Effect of altered hormonal balance in the mother-fetus system on body weight, the adrenal glands, thymus gland and peripheral blood leukocyte composition in the offspring

Pregnant rats have been injected intramuscularly with hydrocortisone-acetate or desoxycorticosterone-acetate (DOCA) for the III or the II-III trimesters of pregnancy. In the last case the animals are given not only greater doses of the hormones, but during a longer period. By the end of the pregnancy the drug dose decreases. In mother-rats after hydrocortisone injections the adrenals mass increases; after DOCA the body mass increases, and that of the adrenals drops. In the offspring hydrocortisone produces decline of the thymus and adrenals mass, as well as neutrophilic leucocytosis, lympho-, eosino-++- and monocytopenia. Just the opposite, DOCA results in lympho- and monocytosis, in increasing morphofunctional activity of monocytes. Common effects in hormonal action is demonstrated as underdevelopment of the adrenals, in decreasing thymus mass, in development of neutrophilic leucocytosis and eosinopenia. With increasing doses and duration of prenatal injections of corticosteroids in rats the mass of the adrenals and thymus drops even greater, stimulating effect of the hormones on the neutrophilic granulocytopoiesis decreases.     

Participation of tissue basophils of the parathyroid glands in the regulation of the morphofunctional state of the organ

By means of cytometry estimation of mitotic index and nucleo-cytoplasmic relation of parathyrocytes, parathyroid glands have been studied in 52 white rats and 28 white mice under conditions of intal inhibition (30 mg/kg of body mass twice a day intraperitoneally) of tissue basophils (TB) secretory activity. For stimulation of the glands, repeated injections of trilon B and hemiparathyroidectomy are used. The experiment lasts for 3.5 days. In the rat parathyroid glands, containing a considerable amount of TB, intal does not produce any important effect of quantitative parameters of the organ's structure in intact animals, nevertheless it prevents development of hypertrophy of the parathyrocytes in the stimulated glands, as well as makes weaker the proliferative response of the cells to hemiparathyroidectomy. In mice, their parathyroid glands containing single TB, under conditions of stimulation of the parathyroid function, intal does not produce any inhibitory effect to growth of middle size parathyrocytes. The results obtained demonstrate that the parathyroid TB actively participate in regulation of the organ's morphofunctional state, intensifying the stimulating effect of hypocalcaemia to the parathyroid parenchyma.     

Effect of total profound hypothermia on the relation between cells in the skin of the white rat at different ages

In experiments performed on 190 white inbred rats of various age, effects of total deep hypothermia on mitotic activity of basal epidermocytes, quantity of non-pigmented granular dendrocytes and lymphocytes of epidermis, tissue dermal basophils has been studied. Age difference in changes of the parameters examined at cold effect have been stated. A suggestion is made on existence in the skin of a local regularity system "basal epidermocyte--non-pigmented granular dendrocyte--intraepidermal lymphocyte--tissue dermal basophils", participating in regulation of protective properties of the organ.     

Effects of estradiol and progesterone on accumulation of relaxin- and carbohydrate-containing granules in endometrial gland cells of the guinea pig

Guinea pigs were spayed and given various regimens of injections of estradiol and progesterone. The following were monitored in each animal: pubic separation (relaxin stimulation), resorption of the vaginal membrane (estrogen priming), and the presence of PAS-positive granules and/or relaxin in endometrial gland cells (EGC). Injections of estradiol alone resulted in resorption of the vaginal membrane, accumulation of PAS-positive granules in EGC of all animals, and accumulation of relaxin in EGC of two of four animals; but they did not cause pubic separation. Progesterone injections did not result in resorption of the vaginal membrane, separation of pubes, or accumulation of PAS-positive granules; however, one of three animals demonstrated a few EGC that contained relaxin. Animals that received both estradiol and progesterone exhibited PAS-positive granules and relaxin in EGC as well as separated pubes, but did not have resorbed vaginal membranes. Upon examination with the electron microscope, EGC from animals that received estradiol alone exhibited remarkable numbers of secretory granules that contained a carbohydrate-rich material. Secretory granules were not prominent in EGC from animals that received progesterone alone. Estradiol and progesterone injections resulted in accumulation of secretory granules in EGC that contained relaxin and a carbohydrate-rich material. The observations that estradiol and progesterone induce relaxin production in EGC support the hypothesis that uterine relaxin plays an important role in pregnancy and/or parturition in the guinea pig.     

Tissue basophils in the thymus (luminescence microscopic and immunologic analysis)

By means of fluorescent method biogenic amines in the thymus tissue basophils have been investigated in intact and immunized rats and, simultaneously, in the peripheral blood of the immunized animals the hemolysins titer has been defined as an index of the immune response of the organism. In the immunized rats the number of the tissue basophils is increasing, there are many degranulated forms among them. Hence, the antigenic stimulation facilitates to the tissue basophils secretory activation, which is morphologically manifested as their degranulation. The hemolysins titer level in the peripheral blood is gradually decreasing, as the time of immunization increases. The content of biogenic amines in the tissue basophils does not undergo similar changes.     

Histochemical studies of acid proteoglycans and glycoproteins and activities of hydrolytic and oxidoreductive enzymes in the skin epidermis of the fish Blennius sanguinolentus pallas (Teleostei: Blenniidae)

In the epidermis of the fish Blennius sanguinolentus the histochemistry of complex carbohydrates and various oxidoreductases has been studied by means of a series of selected light microscopical techniques. The epidermis is endowed with three types of secretory cells namely, the mucous goblet cells, the superficial polygonal cells and the ionocytes, which provide protective functions in view of their involvement in the prevention of the skin epithelium from invading pathogens and in the osmoregulation processes respectively. The secretory substances of mucous goblet cells contain sulfated, carboxylated and neutral complex carbohydrates in addition to a glycoprotein with sialic acid terminal to galactose in its oligosaccharide chains. Activities of SDH, ICDH, MDH and G-6-PDH were studied, to elucidate some aspects of the correlated functions of the ionocytes which play a key role in the performance of the maintenance of the electrolyte pattern in the internal milieu of the skin. Differences in the intensity of various oxidoreductases are correlated with the extent of the activity of the epidermis for related to secretion of a mucous cover over the surface. Activity of the oxidoreductases is confined mainly to the basal and outer epidermal layers and this enzyme zonation is discussed with reference to the existence of the high turnover rate of the epithelial cells and both the proliferative and respiratory capabilities of the skin epithelium.     

Morphofunctional characteristics of microcirculation in the skin of girls 5-17 years of age

In 261 girls year-to-year morphofunctional transformations of spatial composition of the skin microcirculatory bed have been studied at rest and after a dynamic local load. By means of biomicroscopy main regularities in development of the skin capillary network have been revealed in the nail torus in the postnatal ontogenesis. Formation of the microvessels reactivity during various age periods and maturation of mechanisms of the compensatory-adaptive reactions are connected with formation of the definitive composition of the microcirculatory bed, that in girls corresponds to 11-12 years. Qualitative transformations in the skin capillary network bring certain quantitative changes in the structural microcirculatory parameters--increasing diameter of microvessels and increasing density of functioning capillaries.     

Comparative characteristics of villous chorionic polysaccharides during the first 3 months of human uterine and tubal pregnancy]

Qualitative composition of polysaccharides has been studied in 100 chorions of human fetuses, 3.5--12 weeks of embryonic development, 40 of them have been obtained from tubal pregnansies. It has been stated that in villous stroma of the tubal pregnancy at the 4--5th week of gestation, sulphate polysaccharides appear that demonstrates an earlier maturation of villi at this type of pregnancy. It is possible that in the formation of intracellular substance of the villous stroma together with fibroblasts Kastschenko -- Hoffbauer's cells also participate. A characteristic feature for the tubal pregnancy is the appearance of distrophically changed villi beginning from the 4th week of gestation. In plasmodiotrophoblast of such villi the amount of Hale-positive and PAS-positive substances is sharply decreasing, and neutral polysaccharides, chondroitin-4-sulfate, chondroitin-6-sulfate and dermatan-sulfate (small amount) are intensively accumulating in the stroma.     

Secretory leukocyte protease inhibitor is a novel inhibitor of fibroblast-mediated collagen gel contraction

Cultured epithelial cells, including those from the oral epithelium, have been successfully applied in the promotion of scarless wound healing. Factors released from the epithelial cells are thought to contribute significantly to the beneficial effects. In the conditioned medium of human oral epithelial cells, we found a factor that inhibited fibroblast-mediated collagen gel contraction, an in vitro model of wound healing and scar formation. Biochemical analysis identified the factor to be human secretory leukocyte protease inhibitor (SLPI). Fibroblasts transfected with SLPI cDNA showed reduced gel-contracting activity. SLPI purified from the conditioned medium inhibited gel contraction in a dose-dependent manner, and anti-SLPI antibody counteracted this activity. Upon SLPI treatment, human skin fibroblasts in collagen gel became shorter in length and were inhibited in pseudopodia extension. Furthermore, after SLPI treatment, alpha(1)-integrin immunoreactivity decreased, and cyclic AMP levels increased. Excessive gel contraction was observed when fibroblasts treated with TGF-beta1 and fibroblasts from hypertrophic and from keloid scar tissue were cultured in collagen gel. SLPI was also effective in inhibiting gel contraction in the above three models of scar formation. These results suggest that SLPI may be useful in promoting scarless wound healing.     

Developmentally and regionally regulated participation of epidermal cells in the formation of collagen lamella of anuran tadpole skin

We investigated the cellular mechanism of formation of subepidermal thick bundles of collagen (collagen lamella) during larval development of the bullfrog, Rana catesbeiana, using cDNA of alpha1(I) collagen as a probe. The originally bilayered larval epidermis contains basal skein cells and apical cells, and the collagen lamella is directly attached to the basement membrane. The basal skein cells above the collagen lamella and fibroblasts beneath it intensively expressed the alpha1(I) gene. As the skin developed, suprabasal skein cells ceased expression of the gene. Concomitantly, the fibroblasts started to outwardly migrate, penetrated into the lamella and formed connective tissue between the epidermis and the lamella. These fibroblasts intensively expressed the gene. As the connective tissue developed, the basal skein cells ceased to express the gene and were replaced by larval basal cells that did not express the gene. These dynamic changes took place first in a lateral region of the body skin and proceeded to all other regions except the tail. Isolated cultured skein cells expressed the gene and extracellularly deposited its protein as the type I collagen fibrils. Thus, it is concluded that anuran larval epidermal cells can autonomously and intrinsically synthesize type I collagen.     

Changes in the epithelium of the proximal part of the nephron tubule after bile loss by the body

As demonstrate the experiments performed in 45 male rats of Wistar line, a complete bile loss by the organism results in 24-72 h in certain distrophic changes of epithelium in the proximal parts of the nephron canaliculi. The brush border microvilli undergo destruction, the cellular membranes disorganize, cytoplasm of epitheliocytes vacuolizes up to their complete rupture. The PAS-positive reaction of the basal membrane in the canaliculi increases. In cytoplasm of epitheliocytes in the straight portions of the canaliculi of the distal parts of the nephron glycogene is deposited. Alkaline phosphatase, succinate dehydrogenase, cytochromoxydase activity decreases, and that of acidic phosphatase increases. The changes advance with increasing time from the beginning of bile loss.     

Morphological and histochemical studies on a PAS-positive granular leukocyte in blood and connective tissues of Catostomus commersonii Lacépède (teleostei:pisces).

This study was undertaken to identify an ubiquitous granular leukocyte found in Catostomus commersonni Lacépède. The cell contains large, numerous, strongly PAS-positive cytoplasmic granules, an eccentric nucleus and prominent, persistent juxtanuclear space. It develops in the hemopoietic tissue of the kidney, and mature cells are found not only in kidney and peripheral blood but also in areas of connective tissue where mast cells are usually located. Electron microscopy confirms the presence of a large Golgi apparatus, unlamellated cytoplasmic granules and extensive rough-surfaced endoplasmic reticulum. Histochemical studies show that the cytoplasmic granules are alcianophobic, non-metachromatic and unstained by acridine orange. Histamine is detectable spectrophotometrically in kidney tissue, but the PAS-positive granular leukocyte does not consistently degranulate after treatment with histamine liberator 48/80. The authors suggest that while the PAS-positive granular leukocyte is not identical with classical basophils/mast cells, which are absent in C. commersonnii, it may represent an evolutionary precursor of these cells.     

Elevated prolactin redirects secretory vesicle traffic in rabbit lacrimal acinar cells

During pregnancy, lymphocytes infiltrating the rabbit lacrimal gland disperse to the interacinar space from their normal focal concentrations, basal fluid secretion decreases, pilocarpine-induced fluid secretion increases, and stimulated fluid protein concentration decreases. Ductal epithelial cell prolactin (PRL) content increases and redistributes from the apical to the basal-lateral cytoplasm. A replication-incompetent adenovirus vector for rabbit PRL (AdPRL) was used to test the hypothesis that increased intracrine/autocrine PRL signaling alters secretory protein traffic in an ex vivo lacrimal acinar cell model. AdPRL had no discernable influence on microtubules or actin microfilaments or their responses to carbachol (CCh). Endogenous and transduced PRLs exhibited similar, nonpolarized, punctate distributions. Cells secreted PRL consititutively and at increased rates in response to CCh. In contrast, constitutive secretion of beta-hexosaminidase was negligible, suggesting that the constitutive pathway for PRL is relatively inaccessible to typical secretory proteins. AdPRL had no significant effect on total secretion of beta-hexosaminidase or syncollin-green fluorescent protein (GFP), a chimeric secretory protein construct. However, it reversed the polarized distributions of vesicles containing rab3D and syncollin-GFP. Live-cell imaging indicated that AdPRL redirected CCh-dependent syncollin-GFP exocytosis from the apical plasma membrane to the basal-lateral membrane. Elevated concentrations of exogenous rabbit PRL in the ambient medium elicited similar changes. These observations suggest that elevated PRL, as occurs in the physiological hyperprolactinemia of pregnancy, induces lacrimal epithelial cells to express a mixed exocrine/endocrine phenotype that secretes fluid to the acinus-duct lumen but secretes proteins to the underlying tissue space. This phenotype may contribute to the pregnancy-associated immunoarchitecture.     

Structuro-functional organization of the blood-brain barrier

The analysis of the authors and published data allowed to conclude that the blood-brain barrier was a complex structural-functional system. Its basic components are endothelial cells, basal membrane, pericytes, tissue basophils, and astrocytes. The role of these structural elements in the permeability of blood-brain barrier has been studied. Organ-specific peculiarities of the brain tissue basophils have been established in rats, cats, and dogs. The astrocyte-capillary complexes were found to be very reactive, and at the same time, very plastic CNS structures.     

The differential proliferative response of fetal and adult human skin fibroblasts to TGF-β is retained when cultured in the presence of fibronectin or collagen

Background

Transforming growth factor-β is a multifunctional and pleiotropic factor with decisive role in tissue repair. In this context, we have shown previously that TGF-β inhibits the proliferation of fetal human skin fibroblasts but stimulates that of adult ones. Given the dynamic reciprocity between fibroblasts, growth factors and extracellular matrix (ECM) in tissue homeostasis, the present study aims to investigate the role of fibronectin and collagen in the proliferative effects of TGF-β on fetal and adult cells.

Methods

Human fetal and adult skin fibroblasts were grown either on plastic surfaces or on surfaces coated with fibronectin or collagen type-I, as well as, on top or within three-dimensional matrices of polymerized collagen. Their proliferative response to TGF-β was studied using tritiated thymidine incorporation, while the signaling pathways involved were investigated by Western analysis and using specific kinase inhibitors.

Results

Fetal skin fibroblast-proliferation was inhibited by TGF-β, while that of adult cells was stimulated by this factor, irrespective of the presence of fibronectin or collagen. Both inhibitory and stimulatory activities of TGF-β on the proliferation of fetal and adult fibroblasts, respectively, were abrogated when the Smad pathway was blocked. Moreover, inhibition of fetal fibroblasts was mediated by PKA activation, while stimulation of adult ones was effected through the autocrine activation of FGF receptor and the MEK–ERK pathway.

Conclusions

Fetal and adult human skin fibroblasts retain their differential proliferative response to TGF-β when cultured in the presence of fibronectin and unpolymerized or polymerized collagen.

General significance

The interplay between TGF-β and ECM supports the pleiotropic nature of this growth factor, in concordance with the different repair strategies between fetuses and adults. This article is part of a Special Issue entitled Matrix-mediated cell behaviour and properties.     

Prostaglandin F2 alpha output by human endometrium under superfusion and organ culture conditions

Specimens of proliferative and secretory human endometrium were incubated under organ culture or superfusion conditions and the levels of PGF2 alpha in the medium were measured by radioimmunoassay. Basal rates of PGF2 alpha output during short-term superfusions and long-term (1-2 day) batch incubations, performed on the same tissue specimens, were similar. Basal output of PGF2 alpha by proliferative endometrium (230-280 ng/mg protein X d) was significantly higher than that of secretory tissue under both experimental conditions. Estradiol (10(-8) M) increased PGF2 alpha output significantly (4-fold) only in secretory endometrium under organ culture conditions; Progesterone (10(-7) M) decreased it significantly (to 1/2-1/4 of the basal level) in both types of endometria during long-term incubations and in proliferative endometrium during superfusion. Glands isolated from proliferative and secretory endometrium produced PGF2 alpha during superfusion at a rate comparable to that of endometrial tissue under similar conditions. PGF2 alpha output by glands isolated from secretory endometrium increased significantly (3-fold) when estradiol was added to the superfusion medium.     

Poly(L-lactide-co-glycolide) thin films can act as autologous cell carriers for skin tissue engineering

Degradable aliphatic polyesters such as polylactides, polyglycolides and their copolymers are used in several biomedical and pharmaceutical applications. We analyzed the influence of poly(L-lactide-co-glycolide) (PLGA) thin films on the adhesion, proliferation, motility and differentiation of primary human skin keratinocytes and fibroblasts in the context of their potential use as cell carriers for skin tissue engineering. We did not observe visible differences in the morphology, focal contact appearance, or actin cytoskeleton organization of skin cells cultured on PLGA films compared to those cultured under control conditions. Moreover, we did not detect biologically significant differences in proliferative activity, migration parameters, level of differentiation, or expression of vinculin when the cells were cultured on PLGA films and tissue culture polystyrene. Our results indicate that PLGA films do not affect the basic functions of primary human skin keratinocytes and fibroblasts and thus show acceptable biocompatibility in vitro, paving the way for their use as biomaterials for skin tissue engineering.     

Ultrastructure des cellules de Sertoli humaines aux stades embryonnaires précoces de la différenciation gonadique

The morphofunctional development of Sertoli cells defines the testicular differentiation. These somatic cells are mostly of mesonephric origin and can be first recognized in 7 week-old embryos altogether with the formation of testicular cords. The latter organize as primordial germ cells surrounded by pre-Sertoli cells. Due to the great synthetic activity of pre-Sertoli cells the rough endoplasmic reticulum develops. The basal lamina of the cords becomes distinguishable at 7 to 8 weeks of development. Either prespermatogonia and pre-Sertoli cells actively proliferate but the latter greatly outnumber prespermatogonia. Many interdigitations and cytoplasmic processes are observed between neighboring pre-Sertoli cells. Due to the proliferative activity a sort of compartmentalization is established inside the cords in which pre-Sertoli cells tend to localize closer to the basal membrane embracing the prespermatogonia with long and thin cytoplasmic processes. One of the main features typical of differentiating pre-Sertoli cells is the irregular nucleus and the prominent nucleolus. When the embryo is 14 to 20 weeks-old the most significative change is the maximum development of the Leydig cells. Testicular cords do not show lumen at all, so they cannot be termed “tubules”     

Mammary gland development: Role of basal myoepithelial cells

Mammary epithelium is organized as a bilayer with a layer of luminal secretory cells and a layer of basal myoepithelial cells. To dissect the specific functions of these two major compartments of the mammary epithelium in mammary morphogenesis we have used genetically modified mice carrying transgenes or conditional alleles whose expression or ablation were cell-type specific. Basal cells are located in close proximity to mammary stroma and directly interact with the extracellular matrix (basement membrane) during all their lifespan. On the contrary, luminal secretory cells during early stages of the postnatal mammary development have only limited contacts with basement membrane and become exposed to the extracellular matrix only during late developmental stages at the end of pregnancy and in lactation. Consistently perturbation of beta1-integrin function specifically in the luminal layer of the mammary epithelium, did not interfere with mammary morphogenesis until the second part of pregnancy but led to impaired secretory differentiation and lactation. On the contrary, ablation of beta1-integrin gene in the basal mammary epithelial cells resulted in a more precocious phenotype: disorganized branching in young virgin animals and a complete arrest of lobuloalveolar development. Further, a constitutive activation of beta-catenin signaling due to expression of N-terminally truncated (stabilized) beta-catenin specifically in basal myoepithelial cells resulted in accelerated differentiation of luminal secretory cells in pregnancy, precocious postlactational involution, increased angiogenesis and development of mammary tumors. Altogether these data suggest that basal mammary epithelial cells can affect growth and differentiation of luminal secretory cells, have an impact on the epithelium-stroma relationships and, thereby, play an important role in the process of mammary morphogenesis and differentiation.     

Origin of periendothelial cells in microvessels derived from human microvascular endothelial cells

In microvessels, periendothelial cells expressing alpha smooth muscle actin (alphaSMA) interact with the endothelial cells and are essential for vessel maturation and stabilization. In adult tissues, the cellular origin of the periendothelial cells is still not clear, in particular in humans. To determine the origin of human periendothelial cells, we used a recently developed 3D co-culture system that mimics human skin connective tissue. This system is composed of normal human dermal fibroblasts (NHDF), human dermal microvascular endothelial cells (HMEC-1), and a collagen matrix. In this system, "microvessels" composed of an endothelial lumen associated with periendothelial cells develop. Using this co-culture system, we (i) labelled fibroblasts with the vital dye CFDA-SE, cultured them with unlabelled endothelial cells, and observed that only endothelium-associated CFDA-SE-labelled cells express alphaSMA; (ii) infected endothelial cells with a retrovirus stably expressing eGFP, cultured them with unlabelled fibroblasts, and observed that cells expressing alphaSMA did not co-express eGFP, but were associated with the eGFP-expressing endothelial cells of the microvessels. Together, these results indicate that periendothelial cells arise by differentiation from fibroblasts and that they require interaction with endothelial cells to do so.