Biomedical subjects

A Fjose

Publications and source records attributed to A Fjose.

48 records · Page 3Linked to original sources

PubMed1988

Molecular structure and spatial expression of a homeobox gene from the labial region of the Antennapedia-complex.

We report the molecular characterization of a homeobox-containing gene that maps at 84A in the proximal region of the Antennapedia-complex. The structure and complete sequence are presented. Deletion analysis indicates that the cloned gene, F24, most likely corresponds to the labial (lab) gene. Northern blot experiments show a single approximately 3-kb transcript that is expressed at all embryonic stages from cellular blastoderm onwards and during larval development. The homeobox is split by an intron in the region which encodes the putative DNA-binding helix, a splicing position for homeobox-containing genes which is unique so far. The 5' part of the gene contains four M-repeat sequences (CAA/G repeats) in the protein-coding region. In situ hybridization to the transcripts during embryogenesis reveals two domains of expression. The anterior one is located in parts of the developing head, mainly in the hypopharyngeal organ and in anterior parts of the mandibular lobe, and is restricted to the ectoderm. The posterior domain is part of the posterior midgut primordium (endoderm), that invaginates and later contacts the endoderm cells from the anterior midgut invagination.

Amino Acid Sequence↗

PubMed1988

Molecular cloning and characterization of homeo-box-containing genes from Atlantic salmon.

As the most primitive group among vertebrates, fish might serve as a model system when studying the genetic regulation of embryogenesis in higher animals. To identify genes important for early development, we have constructed a genomic library from Atlantic salmon (Salmo salar) and screened it with homeobox-containing probes from Drosophila melanogaster. Five different salmon homeoboxes were isolated. Two of these were located in the same clone, separated by only 7.5 kb. This demonstrates the presence of clustered homeobox genes in fish. The two clustered homeoboxes were sequenced and shown to be closely related to the ANT-C/BX-C class of Drosophila, being about 80% homologous to the Ultrabithorax gene (Ubx) homeobox. One of the clustered genes appears to be the salmon equivalent of the mouse Hox-2.1 gene, indicating that some of the vertebrate homeobox-containing genes are conserved in evolution. A more diverged homeobox that shares only 60% homology with Ubx, was also sequenced. In analogy to Drosophila, therefore, the salmon genome contains more than one class of homeoboxes. In addition, Northern-blot experiments demonstrated that two of the homeobox genes are expressed in salmon embryos, suggesting their importance for proper development.

Amino Acid Sequence↗

PubMed1987

A zebrafish homeobox-containing gene with embryonic transcription.

A genomic library of zebrafish (Brachydanio rerio) was constructed and screened with homeobox-containing probes. One of the strongly cross-hybridizing clones was characterized by DNA sequencing. The deduced amino acid sequence exhibits extensive homology (greater than 80%) relative to the Antennapedia-class of Drosophila homeobox sequences. Characterization of the gene with respect to expression demonstrated that two transcripts of 2.1 and 1.4 kb, respectively, are present in embryonic poly (A+) RNA. The highest concentration of the two RNA species was observed in embryos which have terminated the process of somite formation.

Animals↗

PubMed1985

Isolation of caudal, a Drosophila homeo box-containing gene with maternal expression, whose transcripts form a concentration gradient at the pre-blastoderm stage.

We report the isolation and characterization of caudal (cad), a previously unknown Drosophila homeo box-containing gene from the 38E region on the left arm of the second chromosome. This homeo box has diverged from the prototype sequence in Antennapedia, but contains subregions which are highly homologous. By Northern analysis and in situ hybridization experiments two transcripts of 2.4 kb were found to accumulate in nurse cells and in the oocyte during oogenesis. These transcripts generate a transient concentration gradient along the antero-posterior axis at the syncytial blastoderm stage. At the cellular blastoderm stage transcripts accumulate in a single band from 13-19% egg length at the posterior end. One zygotic transcript of 2.6 kb is detected. At later stages this transcript is localized in ectodermally and endodermally derived tissues such as the proctodeum, the Malpighian tubules and the posterior midgut. The 2.6-kb transcript is detectable until the onset of metamorphosis.

Journal Article↗

PubMed1984

Altered distribution profiles of endoplasmic reticulum subfractions after incubation of Krebs II ascites cells with different concentrations of cytochalasin B.

Information on the interaction between endoplasmic reticulum (ER) membranes and components of the skeletal network of the cell was gained by treating cells with the antimicrofilament agent cytochalasin B prior to cell disruption by nitrogen cavitation. Treatment of Krebs II ascites cells with cytochalasin B (5-10 micrograms ml-1) resulted in an increased yield of three ER membrane subfractions--heavy rough (HR), light rough (LR) and smooth (S) membranes, as judged by 3H-choline incorporation in gradient fractions following discontinuous sucrose gradient centrifugation. The major increase was observed in the HR fraction. These results indicate that the actual yield of the respective ER membrane subfractions after cell disruption is dependent on the degree of direct and/or indirect interaction between individual ER membranes and actin containing filaments of the cytoskeleton in the intact cell.

Actins↗

PubMed1984

Developmental expression of the white locus of Drosophila melanogaster.

We have isolated several cDNA clones of the white locus which are derived from embryonic and pupal transcripts of Drosophila melanogaster. The cDNA sequences map within 7.5 kb (coordinates -3.0 to +4.6) of the genomic DNA and correspond mainly to sequences within the distal region of the gene (coordinates -0.2 to -3.0). A major RNA species of 2.6 kb was detected on Northerns of poly(A) RNA isolated from all developmental stages. The total accumulation of this transcript peaks in the mature third instar larva to a level of 0.003% which is about ten times higher than that observed in embryos. The spatial distribution of white locus transcripts was determined by in situ hybridization to tissue sections. In embryos, hybridization signals are restricted to the cells of the developing Malpighian tubules and the signal strength corresponds with 50 transcripts per cell. Before the termination of the third instar stage, hybridization signals are also detected at a comparable level in the eye antennal disks. At the same stage, a third site of labeling is observed over a small cluster of cells which seems to be associated with the larval photoreceptor organs. Thus, white locus expression is largely restricted to tissues which are known to be involved in the biosynthesis of eye pigments and these different cell types act in a temporally autonomous manner with respect to the induction of the white gene during development.

Journal Article↗

PubMed1983

The nuclear-associated endoplasmic reticulum is an early target for the action of the tumor promoter 12-O-tetradecanoyl phorbol-13-acetate in C3H/10T1/2 fibroblasts.

12-O-Tetradecanoyl-phorbol-13-acetate (TPA) had a dual effect on the cellular membranes of C3H/10T1/2 cells in that it caused both a stimulation of [3H]choline incorporation and an enhancement of the solubilization of choline from prelabelled cells. Subfractionation studies showed that the release of [3H]choline occurred almost exclusively from nuclear-associated endoplasmic reticulum. The release was dependent on the presence of Mg2+ and Ca2+, indicating an enzyme-mediated reaction. In vivo, TPA stimulated the incorporation of [3H]choline into all subcellular fractions. The data indicate that the nuclear-associated endoplasmic reticulum represents an early target for TPA action.

Animals↗

PubMed1983

Time-dependent alteration in endoplasmic reticulum membrane profiles during in vitro incubation of Krebs II ascites cells.

Krebs II ascites cells were harvested from the mouse peritoneum 6-8 days after inoculation and incubated in vitro in roller suspension culture for up to 22 hr. Within 2 hr of incubation a large portion of the cells entered S phase as judged by the incorporation of 3H-thymidine into DNA. The incorporation of radioactive precursors into phospholipid, protein and RNA increased rapidly during in vitro incubation indicating a high degree of macromolecular synthesis. The rates of incorporation were maximal within 4 hr of incubation. When cells labeled with 3H-choline were disrupted by nitrogen cavitation and endoplasmic reticulum (ER) membranes analyzed for subfractions on discontinuous sucrose gradients, it was observed that only small amounts of radioactivity could be detected in the HR region after 1/2 hr incubation while 63.2% of total radioactivity in ER membranes appeared in the LR fraction. Between 8-18 hr there was a considerable increase in the amount of HR membranes. Of the total radioactivity in ER membranes that in the HR fraction increased from 16.9% to 53.9% during this period. There were only small changes in the amounts of radioactivity in LR and S membranes between 8 and 18 hr. The results suggest a time-dependent appearance of ER membrane subfractions during a 22 hr period of in vitro incubation of Krebs II ascites cells.

Animals↗

PubMed1983

The phorbol ester 12-0-tetradecanoyl-phorbol-13-acetate and stimulation of 3H-choline incorporation into endoplasmic reticulum membranes and other subcellular fractions of Krebs II ascites cells during in vitro incubation.

After transfer of Krebs II ascites cells from the mouse peritoneum to suspension culture addition of the phorbol ester 12-0-tetradecanoyl-phorbol-13-acetate (TPA) causes an early stimulation of 3H-choline incorporation into phosphatidylcholine (PC). Choline transport into the treated cells, however, was unaffected. Within 30 min of TPA treatment 3H-choline incorporation was almost 300% above the control level. During a 5 hr period of suspension culture the overall patterns of 3H-choline incorporation were similar in TPA-treated and control cultures though the rate was greatly accentuated by the presence of the phorbol ester. Incubation of cells with cycloheximide prior to incubation with TPA did not result in an inhibition of the TPA-directed 3H-choline incorporation. After 3 hr incubation with TPA there were large increases in radioactivity in all subcellular fractions. At 20 hr, however, the values were not far from those of the control. During the first 3 hr of incubation with TPA the incorporation of 3H-choline into light rough (LR) and smooth (S) membranes was stimulated to levels of 400% and 320% respectively above control values. At later times the profiles of radioactivity in membrane subfractions in TPA-treated and control cultures were similar. The results illustrate an early effect of TPA on PC biosynthesis in Krebs II ascites cells while at later times of incubation the stimulatory effect was virtually abolished.

Animals↗

PubMed1983

The specific appearance of a 65,000 dalton protein in the nuclear associated endoplasmic reticulum of Krebs II ascites cells early after treatment with the phorbol ester TPA.

The protein compositions of subcellular fractions prepared from untreated and 12-O-tetradecanoyl phorbol-13-acetate (TPA)-treated Krebs II ascites cells were examined by SDS-polyacrylamide gel electrophoresis. After 6 h of TPA treatment a 65,000 dalton protein in the nuclear associated endoplasmic reticulum showed a 240% increase when compared with the untreated control cells. The amount of this protein was similar in both untreated and TPA-treated cells up to 4 h of in vitro incubation. Though present in all other subcellular fractions, the amount of the 65,000 dalton protein was the same in both untreated and TPA-treated cells. Small changes were also observed in the amounts of other proteins in the nuclear associated endoplasmic reticulum in TPA-treated cells. The possibility was tested that the increased amount of the protein may have represented increased ornithine decarboxylase activity in response to TPA treatment. The time dependent appearance of the enzyme in untreated and TPA-treated cells, however, and the behaviour and subcellular localization of the 65,000 dalton protein ruled out the possibility that this protein and ornithine decarboxylase are one and the same molecule.

Animals↗

PubMedDate not supplied

Molecular cloning and characterization of five opsin genes from the marine flatfish Atlantic halibut (Hippoglossus hippoglossus).

Most molecular studies on the visual system in fish have been performed on freshwater teleosts such as goldfish and zebrafish where cones and rods appear simultaneously. Many marine fishes have long larval phase in the upper pelagic zone before transformation into a juvenile and a benthic life style. The retina at the larval stages consists of only single cone cells; later during metamorphosis double cones and rods develop. The flatfish Atlantic halibut (Hippoglossus hippoglossus) is a typical example of a marine species with such a two-step retina development. In this study, we have cloned five different opsins from Atlantic halibut larvae and juvenile retinas. Sequence comparisons with other opsins and phylogenetic analysis show that the five genes belong to the opsins of long-wavelength sensitive (L); middle-wavelength sensitive, M(Cone) and M(Rod); and short-wavelength sensitive, S(Blue) and S(Ultraviolet), respectively. In situ hybridization analysis reveals expression in double cone (L and M(Cone)), single cone (S(Blue) and S(Ultraviolet)), and rod (M(Rod)) types of photoreceptor cells in juvenile halibut retina. The visual system in Atlantic halibut seems therefore to have all four types of cone photoreceptors in addition to rod photoreceptors. This work shows for the first time molecular isolation of a complete set of retinal visual pigment genes from a marine teleost and describes the first cloning of an ultraviolet-sensitive opsin type from a marine teleost.

Amino Acid Sequence↗

PubMedDate not supplied

Isolation of a homoeo box-containing gene from the engrailed region of Drosophila and the spatial distribution of its transcripts.

The engrailed locus of Drosophila melanogaster has the characteristics of both a homoeotic gene and a segmentation gene: like a homoeotic gene, it specifies the development of specific compartments of the Drosophila embryo (the posterior compartments of each segment), and, like mutations of segmentation genes, lethal alleles of engrailed affect also the pattern of segmentation of the embryo. Here we report that like many of the homoeotic genes of the bithorax and Antennapedia complexes, engrailed has a 'homoeo box' sequence: also, like the segmentation gene fushi tarazu, the engrailed gene displays a periodic pattern of expression in Drosophila embryos.

Amino Acid Sequence↗