PubMed Health⌕ Search

Biomedical subjects

M Tardy

Publications and source records attributed to M Tardy.

At least 19 recordsLinked to original sources
PubMed1992

Transcriptional regulation of glial fibrillary acidic protein (GFAP)-mRNA expression during postnatal development of mouse brain.

During mouse brain maturation, GFAP-mRNA undergoes a two-step developmental expression. It increases between birth and day 15 (period of astrocytic proliferation) and then decreases until day 55 (period of astrocytic morphological differentiation). We have developed an in vitro transcription procedure, as a mean to study the part of transcriptional control in this biphasic expression. After RNA synthesis by endogenous RNA polymerases in nuclei isolated from mouse brain (of 3 to 55 days and 217 days), the relative rates of GFAP-mRNA transcripts were analysed by hybridization with a specific cDNA probe. As early as 3 days after birth, the rate of GFAP-mRNA transcripts was maximal, whereas unexpectedly, it showed a significant decrease in mice of 15 days and stayed low until the 55th day. Therefore, a transcriptional control may take place early in mouse brain postnatal development by increasing the transcriptional rate of the GFAP gene in astrocytes, and during the transition from proliferation to differentiation phase of astrocytes (that occurs at the 15th day after birth) by decreasing this rate. However, posttranscriptional events may also occur to modulate the level of the cytoplasmic GFAP-mRNA. In older mice (217 days), the low rate of GFAP-mRNA transcripts found is not concordant with the high cytoplasmic level generally observed in gliosis of the aging brain. Our data suggest posttranscriptional events at this age.

Aging↗
PubMed1992

Subcellular localization of two neurotropic drugs in three varieties of central nervous system cells by secondary ion mass spectroscopy (SIMS) microscopy.

The intracellular localization of two neurotropic drugs, flunitrazepam (benzodiazepine) and triflupromazine (phenothiazine), was studied by secondary ion mass spectrometry microscopy (SIMS) in three varieties of cells. The images of the intracellular distributions of the two drugs are easily obtained by selecting the fluorine atom of the molecules. These images show that the drug from the benzodiazepine group is mainly located in the nuclei, whereas the phenothiazine is exclusively located inside the cytoplasm.

Animals↗
PubMed1992

[Localization of two neurotropic molecules in cultured glial cells by ion microscopy].

The intracellular localization of two families of neurotropic drugs: flunitrazepam and flurazepam (benzodiazepine), triflupromazine and trifluoperazine (phenothiazine) has been studied by ion microscopy. The molecules have been incubated with C6 glioblastoma cells from rat origin and with astroglial primary cultures. The images of the intracellular distributions of the two drugs are easily obtained by selecting the fluorine atom of the molecules. The images obtained show that flunitrazepam and flurazepam, two drugs of the benzodiazepine group are mainly located to the nuclei, whereas triflupromazine and trifluoperazine, two phenothiazines are exclusively located inside the cytoplasm.

Animals↗
PubMed1992

Paradoxical sleep deprivation increases glutamine synthetase in rat brain.

The modifications of glutamine synthetase (GS) level, an enzyme mainly located in astrocytes, were investigated in rat after paradoxical sleep deprivation and during recovery. An immunotitration method was used to evaluate the relative level of GS in brain tissue. At the end of a 24 hrs. paradoxical sleep deprivation, a significant increase in GS level was observed both in the frontoparietal cortex and the locus coeruleus area. 4 hrs. later, during recovery, the GS level returned to control level in the cortex but was lower in the locus coeruleus area.

Animals↗
PubMed1991

Primary dissociated cell culture of embryonic rat metencephalon: presence of GABA in serotonergic neurons.

This study was performed to determine whether neurons, where gamma-aminobutyric acid (GABA) and serotonin (5-HT) coexist, represent neuronal entities which can survive in vitro. In dissociated cultures from 18-day-old embryonic rat metencephalon, it was possible to develop glial and neuronal cells. Among the neurons, some of them, which contain glutamate decarboxylase or are capable of accumulating [3H]GABA are GABAergic; others, containing tryptophan hydroxylase or 5-HT are serotoninergic. By combining radioautography and immunocytochemistry, it was possible to observe neurons where 5-HT and GABA coexist. Cultures might be a suitable model to study the functioning (release or synthesis of both neurotransmitters) of neurons where two classical neurotransmitters coexist.

Animals↗
PubMed1991

Isolation and characterization of two different molecular forms of basic fibroblast growth factor extracted from human placental tissue.

Basic fibroblast growth factor (bFGF) was purified to homogeneity from human placental tissue on a semi-large scale. Placental bFGF consists of two proteins of apparent molecular masses 16,000 and 18,000 dalton, as determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis under non-reducing conditions. Microsequence analysis showed that both proteins have the same N-terminal sequence Pro-Ala-Leu-Pro-Glu-Asp-Gly-Gly-Ser-Gly-Ala-Phe..., which is identical with that of (1-146) bFGF extracted from human brain. After reduction by dithiothreitol or mercaptoethanol, placental bFGF appears as a single protein of 16,000 dalton. The reduced protein displays the same ability to stimulate the proliferation of CCL39 fibroblasts as the non-reduced doublet. These data indicate that bFGF extracted from placental tissue consists of two proteins with different apparent molecular masses which do not differ in their N-terminal sequence but in their oxidation state.

Amino Acid Sequence↗
PubMed1991

Effect of thyroid deficiency on glial fibrillary acidic protein (GFAP) and GFAP-mRNA in the cerebellum and hippocampal formation of the developing rat.

The concentrations of glial fibrillary acidic protein (GFAP) and its encoding mRNA in the cerebellum and hippocampal formation were assayed during the development of normal and hypothyroid rats. Neonatal hypothyroidism induced a significant reduction in the GFAP concentration in both regions from day 14. The reduction was especially marked on day 35 in the cerebellum (-43%) and the hippocampal formation (-55%). The immunocytochemical study of vimentin showed that the developmental disappearance of this protein from the Bergmann and internal astrocytes is greatly delayed in the cerebellum of the hypothyroid rats. The reduction in GFAP concentration together with the delayed vimentin-GFAP transition could explain how astrocyte morphogenesis is impaired by neonatal thyroid deficiency. The GFAP-mRNA concentration in the hippocampal formation was reduced throughout the development of thyroid-deficient rats, while the GFAP-mRNA concentration in the cerebellum first increased between birth and day 14 to reach a peak well above the normal value (+78%) and decreased thereafter to reach 53% of the normal value by day 35. This transient increase in the cerebellar GFAP-mRNA concentration may be related to the astroglial hyperplasia that occurs in these animals. The difference between the developmental profile of GFAP and its encoding mRNA, especially under pathological conditions, indicates that two distinct mechanisms control the synthesis or stability of the protein and its messenger RNA, as was previously found in the forebrain of the developing normal rat.

Animals↗
PubMed1991

DBcAMP effect on the expression of GFAP and of its encoding mRNA in astroglial primary cultures.

Short term and chronic dBcAMP effects on the expression of glial fibrillary acidic protein (GFAP) in astroglial primary cultures are investigated. Short (48 h) and long (more than 7 days) treatments with the cAMP derivative induce both cell shape changes and an increase in GFAP immunolabelling. Such effects are only associated with an increase in GFAP and in GFAP-mRNA levels in the long term treatment. These results suggest that the short term effect of dBcAMP induces post-translational modifications of the protein whereas the long term effect is associated with an increase in GFAP mRNA transcription and/or stability.

Animals↗
PubMed1991

In vitro and post-transplantation differentiation of human keratinocytes grown on the human type IV collagen film of a bilayered dermal substitute.

Using human type IV and type I + III collagens and a new, nontoxic cross-linking procedure, we have developed a cell-free bilayered human dermal substitute for organotypic culture and transplantation of human skin keratinocytes. We have studied the formation of the basement membrane, and the differentiation of keratinocytes grown on the type IV collagen layer of this dermal substitute, in vitro and after grafting onto nude mice. These studies demonstrated the formation of essential constituents of the basement membrane in culture: hemidesmosomes and deposition of extracellular matrix on the top of the type IV collagen were observed as early as 6 days after plating of human keratinocytes. Although the keratinocytes formed a well-organized multilayered epithelium, they exhibited limited differentiation when grown submerged in liquid medium. However, the multilayered sheet obtained after 14 days in submerged culture was composed of a regular basal cell layer, several nucleated suprabasal cell layers containing granular cells, and several dense, anucleated cell layers. The grafting experiments have shown a good biocompatibility of the dermal substitute. It is repopulated by fibroblasts, newly synthesized collagen, vessels, and a few mononuclear cells. At Day 14 after grafting, the type IV collagen layer was still present and very dense, and the basement membrane appeared as in culture, with numerous well-structured hemidesmosomes and deposition of extracellular matrix resembling lamina densa. At Day 55 after transplantation, even if the epidermal graft did not exhibit all the characteristics of the normal epidermis in vivo, it was very close to it. At this stage, the basement membrane was complete, with structures clearly indicative of anchoring fibrils. This new dermal substitute offers many advantages. It is stable and easy to handle. Its production is standardized. The oxidation induced by periodic acid led to a nontoxic cross-linked matrix. This dermal substitute is the first one entirely composed of human collagens. The type I + III collagen underlayer is reorganized when grafted. It supports a type IV collagen top layer which offers an excellent substrate for keratinocytes, favors their anchorage, and favors the formation of the basement membrane in vitro. This dermal substitute could be useful for wound coverage or as an in vitro model for toxicological and pharmacological studies.

Basement Membrane↗
PubMed1991

Expression of various microtubule-associated protein 2 forms in the developing mouse brain and in cultured neurons and astrocytes.

A cDNA probe specific to microtubule-associated protein 2 (MAP2) was used to study the expression of the mRNAs encoding the high- and low-molecular-weight MAP2 variants in cultured neurons and astrocytes. The timing and relative abundance of these MAP2 transcripts and of their encoded proteins were also studied in the developing cerebral hemispheres and cerebellum of the mouse. A 9-kb mRNA, known to encode high-molecular-weight MAP2, was expressed in cultured astrocytes, albeit at a lower level than in neurons. The 6-kb transcript, recently shown to encode low-molecular-weight MAP2 (MAP2c), was expressed in neurons and was the predominant MAP2 transcript of the astrocytes. The level of the 9- and 6-kb transcripts decreased at late stages of astroglial and neuronal cell culture. The 9-kb mRNA was detected in the cerebellum and cerebral hemispheres at every developmental stage. Although the levels of this mRNA varied slightly in the cerebral hemispheres, its expression was biphasic in the cerebellum. This might be explained by the differences in timing of development of the various neuronal cell types formed in these two brain areas. The 6-kb transcript was detected only at early developmental stages in the two brain areas. Correlating the temporal expression of the 9-kb mRNA to that of high-molecular-weight MAP2 indicates that the accumulation of this protein is in part regulated at a cytoplasmic level.

Aging↗
PubMed1990

GFAP turnover during astroglial proliferation and differentiation.

The expression and turnover of the glial fibrillary acidic protein (GFAP) were studied in astroglial primary cultures during postnatal proliferation and maturation. 1. Immunocytochemical studies demonstrated that in immature proliferating astrocytes. GFAP was expressed as a filamentous organized crown around the nucleus whereas in the maturating cells, a GFAP labelled network began to radiate throughout the cytoplasm and GFAP was highly expressed in the astroglial processes. 2. GFAP turnover was studied at 3 periods of culture. The decay of radioactivity from prelabelled GFAP was followed from day 4-12 (immature stage), 11-19 (maturing stage) and 21-29 (morphologically differentiated stage). GFAP displayed a biphasic decay kinetic at each considered period. Two pools of GFAP distinctly appeared. The first one was a fast decaying pool with a half life of 16-18 h and of 5-6 days for the stable one. The unstable pool decreased from 70% to 30% of the total incorporated radioactivity from the proliferating stage to the most mature stage, whereas the stable pool increased proportionally.

Animals↗
PubMed1990

Neuron-glia interactions: effect of serotonin on the astroglial expression of GFAP and of its encoding message.

The trophic effect of serotonin on the glial fibrillary acidic protein (GFAP) expression was investigated in rat brainstem astrocytes in primary culture. GFAP immunolabelling decreased and gliofilaments appeared localized in the cytoplasm periphery. GFAP protein level decreased in parallel with a decrease in its encoding message. Serotonin may act as an inhibitor of GFAP expression either on the transcription or on the stability of the GFAP-mRNA.

Animals↗
PubMed1990

Glutamine synthetase modulation in astrocyte cultures of different mouse brain areas.

Astroglial cells from mouse cerebral hemispheres, cerebellum, olfactory bulbs, and medulla oblongata were grown in the presence of either hormones (hydrocortisone, insulin) or cell second messengers (dBcAMP, dBcGMP). Glutamine synthetase (GS) specific activity, GS protein level, and GS translation were investigated under the effect of these factors. Hydrocortisone produced a simultaneous increase in GS translation, GS level, and activity. This increase was observed in the astrocytes cultured from the four brain areas but at a variable magnitude depending on the area. The hydrocortisone effect appeared at the transcriptional level. Inversely, insulin decreased both the GS activity and the in vitro translated GS. This effect was seen only in the olfactory bulbs and the medulla. DBcAMP increased the GS biological activity only in the cerebral hemisphere cultures. It raised, however, the level of translated GS and GS protein in astrocytes from all the areas, suggesting a post-translational effect for intracellular cAMP. DBcGMP only affected GS in the astrocytes from cerebral hemispheres and the medulla modulating either the GS transcription or the messenger RNA stability. These results suggest specific regulation for GS expression, depending on the brain area from which the cells were dissociated or on the astroglial cell population present in these cultures affecting either the transcription, the mRNA stability, or the biological activity of the protein.

Animals↗
PubMed1990

Fibroblast behavior on gels of type I, III, and IV human placental collagens.

Various collagens were extracted and purified from human placenta after partial pepsin digestion. We prepared type III + I (57:43), enriched type I, type III, and type IV collagens on an industrial level, and studied their biological properties with MRC5 fibroblast cells. Using the process of contraction of a hydrated collagen lattice described by Bell, we found tha the contraction rate was dependent on collagen type composition. The contraction was faster and more pronounced with pepsinized type I collagen than with pepsinized type III + I (57:43) collagen; the lowest rate was obtained with the pepsinized type III collagen. Using a new technique of collagen cross-linking, a gel was made with type IV collagen. This cross-linking procedure, based on partial oxidation of sugar residues and hydroxylysine by periodic acid, followed by neutralization, resulted in an increased number of natural cross-link bridges between oxidized and nonoxidized collagen molecules, without internal toxic residues. The fibroblasts were unable to contract type IV/IVox collagen gels. The type IV/IVox collagen gel was transparent and its amorphous ultrastructure lacked any visible striated fibrils. Fibroblast cells exhibited atypical behavior in these type IV/IVox collagen gels as evidenced by optical and electron microscopy. The penetration of fibroblasts could be measured. Fibroblasts penetrated faster in type IV/IVox collagen gels than in untreated type III + I collagen gels. The lowest rate of penetration was obtained with cross-linked type III + I gels. Fibroblast proliferation was similar on untreated or cross-linked type III + I collagen gels and slightly increased on type IV/IVox collagen gels, suggesting that this cross-linking procedure was not toxic.

Cell Division↗
PubMed1990

[Use of human placental type IV collagen in tympanoplasty. Results of initial animal experimentation].

This study presents the use of human type IV collagen film in myringoplasty in dogs. Evaluation criteria were clinical (repeated otoscopies) and histological (histological analysis of cicatrised membranes). This new biomaterial should seem to be a technological advance on temporal bone aponevrosis or perichondria auto-grafts. This study constitutes the necessary preliminary research to any application in man.

Animals↗
PubMed1989

Comparative marker analysis of the ependymocytes of the subcommissural organ in four different mammalian species.

The subcommissural organ (SCO), classified as one of the circumventricular organs, is composed mainly of modified ependymal cells, attributable to a glial lineage. Nevertheless, in the rat, these cells do not possess glial markers such as glial fibrillary acidic protein (GFAP), protein S100, or the enzyme glutamine synthetase (GS). They receive a synaptic 5-HT input and show pharmacological properties for uptake of GABA resembling the uptake mechanism of neurons. In this study, we examine the phenotype of several mammalian SCO (cat, mouse, rabbit) and compare them with the corresponding features of the rat SCO. In all these species, the SCO ependymocytes possess vimentin as an intermediate filament, but never express GFAP or neurofilament proteins. They do not contain GS as do glial cells involved in GABA metabolism, and when they contain protein S100 (rabbit, mouse), its rate is low in comparison to classical glial or ependymal cells. Thus, these ependymocytes display characteristics that differentiate them from other types of glial cells (astrocytes, epithelial ependymocytes and tanycytes). Striking interspecies differences in the capacity of SCO-ependymocytes for uptake of GABA might be related to their innervation and suggest a species-dependent plasticity in their function.

Animals↗