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D Goldman

Publications and source records attributed to D Goldman.

At least 271 records · Page 15Linked to original sources
PubMed1989

Cloning and comparative mapping of a human class III (chi) alcohol dehydrogenase cDNA.

A cDNA encoding human class III (chi ADH5) alcohol dehydrogenase was isolated, sequenced and used to comparatively map this unusual ADH. In their coding sequences, the three major ADH classes were approximately equisimilar, class II and III ADHs sharing the highest sequence identity (67%). A class III-like ADH was mapped to mouse chromosome 3, site of the ADH gene complex, and synteny of ADH5 with four other ADH loci on human chromosome 4 was confirmed. The nearly full-length 1613 nucleotide cDNA contained 433 nucleotides of 3' nontranslated sequence and two possible initiation sites for translation. A protein of 374 amino acid residues could be synthesized using the potential initiation codon at nucleotide 59. However, use of the likely initiation codon at nucleotide 5 would produce a protein of 392 residues with 19 additional N-terminal residues as compared to the known protein sequence. The derived protein sequence also differs at residue 166, where Tyr is found. This difference, due to a single base substitution, could result from cloning artifact, polymorphism, or two expressed class III ADH genes.

Alcohol Oxidoreductases↗
PubMed1989

Coordinate regulation of RNAs encoding two isoforms of the rat muscle nicotinic acetylcholine receptor beta-subunit.

The nicotinic acetylcholine receptor (nAchR) mediates communication between nerve and skeletal muscle. The properties, levels and distribution of these receptors change during development of the neuromuscular junction. These changes may be due, in part, to expression of different gene products. We are using nuclease protection experiments and cDNA cloning to identify the RNA transcripts that encode nAchRs in rat muscle. This analysis has identified two beta-subunit mRNAs. Complementary DNAs corresponding to these two RNAs have been isolated from a rat skeletal muscle cDNA library. Based on nucleotide sequence analysis, these RNAs differ by 9 bases in their 5' coding sequence. The levels of both mRNAs change similarly during muscle development and upon denervation of adult skeletal muscle. These two beta-subunit-RNAs probably result from the use of different exon/intron splice sites in the beta-subunit gene.

Animals↗
PubMed1989

Distribution and possible metabolic role of class III alcohol dehydrogenase in the human brain.

In human brain, the sole alcohol dehydrogenase (ADH) present in significant quantity has been shown to be Class III (chi) ADH and this ADH is ineffective in generating potentially toxic and reactive acetaldehyde from ethanol at concentrations attainable in living brain tissue. We have extended this finding to show that Class I ADH potentially present is undetectable even when concentrated several hundred-fold. Purified Class III ADH from human brain is identical in its pattern of tryptic peptides and in other properties to Class III ADH from human liver. Immunohistochemical staining and western immunoblots using polyclonal antibodies reveal that Class III ADH is widely distributed in brian and most concentrated in the subependymal layer and perivascular areas. Class III ADH closely resembles omega-hydroxyfatty acid dehydrogenase and a possible role for the brain enzyme is in the oxidation of long chain fatty alcohols and omega-hydroxyfatty acids.

Alcohol Oxidoreductases↗
PubMed1989

Spatial and temporal expression of acetylcholine receptor RNAs in innervated and denervated rat soleus muscle.

In adult vertebrate skeletal muscle acetylcholine receptors are localized to the neuromuscular junction. Upon denervation, this distribution changes, with new receptors appearing in extrajunctional regions of the muscle fiber. The location of acetylcholine receptors in innervated or denervated muscle may result, in part, from the distribution of their RNAs. This was tested by assaying for receptor RNAs in junctional and extrajunctional regions of innervated and denervated rat soleus muscle using in situ hybridization and RNAase protection assays. These experiments showed alpha, beta, and delta subunit RNAs concentrated beneath the endplates of innervated muscle fibers. Following denervation, there was an unequal distribution of receptor RNAs along the muscle fiber, with highest levels occurring in extrajunctional regions near the endplate. These data are consistent with a nonuniform pattern of gene expression in adult skeletal muscle fibers.

Animals↗
PubMed1989

A shortened synthesis of 4-(3-aminopropyl) pyrazole, an affinity ligand for alcohol dehydrogenase purification.

The most efficient, specific and rapid procedures for alcohol dehydrogenase (ADH) purification utilize immobilized 4-(3-aminopropyl) pyrazole to which pyrazole sensitive ADHs, i.e. class I isozymes, bind. Because of the length of the reported synthesis of this affinity resin, we synthesized the 4-(3-aminopropyl) pyrazole ligand by a new method in two steps from commercially available nicotinaldehyde. The ligand synthesized by this simplified procedure was directly coupled to the chain-extended support, Activated CH-Sepharose 4B, to yield the same ligand-spacer combination as reported by L.G. Lange and B.L. Vallee (Biochem. 15: 4681-4686, 1976). Human and hamster class I ADHs purified using this resin were homogeneous by SDS-PAGE followed by silver staining. Specific activity and recovery of human class I ADH were comparable to those previously reported.

Alcohol Dehydrogenase↗
PubMed1989

Identification of a novel nicotinic acetylcholine receptor structural subunit expressed in goldfish retina.

A new non-alpha (n alpha) member of the nicotinic acetylcholine receptor (nAChR) gene family designated GFn alpha-2 has been identified in goldfish retina by cDNA cloning. This cDNA clone encodes a protein with structural features common to all nAChR subunits sequenced to date; however, unlike all known alpha-subunits of the receptor, it lacks the cysteine residues believed to be involved in acetylcholine binding. Northern blot analysis shows multiple transcripts hybridizing to the GFn alpha-2 cDNA in goldfish retina but undetectable levels of hybridizable RNA in brain, muscle, or liver. S1 nuclease protection experiments indicate that multiple mRNAs are expressed in retina with regions identical or very similar to the GFn alpha-2 sequence. In situ hybridization shows that the gene encoding GFn alpha-2 is expressed predominantly in the ganglion cell layer of the retina.

Amino Acid Sequence↗
PubMed1989

Genes encoding neuronal nicotinic acetylcholine receptors.

Four genes (alpha 2, alpha 3, alpha 4, and beta 2), which encode proteins homologous to the Torpedo electric organ and vertebrate muscle nicotinic acetylcholine receptors, have been identified by cloning rat brain cDNAs. Injection of transcripts derived from these cDNAs into Xenopus laevis oocytes results in the formation of three nicotinic acetylcholine receptors. Two of these receptors, alpha 3/beta 2 and alpha 4/beta 2, have the characteristics of ganglionic nicotinic receptors. The third (alpha 2/beta 2) exhibits a previously undescribed pharmacology and thus represents a novel subtype that may be expressed in the brain. The wide distribution of alpha 2, alpha 3, alpha 4, and beta 2 transcripts in the brain indicates that neuronal nicotinic acetylcholine receptors are a major neurotransmitter receptor system.

Animals↗
PubMed1988

Purification and characterization of three forms of class III alcohol dehydrogenase.

We have resolved and characterized three forms of human and rat hepatic class III alcohol dehydrogenase. Separations were carried out in narrow immobilized pH gradients. Both in humans and rats the three forms were visualized by enzyme staining with cinnamol, but not with ethanol. They were insensitive to the inhibitory effect of pyrazole. The isoelectric points were approximately from 6.3-6.4, from 5.9-6.0 and 5.6. Each electroeluted enzyme extract, purified further by analytical isoelectric focusing over the pH range from 5-6 or 6-7, revealed a single band by enzyme and silver staining and by Western blotting followed by avidin-biotin staining. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate of each extract revealed a single molecular mass species corresponding to class III alcohol dehydrogenase (ADH). All forms of class III alcohol dehydrogenase were recognized by antisera raised against total class III ADH.

Alcohol Dehydrogenase↗
PubMed1988

Molecular markers for linkage of genetic loci contributing to alcoholism.

Specific locus and random locus linkage approaches to identify markers for genes whose allelic variants predispose to alcoholism or for genes controlling relevant physiological and behavioral phenotypes are discussed. Sib-pair analysis is superior for the direct analysis of complex genetic traits such as alcoholism, but classic family analysis will be useful for transmission and linkage analysis for marker traits whose genetics is less complex. In mice, a large number of inbred strains, recombinant inbred and congenic strains, and specifically selected outbred strains are available. In the human, an intriguing linkage result has emerged between a brain protein variant and alcoholism accompanied by suicide. In the mouse, preliminary linkages have been established to loci controlling ethanol preference and also activation after ethanol. Large panels of random DNA and protein genetic probes and of probes for specific loci will in the future increase the probability of establishing linkage in both species.

Alcoholism↗
PubMed1988

Acetylcholine receptor alpha-, beta-, gamma-, and delta-subunit mRNA levels are regulated by muscle activity.

Denervation of adult skeletal muscle results in increased sensitivity to acetylcholine in extrajunctional regions of the muscle fiber. This increase in acetylcholine sensitivity is accompanied by a large increase in the level of mRNAs coding for the alpha-, beta-, gamma-, and delta-subunits of the acetylcholine receptor. To determine whether muscle activity is sufficient to regulate expression of extrajunctional acetylcholine receptor mRNA levels, denervated muscles were stimulated with extracellular electrodes. Direct stimulation of denervated muscle suppresses both the increase in extrajunctional acetylcholine sensitivity and the expression of mRNA encoding the alpha-, beta-, gamma-, and delta-subunits of the acetylcholine receptor. These results show that muscle activity regulates the level of extrajunctional acetylcholine receptors by regulating the expression of their mRNAs.

Animals↗
PubMed1988

Review of the molecular biology of the human alcohol dehydrogenase genes and gene products.

Using protein and enzymatic methods, a major role in ethanol metabolism was assigned to the alcohol dehydrogenase (ADH) enzymes. Three major classes of ADHs were described on the basis of structure and function, including timing and location of expression. Polymorphic variants, including a common functional variant, were identified. Molecular cloning allowed the demonstration of a high degree of sequence homology between the three class I ADH genes and enabled the definition of ADH variants at the DNA sequence level. The existence of an ADH gene cluster on chromosome 4 and the shared evolutionary roots of these genes suggests that the continued integration of studies of the different ADH genes will yield further insights into alcohol metabolism in humans.

Alcohol Dehydrogenase↗
PubMed1987

Mapping of a putative genetic locus determining ethanol intake in the mouse.

In the mouse, there is evidence that a single genetic locus is a major determinant of differences in ethanol intake between some preferring and non-preferring inbred strains. In this report, we present evidence from two independent experiments indicating that this locus maps to chromosome 1 and that its expressed product is the abundant protein LTW-4 (a 28 kDa, pI 5.6 protein expressed in brain, liver and kidney). The genetic association was found using a panel of 14 polypeptides of mouse brain which were visualized by two-dimensional electrophoresis and which exhibit genetic variation in isoelectric point. Fifteen BXD recombinant inbred strains and the two parental strains were typed for these loci and also tested for ethanol acceptance. Strains exhibiting the basic allele showed significantly higher ethanol acceptance. When 19 distantly related inbred mouse strains were tested for ethanol acceptance and typed for LTW-4, it was again found that strains exhibiting the basic allele showed significantly higher ethanol acceptance.

Alcohol Drinking↗
PubMed1987

Muscle acetylcholine receptor biosynthesis. Regulation by transcript availability.

The expression of the muscle nicotinic acetylcholine receptor (AChR) on the cell surface entails a complex biosynthetic pathway, involving the expression and assembly of four subunits. The amount of AChR on the cell surface changes throughout muscle development and upon muscle denervation. We have examined the regulatory role of transcript levels on surface AChR expression by RNA blot analysis. During myogenesis of the fusing mouse muscle cell line C2, which expresses an embryonic type of receptor, changes in the rate of appearance of cell surface AChR have been assayed by 125I-alpha-bungarotoxin binding. The maximal increase in the rate of appearance of cell surface AChR occurs upon cell fusion, closely following the maximal increase in transcript levels for the alpha-, beta-, gamma-, and delta-AChR subunits. AChR alpha-, beta-, gamma-, and delta-subunit gene transcript levels have also been examined in innervated and denervated rat and mouse muscle. Muscle denervation results in an increase of transcripts for all four subunits. However, the amount of beta-subunit transcript in innervated rat skeletal muscle is high relative to the other subunit transcripts, and increases less than the other subunit transcripts upon denervation. Our results indicate that, during myogenesis and upon denervation, the availability of AChR subunit transcripts for translation plays a regulatory role in surface AChR appearance.

Animals↗
PubMed1987

Members of a nicotinic acetylcholine receptor gene family are expressed in different regions of the mammalian central nervous system.

Nicotinic acetylcholine receptors found in the peripheral and central nervous system differ from those found at the neuromuscular junction. Recently we isolated a cDNA clone encoding the alpha subunit of a neuronal acetylcholine receptor expressed in both the peripheral and central nervous system. In this paper we report the isolation of a cDNA encoding the alpha subunit of a second acetylcholine receptor expressed in the central nervous system. Thus it is clear that there is a family of genes coding for proteins with sequence and structural homology to the alpha subunit of the muscle nicotinic acetylcholine receptor. Members of this gene family are expressed in different regions of the central nervous system and, presumably, code for subtypes of the nicotinic acetylcholine receptor.

Amino Acid Sequence↗
PubMed1987

Further delineation of the McKusick-Kaufman hydrometrocolpos-polydactyly syndrome.

Six cases of the McKusick-Kaufman syndrome (MKS), including two cases that were diagnosed prenatally, were studied. Review of the 54 previously described cases indicates that postaxial polydactyly and hydrometrocolpos in female patients are the hallmark features of this entity. Other manifestations, such as malformations of gastrointestinal, cardiovascular, and ophthalmic structures, occur less consistently. Affected children require careful medical follow-up. Recurrence of hydrometrocolpos following surgical repair may lead to serious sequelae, such as chronic renal failure. We believe that MKS is a distinct panethnic genetic entity, inherited in an autosomal recessive fashion, and that the diagnosis should be made only in female patients with hydrometrocolpos and polydactyly or in male patients with polydactyly who have an affected female relative.

Female↗
PubMed1987

Protein polymorphisms detected by two-dimensional electrophoresis: an analysis of overall informativeness of a panel of linkage markers.

Forty-two independent polymorphic loci are detectable by two-dimensional electrophoresis (2DE) of four peripherally accessible human tissues. Fifteen have been chromosomally mapped and, taken together, these constitute a useful panel of markers for genetic linkage studies in humans. An analysis of the overall informativeness for linkage of this panel of markers is presented, taking into account the effect of varying the number of families or matings studies. Use of 2DE polymorphic markers for linkage of genetically determined behaviour traits in humans and mice is reviewed.

Blood Proteins↗
PubMed1987

A molecular phylogeny of the hominoid primates as indicated by two-dimensional protein electrophoresis.

A molecular phylogeny for the hominoid primates was constructed by using genetic distances from a survey of 383 radiolabeled fibroblast polypeptides resolved by two-dimensional electrophoresis (2DE). An internally consistent matrix of Nei genetic distances was generated on the basis of variants in electrophoretic position. The derived phylogenetic tree indicated a branching sequence, from oldest to most recent, of cercopithecoids (Macaca fascicularis), gibbon-siamang, orangutan, gorilla, and human-chimpanzee. A cladistic analysis of 240 electrophoretic characters that varied between ape species produced an identical tree. Genetic distance measures obtained by 2DE are largely consistent with those generated by other molecular procedures. In addition, the 2DE data set appears to resolve the human-chimpanzee-gorilla trichotomy in favor of a more recent association of chimpanzees and humans.

Animals↗