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Biomedical subjects

C Remacle

Publications and source records attributed to C Remacle.

At least 91 records · Page 5Linked to original sources
PubMed1989

The development of the fetal rat intestine and its reaction to maternal diabetes. II. Effect of mild and severe maternal diabetes.

Diabetes during pregnancy induces specifically structural and functional changes in the fetal endocrine pancreas. Other organs are affected as well. In this study, the fetal intestinal tract which is in close connection with the endocrine pancreas was analysed during diabetic gestation. The disease was induced by two different doses of streptozotocin which led to a mild or severe diabetic state in the mother. In fetuses from mildly diabetic as well as from severely diabetic rats, the time sequence in the appearance of the differentiated cells was identical and similar to that of controls. However, morphometric analysis of the intestine of fetuses from severely diabetic rats revealed a decrease in each of the parameters measured which led to a general hypotrophy of the intestine. In the fetuses from mildly diabetic rats, the values of the morphometric parameters of the duodenal mucosa remained unchanged and comparable to those of the control group. The vascularisation of the duodenum is modified in these fetuses because the volume density of the blood vessels is significantly increased. In conclusion, both diabetic states of the mother induce various alterations in the fetal intestine, including the blood vessels. The nature of the structural changes observed in the intestine could lead to modifications in the function of the entero-pancreatic system in these fetuses.

Animals↗
PubMed1989

High-affinity GABA uptake in a subpopulation of somatostatin cells in rat pancreas.

The aim of this study was to localize the high-affinity uptake of [3H]-GABA in Langerhans islets of rats aged 2.5, 7.5, and 75 days. On high-resolution autoradiography, cells presenting characteristic somatostatin granules were labeled, whereas others containing similar granules appeared nearly devoid of silver grains. Immunogold detection with antisomatostatin antibodies and high-resolution autoradiography suggested that uptake of GABA is indeed performed by somatostatin cells. To test the heterogeneity of uptake frequency in somatostatin cells, a second approach, coupling immunohistochemistry with anti-somatostatin, anti-PP, anti-glucagon, anti-glicentin, and anti-CCK antibodies, and low-resolution autoradiography, was applied on paraffin sections. It demonstrated that the uptake ability is not characteristic of all the somatostatin cells but of only a subpopulation of them. A few cells not immunoreactive to the anti-somatostatin antiserum also appeared to be able to take up GABA. Moreover, except for a rare few, the PP-glucagon-, glicentin-, and CCK-39-immunoreactive cells were not labeled by autoradiography.

Animals↗
PubMed1989

Effects of stimulation of adenylate cyclase and protein kinase-C on cultured fetal rat B-cells.

To study the maturation of fetal pancreatic B-cells, cell suspensions of pancreas from 21.5-day-old fetuses were cultured in RPMI medium containing 10 mM glucose. Forskolin (1 microM), used to stimulate adenylate cyclase, moderately delayed the neoformation of islets, slightly accelerated the proliferation of endocrine cells, and considerably increased insulin release by the cultures. The latter increase was not completely compensated for by the stimulation of insulin biosynthesis, so that the islet insulin content was decreased. The phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA; 25 nM), used to stimulate protein kinase-C, had little effect on the evolution of the cultures, but increased insulin release. This increase was almost compensated for by the stimulation of insulin biosynthesis. After 9-10 days of culture, insulin release in response to 15 mM glucose or 10 mM leucine was studied with perifused islets. In control islets, glucose produced a sustained increase in insulin release, which, however, was 6-fold smaller than that produced by leucine. Addition of forskolin or TPA to the perifusion medium markedly amplified the response to glucose without causing a biphasic pattern of release. In islets cultured with forskolin, the insulin response to glucose or leucine was decreased, largely owing to the lower insulin stores. In islets cultured with TPA, the insulin response to glucose or leucine was also decreased, but these differences cannot be explained simply by changes in insulin content. Neither treatment affected the kinetics of release. In conclusion, acute stimulation of adenylate cyclase or protein kinase-C markedly increased insulin release from fetal islets without causing an adult-like biphasic pattern of secretion. Chronic stimulation did not accelerate maturation of B-cells.

Adenylyl Cyclases↗
PubMed1988

In vitro cytodifferentiation of perinatal rat islet cells within a tridimensional matrix of collagen.

Cell suspensions prepared by collagenase digestion of pancreases obtained from rat fetuses (21.5 d old) and newborns (2.5 d old) were mixed with a collagen solution and inoculated on a collagen base layer. At the onset of the culture, most acinar cells became necrotic, whereas other epithelial cells proliferated. Most of the cell clusters arranged themselves into simple polarized structures composed of epithelial cells forming hollow spheres, and from these budded neoformed endocrine islets. Scarce fibroblasts were located close to these structures. Immunocytochemical localization of insulin and glucagon, as well as ultrastructural characteristics of the cell types revealed an intrainsular distribution similar to the in vivo localization. Tridimensional matrix of collagen offers, to perinatal pancreatic cells in culture, an environment close to the in vivo conditions: cells reorganize themselves in tissuelike structures and cell interactions concerned in the cytodifferentiation of pancreatic islets occur. This system allows for the study of undifferentiated epithelial cells--the presumed stem cells--differentiating and differentiated endocrine cells in the same preparation.

Animals↗
PubMed1988

The aging of the endocrine pancreas of the rat. I. Parameters of cell proliferation.

Morphometrical analysis of the endocrine pancreas of senile 30-month-old rats revealed that the volume density as well as the numeric density of islets of Langerhans were much lower than in 24-month-old rats, which coincided with a much higher percentage of pycnotic nuclei in islet cells. The proportion and localization of the different categories of endocrine cells (A, B, D and PP) remained however unchanged with aging. The apparent problem of cell renewal observed in vivo in the very old age was detected earlier in vitro by tritiated thymidine incorporation. Such experiments showed that 24-month-old islet cells had a decreased labelling index when compared to 3-month-old cells. The proliferation capacity of the old cells could be partially increased by changing the serum concentration or type. Similarly as being more sensitive to serum factors, these cells underwent also more pronounced negative influence of high oxygen pressure on replication. A stereological analysis of the ultrastructure of non-degenerated B-cell nuclei revealed that with age, the relative volume of the condensed chromatin increased progressively at the expense of the dispersed form. This suggests that the still functioning senile B-cells could reduce their transcriptional activity.

Aging↗
PubMed1988

The aging of the endocrine pancreas of the rat. II. Cytoplasmic parameters of the B-cell, including insulin synthesis and secretion.

Comparative ultrastructural stereology of 6 and 24-month-old rat B-cell cytoplasm revealed an increase with age in secondary lysosomes and a decrease in the volume density of RER and Golgi apparatus. The reduction of RER observed in freshly isolated islets could affect (pro)insulin biosynthesis in vitro: if the initial mobilization of precursor molecules for protein synthesis was the same, a delay was noted in their transit to the Golgi apparatus in B-cells of old islets. No further differences were seen in the autoradiographic distribution of radioactive amino-acids. More, the stock of insulin granules was similar in all age groups in both in vivo and in vitro conditions. Neither were any differences observed in the insulin secretion into culture media as well as during a subsequent incubation in supraphysiological glucose concentrations.

Aging↗
PubMed1988

Immunocytochemical localisation of GABA in endocrine cells of the rat entero-pancreatic system.

The occurrence of GABA-containing cells in the rat entero-pancreatic system was investigated by using anti-GABA-glutaraldehyde antibodies at the light and electron microscope level. In the pancreas, the B cells showed intense immunoreactivity, contrary to non-B and exocrine cells. Moreover, post-embedding immunogold staining was localised mostly in mitochondria, close to rough endoplasmic reticulum and in the nucleus. The insulin granules appeared nonsignificantly stained, which suggests the lack of cosecretion of GABA together with insulin. In the duodenum, GABA immunoreactivity was detected in certain endocrine cell types, suggesting a possible interaction with this amino acid. The well established GABAergic innervation in the enteric system was also confirmed by immunolabelling.

Animals↗
PubMed1987

Localization of high-affinity GABA uptake and GABA content in the rat duodenum during development.

The localization of high-affinity uptake sites for 3H gamma-aminobutyric acid (3H-GABA) was investigated in the rat duodenum during ontogenesis and also at the adult stage (from 15.5 days of fetal life up to 105 days post natum) by means of low- and high-resolution autoradiography. At all stages studied, specific endocrine cell types of the epithelium were labelled and an intense uptake was detected in the nervous tissue, especially in glial cells but also in scarce neurones. When the incubation medium was supplemented with beta-alanine (1 mM), a blocker of the glial uptake for GABA, the labelling persisted only in endocrine cells and in few neurones. The intensity and the frequency of the labelling decreased at later periods compared to the earlier developmental stages. The GABA content of the duodenum as measured by a new ion-exchange column chromatography-HPLC-coupled method was higher in the early postnatal period compared to later stages. These observations suggest that GABA, in addition to being a neurotransmitter, may play an important role during development of the duodenum.

Aging↗
PubMed1987

Ultrastructural aspects of streptozotocin cytotoxicity on rat pancreatic islets in vitro. Test of a protective effect of zinc.

Pancreatic islets, newly formed in vitro were incubated in the presence of streptozotocin (STZ; 0.4 mM) for up to 6 h. Ultrastructural changes first appeared between 2 and 4 h; heterochromatization, was followed by swelling of nuclear and reticular membranes, vesiculation of the Golgi apparatus, fragmentation of cell membranes and finally mitochondrial destruction. At the end of the experiment all the B cells were destroyed, whereas the other cell types remained intact. Exogenous ZnSO4 was added during preincubation periods to increase the intrainsular zinc content and to determine any protective effect against STZ-cytotoxicity. Since the addition of zinc had no obvious effect, it is suggested that STZ cytotoxicity on B cells cannot be attributed to competition for zinc between copper-zinc superoxide dismutase (Cu-Zn-SOD) and the crystallization of insulin.

Animals↗
PubMed1987

Effect of aging on the morphology of epididymal adipose tissue in the rat.

Quantitative morphometrical parameters were compared in epididymal adipose tissue of adult (6 months old) and old (24 months old) Wistar rats, using light and electron microscopy of embedded material and freeze-etch replicas, and taking into account the functional unit of adipose tissue: the capillary-adipocyte. Despite an insignificant reduction of the adipocyte number and size in old rats when the whole population was sampled, the confounding factor of size of adipocytes could be excluded from morphometric computations in adult and old rats. Morphometric measures were performed on the whole transit from capillaries to adipocytes. They revealed that the plasma membranes, as seen in freeze etching, and the thicknesses of endothelial and adipocyte cytoplasms, as seen in ultra-thin sections, remained unaltered with aging. By contrast, the basement membranes were changed but differently around capillaries and adipocytes. The capillary-adipocyte distances were shorter and the vascularization density was higher in old rats.

Adipose Tissue↗
PubMed1987

Lessons from the pathology of the diabetic pancreas.

Different events such as virus infections, toxins, nutritional factors, antiinsulin antibodies may be rendered responsible for the pathological changes in the pancreatic B-cell and result in a diabetic state in the postnatal, adolescent or adult age. These different interferences which may lead to the diabetic state need full consideration and assessment if prevention and cure are to be considered.

Adult↗
PubMed1986

Morphology and fibrinolytic activity of canine autogenous mesothelium used as venous substitute.

Autogenous mesothelium was used as venous substitute in ten dogs. Patches of mesothelium of three different origins were grafted into the anterior wall of the common iliac veins (CIV): peritoneum taken from and including the posterior rectus sheath (PRS), simple peritoneum (P) and mesentery (M). Animals were killed after 2, 4, 8, and 16 days and after 3 months. The segments of CIV, including the patches, were removed for study. On light microscopy, the PRS grafts showed a normal mesothelium but marked submesothelial fibrosis. The M and P grafts showed normal mesothelium and only mild fibrous thickening. On scanning electron microscopy, there was a perfect continuity of the mesothelial cells and the normal endothelium at the suture line. In the center of the graft, the cells had become elongated along the axis of blood flow. Fibrinolytic activity (FA) was measured by a standardized fibrin plate technique and quantitated in tissue activator units per gram of tissue (TAU/g). The mean FA of iliac vein specimens was 1101.7 +/- 133.3 TAU/g (mean +/- SEM). The mean FA determined before grafting for each kind of mesothelium was the following: PRS = 418.8 +/- 26.9 TAU/g; P = 873.0 +/- 107.1 TAU/g; M = 1142.3 +/- 91.4 TAU/g where only PRS showed values significantly lower than iliac vein mean FA (P less than 0.001). Postoperatively, the mesothelial FA, after an initial reduction, increased on day 4 and reached values significantly higher than the control values (1445.7 +/- 204.1 TAU/g tissue vs 853.1 +/- 62.3 TAU/g tissue; P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Cell interactions during the in vitro neoformation of fetal rat pancreatic islets.

As shown by scanning electron microscopy, transmission electron microscopy and membrane labeling analysis, the in vitro neoformation of rat pancreatic islets arose from two main processes: a budding from explants containing duct cells, and a competition between endocrine monolayers and fibroblasts on the culture substratum. The stronger cytoskeleton of fibroblasts and their higher adhesive properties, probably related to their more homogeneous distribution of membrane charges, may explain the spherization of the islets. The pure endocrine cell population of neoformed islets was composed mainly of insulin-secreting cells, and the other types of endocrine cells were distributed in the periphery. Preformed extracellular matrices of osmotically disrupted fibroblasts enhanced the yield of the cultures by increasing the anchorage of endocrine cells and slowing down the fibroblastic growth.

Animals↗
PubMed1986

Ultrastructural analysis of the in vitro differentiation of female rat preadipocytes.

In an attempt to characterize the preadipocytes of the adipose tissue of female rat, we studied by electron microscopy the differentiation of the cells into mature adipocytes in in vitro cultures. The preadipocytes arose from the stroma-vascular fraction of perirenal and perigenital adipose tissue. Culture of the preadipocytes in an enriched medium consisting of Dulbecco's medium supplemented with 10% fetal calf serum, antibiotics, rat triglycerides (0.5%), insulin (290 nM) and Tween 80 (0.1 mg/ml) induced their adipose conversion. The morphology of preadipocytes changed progressively. They accumulated fat granules, droplets and finally globules, which fused together. The cell organelles featured qualitative and quantitative modifications. The nucleus migrated with most mitochondria and a part of the Golgi system towards the cell periphery; the rough endoplasmic reticulum, dilated at the initial stage of differentiation became less and less conspicuous; the perinuclear Golgi system was dispersed between lipid droplets during fat accumulation; thick bundles of microfilaments, localized beneath the plasma membrane disappeared; large lipid droplets were surrounded by a network of microfilaments; many microvesicles and some "rosettes" typical of mature adipocytes could be observed. Nevertheless, the ultrastructural criteria did not allow to clearly discriminate the undifferentiated cells: early preadipocytes (without lipid droplets), adipoblasts and fibroblasts, all of these being probably present in the culture system.

Adipose Tissue↗