Two different sets of cis elements regulate scute to establish two different sensory patterns

Summary We have analysed the role of the achaete-scute gene complex in the development of the pattern of campaniform sensilla on the wing blade of Drosophila. We show that the complete pattern results from the superimposition of two independent subpatterns, one of which depends on the achaete gene and the other on scute. The scute subpattern comprises several clusters of sensilla, most of which seem to require the presence of control regions located upstream of the transcribed region. This is in contrast with the pattern of scute-dependent bristles, most of which depends on control elements located downstream of the transcribed region.     

The plasma membrane of myxosporidian valve cells: freeze fracture data

Freeze fracturing of Myxosporidian spores reveals the occurrence of a continuous layer of transmembrane particles all over the surface area of the valve cells which form the spore envelope. These particles are densely packed all over the P face membrane. Due to their polygonal outline, their diameter (6-7 nm) and their central core, they resemble the particles forming the connections of gap junctions which metabolically couple the neighboring cells in animal tissues. In the present report, the role of the transmembrane particles is still hypothetical. However, they might represent a membrane structural specialization of the spores which are submitted to osmotic variations of the fluid external medium. Furthermore similar transmembrane particles are observed at the level of the septate junction which seals the valve cells. In this occurrence, they are arranged in a series of 40 double rows parallel to the suture of the spore envelope. These findings support the view that Myxosporidia are Metazoa and raise the problem of their origin.     

Barium- and calcium-permeable channels open at negative membrane potentials in rat ventricular myocytes

Summary Ca²⁺- and Ba²⁺-permeable channel activity from adult rat ventricular myocytes, spontaneously appeared in the three single-channel recording configurations: cell-attached, and excised inside-out or outside-out membrane patches. Single-channel activity was recorded at steady-state applied membrane potentials including the entire range of physiologic values, and displayed no rundown in excised patches. This activity occurred in irregular bursts separated by quiescent periods of 5 to 20 min in cell-attached membrane patches, whereas in excised patch experiments, this period was reduced to 2 to 10 min. During activity, a variety of kinetic behaviors could be observed with more or less complex gating patterns. Three conductance levels: 22, 45 and 78 pS were routinely observed in the same excised membrane patch, sometimes combining to give a larger level. These channels were significantly permeable to divalent cations and showed little or no permeability to potassium or sodium ions. The inorganic blockers of voltage-gated Ca channels, cobalt (2mm), cadmium (0.5mm) or nickel (3mm), had no apparent effect on these spontaneous unitary currents carried by barium ions. Under 10^–5 m bay K 8644 or nitrendipine, the activity was clearly increased in about half of the tested excised inside-out membrane patches. Both dihydropyridines enhanced openings of the larger conductance level, which was only very occasionally seen under control conditions. When the single-channel activity became sustained under 5×10^–6 m Bay K 8644, it was possible to calculate the mean unitary current at different membrane potentials and show that the mean current value increased with membrane potential.     

Different Hoechst 33342 and DAPI fluorescence of the human Y chromosome in bivariate flow karyotypes

Summary The staining properties of AT-specific dyes Hoechst 33342 and DAPI as revealed by Hoechst 33342/mithramycin and mithramycin/DAPI bivariate human flow karyotype patterns are different for chromosomes rich in heterochromatin. The peak corresponding to chromosome Y of a given cell line is higher on the A/T axis with mithramycin/DAPI staining than with mithramycin/Hoechst. The chromosome 1 was found slightly more fluorescent in mithramycin/DAPI than in mithramycin/Hoechst as judged by the slight displacement of its area on the Hoechst-DAPI axis. The other peaks did not show major differences. On the same flow karyotypes, chromosomal rearrangements were detected.     

Protein engineering by cDNA recombination in yeasts: shuffling of mammalian cytochrome P-450 functions

We have constructed, in the yeast Saccharomyces cerevisiae, a mosaic assembly of genes by in vivo recombination of partially homologous sequences. The approach was tested on cDNAs encoding functionally distinct mammalian cytochromes P-450 (P-450). The selection for recombinant cDNAs used the transformation of yeast cells, which required the recircularization of a linearized plasmid by recombination of two partially homologous cDNAs. Libraries of mosaic genes with bipartite or tripartite structures were generated by intramolecular and intermolecular recombination events. The presence of yeast promoter and terminator sequences on the flanking sides of the recombined cDNAs has allowed the synthesis of encoded mosaic proteins. A library of yeast clones producing recombinant mouse P-450 P1 and rabbit P-450 LM4 was screened using functional criteria to identify chimeras with shuffled substrate specificity. Restriction mapping of mosaic genes, biochemical analysis of the synthesized proteins, comparison of chimeric enzymes, and the alignment of sequences with bacterial P-450 camphor hydroxylase of known three-dimensional structure, all suggest that the P-450 P1 amino acid residues 203-238 play a major role in the control of cytochrome activity toward carcinogenic polycyclic aromatic hydrocarbons. Similar approaches to structure-function analysis are believed to be applicable to other protein families.     

A revision of coregonine fish distribution and abundance in eastern James-Hudson Bay

Synopsis Recent sampling programs conducted in the estuaries of the Eastmain and La Grande rivers (James Bay) and the Great Whale, Little Whale, Innuksuac and Povungnituk rivers (Hudson Bay) revealed patterns of coregonine fish distribution that differ from previous observations. The relative abundance of cisco, Coregonus artedii, and lake whitefish, C. clupeaformis, varied among rivers but did not reveal a latitudinal cline. Previous sampling programs underestimated the abundance of cisco in the Little Whale River. In addition, cisco was the third most abundant species captured in the Povungnituk River, situated 200 km to the north of the previously proposed northern limit at Innuksuac River. As such, the low abundances of cisco in the Great Whale and Innuksuac rivers cannot be attributed to a physiological inability to cope with a reduced growing season. Immature cisco were almost totally absent from the estuaries of the Hudson Bay rivers following spring breakup whereas immature lake whitefish made up 100% of the catch in the Innuksuac River at the same time of year. Species-specific migration patterns in Hudson Bay that differ from those observed in James Bay and the existence of unique juvenile overwintering rivers are 2 hypotheses proposed to explain the discontinuous age-class distribution of cisco and lake whitefish observed in Hudson Bay.Contribution to the program of GIROQ (Groupe Interuniversitaire de Recherche Océanographique du Québec).     

Interaction of antimicrobial dermaseptin and its fluorescently labeled analogues with phospholipid membranes.

Dermaseptin, a 34 amino-acid residue antimicrobial polypeptide [Mor, A., Nguyen, V. H., Delfour, A., Migliore-Samour, D., & Nicolas, P. (1991) Biochemistry 30, 8824-8830] was synthesized and selectively labeled at its N-terminal amino acid with either 7-nitrobenz-2-oxa-1,3-diazole-4-yl (NBD), rhodamine, or fluorescein. The fluorescent emission spectra of the NBD-labeled dermaseptin displayed a blue-shift upon binding to small unilamellar vesicles (SUV), reflecting the relocation of the fluorescent probe to an environment of increased apolarity. Titrations of solutions containing NBD-labeled dermaseptin with SUV composed of zwitterionic or acidic phospholipids were used to generate binding isotherms, from which were derived surface partition constants of (0.66 +/- 0.06) x 10(4) M-1 and (2.8 +/- 0.3) x 10(4) M-1, respectively. The shape of the binding isotherms, as well as fluorescence energy transfer measurements, suggests that some aggregation of membrane-bound peptide monomers occurs in acidic but not in zwitterionic vesicles. The preferential susceptibility of the peptide to proteolysis when bound to zwitterionic but not to acidic SUV suggests that these aggregates might then penetrate a relatively short distance into the hydrophobic region of the acidic membrane. Furthermore, the results provide good correlation between the peptide's strong binding and its ability to permeate membranes composed of acidic phospholipids, as revealed by a dissipation of diffusion potential and a release of entrapped calcein from SUV.     

Transcobalamin II--cobalamin binding sites are present on rabbit germ cells.

Specific binding sites for rabbit transcobalamin II have been found on isolated adult rabbit germ cells. Scatchard analysis revealed a single class of binding sites for [57Co]cyanocobalamin-transcobalamin II with an association constant (Ka) of 1.3 x 10(10) M-1 and 700 sites per cell. Binding was reversible, saturable and calcium dependent. Electron microscope radioautography following incubation with iodinated transcobalamin II at 4 degrees C led to a detectable labeling mainly restricted to the plasma membrane.     

Redescription de Proteocephalus paraguayensis (Rudin, 1917) (Cestoda: Proteocephalidae) parasite de Hydrodynastes gigas (Dum., Bibr. & Dum., 1854) du Paraguay

Redescripción de Proteocephalus paraguayensis (Rudin, 1917) (Cestoda: Proteocephalidae), parásito de Hydrodynastes gigas (Dum., Bibr. & Dum., 1854) de Paraguay. Se describe esta especie notable, considerada por Freze (1965) como species inquirenda. Si comparamos esta especie con otras especies de Proteocephalidea, veremos que ella presenta caracteres anatomo-morfológicos propios. Esta especie se caracteriza por: vitelógenos de posición dorsal, adosados a la musculatura interna longitudinal, desbordando en parte sobre el córtex y la médula; formación particular del útero; posición cortical del tronco uterino y medular de los divertículos uterinos; esfínter vaginal proximal; cirro muy alargado; glándula de Mehlis muy desarrollada. Consideramos P. paraguayensis como una especie válida y presumimos que el huesped-tipo es Hydrodynastes gigas (Dum., Bibr. & Dum., 1854). A pesar de las características que posee P. paraguayensis, pensamos que no es oportuno clasificar esta especie en un nuevo género monotípico. Proteocephalus paraguayensis, considered by Freze (1965) as a species inquirenda, is redescribed and figured. When compared with other members of the Proteocephalidea, the species shows the following morpho-anatomical characters: the vitelline follicles in a dorsal position, attached to the internal longitudinal musculature and extending into both the cortex and medulla; a characteristic formation of the uterus; the uterine stem in a cortical position and uterine branches in a medullary position; a proximal vaginal sphincter; a very elongate cirrus; and very large Mehlis' glands. The specific status of Proteocephalus paraguayensis is confirmed. Our specimens were taken from Hydrodynastes gigas (the host according to Rudin was Coluber sp.). Even though the species differs significantly from other proteocephalideans, its systematic and phylogenetic position is not yet clearly demonstrated, and it is decided to refrain from attributing it to a new genus.