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Biomedical subjects

J F Powell

Publications and source records attributed to J F Powell.

At least 91 records · Page 5Linked to original sources

Isolation and characterization of a candidate gene for Norrie disease.

Previous analysis has refined the location of the gene for Norrie disease, a severe, X-linked, recessive neurodevelopmental disorder, to a yeast artificial chromosome subfragment of 160 kilobases (kb). This fragment was used to screen cDNA libraries from human fetal and adult retina. As a result, we have identified an evolutionarily conserved cDNA, which is expressed in fetal and adult brain and encodes a predicted protein of 133 amino acids. The cDNA detects genomic sequences which span a maximum of 50 kb, and which are partly deleted in several typical Norrie disease patients. An EcoRI polymorphism with a calculated heterozygosity value of 43% was observed. The locus identified is a strong candidate for the Norrie disease gene.

Amino Acid Sequence↗

Peopling of the Americas: paleobiological evidence.

A subjective and bivariate analysis of 8500-10,000-year-old human fossil remains from North America substantiates that the fossils' closest affinities are with Asian populations. Within North American prehistoric Indian populations, increasing brachycephalization and the possible development of a larger, broader face are two structural trends that can be identified. In those respects where Paleo-Indian specimens differ from modern northern Asians and North American Indians, they tend to resemble southern Asian and European populations. These assessments generally support the inference that populations entered the New World relatively recently but before the modern northern Asian and North American features were fully developed. Based on the data examined, no date can be specified for time of entrance of the first populations, nor can the number of founding populations be discerned.

Anthropology, Physical↗

Structure of the human gene for monoamine oxidase type A.

Monoamine oxidases, type A and type B, are principal enzymes for the degradation of biogenic amines, including catecholamines and serotonin. These isozymes have been implicated in neuropsychiatric disorders. Previously, cDNA clones for both MAO-A and MAO-B have been sequenced and the genes encoding them have been localized to human chromosome Xp11.23-Xp11.4. In this work, we isolated human genomic clones spanning almost all the MAOA gene from cosmid and phage libraries using a cDNA probe for MAO-A. Restriction mapping and sequencing show that the human MAOA gene extends over 70 kb and is composed of 15 exons. The exon structure of human MAOA is similar to that described by others for human MAOB. Exon 12 (bearing the codon for cysteine, which carries the covalently bound FAD cofactor) and exon 13 are highly conserved between human MAOA and MAOB genes (92% at the amino acid level). Earlier work revealed two species of MAO-A mRNA, 2.1 kb and 4.5-5.5 kb. We now report on further cDNA isolation and sequencing, which demonstrates that the longer message has an extension of 2.2 kb in the 3' noncoding region. This extended region is contained entirely within exon 15. The two messages therefore appear to be generated by the use of two alternative polyadenylation sites. Results from the present work should facilitate the mutational analysis of functional domains of MAO-A and MAO-B. Knowledge of the gene structure will also help in evaluating the role of genetic variations in MAO-A in human disease through the use of genomic DNA, which is more accessible than the RNA, as a template for PCR-amplification and sequencing.

Amino Acid Sequence↗

Atlanto-axial subluxation in Down's syndrome.

A case of atlanto-axial subluxation in a patient with Down's syndrome is described. The gradual deterioration in the patient's locomotor ability caused a delay in diagnosis. Patterns of presentation of this condition are discussed.

Adolescent↗

The primary structure of bovine monoamine oxidase type A. Comparison with peptide sequences of bovine monoamine oxidase type B and other flavoenzymes.

We have isolated cDNA clones believed to encompass the full-length coding sequences for a subunit of bovine monoamine oxidase type A (MAO-A). The clones code for an apoprotein of 527 amino acid residues corresponding to a molecular mass of 59,806 Da. The inferred protein sequences show an overall similarity of 68% with partial amino acid sequences of bovine type B MAO (about 41% of the total sequence), as well as a greater similarity (greater than 90%) with some regions including that for the published sequence of the flavin-binding region. Sequence comparisons indicate that these two forms of MAO are encoded by distinct genes. Comparison of this sequence with other flavoenzymes showed similarity with regions associated with non-covalent flavin-binding sites. Analysis of mRNAs coding for MAO enzymes showed a heterogeneity of transcripts consistent with several different forms of monoamine oxidase.

Amino Acid Sequence↗

Localization of human monoamine oxidase-A gene to Xp11.23-11.4 by in situ hybridization: implications for Norrie disease.

A cDNA for the neurotransmitter-degrading enzyme monoamine oxidase-A (MAO-A) has been assigned by in situ hybridization to the human X chromosome and subregionally localized to Xp11.23-11.4. As restriction fragments detected by this probe are deleted in some patients with Norrie disease, this assignment provides confirmation of the localization of the disease.

Chromosome Banding↗

Monoamine oxidase deficiency in males with an X chromosome deletion.

Mapping of the human MAOA gene to chromosomal region Xp21-p11 prompted our study of two affected males in a family previously reported to have Norrie disease resulting from a submicroscopic deletion in this chromosomal region. In this investigation we demonstrate in these cousins deletion of the MAOA gene, undetectable levels of MAO-A and MAO-B activities in their fibroblasts and platelets, respectively, loss of mRNA for MAO-A in fibroblasts, and substantial alterations in urinary catecholamine metabolites. The present study documents that a marked deficiency of MAO activity is compatible with life and that genes for MAO-A and MAO-B are near each other in this Xp chromosomal region. Some of the clinical features of these MAO deletion patients may help to identify X-linked MAO deficiency diseases in humans.

Adolescent↗

Insulin-like growth factor I gene expression in the rat ovary is confined to the granulosa cells of developing follicles.

The in vivo intraovarian synthesis of insulin-like growth factor-I (IGF-I) has been studied in the rat by Northern blot, dot blot hybridization, and in situ hybridization histochemistry. Ovarian IGF-I mRNA transcript sizes (7.0 kilobases (kb), 1.6 kb, and a group from 0.4-0.9 kb) were similar to those in liver and other tissues. The proportion of ovarian IGF-I to tubulin messenger RNA (mRNA) was increased to 176% of control values by treatment with diethylstilbestrol, while the ratio in liver was decreased to 64.4%. In situ hybridization identifies the major in vivo site of IGF-I synthesis in the ovary as the granulosa cells of developing follicles. IGF-I mRNA was present in the granulosa cells of developing preantral and antral follicles, but was not seen in atretic follicles or corpora lutea. In preovulatory follicles high levels of IGF-I mRNA were confined to the antral cell layers and to the cells of the cumulus oophorus. High levels of tubulin gene expression within follicles were seen in a similar distribution to that for IGF-I but 80-90% of corpora lutea also strongly expressed the tubulin gene. Interstitial cells, including thecal cells, express the tubulin gene at low levels but do not express the IGF-I gene. The distribution of IGF-I mRNA is the same as that previously observed for mitotically active granulosa cells, and therefore offers strong support for the view that IGF-I in the ovary acts by an autocrine-paracrine mechanism to promote granulosa cell replication.

Animals↗

Human monoamine oxidase gene (MAOA): chromosome position (Xp21-p11) and DNA polymorphism.

An essentially full-length cDNA clone for the human enzyme monoamine oxidase type A (MAO-A) has been used to determine the chromosomal location of a gene encoding it. This enzyme is important in the degradative metabolism of biogenic amines throughout the body and is located in the outer mitochondrial membrane of many cell types. Southern blot analysis of PstI-digested human DNA revealed multiple fragments that hybridized to this probe. Using rodent-human somatic cell hybrids containing all or part of the human X chromosome, we have mapped these fragments to the region Xp21-p11. A restriction fragment length polymorphism (RFLP) for this MAOA gene was identified and used to evaluate linkage distances between this locus and several other loci on Xp. The MAOA locus lies between DXS14 and OTC, about 29 cM from the former.

Alleles↗

Structural features of human monoamine oxidase A elucidated from cDNA and peptide sequences.

Monoamine oxidase (MAO), an important enzyme for the degradation of amine neurotransmitters, has been implicated in neuropsychiatric illness. The amino acid sequence for one form of the enzyme, MAO-A, has been deduced from human cDNA clones and verified against proteolytic peptides. The covalent binding site for the flavin adenine dinucleotide (FAD) cofactor is near the C-terminal region. The presence of features characteristic of the ADP-binding fold suggests that the N-terminal region is also involved in the binding of FAD. These cDNAs should facilitate the study of the structure, function, and intracellular targeting of MAO, as well as the analysis of its expression in normal and pathological states.

Adenosine Diphosphate↗

An antibody purified with a lambda GT11 fusion protein precipitates enkephalinase activity.

An antiserum was raised against the neutral endopeptidase "enkephalinase" in guinea pig and used to probe a rabbit kidney cDNA expression library. A positive clone has been isolated and sequenced. The identity of the corresponding fusion protein was ascertained by its ability to select, from the crude antiserum, antibodies which specifically immunoprecipitate neutral endopeptidase enzymatic activity. This approach eliminates the uncertainty inherent to clone identification obtained from oligonucleotide probe derived from a partial sequence of the protein.

Amino Acid Sequence↗

The effects of acid exposure on the ion regulation and seawater adaptation of coho salmon (Oncorhynchus kisutch) parrs and smolts.

Smolts exhibited decreases in plasma Na+ levels after 7 days and lower Na+, K+-ATPase activities 14 days after acid exposure. Parrs exhibited decreased plasma Na+ after 24 hr acid exposure. Plasma Na+ increased and Na+, K+-ATPase decreased in smolts after transfer to seawater. Parrs exhibited increased plasma Na+ as well as Na+, K+-ATPase activity immediately after transfer to seawater. It was concluded that acid exposure prior to entry into seawater was detrimental to coho salmon with regard to the length of acid exposure and stage of development. A possible mechanism by which fish die from acid stress is inhibition of gill Na+, K+-ATPase concomitant with decreases in plasma Na+ levels.

Acids↗

GABA axons in synaptic contact with dopamine neurons in the substantia nigra: double immunocytochemistry with biotin-peroxidase and protein A-colloidal gold.

Two different antigens in the same ultrathin section of brain tissue can be revealed by 'double immunocytochemistry' in which one antigen is detected by horseradish peroxidase and the other by silver intensification of colloidal gold (SIG) adsorbed to Protein A. By means of this procedure it has been possible to show that GABAergic axon terminals (containing glutamate decarboxylase) are in synaptic contact with the cell bodies and dendrites of dopaminergic neurons (containing tyrosine hydroxylase) in the substantia nigra of the rat. Thus, several of the physiological and pharmacological effects of GABA and GABAergic drugs in this part of the brain are likely to be mediated by a direct action via postsynaptic GABAergic receptors located on dopaminergic nigrostriatal neurons.

Animals↗

Glutamate decarboxylase-immunoreactive structures in the rat neostriatum: a correlated light and electron microscopic study including a combination of Golgi impregnation with immunocytochemistry.

An antibody to glutamate decarboxylase has been used in a light and electron microscopic study of the neostriatum of rats that had received intracerebral injections of colchicine. In the light microscope, neuronal perikarya and small punctate structures that displayed immunoreactivity were found. The perikarya could be divided into two classes based on their sizes: small-to-medium-sized and large. Proximal dendrites, axon initial segments, and axon collaterals were occasionally stained. When the nuclei of the neurons were visible, they possessed indentations. The immunoreactive punctate structures were spread evenly throughout the neostriatum but occasionally were associated with immunoreactive and nonimmunoreactive perikarya. When the same sections were examined in the electron microscope, the small-to-medium-sized immunoreactive perikarya were found to be similar in morphology and synaptic input to a class of Golgi-impregnated neuron that has been previously shown to accumulate locally administered, radiolabelled gamma-aminobutyric acid. Neurons with the ultrastructural characteristics of typical striatonigral neurons did not display immunoreactivity. As neurons in this pathway probably contain gamma-aminobutyric acid, it is possible that our procedure or our antibody does not stain all gamma-aminobutyric-acid-containing structures in the neostriatum. A total of 404 immunoreactive punctate structures were examined by correlated light and electron microscopy or by electron microscopy alone. They were identified as immunoreactive axonal boutons and each of them, when examined in serial sections, displayed typical synaptic specialisations. Membrane specialisations were always of the symmetrical type. At least five distinct targets of the immunoreactive terminals were identified: neurons that were themselves immunoreactive for glutamate decarboxylase; the immunoreactive terminals made synaptic contact with all parts of the neurons examined, i.e., perikarya, proximal dendrites, and axon initial segments. Neurons identified by Golgi impregnation of the same sections as medium-sized and densely spiny; the immunoreactive terminals made contact predominantly with the perikarya and dendritic shafts. Large neurons found only in the ventral caudate-putamen, whose somata and dendrites were ensheathed in immunoreactive terminals. Medium-sized nonimmunoreactive perikarya that possessed nuclear indentations. Large nonimmunoreactive perikarya that had the typical structural features of striatal cholinergic neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗