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M V Bell

Publications and source records attributed to M V Bell.

At least 19 recordsLinked to original sources
PubMed2003

Pyloric ceca are significant sites of newly synthesized 22:6n-3 in rainbow trout (Oncorhynchus mykiss).

In this pulse-chase study, rainbow trout fed a diet containing deuterated (D5) (17,17,18,18,18)-18:3n-3 ethyl ester accumulated D5-22:6n-3 in pyloric ceca to a greater extent than in liver 2 d post-dose. The ratio of newly synthesized D5-22:6n-3 in ceca to that in liver 2 d after feeding D5-18:3n-3 was 4.7 +/- 1.2 when expressed as per mg tissue and 5.2 +/- 2.4 when expressed as per mg protein. The amount of D5-22:6n-3 in ceca then declined whereas that in liver and blood increased, with the ratio of ceca to liver falling to 1.7 and 1.4, respectively, by day 5 and approaching unity by day 9. A crude cecal mucosa fraction contained 123 +/- 50 ng D5-22:6n-3/mg protein/mg D5-18:3n-3 eaten 2 d after feeding the tracer, compared with 35 +/- 21 ng D5-22:6n-3/mg protein/mg D5-18:3n-3 eaten in liver. Three days later the amount in cecal mucosa had fallen by one-third and that in liver had increased threefold. Most of the D5-18:3n-3 was catabolized very rapidly. The ratio of D5-18:3n-3 to 21:4n-6 (a relatively inert FA marker) in the diet was 4.0, but this fell to 0.30 in ceca and ca. 0.8 in liver, blood, and whole carcass one day after feeding. These results indicate that ceca are active in the synthesis of 22:6n-3 and the oxidation of 18:3n-3.

5,8,11,14-Eicosatetraynoic Acid↗
PubMed2001

Biosynthesis of eicosapentaenoic acid in the sea urchin Psammechinus miliaris.

The sea urchin Psammechinus miliaris (Gmelin) (Echinodermata: Echinoidea) was shown by using a deuterated tracer (D5-18:3n-3) and quantitation by negative chemical ionization gas chromatography-mass spectrometry to convert 18:3n-3 to 20:5n-3. The rate of conversion was very slow, corresponding to 0.09 microg/g tissue/mg 18:3n-3 eaten over 14 d. Deuterated arachidonic acid (D8-20:4n-6) was also included in the diet to give a measure of the relative amounts of diet eaten by the different animals. The recovery of this fatty acid in tissue lipids was 33.7% compared with only 0.95% recovery of D5-18:3n-3 and its anabolites, indicating that the majority of the D5-tracer was catabolized. Considerable elongation of D5-18:3n-3 into 20:3n-3 and a trace of 22:3n-3 was found, and these were accompanied by minor amounts of the intermediates 18:4n-3 and 20:4n-3. No deuterated 22:6n-3 was found.

Animals↗
PubMed2001

Biosynthesis and tissue deposition of docosahexaenoic acid (22:6n-3) in rainbow trout (Oncorhynchus mykiss).

Rainbow trout (Oncorhynchus mykiss) weighing ca. 5 g and previously acclimated for 8 wk on a diet comprising vegetable oil (11%), fish meal (5%), and casein (48%) as the major constituents were fed a pulse of diet containing deuterated (D5) (17,17,18,18,18)-18:3n-3 ethyl ester. The synthesis and tissue distribution of D5-22:6n-3 was determined 3, 7, 14, 24, and 35 d after the pulse. The whole-body accumulation of D5-22:6n-3 was linear over the first 7 d, corresponding to a rate of 0.54 +/- 0.12 microg D5-22:6n-3/g fish/mg D5-18:3n-3 eaten/d. Maximal accretion of D5-22:6n-3 was 4.3 +/- 1.2 microg/g fish/mg of D5-18:3n-3 eaten after 14 d. The amount of D5-22:6n-3 peaked in liver at day 7, in brain and eyes at day 24, and plateaued after day 14 in visceral and eye socket adipose tissue and in the whole fish. The majority of D5-22:6n-3 was found in the carcass (remaining tissues minus the above tissues analyzed separately) at all times. On a per milligram lipid basis, liver and eyes had the highest concentration of D5-22:6n-3. The experimental diet also contained 21:4n-6 ethyl ester as a marker to estimate the amount of food eaten by individual fish. From such estimates it was calculated that the great majority of the D5-tracer was catabolized, with the combined recovery of D5-18:3n-3 plus D5-22:6n-3 being 2.6%. The recovery of 21:4n-6 was 57.6%. The concentration of 22:6n-3 in the fish decreased during the 13-wk period, and the amount of 22:6n-3 synthesized from 18:3n-3 was only about 5% of that obtained directly from the fish meal in the diet.

Adipose Tissue↗
PubMed2000

Nociceptin/orphanin FQ (N/OFQ) induces a quasi-morphine abstinence syndrome in the rat.

Nociceptin/orphanin FQ (N/OFQ) is a neuropeptide that exerts antiopiate effects under some circumstances, and there is evidence that it contributes to opiate tolerance. This raises the question, might N/OFQ also contribute to opiate dependence and abstinence? Twenty-two male Sprague-Dawley rats were cannulated in the third ventricle and challenged 7 days later by third ventricle injection of 50, 200 or 1,000 ng N/OFQ or saline alone. Each rat was observed under "blind" conditions for 30 min beginning 15 min after onset of the third ventricle injection. There was a significant positive linear trend of signs as a function of N/OFQ dose. Subjects receiving saline had 18.0+/-2.0 (mean+/-SEM) overall abstinence-like signs, whereas subjects receiving 50, 200 or 1000 ng N/OFQ had 35.2+/-3.6, 49.8+/-2.6 and 63.5+/-9.7 signs, respectively. In 16 additional rats, abstinence-like signs induced by 1000 ng N/OFQ were significantly attenuated by low SC doses of morphine or clonidine. These results raise the possibility that N/OFQ might contribute to opiate dependence and subsequent abstinence syndrome. On the other hand, N/OFQ over a wide dose range induced abstinence signs with similar potency in morphine dependent and non-dependent rats.

Animals↗
PubMed1999

v-Myb can transform and regulate the differentiation of melanocyte precursors.

The activity of the c-Myb transcription factor is essential for the development of definitive multi- and uni-lineage progenitors of the haemopoietic system. Reflecting this requirement, c-Myb has been oncogenically activated by transduction in the E26 avian retrovirus which elicits an acute leukaemia by transforming haemopoietic progenitors. Here, we report the novel finding that Myb in cooperation with EGF receptor signalling can be used to generate clonally expanded populations of transformed cells which have the phenotype of melanocyte precursors. Through the use of a conditional temperature sensitive mutant of Myb, we show that in the transformed cells Myb regulates commitment to melanocyte differentiation and possibly proliferation. These results add to our understanding of the roles of c-Myb beyond the haemopoietic system and to our knowledge and means of investigating the importance of transcription factors in the melanocyte lineage.

Animals↗
PubMed1999

Low C18 to C20 fatty acid elongase activity and limited conversion of stearidonic acid, 18:4(n-3), to eicosapentaenoic acid, 20:5(n-3), in a cell line from the turbot, Scophthalmus maximus.

The TF cell line, derived from a top predatory, carnivorous marine teleost, the turbot (Scophthalmus maximus), is known to have a limited conversion of C18 to C20 polyunsaturated fatty acids (PUFA). To illuminate the underlying processes, we studied the conversions of stearidonic acid, 18:4(n-3), and its elongation product, 20:4(n-3), in TF cells and also in a cell line, AS, derived from Atlantic salmon (Salmo salar), by adding unlabelled (25 microM), U-14C (1 microM) or deuterated (d5; 25 microM) fatty acids. Stearidonic acid, 18:4(n-3), was metabolised to 20:5(n-3) in both cells lines, but more so in AS than in TF cells. Delta5 desaturation was more active in TF cells than in AS cells, whereas C18 to C20 elongation was much reduced in TF as compared to AS cells. Only small amounts of docosahexaenoic acid (22:6(n-3)) were produced by both cell lines, although there was significant production of 22:5(n-3) in both cultures, especially when 20:4(n-3) was supplemented. We conclude that limited elongation of C18 to C20 fatty acids rather than limited fatty acyl Delta5 desaturation accounts for the limited rate of conversion of 18:3(n-3) to 20:5(n-3) in the turbot cell line, as compared to the Atlantic salmon cell line. The results can account for the known differences in conversions of C18 to C20 PUFA by the turbot and the Atlantic salmon in vivo.

Acetyltransferases↗
PubMed1998

Stable-carbon-isotope composition of Fatty acids in hydrothermal vent mussels containing methanotrophic and thiotrophic bacterial endosymbionts.

Fatty acid biomarker analysis coupled with gas chromatography-isotope ratio mass spectrometry was used to confirm the presence of methanotrophic and thiotrophic bacterial endosymbionts in the tissues of a hydrothermal vent mussel (Bathymodiolus sp.), collected from the Menez Gwen vent field on the mid-Atlantic ridge. Monounsaturated (n-8) fatty acids, which are diagnostic of methanotrophic bacteria, were detected in all three types of tissues examined (gill, posterior adductor, and mantle), although levels were highest in gill tissues where the bacteria were found. Stable-carbon-isotope compositions (delta-C per mille relative to that of Peedee belemnite) of fatty acids for all three tissues ranged from -24.9 to -34.9 per thousand, which encompasses the range predicted for both thiotroph- and methanotroph-based nutrition. The data suggest that these thio- and methanotrophic bacterial endosymbionts are equally important in the nutrition of the vent mussel at this particular vent site.

Journal Article↗
PubMed1998

Influence of intron length on alternative splicing of CD44.

Although the splicing of transcripts from most eukaryotic genes occurs in a constitutive fashion, some genes can undergo a process of alternative splicing. This is a genetically economical process which allows a single gene to give rise to several protein isoforms by the inclusion or exclusion of sequences into or from the mature mRNA. CD44 provides a unique example; more than 1,000 possible isoforms can be produced by the inclusion or exclusion of a central tandem array of 10 alternatively spliced exons. Certain alternatively spliced exons have been ascribed specific functions; however, independent regulation of the inclusion or skipping of each of these exons would clearly demand an extremely complex regulatory network. Such a network would involve the interaction of many exon-specific trans-acting factors with the pre-mRNA. Therefore, to assess whether the exons are indeed independently regulated, we have examined the alternative exon content of a large number of individual CD44 cDNA isoforms. This analysis shows that the downstream alternatively spliced exons are favored over those lying upstream and that alternative exons are often included in blocks rather than singly. Using a novel in vivo alternative splicing assay, we show that intron length has a major influence upon the alternative splicing of CD44. We propose a kinetic model in which short introns may overcome the poor recognition of alternatively spliced exons. These observations suggest that for CD44, intron length has been exploited in the evolution of the genomic structure to enable tissue-specific patterns of splicing to be maintained.

Alternative Splicing↗
PubMed1997

Molecular speciation of fish sperm phospholipids: large amounts of dipolyunsaturated phosphatidylserine.

The molecular species compositions of the main diacyl phosphoglyceride classes and ether-linked subclasses from sperm of three species of fish, sea bass Dicentrarchus labrax, Atlantic salmon Salmo salar and Chinook salmon Onchorhynchus tsawytscha, were determined. The phospholipids from sperm were highly unsaturated, dipolyunsaturated fatty acid (diPUFA) molecular species comprised 64.6 to 71.8% of phosphatidylserine (PS), 10.1 to 17.4% of phosphatidylethanolamine (PE), and 3.3 to 10.1% of phosphatidylcholine (PC). In sea bass sperm, di22:6n-3 phospholipid was the predominant diPUFA molecular species, but in both salmon species 22:5n-3/22:6n-3 was also a major constituent of PS. Phospholipids containing 22:6n-3 dominated in sea bass sperm with 16:0/22:6n-3 as a major component of PC and PE, and 18:0/22:6n-3 of PE and PS in addition to di22:6n-3 in the latter two classes. In contrast, both salmon species contained much more 20:5n-3 and less 22:6n-3 so that saturated/20:5n-3 and monounsaturated/20:5n-3 molecular species were more abundant than the corresponding molecules containing 22:6n-3. Ether-linked lipids comprised 11.3-36.3% of choline and ethanolamine phosphoglycerides in each fish species. Molecular species containing 22:6n-3 were the major components of 1-O-alkyl-2-acyl-glycerophosphocholine, especially 16:0a/22:6n-3 in sea bass and 18:1a/22:6n-3 in the two salmon species, while in 1-O-alk-1'-enyl-2-acyl-glycerophosphoethanolamine, 16:0a/22:6n-3 was the major component in both salmon and 18:0a/22:6n-3 in sea bass with 18:1a/22:6n-3 abundant in all three species. In Atlantic salmon 1-O-alkyl-2-acyl-glycerophosphoethanolamine comprised 24.6% of ethanolamine glycerophospholipids which were predominantly 16:0a/22:6n-3 and 18:1a/22:6n-3. Phosphatidylinositol from sperm was dominated by stearoyl/C20 PUFA molecular species, in sea bass overwhelmingly 18:0/20:4n-6, while in both salmon species 18:0/20:4n-6 and 18:0/20:5n-3 were equally abundant.

Animals↗
PubMed1997

Fatty acid composition, eicosanoid production and permeability in skin tissues of rainbow trout (Oncorhynchus mykiss) fed a control or an essential fatty acid deficient diet.

Rainbow trout (Oncorhynchus mykiss) were fed either a control diet containing fish oil or an essential fatty acid (EFA) deficient diet containing only hydrogenated coconut oil and palmitic acid as lipid source (93.4% saturated fatty acids) for 14 weeks and the fatty acid compositions of individual phospholipid classes from skin and opercular membrane (OM) determined. The permeability of skin and OM to water and the production of eicosanoids in skin and gills challenged with the Ca2+ ionophore A23187 were also measured. Phospholipid (PL) fatty acid compositions were substantially modified in EFA-deficient fish, with increased saturated fatty acids and decreased polyunsaturated fatty acids (PUFA), especially arachidonic acid (AA) and eicosapentaenoic acid (EPA), while docosahexaenoic acid (DHA) was largely retained. The onset of EFA deficiency was shown by the appearance of n-9 PUFA, particularly 20:3n-9. The main effects of EFA deficiency on phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were to increase saturated fatty acids and monoenes, especially 16:1 and 18:1, and to decrease EPA and DHA. The content of DHA in phosphatidylserine (PS) was high in control animals (40% in skin and 35% in opercular membrane) and was mostly retained in EFA deficient animals. Arachidonic acid (AA) was the most abundant PUFA esterified to phosphatidylinositol (PI) and was significantly reduced in EFA deficient animals (from 31% to 13% in skin), where a large amount of 20:3n-9 (9% in skin) was also present. Influxes and effluxes of water through skin and opercular membrane were measured in vitro. No differences were detected between rainbow trout fed the control or the EFA deficient diet. 12-Hydroxyeicosatetraenoic acid (12-HETE), 12-hydroxyeicosapentaenoic acid (12-HEPE) and 14-hydroxydocosahexaenoic acid (14-HDHE) could not be detected in skin from control or EFA deficient fish. There was no difference between control and EFA deficient trout in the levels of leukotriene C4 (LTC4) and leukotriene C5 (LTC5) in skin cells challenged with the calcium ionophore A23187, and of prostaglandin F2alpha (PGF2alpha), 12-HETE and 12-HEPE in gill cells challenged similarly. Prostaglandin F3alpha (PGF3alpha) production by ionophore stimulated gill cells was significantly reduced in fish fed the EFA-deficient diet. 14-HDHE produced by gill cells was 3.3 fold higher in EFA deficient fish compared to controls.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1995

Identification and characterization of three members of the human SR family of pre-mRNA splicing factors.

SR proteins have a characteristic C-terminal Ser/Arg-rich repeat (RS domain) of variable length and constitute a family of highly conserved nuclear phosphoproteins that can function as both essential and alternative pre-mRNA splicing factors. We have cloned a cDNA encoding a novel human SR protein designated SRp30c, which has an unusually short RS domain. We also cloned cDNAs encoding the human homologues of Drosophila SRp55/B52 and rat SRp40/HRS. Recombinant proteins expressed from these cDNAs are active in constitutive splicing, as shown by their ability to complement a HeLa cell S100 extract deficient in SR proteins. Additional cDNA clones reflect extensive alternative splicing of SRp40 and SRp55 pre-mRNAs. The predicted protein isoforms lack the C-terminal RS domain and might be involved in feedback regulatory loops. The ability of human SRp30c, SRp40 and SRp55 to modulate alternative splicing in vivo was compared with that of other SR proteins using a transient contransfection assay. The overexpression of individual SR proteins in HeLa cells affected the choice of alternative 5' splice sites of adenovirus E1A and/or human beta-thalassemia reporters. The resulting splicing patterns were characteristic for each SR protein. Consistent with the postulated importance of SR proteins in alternative splicing in vivo, we demonstrate complex changes in the levels of mRNAs encoding the above SR proteins upon T cell activation, concomitant with changes in the expression of alternatively spliced isoforms of CD44 and CD45.

Alternative Splicing↗
PubMed1995

Dietary deficiency of docosahexaenoic acid impairs vision at low light intensities in juvenile herring (Clupea harengus L.).

In the retina of herring (Clupea harengus L.), rods are recruited from about 8 wk after hatching, and from this time there is a linear relationship between the number of rods in the photoreceptor cell population and the content of di22:6n-3 molecular species of phospholipids. Juvenile herring were reared from four weeks' post-hatching for 15 wk on either Artemia nauplii deficient in 22:6n-3 or on enriched Artemia nauplii containing 4.3% 22:6n-3. The visual performance of the fish was then determined at three light intensities (0.01, 0.1, and 1.0 lux) by observing their frequency of striking at live Artemia nauplii using infrared video recording. Herring reared on the diet containing no 22:6n-3 were less active predators, especially at the lowest light intensity where very few strikes were observed. The eyes of these fish contained greatly reduced levels of di22:6n-3 molecular species of total phospholipid, 2.1% vs. 12.0% in fish supplemented with 22:6n-3. The contribution of saturated and monounsaturated fatty acids in the molecular species of phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylcholine (PC) was virtually unchanged, while 20:5n-3 and 22:5n-3 largely replaced 22:6n-3. There was an almost complete disappearance of di22:6n-3 PC, while the amounts of di22:6n-3 PE and PS fell by 18.1 and 20.6% to 2.7 and 7.6%, respectively. The dipolyunsaturated molecular species di20:5n-3, 20:5n-3/22:5n-3, and di22:5n-3 made up a substantial part of the deficit. We conclude that a dietary deficiency of 22:6n-3 during the period early in rod development impairs visual performance such that the fish can no longer feed at low light intensities.

Animals↗
PubMed1995

The small cell lung cancer antigen cluster-4 and the leukocyte antigen CD24 are allelic isoforms of the same gene (CD24) on chromosome band 6q21.

Cluster-4 and CD24 cDNA's have recently been cloned from the small cell lung carcinoma (SCLC) cell line SW2 and from the erythroleukemia cell line K562, respectively. The only difference in the coding sequence, between cluster-4 and CD24 antigens is the substitution of a single base pair leading to a substitution of Val by Ala near the putative glycosylphosphatidylinositol (GPI) anchorage sites of the mature protein. Here we demonstrate that the nucleotide substitution which distinguishes the cluster-4 and CD24 antigen genes is due to an allelic polymorphism on chromosome band 6q21. In addition, we identified by Southern blotting and PCR of DNA from somatic human x hamster hybrid cell lines homologues of cluster-4/CD24 on the Y chromosome and chromosome 15. We suggest, however, that the gene on 6q21 is the active locus since the mRNA of cell lines always represents the allelic variants found on chromosome 6. The distribution pattern of this allelic polymorphism in SCLC cell lines and leukocytes of healthy donors did not reveal any obvious relationship with disease. However, it is noteworthy that homozygosity for cluster-4 was found in only one case whereas heterozygosity and homozygosity for CD24 both contribute up to 50% of the samples examined.

Alleles↗
PubMed1994

CD44 isoform expression mediated by alternative splicing: tissue-specific regulation in mice.

CD44 is a widely distributed cell surface glycoprotein which shows heterogeneity in molecular expression as a result of post-translational modification as well as alternative splicing of CD44 mRNA. Functional studies have indicated that CD44 plays a role as an adhesion molecule and that different CD44-expressing cells differ in their capacities for CD44-dependent ligand binding. These observations have raised the possibility that structural modifications of CD44, including those resulting from alternatively spliced mRNA isoforms, are involved in the functional heterogeneity of CD44. To assess the expression of CD44 isoforms in the mouse, we examined CD44 cDNA by reverse transcription polymerase chain reaction (RT-PCR). Southern blotting of PCR products with a CD44 cDNA probe or with internal oligonucleotides revealed the expression in mouse tumor cell lines and normal tissues of multiple CD44 mRNA products which are larger than that observed in the absence of variable exon expression. Interestingly, different mouse tissues, including lymphoid cells, showed unique patterns of alternative CD44 mRNA in Southern blotting analysis. The use of exon-specific primers allowed detection of multiple alternatively spliced mRNA species involving expression of at least seven variable exons. Cloning and sequencing of these PCR products revealed sequence identity with recently identified genomic CD44 sequences and confirmed that the PCR products correspond to mature mRNA expressing alternatively spliced CD44 exons. Taken together, these findings demonstrate that the mouse expresses multiple variably spliced CD44 isoforms and that expression is regulated in a tissue- and cell-type specific manner.

Alternative Splicing↗
PubMed1993

The identification of a new alternative exon with highly restricted tissue expression in transcripts encoding the mouse Pgp-1 (CD44) homing receptor. Comparison of all 10 variable exons between mouse, human, and rat.

The human CD44 cell surface glycoprotein family which has been implicated in lymphocyte homing, tumor metastasis, and extracellular matrix attachment consists of a large number of related isoforms that derive from the differential splicing of a single CD44 gene transcript. Recent mapping of the CD44 locus in man indicated the presence of at least nine alternative exons within the region of the gene encoding the variable membrane proximal extracellular domain. Here we report the identification of a 10th alternative exon (termed V1) in the mouse, human, and rat CD44 genes. We demonstrate tissue-specific patterns of expression for transcripts containing exons V1-V10 in the mouse and a highly restricted usage of exon V1 in transcripts from mouse gastric tissue. Close sequence homology between exons V1-V10 from mouse rat and human points to a specific functional role rather than a purely structural role for the membrane proximal extracellular domain of the CD44 molecule.

Alternative Splicing↗
PubMed1993

CD44 and cancer.

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Humans↗
PubMed1993

Molecular species composition of glycerophospholipids from white matter of human brain.

The molecular species composition of the major glycerophospholipids from white matter of human brain were determined by high-performance liquid chromatography of the 3,5-dinitrobenzoyl derivatives of the corresponding diradylglycerols. In phosphatidylcholine (PC) and phosphatidylserine (PS), molecular species containing only saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) comprised 85.7 and 82.4% of the respective totals, with 18:0/18:1 predominant in PS and 16:0/18:1 in PC. These molecular species were also abundant in phosphatidylethanolamine (PE), but in this phospholipid species containing polyunsaturated fatty acids (PUFA), largely 18:0/22:6n-3 and 18:0/20:4n-6, accounted for over half the total; 18:1/18:1 was also abundant in PE. In contrast, 1-O-alk-1'-enyl-2-acyl sn-glycero-3-phosphoethanolamine (GPE) had much more SFA- and MUFA-containing species, predominantly 16:0a/18:1, 18:0a/18:1 and 18:1a/18:1, with low amounts of species containing 20:4n-6 and 22:6n-3. In alkenylacyl GPE, 22:4n-6 was the major PUFA and 16:0a/22:4n-6 and 18:1a/22:4n-6 the main PUFA-containing species. There was six times more 22:6n-3, twice as much 20:4n-6 and half the amount of 22:4n-6 in PE as compared to alkenylacyl GPE.

Aged↗