Evolution of the glycogen content and of glucose-6-phosphatase activity in the liver of Salmo gairdneri during development.

The evolution of the liver's glycogenic content, cytochemical characterization of the glycogen and glucose-6-phosphatase activity enable us to define three successive phases up to stage 36, just before the first feed. The grade which was low up to stage 24 is then due to beta-particles of ovule origin. Then, up to stage 27, there is a storage phase: alpha-particles appear and accumulate while the enzymatic activity remains non-existent. From the stage 28 to 36 the grade is progressively increasing, the enzymatic activity appears and increases. When the phase ends the liver is able to ensure glycemic regulation and to deal with exogenous nutritional contributions.     

Creation of an HLA-A2/HLA-Aw69 alloantigenic determinant on an HLA-A3 molecule by site-directed mutagenesis

The HLA-A2 and HLA-Aw69 molecules share an antigenic determinant not expressed by HLA-Aw68 and HLA-A3. Comparison of the amino acid (aa) sequences of these molecules and previous studies of the antigenic determinant expressed by different HLA-A2 X HLA-A3 hybrid molecules had established that three aa at positions 95, 97, and 107 were possibly involved in the formation of this determinant. The HLA-A3 gene was therefore mutagenized to replace successively at these positions the HLA-A3-specific aa by the HLA-A2 residues. A single substitution at position 107 of a glycine by a tryptophan residue is sufficient for full expression by HLA-A3 molecules of the HLA-A2/Aw69 shared antigenic determinant without modification of the other serological reactivities characteristic of the HLA-A3 molecules. Previous studies of ethyl methanesulfonate mutants having shown the involvement of aa 161 in this determinant, we assume that the two aa residues 107 and 161 are close to each other.     

The same cell lineage is involved in scale formation and regeneration in the teleost fish Hemichromis bimaculatus

The elasmoid scales of the cichlid fish, Hemichromis bimaculatus, are localized within dermal pockets, the floors of which are separated from the stratum compactum by uninterrupted cellular sheets, the scale-pocket linings (SPL). TEM study of the fry skin shows that the SPL cells originate from the cell population constituting the dermal papilla of the scale. The upper-layer cells of the papilla, close to the epidermal-dermal junction, differentiate into scleroblasts that, subsequently, form the scale-bag, while the inner-layer cells, close to the stratum compactum, constitute a bi-layered sheet, the SPL. The SPL cells are joined one to another by numerous desmosomes and their cytoplasm is filled principally by microfilaments and free ribosomes. The SPL is also characterized by the presence of a basement membrane on its two faces. When a scale is experimentally pulled off, the scale-forming cells are removed with the dermis and the epidermis covering the free region of the scale, but the SPL is not damaged and epidermal fragments remain at the posterior edge of each scale-pocket. The epithelial cells migrate, from the epidermal fragments, on an extracellular matrix situated on the surface of the SPL, and the wound is closed from 3 to 6 h after scale removal. The scale-regenerating cells differentiate from the upper-layer cells of the SPL, initially in the central region of the scale-pocket where epithelial cells first contacted the SPL surface. Consequently, it is shown that scale-forming cells and scale-regenerating cells are derived from the same ontogenetic population, the dermal papilla.     

A study of functionally active amino acids involved in the interaction of HLA-A2 or HLA-A3 molecules with cytolytic T lymphocytes

A large series of HLA-A2/HLA-A3 recombinant genes were generated by using the in vivo recombination technique. These genes have each been modified in the last two-thirds of the third exon such that one or several HLA-A2-specific substitutions have been made in the HLA-A3 gene and vice versa. The recombinant genes were transfected into the murine cell line P815 and the transfectants were used as targets for a series of 20 human CTL lines or clones specific for HLA-A2 or HLA-A3, or restricted by HLA-A2 and specific for influenza A. Several patterns of anti-HLA-A2, anti-HLA-A3, and HLA-A2-restricted anti-influenza CTL activity were observed and when uncloned cell lines were studied, a progressive selection of some clones with a similar pattern of activity was regularly found. From the comparison of these different patterns the following conclusions can be drawn: 1) In most but not all cases both domains of the class I molecule were essential for CTL recognition, but residue 152 was critically important for the majority of CTL tested; 2) amino acids 114/116 were also critical in most cases, and their position close to amino acid 152 in the tertiary structure of the molecule may have some functional significance; and 3) amino acid 161, although highly conserved, plays an unexpected but very important role in CTL function.     

Changes of fatty acid composition of phospholipids and lipid structural order in rat liver mitochondrial membrane subsequent to galactosamine intoxication. Effect of clofibrate

Since it has been earlier reported that D-galactosamine induces an inhibition of palmitoylcarnitine transferase I and a depletion of mitochondrial phospholipids which were both prevented by clofibrate, an evaluation of the effects of these drugs on mitochondrial fatty acid composition was made. Galactosamine does not alter the fatty acid pattern of these fatty acids whereas clofibrate induces a 2-fold increase in monounsaturated/saturated fatty acids ratio and a 10-fold decrease of the 20:4 (n - 6)/20:3 (n - 6) ratio in phosphatidylcholine. These alterations suggest an increase of delta 9-desaturation and a decrease of delta 5-desaturation. To determine whether the drug-induced changes in mitochondrial phospholipids has an effect on the physical properties of the membrane, the lipid structural order of mitochondrial preparations was studied using the lipophilic probes DPH and TMA-DPH. Mitochondrial isolated either from galactosamine- or clofibrate-treated rats showed a decrease in fluorescence polarization, indicating an overall decrease in lipid structural order. This alteration is more drastic when both drugs are administered. This phenomenon suggests drastic changes in the bulk phase of inner mitochondrial membrane lipids after treatments and could explain the altered kinetic properties of palmitoylcarnitine transferase I.     

Is the Golgi apparatus the obligatory final step for lipoprotein secretion by intestinal cells?

It is currently admitted that the synthesis and excretion of triglyceride-rich lipoproteins (chylomicrons and 'small chylomicrons') by intestinal epithelial cells involves the Golgi apparatus as an obligatory final step before exocytosis. The cells of the proximal intestine of the trout are an excellent model for investigating functional compartmentalization in the course of lipid absorption. Using this model, our data invalidate morphological data which were the basis for considering the Golgi apparatus as the mandatory final stage for their secretion. In particular, we show that triglyceride-rich particles can be transported directly from the endoplasmic reticulum to the intercellular space. Two pathways of intestinal lipoprotein excretion appear to coexist. One follows the classical export route, the second functions in a manner that bypasses the Golgi apparatus. The arguments used to affirm the requirement for the Golgi apparatus as a final step (glycosylation of apoprotein B, membrane vehicle for exocytosis) are discussed.     

Ultrastructural observations on chondroid bone in the teleost fish Hemichromis bimaculatus

This paper presents transmission electron microscopical observations on the chondroid bone (CB) supporting the neurocraniad articulation facet of the upper pharyngeal jaws of juvenile specimens of Hemichromis bimaculatus (an acellular-boned teleost fish). Chondroid bone, a skeletal tissue morphologically intermediate between cartilage and bone, is composed of a dense mineralized collagenous matrix, resembling that of woven-fibred bone, and large chondrocyte-like cells. The latter vary considerably in their morphological features (functional cells, cells containing a large vacuole and degenerating cells). The CB is mineralized except for its upper layer. Mineralization is initiated in matrix vesicles. Clusters of apatite crystals coalesce at the mineralization front. Distally, the tissue grows by incorporation of cells which exhibit the features of osteoblasts, and which derive from less differentiated fibroblast-like cells located in the outermost layer of the tissue. Proximally, the CB is subjected to erosion by multinucleated clastic cells. The deposition of bone against the wall of lacunae which have been opened by clastic resorption may suggest a possible active involvement of the CB cells. Further studies should point out whether this bone substantially contributes to the acellular dermal dentigerous bone located below.