in the isolated neurophypophysis activation of D-1 receptors facilitates, while activation of D-2 receptors inhibits, release of vasopressin. Further work will be required to determine whether the regulation by D-1 and D-2 receptors of the protein phosphorylation in the neurohypophysial peptidergic terminals is related to the regulation by those receptors of the neurohypophysial hormone secretion.

1-Methyl-3-isobutylxanthine↗
PubMed1983

3',5'-cyclic adenosine monophosphate- and Ca2+-calmodulin-dependent endogenous protein phosphorylation activity in membranes of the bovine chromaffin secretory vesicles: identification of two phosphorylated components as tyrosine hydroxylase and protein kinase regulatory subunit type II.

Membranes of the secretory vesicles from bovine adrenal medulla were investigated for the presence of the endogenous protein phosphorylation activity. Seven phosphoprotein bands in the molecular weight range of 250,000 to 30,000 were observed by means of the sodium dodecyl sulphate electrophoresis and autoradiography. On the basis of the criteria of molecular weight, selective stimulation of the phosphorylation by cyclic AMP (as compared with cyclic GMP) and immunoprecipitation by specific antibodies, band 5 (molecular weight 60,300) was found to represent the phosphorylated form of the secretory vesicle-bound tyrosine hydroxylase. The electrophoretic mobility, the stimulatory and inhibitory effects of cyclic AMP in presence of Mg2+ and Zn,2+ respectively, and immunoreactivity toward antibodies showed band 6 to contain two forms of the regulatory subunits of the type II cyclic AMP-dependent protein kinase, distinguishable by their molecular weights (56,000 and 52,000, respectively). Phosphorylation of band 7 (molecular weight 29,800) was stimulated about 2 to 3 times by Ca2+ and calmodulin in the concentration range of both agents believed to occur in the secretory tissues under physiological conditions.

Adrenal Medulla↗
PubMed1983

Calmodulin binding to secretory granules isolated from bovine neurohypophyses.

Secretory granules, isolated from bovine neurohypophyses on isoosmolar Percoll-sucrose-EGTA gradients had a calmodulin content of 0.09 +/- 0.01 micrograms/mg protein (SE, n = 6). The distribution of calmodulin on the gradient showed that it did not copurify with the granules. Specific binding sites for calmodulin with a high affinity (Kd = 2.43 +/- 0.27 X 10(-9) M (SE, n = 5] and a maximum binding capacity of 1.3 +/- 0.4 pmol/mg protein (SE, n = 5) could be demonstrated when such secretory granules were incubated with 125I-calmodulin.

Animals↗
PubMed1980

Complex phosphorylation activity in neurosecretosomal membranes isolated from ox neurohypophyses.

Homogenates of neural lobes of bovine pituitary glands were fractionated on Ficoll gradients to yield neurosecretosomes (nerve endings). The neurosecretosomes were lysed in a hypo-osmotic buffer and the membranes were separated from the soluble components by centrifugation. On incubation with [gamma-32P]ATP this membrane preparation showed an endogenous phosphorylation activity, which was studied by means of gel electrophoresis in the presence of sodium dodecyl sulphate, and subsequent autoradiography. The major part of the [32P]Pi detected on the gel was shown to be incorporated into three protein bands, termed A, B and C, with minimal mol.wts. of 83 000, 59 000 and 47 000 respectively. The phosphorylation of these three proteins was studied under a variety of experimental conditions. The patterns obtained were partly similar. However, important individual differences were noted, particularly with respect to the effects of cyclic AMP, Mg2+ and Ca2+. On the basis of these differences, it is suggested that in this system the phosphorylation activity is heterogenous, bands A, B and C each reflecting the presence of a different site of phosphate turnover. The relationship of bands A, B and C to several of the previously described phosphoproteins in the brain is discussed.

Adenosine Triphosphate↗
PubMed1980

Isolation and characterization of secretory vesicles from bovine neurohypophyses.

A procedure is described for the isolation of secretory vesicles from bovine neurohypophyses by differential centrifugation followed by density gradient centrifugation on iso-osmolal gradients of percoll/sucrose. Only negligible contamination of the secretory vesicle fraction with markers for mitochondria, microsomes and plasma membranes could be detected. The amount of Ca2-ATPase in the isolated neurohypophysial secretory vesicles was of the same low order of magnitude as that of (Na, K)-ATPase. Thin-section electromicrographs confirmed the high purity of the isolated secretory vesicle fractions, In freeze-fracture electronmicrographs, vesicle fusion was demonstrated after incubation with Ca2. As shown in dodecyl sulfate-gel electrophoresis and subsequent autoradiography secretory vesicles exhibited an endogenous phosphorylation activity. The secretory vesicles contained an average of 23.1 microgram vasopressin/mg of protein. On incubation in media differing in ionic strength, pH and Ca2 concentration the vesicles were stable for at least 1 h.

Animals↗
PubMedDate not supplied

The distribution of the endoplasmic reticulum in living pancreatic acinar cells.

Studies on pancreatic acinar cells provided the original evidence for the Ca(2+) releasing action of inositol 1,4,5-trisphosphate (IP(3)). Ironically, this system has presented problems for the general theory that IP(3) acts primarily on the endoplasmic reticulum (ER), because the IP(3)-elicited Ca(2+) release occurs in the apical pole, which is dominated by zymogen granules (ZGs) and apparently contains very little ER. Using confocal and two-photon microscopy and a number of different ER-specific fluorescent probes, we have now investigated in detail the distribution of the ER in living pancreatic acinar cells. It turns out that although the bulk of the ER, as expected, is clearly located in the baso-lateral part of the cell, there is significant invasion of ER into the granular pole and each ZG is in fact surrounded by strands of ER. This structural evidence from living cells, in conjunction with recent functional studies demonstrating the high Ca(2+) mobility in the ER lumen, provides the framework for a coherent and internally consistent theory for cytosolic Ca(2+) signal generation in the apical secretory pole, in which the primary Ca(2+) release occurs from ER extensions in the granular pole supplied with Ca(2+) from the main store at the base of the cell by the tunnel function of the ER.

Animals↗