Epidermal growth factor stimulates DNA-synthesis of astrocytes in primary cerebellar cultures

Summary The capability of epidermal growth factor (EGF) to stimulate DNA synthesis in neural cells was investigated in primary cultures of early postnatal mouse cerebellum. At concentrations of 10^-8M, EGF stimulates DNA synthesis in astrocytes, which were identified immunocytologically by the cell type-specific marker, glial fibrillary acidic (GFA) protein. Astrocytes express cell-surface receptors for EGF as can be shown by binding of [¹²⁵I]-labeled EGF to live monolayer cultures. In the presence of 10% horse serum, EGF stimulates DNA-synthesis by a factor of about two-fold. Stimulation by EGF over control values is approximately 4-fold in the presence of 1% serum and 6 to 10-fold in the absence of serum. Absolute numbers of astrocytes are increased after more prolonged action of EGF. DNA-synthesis in neurons or oligodendroglia is not significantly stimulated by EGF. EGF enhances cell survival of serum-deprived cerebellar cultures. Fibroblast growth factor does not increase DNA-synthesis in astrocytes under the conditions used in this study.Abbreviations BME basal medium, Eagle's - BME-BSA basal medium, Eagle's containing 0.1% bovine serum albumin - EDTA ethylene-diamino-N, N-tetraacetic acid - EGF epidermal growth factor - FGF fibroblast growth factor - GFA glial fibrillary acidic - HS horse serum - [³ H] TdR tritium-labeled thymidine - PAP peroxidase-anti-peroxidase - PBS phosphate-buffered saline - SDS sodium dodecyl sulfate - TCA trichloroacetic acid     

The hypothalamic magnocellular system in the domestic fowl

Summary In the rostral hypothalamus of the domestic fowl, the magnocellular neurosecretory nuclei show a peculiar differentiation. Golgi studies of the supraoptic and paraventricular nuclei of the fowl reveal at least two major cell types: 1) large multipolar neurons, and 2) small interneurons. Golgi impregnations provide a detailed cytoarchitectural picture of the large-sized cells; the latter may well correspond to the neurosecretory cells demonstrated in the same regions by selective staining, and immunocytochemical and electron microscopical techniques.Electron microscopically, neuronal perikarya are observed to contain variable amounts of neurosecretory granules (100–200 nm in diameter; mean diameter of 160 nm) scattered throughout the cytoplasm. The diameters of these granules do not differ statistically in the two principal nuclear areas examined. The perikarya of these neurons display only a few axosomatic synapses containing electron-lucent and dense-cored vesicles (70–90 nm in diameter). Numerous nerve terminals of this type also end on the dendritic ramifications in the surrounding neuropil.     

Establishment of regional differences in brush border enzymatic activities during the development of fetal mouse small intestine

Summary In order to study the establishment of regional differences in brush border enzymic activities during the development of fetal mouse small intestine we have followed (1) the differentiation of microvilli by morphometry, and (2) the developmental pattern of three brush border enzymes (lactase, glucoamylase and alkaline phosphatase). From day 16 to day 19 of gestation, the height of duodenal microvilli increases 2.4 times on the absorptive cells located near the tip of the villi. During the same period in the upper half of the duodenal villi, the number of microvilli per square m rises by a factor of 2.4 and the microvillous surface area increases by a factor of 5.2. The differentiation of ileal microvilli follows a similar pattern but they are always shorter and less numerous than those of the duodenum. Lactase activity appears at 18 days of gestation; the other two brush border enzymes are first detected at 16 days of gestation. Afterwards all three enzyme activities increase rapidly and a decreasing gradient of activity is established from the proximal to the distal segment of the small intestine. Hence, the structural development of the microvilli and the appearance of brush border enzyme activities occur simultaneously and a proximo-distal gradient is already established at 16 days of gestation.Supported by MRC of Canada research grant MA-6069Mr. D. Malka was supported by a studentship from the F.C.A.C.Dr. D. Ménard is a chercheur boursier du Conseil de la Recherche en Santé du Québec     

Moulting hormone,juvenile hormone and the ultrastructure of the fat body of adult Sarcophaga bullata (Diptera)

Summary In the ovoviviparous fly, Sarcophaga bullata, vitellogenesis is cyclic; a process reflected in ultrastructural changes in the fat body cells and oenocytes. At eclosion the larval fat body has not yet completely disappeared. During vitellogenesis the fat body cells are specialized for intensive protein synthesis showing a very extensive RER and numerous invaginations of the plasma membrane. These features disappear when the eggs descend into the oviducts to complete embryogenesis. The predominant feature of the oenocytes is their very prominent SER. The fat body cells of the males are never as specialized for protein synthesis as those of the females. Feeding of ecdysterone to males for 3 or more days induces a rather extensive subcellular apparatus for protein synthesis, i.e., invaginations of the plasma membrane and an extensive RER. Juvenile hormone is completely ineffective in this respect. Both ecdysterone and juvenile hormone have pronounced but different effects on the oenocytes of males.     

Morphological studies on neuroglia

Summary The silver-impregnation procedure of Tsujiyama is suitable for demonstration of all three classical types of neuroglial cells; in the present study it was used for electron microscopic identification of neuroglial cells in the brain of the cat. The aim of the present study was 1) to determine impregnated structural correlates of neuroglial cells at the light- and electron-microscopic levels, and 2) to determine whether the method of Tsujiyama is applicable for the electron microscopic identification of the single types of neuroglial cells. Silver deposits were observed over the cytoplasm and processes of astrocytes where numerous glial filaments were present. Oligodendrocytes and microglial cells may be precisely differentiated by use of Tsujiyama's silver impregnation method at the electron microscopic level due to the pattern of silver-deposition in these two basic types of cells. This silver-impregnation method combined with electron microscopy is thus suitable for a precise identification of neuroglial cells; the technique may prove to be very helpful in identification of such categories of neuroglial cells that encompass also the images of cells which cannot be classified by use of the standard methods.Supported by a grant (No. 437002) from the Ministry of Education, Science and Culture, Japan     

Morphological investigations on follicular atresia in canine ovaries

Summary Morphologically, canine ovaries show two types of regression pattern in secondary follicles. In type A necrotic changes of the oocytes and the zona pellucida dominate, whereas in type B degeneration, necrobiosis and necrosis of the granulosa prevail. The atretic course of type B regression results in a pseudoantrum and leads, by pseudogrowth, to a structure imitating tertiary follicles. In both type A and type B regression, four consecutive stages of atresia are distinguished by light- and electron microscopy.True tertiary follicles display only one regression pattern which resembles type B of secondary follicles. Early, advanced, late, and terminal stages of atresia are again described.     

Calcium and magnesium levels in isolated cardiac mitochondria from mice injected with isoproterenol

Summary Calcium (Ca) and Magnesium (Mg) are determined by atomic absorption flame spectrometry in isolated cardiac mitochondria from mice receiving subcutaneous injections of DL-isoproterenol HC1 (ISO), and in mitochondria of untreated controls. In the controls, mitochondria were isolated in the presence or absence of ruthenium red. On the absence of ruthenium red in the isolation medium, mitochondrial Ca levels increase by about 300%, while levels of Mg remain unchanged. Focal myocardial necrosis following a single ISO-injection is shown by electron microscopy. Ca and Mg levels are largely unaffected by a single dose of ISO until 24 h after the injection. A slight increase in Ca occurs in the 48 h samples. When multiple injections of ISO are given every 12th hour for 48 h, 72 h and 96 h, respectively, endogenous Ca and Mg increase significantly. It is suggested that this increase might be associated with ISO-induced cardiac hypertrophy rather than with the pharmacological effects of ISO per se.This work was supported by grants from The Norwegian Council on Cardiovascular Disease and from The Norwegian Research Council for Science and the Humanities     

Development of the bovine acrosome

Summary In the present study the development of the bovine acrosome was investigated using conventional electron-microscopical techniques as well as the phosphotungstic-acid (PTA) technique (Rambourg 1967) including enzymatic digestion experiments. As in other species and in accordance with previous light-microscopical studies (Clermont and Leblond 1955) four phases of acrosomal differentiation can be discerned: the Golgi-phase, cap-phase, acrosome-phase, and maturation-phase.In the bull no internal pattern of the acrosomal content can be observed, either with conventional uranyl acetate-lead citrate staining or with the PTA-techniques. Our results support the observation in other species (Fawcett et al. 1971) that no intrinsic polymerization or crystallization process of the acrosomal content is responsible for acrosomal shaping. Some of our results suggest the influence of external forces on acrosomal development in the bull. During the cap-phase and the acrosome-phase accumulations of smooth endoplasmic reticulum and a layer of fine filaments can be observed in the Sertoli-cell cytoplasm, immediately adjacent to the developing acrosome. A temporary influence of these structures on acrosomal development seems possible. The PTA-positive staining of the developing bovine acrosome is probably due to the presence of acrosomal glycoproteins; however, our results do not exclude the possibility that molecules other than glycoproteins contribute to the positive PTA-staining of the developing acrosome.Supported by a grant from the Deutsche Forschungsgemeinschaft     

The isolated perfused rabbit ovary — A model for studies of ovarian function

Summary In the present investigation the ultrastructure of isolated rabbit ovaries, perfused with different media for various time periods, was studied. The steroid hormone production by the perfused ovary was also determined. Perfusion with Medium 199 results in prominent interstitial ovarian oedema which increases with perfusion time. Even after the addition of 6–10 % Dextran T40, oedema appears in the interstitial tissue of the ovary. Perfusion solutions with osmotically active colloid particles of large molecular size (Dextran T70; average molecular weight 70,000 and bovine serum albumin), cause less distortion in the ovarian structure, and ultrastructurally the ovarian tissues appear essentially the same as in the control ovaries.The results indicate that the perfused rabbit ovary, under strictly controlled conditions, can be used as an experimental model for studies of various aspects of ovarian function, including follicular rupture.     

Ultrastructural modifications of the glandular epithelium of the receptaculum seminis in Thermobia domestica (Insecta: Thysanura) during the moulting period

Summary The wall of the receptaculum seminis of Thermobia domestica is composed of numerous glandular units, each with four enveloping cells (denoted 1 to 4) separated by ordinary epithelial cells and associated with a cuticular apparatus. During the moulting periods, which continue to occur in the adult stage, these cells undergo a series of transformations. Just before apolysis there is a dedifferentiation of numerous cytoplasmic organelles, but no mitosis has been observed. When the intima lifts off, the apical system of each glandular unit, i.e. the distal parts of the C2 and C3 cells surrounding the end apparatus, is also eliminated. Then at the apex of each glandular unit, a new ductule is formed in the cavity of which a long ciliary process grows up from cell C1. Finally comes the phase of cuticle formation, i.e., epicuticle for the ductules, epi-and endocuticle for the intima lining the central cavity of the receptaculum. Various cell types participate in secretion of cuticle, the ciliary cells (C1) being responsible for the formation of the porous end apparatus. At ecdysis almost all of the new intima has been secreted and the apical systems are once more differentiated. These transformations are compared with those recently described in other exocrine glands of arthropods, e.g., tegumentary glands and accessory glands of the genital ducts.