Large Glycogen Deposits Research Articles

The ultrastructural features of the infundibulum of the green iguana, Iguana iguana.

The purpose of this study is to describe, in detail, the ultrastructure of the infundibulum of the sexually mature and active female green iguana, Iguana iguana. The infundibulum of five iguanas was remarkably distinct from the uterus, and was also clearly demarcated into cranial (expanded v-shaped) and caudal (tubular) divisions. Tissue samples obtained from five portions (three from the cranial division and two from the caudal division) of the infundibulum were processed conventionally for light and electron microscopy. The epithelial lining of the most anterior, middle, and posterior, parts of the cranial division displayed nonciliated cells predominantly, and occasionally ciliated cells. The numerous secretory granules in nonciliated type 1 cell found in the fimbrial aspect of the infundibulum were homogenous and deeply electron-dense, but those in the other two regions were variants of this cell type because they contained variably electron-dense secretory granules. Two main types of nonciliated cells (type 2 and its variant, type 3, as well as type 4) occurred in the epithelial lining of the caudal division of the infundibulum, but they, clearly, showed no dense secretory granules. Whereas the nonciliated type 2 cell and its variant (type 3 cell) contained large glycogen deposits, the type 4 cell lacked these deposits but its apical part contained large lipid-like droplets and, remarkably, blebbed into the duct lumen. The nonciliated cells lining the mucosal tubular glands contained highly electron-dense secretory granules, which were similar to those found in the nonciliated type 1 cell in the epithelial lining of the fimbrial part of the cranial division of the infundibulum.

Live Cell Imaging in Microfluidic Device Proves Resistance to Oxygen/Glucose Deprivation in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.

Analyses of cellular responses to fast oxygen dynamics are challenging and require ad hoc technological solutions, especially when decoupling from liquid media composition is required. In this work, we present a microfluidic device specifically designed for culture analyses with high resolution and magnification objectives, providing full optical access to the cell culture chamber. This feature allows fluorescence-based assays, photoactivated surface chemistry, and live cell imaging under tightly controlled pO2 environments. The device has a simple design, accommodates three independent cell cultures, and can be employed by users with basic cell culture training in studies requiring fast oxygen dynamics, defined media composition, and in-line data acquisition with optical molecular probes. We apply this technology to produce an oxygen/glucose deprived (OGD) environment and analyze cell mortality in murine and human cardiac cultures. Neonatal rat ventricular cardiomyocytes show an OGD time-dependent sensitivity, resulting in a robust and reproducible 66 ± 5% death rate after 3 h of stress. Applying an equivalent stress to human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) provides direct experimental evidence for fetal-like OGD-resistant phenotype. Investigation on the nature of such phenotype exposed large glycogen deposits. We propose a culture strategy aimed at depleting these intracellular energy stores and concurrently activate positive regulation of aerobic metabolic molecular markers. The observed process, however, is not sufficient to induce an OGD-sensitive phenotype in hiPS-CMs, highlighting defective development of mature aerobic metabolism in vitro.

Sperm ultrastructure and spermatodesm morphology of the spittle bug Locris transversa (Thunberg 1822) (Hemiptera: Cercopidae)

The structure of the spermatozoon and spermatodesm of the spittlebug Locris transversa (Thunberg 1822) was investigated by light and transmission electron microscopy. Males produced only one size class of sperm, which was 93–106 μm long. During spermatogenesis groups of spermatozoa are arranged around, and attached by their acrosomes to, a small central extracellular matrix to form a ball-shaped spermatodesm. Spermatodesmata were found in the testis, vas deferens and seminal vesicle of males and spermatheca of females. The sperm are filiform, each consisting of a ~15-μm-long head containing an anteriorly positioned conical ~2-μm-long acrosome and ~13-μm-long nucleus, and a midpiece and tail with a 9+9+2 axoneme. The acrosome, which has two posterior extensions that lie along one side of the anterior region of the nucleus, contains longitudinally orientated microfilaments. The nucleus has two unequal anterior extensions, whereas posteriorly it is flattened laterally to accommodate a putative centriolar adjunct and anterior ends of the two mitochondrial derivatives. The basal body lies posterior to the nucleus. The mitochondrial derivatives are elongated and extend for almost the entire length of the tail. The tail has glycogen within its centre, and towards its terminal end large deposits of glycogen surround the mitochondrial derivatives and axoneme. Compared to other members of the Cicadomorpha, the spermatozoa and spermatodesms of cercopids so far studied have some structural features in common with cicadellids and others in common with cicadas; therefore, if sperm features are to be useful in phylogenetic studies of cicadomorphs, attention must be given to examining key taxa to establish which character states are plesiomorphic or apomorphic.

A HISTOLOGICAL STUDY ON HEPATIC STRUCTURE OF Lyciasalamandra arikani (URODELA: SALAMANDRIDAE)

The liver is one of the most important internal organ in the body. It plays a prominent role in many processes in the body, particularly those concerned with its metabolism (protein synthesis, storage metabolites, bile secretion and detoxification). This report represents the histological characterization of liver Lyciasalamandra arikani, endemic salamander to Turkey. Hematopoietic tissue structures were examined in the connective tissue of the perihepatic regions. The liver of L. arikani probably possesses immunologic capabilities due to the presence of lymphocytes in the perihepatic regions. The hepatic lobules consisted of both hepatocytes and sinusoidal blood capillary networks, in which hepatocyte-sinusoidal structures were formed. The hepatocytes were polygonal and had a rounded nucleus. There is a lot of differing content of melanin containing cells in the hepatic parenchyma. Periodic acid-Schiff (PAS) staining method showed large glycogen deposits in the clusters of melanin granules. These melano-macrophage centers probably act significant role in organs of heterothermic vertebrates including providing energy and protection against pathogens due to fact that heterothermic vertebrates can produce energy slowly and so are torpid related the blood flow rate at low temperatures.

Amorimia exotropica poisoning as a presumptive cause of myocardial fibrosis in cattle

The current study reports the investigation on the cause of sudden deaths associated with cardiac fibrosis in cattle in northern Rio Grande do Sul, Brazil. The exclusion of known causes of bovine cardiac fibrosis as well as the absence of the plants in that region whose consumption has already been linked to the disorder motivated this investigation. The condition, which was attributed to the consumption of Amorimia exotropica, affected draft oxen, most of which died suddenly without showing any clinical signs during usual management or work. Globular hearts with white foci at their cut surfaces were the main gross findings, which corresponded microscopically from multifocal to coalescent areas of myocardial fibrosis. To confirm the condition, A. exotropica from the ranches where cattle died from the disease was dosed to rabbits, which showed similar lesions to those seen in dead cattle after receiving 10 doses of 3.6 g/kg at 4-day intervals. Electron microscopy on rabbit tissues revealed severe tumefaction of the cardiomyocytes associated with mitochondrial swelling, displacement, and rupture of the mitochondrial crests, and of the bundles of myofibrils, apart from large glycogen deposits within the sarcoplasm. It is suggested that mitochondrial changes triggered alterations that lead to cardiac fibrosis and that all of these changes were induced by A. exotropica cardiotoxicity.

Cardiomyocytes derived from human embryonic and induced pluripotent stem cells: comparative ultrastructure

Induced pluripotent stem cells (iPSC) are generated from fully differentiated somatic cells that were reprogrammed into a pluripotent state. Human iPSC which can be obtained from various types of somatic cells such as fibroblasts or keratinocytes can differentiate into cardiomyocytes (iPSC-CM), which exhibit cardiac-like transmembrane action potentials, intracellular Ca2+ transients and contractions. While major features of the excitation-contraction coupling of iPSC-CM have been well-described, very little is known on the ultrastructure of these cardiomyocytes. The ultrastructural features of 31-day-old (post-plating) iPSC-CM generated from human hair follicle keratinocytes (HFKT-iPSC-CM) were analysed by electron microscopy, and compared with those of human embryonic stem-cell-derived cardiomyocytes (hESC-CM). The comparison showed that cardiomyocytes from the two sources share similar proprieties. Specifically, HFKT-iPSC-CM and hESC-CM, displayed ultrastructural features of early and immature phenotype: myofibrils with sarcomeric pattern, large glycogen deposits, lipid droplets, long and slender mitochondria, free ribosomes, rough endoplasmic reticulum, sarcoplasmic reticulum and caveolae. Noteworthy, the SR is less developed in HFKT-iPSC-CM. We also found in both cell types: (1) ‘Ca2+-release units’, which connect the peripheral sarcoplasmic reticulum with plasmalemma; and (2) intercellular junctions, which mimic intercalated disks (desmosomes and fascia adherens). In conclusion, iPSC and hESC differentiate into cardiomyocytes of comparable ultrastructure, thus supporting the notion that iPSC offer a viable option for an autologous cell source for cardiac regenerative therapy.

Unlocking the ultrastructure of colorectal cancer cellsin vitrousing selective staining

To characterise differences between three widely used colorectal cancer cell lines using ultrastructural selective staining for glycogen to determine variation in metastatic properties. Transmission electron microscopy was used in this investigation to help identify intracellular structures and morphological features which are precursors of tumor invasion. In addition to morphological markers, we used selective staining of glycogen as a marker for neoplastic cellular proliferation and determined whether levels of glycogen change between the three different cell lines. Ultrastructural analysis revealed morphological differences between the cell lines, as well as differentiation into two sub-populations within each cell line. Caco-2 cells contained large glycogen deposits as well as showing the most obvious morphological changes between the two sub-populations. SW480 cells also contained large glycogen stores as well as deep cellular protrusions when grown on porous filter membranes. HT-29 cells had trace amounts of glycogen stores with few cellular projections into the filter pores and no tight junction formation. Morphology indicative of metastatic properties coincided with larger glycogen deposits, providing strong evidence for the use of selective staining to determine the neoplastic properties of cells.

Control of glycogen synthase through ADIPOR1-AMPK pathway in renal distal tubules of normal and diabetic rats

Diabetic nephropathies are characterized by glycogen accumulation in distal tubular cells, which eventually leads to their apoptosis. The present study aims to determine whether adiponectin and AMPK are involved in the regulation of glycogen synthase (GS) in these structures. Western blots of isolated distal tubules revealed the presence of adiponectin receptor ADIPOR1, catalytic AMPK subunits alpha(1) and alpha(2), their phosphorylated active forms, and the glycogen-binding AMPK subunit beta(2). ADIPOR2 was not detected. Expression levels of ADIPOR1, AMPKalpha(1), AMPKalpha(2), and AMPKbeta(2) were increased in streptozotocin-treated diabetic rats, whereas phosphorylated active AMPK levels were strongly decreased. Immunohistochemistry revealed the presence of ADIPOR1 on the luminal portion of distal tubules and thick ascending limb cells. Catalytic subunits alpha(1) and alpha(2), their phosphorylated active forms, and the glycogen-binding subunit beta(2) were also found in the same cells, confirming immunoblot results. In vitro, 5-aminoimidazole-4-carboxamide-1-beta-d-ribofuranoside (AICAR; 2 mM) and globular adiponectin (10 mug/ml) activated catalytic AMPK in distal tubules isolated from kidneys of normal rats but much more weakly in those from diabetic rats. GS inhibition paralleled AMPK activation in both groups of animals: active GS levels were low in control animals and elevated in diabetic ones. Finally, glucose-6-phosphate, an allosteric activator of GS, was also increased in diabetic rats. These results demonstrate that in distal tubular cells, adiponectin through luminal ADIPOR1 activates AMPK, leading to the inhibition of GS. During hyperglycemia, this regulation is altered, which may explain, at least in part, the accumulation of large glycogen deposits.

Morphologic, cytometric and functional characterisation of abalone ( Haliotis tuberculata) haemocytes

This work presents the first detailed microscopic and functional analysis of the haemocytes of an abalone; the European Haliotis tuberculata. It is shown that in contrast to the situation in bivalves, only very few basophilic “granulocytes” could be found and exclusively with a histological stain. Neither flow cytometry, phase contrast observation nor transmission electron microscopy were able to detect any granular cells. The large majority of cells was constituted of “hyalinocytes”, which could be sorted by flow cytometry, for the first time, into small (blast-like) and large cells. This permits a detailed analysis of haemocytes and especially of the lowly represented blast-like cells. The differences in haemolymph cell composition between bivalves and gastropods is reviewed in depth and discussed in view of the new data we present. Most of the abalone haemocytes analysed harbour many vacuoles, large glycogen deposits, lipid inclusions and acidic compartments. However, although the number of these “inclusions” was rather variable in between individual hyalinocytes, these experiments did not allow to discern subpopulations using these criteria, and the population appears more as a “differentiation continuum”. Haemocytes adhere very rapidly and are immunologically active as they quickly phagocytose latex beads and zymozan particles. This study is the first step towards understanding the H. tuberculata immune system by adapting new tools to gastropods and in providing a first detailed morpho-functional study of their haemocytes.

AMPK ACTIVATION BY ADIPONECTIN IN RENAL TUBULES OF CONTROL AND DIABETIC RATS

Adiponectin and AMPK regulate glucose metabolism and apoptosis. We have examined their role in renal tubules that accumulate glycogen and undergo apoptosis in diabetic nephropathy. Rat kidneys were processed for light microscopy and biochemical determinations. Renal tubules were isolated and incubated in vitro. Western blots were carried out with semi-quantitative evaluations. Adiponectin receptors ADIPOR1 is present on the luminal side of epithelial cells of distal tubules and thin ascending limb (TAL) in normal and hyperglycaemic rats. ADIPOR2 was not detected. The same cells were positive for catalytic sub-units α1 and α2, the regulatory sub-unit β2 and the phosphorylated active form of AMPK. Interestingly, glycogen-filled tubular cells in tissues of diabetic rats displayed a much lighter staining for active AMPK while ADIPOR1 fluorescence was markedly increased. In vitro incubation of tubules showed that levels of activated AMPK were strongly decreased in hyperglycaemic conditions while total AMPK and ADIPOR1 levels were increased. AICAR and adiponectin potently activated AMPK in tubules from control rats but weakly in those of hyperglycaemic rats. In summary, ADIPOR1 and AMPK are expressed by distal tubules and TAL. Basal and stimulated AMPK activities are decreased in hyperglycaemic conditions. These results suggest that adiponectin resistance develops in renal tubules, which may lead to the accumulation of large glycogen deposits and apoptosis in diabetic nephropathy. (Supported by CIHR).

Mice with elevated muscle glycogen stores do not have improved exercise performance

Skeletal muscle glycogen is considered to be an important source of energy for contraction and increasing the level of the glucose polymer is generally thought to improve exercise performance in humans. A genetically modified mouse model (GSL30), which overaccumulates glycogen due to overexpression of a hyperactive form of glycogen synthase, was used to examine whether increasing the level of the polysaccharide enhances the ability of mice to run on a treadmill. The skeletal muscle of the GSL30 mice had large deposits of glycogen. There were no significant increases in the work performed by GSL30 mice as compared to their respective wild type littermates when exercised to exhaustion. The amount of muscle glycogen utilized by GSL30 mice, however, was greater, while the amount of liver glycogen consumed during exhaustive exercise was less than wild type animals. This result suggests that increased muscle glycogen stores do not necessarily improve exercise performance in mice.

Ultrastructure of the body wall of infective larvae of Cystidicoloides ephemeridarum (Nematoda, Cystidicolidae) from mayflies

Scanning and transmission electron microscopic examinations of tissue-dwelling third-stage larvae of the nematode Cystidicoloides ephemeridarum from the intermediate host (Ephemera danica) were carried out with respect to the morphological changes in the body wall associated with the transfer from the intermediate to the definitive host. The ultrastructure of the cephalic end and zonation of the somatic cuticle of infective larvae basically correspond with those of adults. The somatic cuticle is composed of a fuzzy epicuticle, an outer and inner cortical zone, a median zone and a basal zone consisting of three subzones. Globular bodies are absent from the median zone of the infective larvae. The lateral hypodermal cords of the infective larvae are cellular, consisting of a median cell enclosed by two sublateral cells. The excretory canal is present within each of the lateral cord in both the infective and adult stages. Walls of the excretory canals contain Golgi-derived vesicles which communicate with the lumen of the canal. Large deposits of glycogen, suggesting anaerobic respiration, are present in the hypodermal cords and noncontractile parts of the muscle cells of the third-stage larvae within the intermediate host.

Ultrastructural evidence for nutritional exchange between brooding unionid mussels and their glochidia larvae

Abstract. The life history of unionid bivalve molluscs includes retention of developing embryos within the gills of parental mussels. This brooding behavior may facilitate nutrient transfer to the glochidia larvae, i.e., matrotrophy. To address this possibility, morphological relationships between brood chambers and developing larvae of Pyganodon cataracta and Utterbackia imbecillis were examined with TEM, and larval shells were observed with SEM, for features that could be associated with the uptake of dissolved materials. Early in brooding, glochidia are enclosed in a vitelline membrane that physically contacts numerous cilia and microvilli of the epithelial cells lining the brood chamber (marsupium). The vitelline membrane subsequently disappears. Lamellar tissues of parental mussels initially have large deposits of glycogen that diminish during the course of brooding. Septa separating brood chambers from adjacent secondary water tubes have numerous mitochondria and microvilli, suggesting the potential for active transport of materials into or out of the marsupia. Since punctae (pores) in the larval shells become filled with an organic matrix early in brooding, they are unlikely to be involved in nutrient exchange. Ultrastructure of the brood chamber and physical contact between the parental mussel and larvae are consistent with a nutritive role for retention of glochidia in the marsupia.

Functional and ultrastructural evidence of myocardial stunning after acute carbon monoxide poisoning

To study human myocardial ultrastructural changes after carbon monoxide (CO) poisoning inducing reversible cardiac failure. clinical, functional and morphologic findings. Public university-affiliated hospital and electron microscopy laboratory. A 25-yr-old woman with functional evidence of cardiac failure after acute CO poisoning. Hyperbaric and intensive care treatment over 10 days. Scintigraphic and cardiac angiography with endomyocardial biopsy. Scintigraphy with 99mTc hexakis 2-methoxy-2-isobutyl isonitrile (sestaMIBI) showed an uptake defect in the left anterior descending artery territory. The cardiac angiography demonstrated a slight hypokinesis of the superior two thirds of the anterior wall and of the septal region with completely normal coronary angiograms. Electron microscopy of left ventricular biopsies showed slight ultrastructural changes in the myocytes. In addition, large glycogen deposits were mostly associated with swollen mitochondria. The patient was discharged in good clinical condition on day 10. Presence of glycogen deposits associated with abnormal mitochondria may be signs of the incapability of myocardial cells in utilizing energy substrata. In the presence of normal myocardial perfusion, our findings are consistent with the presence of a stunned myocardium-like syndrome. Early recognition and treatment of this clinical syndrome allow the prevention of myocardial infarction.

Generalized glycogen storage and cardiomegaly in a knockout mouse model of Pompe disease.

Glycogen storage disease type II (GSDII; Pompe disease), caused by inherited deficiency of acid alpha-glucosidase, is a lysosomal disorder affecting heart and skeletal muscles. A mouse model of this disease was obtained by targeted disruption of the murine acid alpha-glucosidase gene (Gaa) in embryonic stem cells. Homozygous knockout mice (Gaa -/-) lack Gaa mRNA and have a virtually complete acid alpha-glucosidase deficiency. Glycogen-containing lysosomes are detected soon after birth in liver, heart and skeletal muscle cells. By 13 weeks of age, large focal deposits of glycogen have formed. Vacuolar spaces stain positive for acid phosphatase as a sign of lysosomal pathology. Both male and female knockout mice are fertile and can be intercrossed to produce progeny. The first born knockout mice are at present 9 months old. Overt clinical symptoms are still absent, but the heart is typically enlarged and the electrocardiogram is abnormal. The mouse model will help greatly to understand the pathogenic mechanism of GSDII and is a valuable instrument to explore the efficacy of different therapeutic interventions.

Immunocytochemical investigation of insulin secretion by pancreatic beta-cells in control and diabetic Psammomys obesus.

Hyperproinsulinemia is a characteristic feature of non-insulin-dependent diabetes mellitus (NIDDM) caused by pancreatic beta-cell dysfunction through a secretion-related alteration or impaired proinsulin processing. We have investigated the insulin processing and secretion in Psammomys obesus fed with low- and high-energy diets, which represent a model for diet-induced NIDDM. With a high-energy diet the animals develop hyperglycemia and hyperinsulinemia, whereas those maintained on a low-energy diet remain normoglycemic. Although a large amount of insulin immunoreactivity was detected in beta-cells of the normoglycemic compared to hyperglycemic animals, in situ hybridization for insulin mRNA demonstrated a particularly high signal in the beta-cells of the hyperglycemic animals. By electron microscopy, the beta-cells of normoglycemic animals displayed large accumulations of secretory granules, whereas those of the hyperglycemic animals contained very few granules and large deposits of glycogen. These results reflect a secretory resting condition for the cells of the normoglycemic animals in contrast to stimulated synthetic and secretory activities in the cells of the hyperglycemic ones. Using colloidal gold immunocytochemistry at the electron microscopic level, we have examined subcellular proinsulin processing in relation to the convertases PC1 and PC2. Immunolabeling of proinsulin, insulin, C-peptide, PC1, and PC2 in different cell compartments involved in beta-cell secretion were evaluated. Both PC1 and PC2 antigenic sites were detected in beta-cells of hyperglycemic Psammomys, but their labeling intensity was weak compared to the cells of normoglycemic animals. In both groups of animals, higher levels of PC2 were found in the Golgi apparatus than in the immature granules. Major decreases in proinsulin, insulin, PC1, and PC2 immunoreactivity were recorded in beta-cells of the hyperglycemic Psammomys. In addition, all these antigenic sites were detected in lysosome-like structures, revealing a major degradation process. These results suggest that the insulin-secreting cells in hyperglycemic Psammomys obesus are in a chronic secretory state during which impaired processing of proinsulin appears to take place.

Ultrastructure of Conidia and the Conidium Aging Process in the Plant Pathogenic Fungus Colletotrichum graminicola

The influence of age on conidial ultrastructure was investigated in Colletotrichum graminicola. The cytoplasmic volume of young conidia-those harvested from 5-7-d-old cultures-was dominated by lipid bodies. These conidia also were characterized by numerous small vacuoles with electron-dense contents. Other structures present included mitochondria, microbodies, ribosomes, elements of tubular ER, short strands of rough ER, multivesicular bodies, simple cisternal elements, Woronin bodies, microtubules, and large amounts of what appeared to be glycogen. Older conidia harvested from 30-35-d-old cultures exhibited a significantly different ultrastructure. They contained fewer lipid bodies, lacked the large glycogen deposits, were very highly vacuolate, and contained sheetlike strands of ER and extensive aggregations of tubular ER. Structures thought to be autophagic vacuoles also were present. The conidial wall appeared similar in conidia of both ages and consisted of a thicker inner electron-transparent layer ...

Microanatomy of Retia Mirabilia of Bowhead Whale Foramen Magnum and Mandibular Foramen

The morphology of vessels of the cervical rete mirabile near the region of the foramen magnum of the bowhead whale, Balaena mysticetus, as well as of retial vessels of the mandibular foramen was studied by light and transmission electron microscopy. A comparison of arterial and venous components of the mandibular rete revealed considerable arterial branching and arteriovenous anastomoses. Although the small arteries of bowhead retia contained the same layers found in nonretial arteries, several distinctive morphologic features were evident. These included wide separation by collagenous connective tissue and small size of individual smooth muscle cells of the tunica media. These myocytes also contained considerable perinuclear glycogen and communicated minimally with adjacent myocytes by small foot-like cytoplasmic appendages as well as branching of basement membranes. Ganglia-like neural plexuses were observed within the tunica media of arterial retia. Endothelial cells lining the intima demonstrated loose peripheral edges which frequently projected into the vessel lumen, and these cells rested upon a subjacent layer of reticular fibers. It is known for some species that the retial supply to the brain in whales largely supplants an internal carotid arterial supply and that the spinal cord is supplied by retia. The physiologic role of the bowhead mandibular retia remains unknown. The retia mirabilia of cetaceans are considered as one of the morphologic adaptations which modulate hemodynamics during diving and resurfacing. The morphologic features of these vessels in the bowhead whale, as revealed by electron microscopy, appear to reflect the capability to respond in a slow but sustained manner to diving, and the large glycogen deposits may help sustain vascular myocyte metabolism during prolonged submersion.

Fetal membranes and placenta of the African green monkey (Cercopithecus aethiops).

This study examined developmental changes in fetal membranes and placenta of Cercopithecus aethiops from a Carnegie developmental stage 12 embryo to near-term fetuses. Ultrastructurally, yolk sac cells (endoderm and mesothelium) were similar to comparable stages in other primates. Endodermal cells had few apical microvilli, abundant rough-endoplasmic reticulum, electron dense mitochondria and dense bodies. In contrast, mesothelial cells were squamous with numerous microvilli, small mitochondria and a few short strands of rough endoplasmic reticulum. Amnion cells early in gestation were squamous with few microvilli, large glycogen deposits and poorly developed cytoplasmic components. Tight junctions and desmosomes held adjacent cells together. The basal surface was smooth and the basal lamina was distinct. As development proceeded the amniotic cells became cuboidal and possessed numerous microvilli. Cytoplasmic organelles were better developed and glycogen deposits increased by mid-gestation. A thick layer of microfibrils and collagen fibers was prominent below the basal lamina. Near-term, the glycogen had virtually disappeared and the amount of lipid droplets increased. Basal infoldings and podocytic processes and the extracellular matrix had increased. The smooth chorion consisted of pseudostratified columnar cells. Cells had short microvilli, numerous granules and vesicles of variable size and electron density in early gestation. With increasing age, amounts of granules and vesicles decreased, as the endoplasmic reticulum became prominent. The chorionic trophoblast was a continuous layer in mid-pregnancy and its cells had well-developed organelles and inclusions. Late in gestation, the trophoblastic layer became discontinuous and wide intercellular spaces and channels were present. In the placenta, the trophoblastic elements showed features characteristic of primate placenta.

20-Hydroxyecdysone induces glycogen accumulation within the testicular sheath during in vitro spermatogenesis renewal in diapausing codling moths ( Cydia pomonella)

In codling moths, as in other Lepidoptera, spermatogenesis discontinues during larval diapause. In vitro addition of 20-hydroxyecdysone to cultures of diapausing testes induces renewal of spermatogenesis. It was proposed (1) that in vitro, the discontinuity results from the establishment in the testicular sheath of a barrier against haemolymph factors promoting spermatogenesis and (2) that in vitro, 20-hydroxyecdysone induces spermatogenesis renewal by restoring sheath penetrability to the haemolymph factors. This proposed effect of 20-hydroxyecdysone in vitro was studied at the ultrastructural level and compared with the corresponding situation in vivo, during spermatogenesis renewal after diapause. The results indicate that 20-hydroxyecdysone promotes in vitro spermatogenesis renewal by sustaining functional integrity of the sheath cells and by enhancing their glycogen metabolism. Thus, diapausing testes cultured in presence of 20-hydroxyecdysone display sheath cells having a structure compatible with active functioning and containing large glycogen deposits. But absence of 20-hydroxyecdysone from the cultures results in sheath cells showing clear indications of decay and containing few, dispersed glycogen particles. Likewise, corresponding non-diapausing testes cultured in absence of 20-hydroxyecdysone show sheaths depleted of glycogen. In vivo, testicular sheaths show accordingly scarce glycogen particles during diapause but large glycogen deposits during post diapause.