L'hématoxyline au plomb permet-elle l'identification des cellules corticotropes de l'hypophyse des téléostéens?

Conclusion La technique à l'hématoxyline au plomb de Mac Conaill permet l'identification d'un type cellulaire situé dans la pars distalis rostrale, en bordure des ramifications de la neurohypophyse dans l'hypophyse de plusieurs Poissons Téléostéens; il est également colorable par le bleu d'alizarine d'Herlant; toutefois, l'intensité de ces deux réactions tinctoriales est très variable selon les espèces. Chez l'Anguille, les Salmonidés et le Cyprin, la nature corticotrope de ces cellules est établie expérimentalement. La colorabilité par l'hématoxyline d'une autre catégorie cellulaire dans la pars intermedia laisse supposer l'existence de constituants communs aux deux types cellulaires, et peut-être même à leurs produits d'élaboration. Cette communauté de certains aminoacides entre l'ACTH et l'intermédine a déjà été démontrée chez divers Mammifères.     

Réactions observées chez l'Anguille maintenue dans un milieu privé d'électrolytes,en particulier au niveau du système hypothalamo-hypophysaire

Résumé 45 Anguilles sont maintenues (32 h. à 46 jours) en eau déminéralisée (désionisée ou distillée). On note un assombrissement de la pigmentation, une hypertrophie importante de la vésicule biliaire et des modifications du système hypothalamo-hypophysaire.Dans la pars distalis rostrale, les cellules à prolactine subissent une régression, ne semblant plus élaborer de granulations. Les cellules corticotropes sont faiblement stimulées, partiellement dégranulées, en accord avec la légère activation de l'interrénal. Les cellules thyréotropes apparaissent souvent peu actives après 46 jours; cependant, l'examen préliminaire des thyroïdes ne montre pas d'hypoactivité. Dans la pars distalis proximale, les cellules gonadotropes ne sont pas modifiées; les cellules somatotropes sont stimulées: hypertrophie nucléaire et nucléolaire, développement de l'ergastoplasme, sans dégranulation complète. Leur rôle dans l'osmorégulation est discuté. Dans le lobe intermédiaire, les cellules du type 1, hématoxyline au plomb positives, prédominent chez les témoins; elles subissent une stimulation avec dégranulation partielle correspondant peut-être à l'assombrissement de la pigmentation. Le type 2, PAS positif, s'hypertrophie et s'hyperplasie rapidement en eau déminéralisée. Cette hyperactivité indiquerait un rôle dans l'osmorégulation. Le neuroséorétat s'accumule, puis tend à diminuer, la neurohypophyse étant de volume réduit. Le noyau préoptique paraît peu actif. L'hypothèse d'une faible utilisation des produits élaborés par l'axe hypothalamo-neurohypophysaire est envisagée. En plus de l'interrénal et des corpuscules de Stannius, l'hypothalamus et l'hypophyse interviennent donc dans la réponse de l'Anguille à une déminéralisation du milieu ambiant.

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Summary 45 male silver eels were kept from 32 hours to 46 days in demineralized (distilled or deionized) water. They show a darkening of the skin, a hypertrophy of the gall-bladder and some modifications of the hypothalamo-neuro-adenohypophysial system.In the rostral pars distalis, the prolactin cells regressed and appeared chromophobic. The corticotrophic cells are slightly stimulated and partly degranulated, in correlation with some activation of the interrenal, previously described. The thyrotrophic cells do not appear active at the end of the experiment, however the thyroid glands are not often inactive.In the proximal pars distalis, the gonadotrophic cells remain undifferentiated. The somatotrophic cells are strongly stimulated: a cellular, nuclear and nucleolar hyperactivity, without degranulation and a development of the ergastoplasm are observed. Their role in osmoregulation and metabolism of potassium is discussed.The pars intermedia is composed of two cell types: one of which is predominant, leadhematoxylin positive. It could elaborate the MSH (intermedin). In demineralized water, it is partly degranulated and progressively stimulated, perhaps in correlation with the darkening of the pigmentation. The other cell type is PAS positive, and not abundant in the control fish. In the treated fish, rapid hyperplasia and hypertrophy occur, with cytoplasmic degranulation and mitotic activity, so that it becomes predominant in this lobe after a month or more in demineralized water. Cytologic criteria indicate a great hyperactivity which seems to play a role in the processes of the osmoregulation; this hypothesis is discussed.The neurosecretory material tends to increase, as a storage, then to decrease in the neurohypophysis which is of a reduced volume. The preoptic nucleus does not appear active. In addition to the modifications of the interrenal and the corpuscles of Stannius previously described, the changes in the hypothalamus and the pituitary constitute evidence of their important role in the reaction of the eel to a demineralized environment.

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Ce travail a été réalisé avec la collaboration technique de Mademoiselle Jacqueline Olivereau, du C.N.R.S., que nous remercions bien vivement: elle nous a beaucoup aidée au cours de ces expériences, en particulier lors des changements quotidiens de l'eau distillée ou désionisée, et a réalisé toutes les préparations histologiques et les microphotographies illustrant cet article.     

A dimorphic Alu Sb-like insertion in COL3A1 is ethnic-specific

Alu elements are a class of repetitive DNA sequences found throughout the human genome that are thought to be duplicated via an RNA intermediate in a process termed retroposition. Recently inserted Alu elements are closely related, suggesting that they are derived from a single source gene or closely related source genes. Analysis of the type III collagen gene (COL3A1) revealed a polymorphic Alu insertion in intron 8 of the gene. The Alu insertion in the COL3A1 gene had a high degree of nucleotide identity to the Sb family of Alu elements, a family of older Alu elements. The Alu sequence was less similar to the consensus sequence for the PV or Sb2 subfamilies, subfamilies of recently inserted Alu elements. These data support the observations that at least three source genes are active in the human genome, one of which is distinct from the PV and Sb2 subfamilies and predates either of these two subfamilies. Appearance of the Alu insertion in different ethnic populations suggests that the insertion may have occurred in the last 100,000 years. This Alu insert should be a useful marker for population studies and for marking COL3A1 alleles.     

Interaction between cells and elastin fibers: An ultrastructural and immunocytochemical study

Mesenchymal cells (fibroblasts, smooth muscle cells) and endothelial cells were shown to interact with elastin fibers. The strong adhesion of elastin fibers to these cells is mediated by a cell membrane complex with a major glycoprotein component of 120 kDa designated as elastonectin. This interaction was studied by transmission electron microscopy (TEM) and immunocytochemical techniques using antibodies raised against the elastin adhesive proteins. When fibroblasts and smooth muscle cells were cultured in presence of elastin fibers, TEM showed an adhesion mechanism that takes place over several sites along the plasma membrane of these cells. Endothelial cells showed a very close association with elastin, emitting “pseudopodia” that embody the fibers. TEM, indirect immunofluorescence, immunoperoxidase, and confocal microscopy showed the presence and localization of cell membrane components synthesized in large quantities when cells were incubated in presence of elastin. Cells without elastin fibers barely revealed the adhesive membrane complex. These results confirm and extend previous findings concerning the presence of an inducible cell membrane complex that mediates the adhesion of elastin fibers to these cell types. © 1994 Wiley-Liss, Inc.     

Drosophila melanogaster acetylcholinesterase: Identification and expression of two mutations responsible for cold- and heat-sensitive phenotypes

Ace ^IJ29 and Ac ^IJ40 are cold- and heat-sensitive variants of the gene coding for acetylcholinesterase in Drosophila melanogaster. In the homozygous condition, these mutations are lethal when animals are raised at restrictive temperatures, i.e., below 23° C for Ace ^IJ29 or above 25° C for Ace ^IJ40. The coding regions of the gene in these mutants were sequenced and mutations changing Ser³⁷⁴ to Phe in Ace ^IJ29 and Pro⁷⁵ to Leu in Ace ^IJ40 were found. Acetylcholinesterases bearing these mutations were expressed in Xenopus oocytes and we found that these mutations decrease the secretion rate of the protein most probably by affecting its folding. This phenomenon is exacerbated at restrictive temperatures decreasing the amount of secreted acetylcholinesterase below the lethality threshold. In parallel, the substitution of the conserved Asp²⁴⁸ by an Asn residue completely inhibits the activity of the enzyme and its secretion, preventing the correct folding of the protein in a non-conditional manner.     

Ozone-induced microscopical changes and quantitative carbohydrate contents of hybrid poplar (Populus × euramericana)

Summary Cuttings of hybrid poplar (Populus × euramericana var. Dorskamp) were exposed to ozone (80 g/m³ from 2100 hours to 0700 hours, 180 g/m³ from 0700 hours to 2100 hours) for 3 months. Ozone reduced the starch content in leaves and stem bark, whereas starch granules accumulated in bundle sheath cells along small leaf veins. At the same time, sucrose and inositol content increased in the leaves. Mesophyll cells in the vicinity of the stomata were injured first, and droplet-like material appeared on their walls. In the sieve plates of fumigated trees, the pores showed a higher degree of narrowing than those of the control treatment. Cell collapse in the leaves was accompanied by water loss and an increase in air space. In the stems, the ozone treatment led to a reduced radial width, particularly in the xylem tissue. These results are discussed in relation to reduced or inhibited phloem loading and ozone-induced drought stress. The plants injured by ozone showed quite distinct patterns of metabolite responses as well as enzyme activities (PEP- and RubP-carboxylase) in the leaves from the top to the bottom. There were also remarkable differences in the reaction of sucrose and inositol between leaves and stem bark. Future research should therefore increasingly follow a whole-plant approach for a better understanding of complex plant reactions.     

Metabolite effectors for short-term nitrogen-dependent enhancement of phosphoenolpyruvate carboxylase activity and decrease of net sucrose synthesis in wheat leaves

It has been demonstrated previously that field pea (Pisum sativum L. cv. Express) grown in hydroponic culture on a complete nutrient solution with low NH₄⁺ concentrations (<0.5 mM) will produce a larger than normal proliferation of nodules. Peas grown in the absence of mineral N in hydroponic culture have been shown to rapidly autoregulate nodulation, forming a static nodule number by 14 to 21 days after planting. The present study further characterizes the effect of NH₄⁺ concentration in hydroponic culture on nodulation and nodule growth. Peas were grown continually for 4 weeks at NH₄⁺ concentrations that were autoregulatory (0.0 mM), stimulatory (0.2 mM) or inhibitory (1.0 mM), or peas were transferred between autoregulatory or NH₄⁺ inhibited and stimulatory solutions after 2 weeks. The peas nodulated as expected when grown under constant autoregulatory, stimulatory or inhibitory concentrations of NH₄⁺. When peas were transferred from the inhibitory (1.0 mM) to the stimulatory solution (0.2 mM) a massive proliferation of nodule primordia over the entire root system was observed within 3 days of the transfer. When they were transferred from the autoregulatory (0.0 mM) to the stimulatory (0.2 mM) solution a 10-day delay occurred before a proliferation in nodule primordia occurred at distal regions of the root system. These findings support our hypothesis that low concentrations (<1.0 mM) of NH₄⁺ in hydroponic culture cause a suppression of autoregulation in pea. In addition, the temporal and spatial differences in nodule proliferation between transfer treatments demonstrate at a whole plant level that autoregulation and NH₄⁺ inhibition suppress early nodule development via different mechanisms.     

Two genes present on a transposon-like structure in Lactococcus lactis are involved in a Clp-family proteolytic activity

The lactose-protease plasmid pUCL22 of Lactococcus lactis subsp. lactis strain CNRZ270 contained two inverted copies of IS 1076 flanking a region of 3.7 kb. This internal region was sequenced and found to contain two large open reading frames, ORF1 and ORFP in opposite orientations. ORF1 consists of 2289 bp; the deduced 763-amino-acid sequence is similar to the ATPases of the ClpA family. It contains two well-conserved consensus ATP-binding sites. It was named ClpL. ORFP consists of 930 bp encoding a protein of 310 amino acids. No similarity with any known protein was found in GenBank data for ORFP. Increased ATP-dependent proteolytic activity was detected in extracts from Escherichia coli cells expressing the clpL and ORFP genes.     

Differentiation and structural decline in the leaves and bark of birch (Betula pendula) under low ozone concentrations

Summary Leaf and bark structure of a birch clone (Betula pendula Roth) continuously exposed to charcoal-filtered air or charcoal-filtered air plus ozone (0.05, 0.075, 0.1 l 1^-1) was investigated throughout one growing season. Increasing ozone dose influenced leaf differentiation by reducing leaf area and increasing inner leaf air space, density of cells developing into stomata, scales and hairs. When approximately the same ozone dose had been reached, macroscopical and microscopical symptoms appeared irrespective of the ozone concentration used during treatment. Structural decline began in mesophyll cells around stomatal cavities (droplet-like exudates on the cell walls), continued with disintegration of the cytoplasma and ended in cell collapse. Epidermal cells showed shrinkage of the mucilaginous layer (related to water loss). Their collapse marked the final stage of leaf decline. When subsidiary cells collapsed, guard cells passively opened for a transitory period before collapsing and closing. With increasing ozone dose starch remained accumulated along the small leaf veins and in guard cells. IIK-positive grains were formed in the epidermal cells. This contrasted with the senescent leaves, where starch was entirely retranslocated. Injury symptoms in stem and petiole proceeded from the epidermis to the cambium. Reduced tissue area indicated reduced cambial activity. In plants grown in filtered air and transferred into ozone on 20 August, injury symptoms developed faster than in leaves formed in the presence of ozone. Results are discussed with regard to O₃-caused acclimation and injury mechanisms.     

Molecular analysis of the Lactococcus lactis subspecies lactis CNRZ270 bidirectional theta replicating lactose plasmid pUCL22

pUCL22 is the lactose protease plasmid of Lactococcus lactis ssp. lactis CNRZ270. The nucleotide sequence of its replication region Rep22 contains a non-transcribed region, the replication origin, followed by a gene encoding a putative 388-amino-acid protein named Rep22A. The promoter regions of the rep22A and pC194 cat genes share strong similarities and the pUCL22 replicon exerted trans or cis negative control on the pC194 cat gene expression in L. lactis. We suggest that Rep22A binds to its own promoter as well as to the pC194 cat promoter and thus is autoregulated. We show that pUCL22 replicates mainly by a bidirectional theta mechanism in L. lactis, and is representative of a widely distributed replicon family, members of which could be co-resident. We propose that compatibility between these closely related replicons results from minor replication protein modifications coupled with base changes in their respective binding sites, supporting the co-existence of numerous related replicons in lactococcal strains.