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Biomedical subjects

D Peter

Publications and source records attributed to D Peter.

At least 37 records · Page 2Linked to original sources
PubMed1995

Identification of residues involved in substrate recognition by a vesicular monoamine transporter.

To identify the residues involved in substrate recognition by recently cloned vesicular monoamine transporters (VMAT1 and VMAT2), we have mutagenized the conserved residues in a cytoplasmic loop between transmembrane domains two and three of VMAT2. Although studies of related bacterial antibiotic resistance proteins indicate an important functional role for this region, we found no effect of these mutations on VMAT2 activity. However, replacement of aspartate 33 in the first predicted transmembrane domain with an asparagine (D33N) eliminates transport. D33N shows normal levels of expression and normal binding at equilibrium to the potent inhibitor reserpine. However, in contrast to wild-type VMAT2, serotonin inhibits reserpine binding to D33N very poorly, indicating a specific defect in substrate recognition. Replacement of three serine residues in transmembrane domain three with alanine (Stmd3A) shows a similarly selective but even more profound defect in substrate recognition. The results suggest that by analogy to receptors and plasma membrane transporters for monoamines, the cationic amino group of the ligand interacts with an asparte in the first transmembrane domain of VMAT2 and hydroxyl groups on the catechol or indole ring interact with a group of serines in the third transmembrane domain. Importantly, D33N and Stmd3A retain coupling to the proton electrochemical gradient as measured by the delta microH(+)-induced acceleration of reserpine binding. This indicates that substrate recognition can be separated from coupling to the driving force.

Amino Acid Sequence↗
PubMed1995

The vesicular monoamine transporter 2 is present in small synaptic vesicles and preferentially localizes to large dense core vesicles in rat solitary tract nuclei.

In central neurons, monamine neurotransmitters are taken up and stored within two distinct classes of regulated secretory vesicles: small synaptic vesicles and large dense core vesicles (DCVs). Biochemical and pharmacological evidence has shown that this uptake is mediated by specific vesicular monamine transporters (VMATs). Recent molecular cloning techniques have identified the vesicular monoamine transporter (VMAT2) that is expressed in brain. This transporter determines the sites of intracellular storage of monoamines and has been implicated in both the modulation of normal monoaminergic neurotransmission and the pathogenesis of related neuropsychiatric disease. We used an antiserum against VMAT2 to examine its ultrastructural distribution in rat solitary tract nuclei, a region that contains a dense and heterogeneous population of monoaminergic neurons. We find that both immunoperoxidase and immunogold labeling for VMAT2 localize to DCVs and small synaptic vesicles in axon terminals, the trans-Golgi network of neuronal perikarya, tubulovesicles of smooth endoplasmic reticulum, and potential sites of vesicular membrane recycling. In axon terminals, immunogold labeling for VMAT2 was preferentially associated with DCVs at sites distant from typical synaptic junctions. The results provide direct evidence that a single VMAT is expressed in two morphologically distinct types of regulated secretory vesicles in central monoaminergic neurons.

Animals↗
PubMed1995

The transport of neurotransmitters into synaptic vesicles.

Using selection in the neurotoxin MPP+, we have isolated a cDNA encoding vesicular amine transport. The transporter protects against MPP+ by sequestering the toxin in vesicles, away from its primary site of action in mitochondria. Unexpectedly, two distinct but highly related genes encode vesicular amine transport in the adrenal gland and the central nervous system. The sequence of both predicts twelve transmembrane domains and weak homology to a class of bacterial antibiotic resistance proteins. The two human genes occur on different chromosomes. In addition, the two transporters show a number of differences in function, including substrate specificity and the interaction with one inhibitor and the amphetamines.

1-Methyl-4-phenylpyridinium↗
PubMed1995

Differential expression of two vesicular monoamine transporters.

Specific transport proteins package classical neurotransmitters into vesicles so that their release can be regulated by neural activity. Previous studies have suggested that a single activity mediates the vesicular transport of monoamines in the adrenal gland, brain, and other tissues such as mast cells and platelets. However, molecular cloning has recently identified two vesicular transporters for monoamines. Although the predicted proteins are closely related in sequence, they show a range of differences in their physiologic and pharmacologic properties. To clarify further the biological significance of the observed functional differences, we have generated anti-peptide antibodies to the C-termini of the two transporters and used them to determine the distribution and localization of the proteins in the rat. We have detected expression of vesicular monoamine transporter 1 (VMAT1) in adrenal chromaffin cells but not in neural cells. Interestingly, some adrenal chromaffin cells also express VMAT2 but the amount of VMAT2 relative to VMAT1 appears much lower than in the bovine adrenal gland. In contrast, sympathetic ganglion cells express only VMAT2, as do enteric neurons and enterochromaffin-like cells of the stomach. Thus, although adrenal chromaffin cells, sympathetic and enteric neurons derive from the neural crest, they express different vesicular amine transporters. In the CNS, dopamine, norepinephrine, epinephrine, 5-HT, and histamine cell groups all express VMAT2. These findings are consistent with the functional characteristics of VMAT1 and VMAT2 and help to explain several classic pharmacological observations. VMAT2-immunoreactivity is generally stronger in cell bodies, proximal dendrites and axonal processes, indicating the potential for monoamine storage at each of these sites. Surprisingly, dopaminergic interneurons in the olfactory bulb show no detectable immunoreactivity for either VMAT1 or VMAT2.

Adrenal Glands↗
PubMed1994

The chromaffin granule and synaptic vesicle amine transporters differ in substrate recognition and sensitivity to inhibitors.

Classical studies using bovine chromaffin granules have defined the physiologic and pharmacologic properties of the vesicular amine transporter that packages monoamine transmitters into intracellular vesicles for subsequent regulated release. The recent isolation of two distinct but closely related cDNA clones encoding vesicular amine transport suggests that the activity expressed in the brain (synaptic vesicle amine transporter or SVAT) may differ significantly from the previously described adrenal gland activity (chromaffin granule amine transporter or CGAT). A direct comparison of the two transporters now shows that SVAT has a higher affinity than CGAT for monoamine substrates, in particular for histamine. In addition, SVAT shows approximately 10-fold greater sensitivity to tetrabenazine than CGAT. [3H]Dihydrotetrabenazine shows no detectable binding to CGAT but does bind to SVAT, accounting for the differential sensitivity. Furthermore, methamphetamine preferentially inhibits transport by SVAT relative to CGAT, apparently by competing at the site of amine recognition rather than by disrupting the vesicular pH gradient. These previously unsuspected differences in the storage of monoamine transmitter in the central nervous system and the adrenal gland may help to account for several classic pharmacological observations.

1-Methyl-4-phenylpyridinium↗
PubMed1993

Chromosomal localization of the human vesicular amine transporter genes.

The physiologic and behavioral effects of pharmacologic agents that interfere with the transport of monoamine neurotransmitters into vesicles suggest that vesicular amine transport may contribute to human neuropsychiatric disease. To determine whether an alteration in the genes that encode vesicular amine transport contributes to the inherited component of these disorders, we have isolated a human cDNA for the brain transporter and localized the human vesicular amine transporter genes. The human brain synaptic vesicle amine transporter (SVAT) shows unexpected conservation with rat SVAT in the regions that diverge extensively between rat SVAT and the rat adrenal chromaffin granule amine transporter (CGAT). Using the cloned sequences with a panel of mouse-human hybrids and in situ hybridization for regional localization, the adrenal CGAT gene (or VAT1) maps to human chromosome 8p21.3 and the brain SVAT gene (or VAT2) maps to chromosome 10q25. Both of these sites occur very close to if not within previously described deletions that produce severe but viable phenotypes.

Adrenal Glands↗
PubMed1992

A cDNA that suppresses MPP+ toxicity encodes a vesicular amine transporter.

Classical neurotransmitters are transported into synaptic vesicles so that their release can be regulated by neural activity. In addition, the vesicular transport of biogenic amines modulates susceptibility to N-methyl-4-phenylpyridinium (MPP+), the active metabolite of the neurotoxin N-methyl-1,2,3,6-tetrahydropyridine that produces a model of Parkinson's disease. Taking advantage of selection in MPP+, we have used gene transfer followed by plasmid rescue to identify a cDNA clone that encodes a vesicular amine transporter. The sequence predicts a novel mammalian protein with 12 transmembrane domains and homology to a class of bacterial drug resistance transporters. We have detected messenger RNA transcripts for this transporter only in the adrenal gland. Monoamine cell populations in the brain stem express a distinct but highly related protein.

1-Methyl-4-phenylpyridinium↗
PubMed1992

Application of models to determination of nutrient requirements: experimental techniques employing tracers.

This paper concerns some issues involved in the design of tracer experiments for the development of whole-body compartmental models of nutrient metabolism. It focuses on tracer administration protocol, use of multiple tracers, sampling strategy, measurement types and experiment duration in a pragmatic approach to obtaining data suitable for analysis and interpretation with use of models.

Animals↗
PubMed1992

[The first interview and construction of the psychodynamic hypothesis].

The development of psychoanalitic short term therapy confirms the importance of the first interview for estimating the expectations of the patient and elaborating the first hypothesis. The therapist is immediately caught in the dynamics of pretransference and precountertransference. He depends upon the comprehension of his own emotional reactions to understand the actual affection and the nodal conflict. The psychodynamic hypothesis should be inferred from what both the patient and the therapist are saying and acting.

Adult↗
PubMed1975

Failure of balance measurements to predict actual retention of magnesium and calcium by rats as determined by direct carcass analysis.

Magnesium (Mg) and calcium (Ca) balances as well as determinations of atomic absorption analysis of atomic absorption analysis of total absolute contents of Mg and Ca fat-free dry carcasses were done on gnotobiotic male Sprague-Dawley rats, kept at 28 degrees or 6 degrees, for 69, 240, or 517 days, and fed either a commercial stock diet or a semipurified diet containing a normal or a subnormal amount of Mg. The often-reported observation of negative balances when Mg intake is subnormal has been confirmed. Total Mg and Ca accumulation in the carcass estimated from balance values was totally inconsistent with the actual content of the carcass as determined by direct analysis. The large discrepancy between the actual analysis of the carcass and the estimated content of Mg and Ca in the carcass derived from balance data demonstrates clearly that balance of trace minerals cannot reflect chronic retentions or losses of these minerals. Possible explanation of this discrepancy are dissussed.

Animals↗
PubMed1975

Effect of a chronic suboptimal intake of magnesium on magnesium and calcium content of bone and on bone strength of the rat.

Sprague-Dawley rats were kept at 28 degrees C from 21 to 517 days age and fed one of the two following diets: a semi-purified diet containing 502 p.p.m. of Mg (control) or the same diet containing only 120 p.p.m. (mg/kg) (low-Mg). The chronic suboptimal intake of Mg by rats fed the low-Mg diet did not result in overt signs of Mg deficiency even when Mg levels were greatly reduced in carcass, plasma, and tibia, but it significantly decreased bone strength. It is suggested that Mg deficiency in man could be a factor in the weakening of bone, commonly observed in old age, even when there are no visible signs of Mg deficiency. Studies of the human situation would be of interest.

Animal Feed↗