The ultrastructure of normal fetal and neonatal pig testis germ cells and the influence of fetal decapitation on the germ cell development

The development of germ cells in the male pig was investigated ultrastructurally in normal and decapitated fetuses. The age ranged respectively from 30 days p.c. till one month after birth and from 52 days p.c. until birth. The ultrastructural organization of the germ cells changes dramatically between 30 days p.c. and 52 days p.c. which coincides with the formation of 'true' sex cords. From 52 days p.c. onwards the morphology is rather stable: cells show a 'hydrated' appearance and typical cell bridges. There is no obvious difference in the ultrastructure of germ cells in decapitated animals, their normal littermates and control animals. Therefore germ cell development in the pig is likely to be insensitive to gonadotropins during the fetal period. The development of pig germ cells follows closely the pattern described for several species. Quantitatively there is an increase in the ratio of germ cell/Sertoli cell per cross sectional diameter in the decapitated animals.     

The ultrastructure of the Sertoli cell of the vervet monkey, Chlorocebus aethiops

The ultrastructure of the Sertoli cell of the vervet monkey was studied using both scanning and transmission electron microscopic techniques. SEM micrographs revealed perforated sleeve-like processes which encased mature elongated spermatids which are ready for spermiation. TEM micrographs showed a large Sertoli cell nucleus characterized by many lobes (4–5) and consisting of a homogenous nucleoplasm and a distinctive nucleolus. The nucleus occupies a significant portion of the basal region of the cell. The distribution of chromatin clearly shows high activity of these cells. Lipid droplets and free ribosomes are also found scattered throughout the cytoplasm. Well-developed Golgi apparatus is found in the basal region of the cell. There is phagocytic activity in the Sertoli cells as revealed by the presence of numerous phagosomes. Numerous mitochondria with well-developed tubular cristae are found on the basal side of the nucleus, whereas few mitochondria are located on the apical side of the nucleus. Distinct desmosomes are located between cells. A well-developed smooth endoplasmic reticulum and granular endoplasmic reticulum are frequently found in the cytoplasm of the Sertoli cells. The results of this investigation showed that Sertoli cells of the vervet monkey are almost similar to those of humans and show many similarities with other mammalian species.     

Quantitative analysis of intramembranous particles in the membranous system of rat liver cells at different stages of development and aging

The density of intramembranous protein particles was studied by freeze-fracture. Particle density on the fracture faces of the plasmalemma and the rough endoplasmic reticulum (RER), as well as the outer and inner membranes of the nucleus and the mitochondria in rat hepatocytes were quantified. Comparison among different age groups sampled days postcoitum (dpc), days postpartum (dpp), and months postpartum (mpp) shows age-related changes in particle density in each membrane system. With the exception of the RER, particle densities increased after the 16th dpc, reached a maximum at birth, and then decreased with increasing age. Simultaneously, the number nuclear pores shows a positive correlation with the particle density of the nuclear membranes. The particle density on the membranes of the RER shows a maximum on the 16th dpc, and on the 6th dpp. Thereafter, the density of the RER decreases slightly. In all membrane systems, the density of the particles on the external fracture faces is more variable than the density of the particles on the protoplasmic fracture faces.     

薏苡胚乳传递细胞的超微结构

传粉后１０ ｄ,薏苡（Ｃｏｉｘ ｌａｃｒｙｍ ａ－ｊｏｂｉＬ．）颖果基部胚乳最外层传递细胞具长而多的壁内突,第二层细胞的壁内突较第一层的短而少,均具瓣裂的细胞核、丰富的线粒体、粗糙内质网、核糖体、产生小泡的高尔基体及与壁内突质膜相连的、含深色物质的囊泡。线粒体分布于壁内突附近或其间。授粉后２５ ｄ,第一、二层细胞壁内突发达,几乎充满了细胞,但细胞器可见。第四层传递细胞具树枝状及网状的壁内突,大量线粒体、具质体膜的淀粉粒、脂体存在壁内突附近或壁内突的间隙内。高尔基体常见,仅见很少的片段内质网。第五层传递细胞具短的壁内突、较大的淀粉粒及许多小蛋白质体。两个时期的第一、二层细胞内均未观察到胞间连丝。授粉后２５ ｄ,第四层及以上的传递细胞的细胞壁和呈网状的壁内突均含有胞间连丝。还讨论了各种细胞器的作用及各层传递细胞的功能     

Ultrastructure of Transfer Cells in Endosperm of Coix lacryma-jobi

Special attention was paid to the ultrastructure of transfer cells (TCs) in different locations of basal endosperm in Coix lacryma-jobi at 10 and 25 days after pollination. At 10 days after pollination. TCs of the outermost layer had long wall ingrowths (WIs) whereas those of the second layer possessed fewer and shorter Wis. In both layers TCs had a lobed nucleus, abundant mitochondria, rough endoplasmic reticulum (RER), ribosomes, and a certain number of dictyosomes and vesicles which contained dense substance connected with plasma membrane of WIs. Mitochondria were located near or between WIs. The distribution of organelles in TCs of the second layer was similar to that of the outermost layer. Mitochondria had well defined cristae and dictyosomes and RER seemed more numerous than in TCs of the outermost layer. At 25 days after pollination, TCs of the outermost and the second layer were almost filled with Wis but the organelles were recognizable. TCs of the fourth layer had branched and network-like WIs, many mitochondria, starch grain within plastids and lipids locating near WIs and in the interstices of WIs. Dictyosomes were frequently found but less RER fragments were seen. TCs of the fifth layer with short WIs contained large starch grains and small protein bodies. Plasmodesmata were not observed in the walls of TCs of the outermost and second layer at both 10 and 25 days after pollination but were found in the walls of TCs of the fourth and upper layers and also in the network-like WIs at 25 days after pollination. The roles of the organelles and functions of TCs of different layers were discussed.     

Fine structure of the corpuscles of stannius in the toadfish.

The micro-anatomy of the corpuscles of Stannius of the toadfish, Opsanus tau, an aglomerular marine teleost, has been studied by light and electron microscopy. The corpuscles are composed of extensively anastomosed cords of epithelial cells which maintain intimate contact with blood capillaries. Most of the epithelial cells contain acidophilic granules which also show a positive reaction with the periodic acid-Schiff technique and aldehyde fuchsin. On the basis of fine structural criteria, three cell types can be recognized. The granular cells contain abundant quantities of granular endoplasmic reticulum, ribosomes, Golgi apparatus with prosecretory granules, coated vesicles, polymorphic mitochondria with lamellar cristae, filaments, microtubules, a cilium, a variety of lysosome-like dense bodies, glycogen particles, lipid droplets, secretory granules and intranuclear lipid-like inclusions. One variety of agranular cell (type I) is characterized by the total absence of secretory granules, but it contains large amounts of granular endoplasmic reticulum and ribosomes, conspicuous profiles of Golgi apparatus, coated vesicles and sometimes an abundance of glycogen. Another variety of agranular cell (type II) has poorly developed cytoplasmic organelles. The perivascular space between the capillary and parenchyma contains connective tissue cells and abundant nerve fibers. The different types of epithelial cells observed in the corpuscles of Stannius of this fish may represent functional stages of the secretory cycle in a single cell type.     

Fine structure of the corpus Luteum of the raccoon during pregnancy

Summary Ultrastructure of the granulosa lutein cells of the raccoon from throughout pregnancy has been described. The lutein cells often from epithelial cords which are separated by the connective tissues, capillaries and lymphatics. Based on the arrangements and modifications of the cytoplasmic organelles and inclusions, three types of lutein cells have been recognized. The type I lutein cells predominantly contain tubular, agranular endoplasmic reticulum, juxtanuclear Golgi complexes, a few round to rod-shaped mitochondria, some free ribosomes, and occasional lipid droplets. Occasionally the tubular cristae of mitochondria and tubular smooth endoplasmic reticulum appear contiguous. The type II cells contain abundant lace-like and/or stacked fenestrated endoplasmic reticulum cisternae that frequently form membranous whorls, some tubular, agranular endoplasmic reticulum, mitochondria, and lipid droplets. Mitochondria are usually small, but unusual large ones also occur. The small, rod-to round-shaped mitochondria usually have tubular cristae; but the large, oval, elongate, and cup shaped mitochondria possess tubular, lamellar, plate like, and whorl-like cristae. The plasma membranes of the cells are complexly elaborated and folded, especially when apposing each other. In favorable sections, strands of fenestrated cisternae appose the folds of the plasma membranes. In general, the amount of cytoplasmic organelles and inclusions vary greatly in the cells. The type III cells predominantly contain lipid droplets and sparse cytoplasmic organelles. The type I and II cells are found throughout pregnancy, but the type III cells are observed from mid gestation to term. The cytological features of type I and II cells suggest that they probably secrete most of the steroids, whereas the type III cells primarily store lipids.This research was supported by UPSHS grant AM-11376 and NIH contract 69-2136.     

Structure of taste buds in foliate papillae of the rhesus monkey, Macaca mulatta

Taste buds in foliate papillae of the rhesus monkey were examined by electron microscopy. Three distinct cell types were identified. Type I cells were narrow elongated cells containing an oval nucleus, bundles of intermediate filaments, several Golgi bodies, and characteristic apical membrane-bounded dense granules. These cells exhibited morphological variations: some had a moderately dense cytoplasm, perinuclear free ribosomes, and flattened sacs of rough endoplasmic reticulum; others had a more lucent cytoplasm, dilated irregular rough endoplasmic reticulum, lysosome-like dense bodies, and lipid droplets. Type II cells typically contained a spherical, pale nucleus, a prominent nucleolus, supranuclear and infranuclear Golgi bodies, mitochondria with tubular cristae, and one or two centrioles. This cell type, too, showed some variation in the relative amounts of ribosomes and smooth endoplasmic reticulum, which varied inversely with each other. Type III cells were characterized by a clear apical cytoplasm essentially devoid of ribosomes and containing microtubules. In a few type III cells, the peri- and infranuclear regions contained many ribosomes and some rough endoplasmic reticulum. In most Type III cells, there were large numbers of dense and clear vesicles in the peri- and infranuclear regions; some of the vesicles were grouped in synapse-like arrangements with adjacent nerves. The morphological variations exhibited by all three cell types could be accounted for by age differences in each of the cells. This would be consistent with the notion that cell renewal occurs in each of the three cell populations.     

COTTON EMBRYOGENESIS: THE EARLY DEVELOPMENT OF THE FREE NUCLEAR ENDOSPERM

An electron microscope study was made of the central cell and the development of the free nuclear endosperm surrounding the zygote and synergids during the first three days after pollination. The cytoplasm of the central cell, concentrated around the partially-fused polar nuclei, contains many ribosomes, mitochondria and large, dense, starch-containing plastids, some dictyosomes and lipid bodies, and long, single cisternae of rough endoplasmic reticulum (RER) that frequently terminate in whorls. Dense, core-containing microbodies are closely associated with the RER. After fertilization the cytoplasm of the 2-and 4-nucleate endosperm shows an increase in number of dictyosomes, and in amount of RER which becomes stacked in arrays of parallel cisternae. Cup-shaped plastids are associated with many long, helical polysomes. Perinuclear aggregates of dense, granular material also appear after fertilization. Granular aggregates and helical polysomes disappear after the first few divisions of the primary endosperm nucleus. During the second and third days of development there is an increase in dictyosome number and RER proliferation, and endosperm nuclei become deeply lobed. Concurrently, there is a sharp decline in the starch and lipid reserves of the central cell and elaborate transfer walls are formed at the micropylar end of the embryo sac and on the outer surface of the degenerating synergid. The transfer walls contain groups of small, membrane-bound vesicles, and are associated with large numbers of mitochondria and with the smooth endoplasmic reticulum.     

Effect of ethionine on the rough endoplasmic reticulum from male and female rat liver

Ethionine causes a decrease in the amount of rough endoplasmic reticulum in rat liver, the effect being greater in female than in male rats. Rough endoplasmic reticulum isolated from rat liver 24 hr after ethionine injection and stripped of its ribosomes partially lost itsin vitro ribosome binding capacity. However, no differences were detected between the binding affinities of ribosomes, isolated from either untreated animals or intoxicated rats, to stripped rough membranes derived from normal rats. Structural changes occur in the rough endoplasmic reticulum of the ethionine treated rats, while the ribosomes are still bound to the membrane.     

Juvenile hormone-controlled vitellogenin synthesis in Locusta migratoria fat body: Cytological development

Cytological development in the fat body of adult female Locusta migratoria, related to vitellogenin synthesis, has been studied by light and electron microscopy. In the newly-emerged adult, the cells are filled with lipid droplets, which indent the nucleus, and with fields of glycogen, while ribosomes and endoplasmic reticulum are scarce. Correlated with the onset of vitellogenin synthesis, about day 8 of adult life, the nucleus enlarges, lipid droplets and glycogen decrease, and rough endoplasmic reticulum and Golgi complexes become the most abundant organelles. These changes reflect a conversion of the principal role of the fat body from nutrient storage to the synthesis and secretion of protein. They are prevented by allatectomy, and restored by subsequent treatment with the juvenile hormone analogue, ZR-515. Late in the first gonotrophic cycle, about day 20, dense bodies, vesicle-containing bodies and lysosomes are seen, indicating recycling of cellular materials. Five days after ZR-515 treatment, when protein synthesis has declined, the rough endoplasmic reticulum appears in arrays adjacent to lipid droplets, possibly awaiting reactivation. By the use of ferritin-labelled antivitellin immunoglobulin, vitellogenin has been localized intracellularly in the RER saccules and Golgi vesicles, and extracellularly in channels between the folded plasma membranes, showing sites of accumulation and secretion of this protein.     

Ultrastructural study on degeneration of tapetum in anther of snap bean (Phaseolus vulgaris L.) under heat stress

Pollen sterility was induced by heat stress applied about 10 days before flowering in the snap bean Phaseolus vulgaris L. Cytohistological changes in the tapetum during early development of the anther were studied to identify the tissues most sensitive to high temperature stress. The first distinct structural abnormalities were detected in the distribution pattern of the rough endoplasmic reticulum (RER) in the tapetum at the early microspore stage under high temperature conditions. Stacks of RER were frequently observed in the tapetum under optimal conditions, but rarely occurred under high temperature conditions. Various patterns of endoplasmic reticulum (ER) arrangement – linear, wavy, looped or circular – were observed in the tapetum. Two types of circular ER were observed at the microspore stage under high temperature conditions, RER with ribosomes on the surface and smooth endoplasmic reticulum (SER) lacking ribosomes. The tapetum underwent degenerative changes under high temperature conditions earlier than under optimal conditions. The structural abnormalities of the microspore were associated with tapetal degeneration. We concluded that high air temperature affected the ER structure and blocked its function in the tapetum, and then induced earlier than usual degeneration of tapetum. Pollen sterility is associated with tapetal degeneration. Received: 12 October 2000 / Revision accepted: 7 March 2001     

Laminin ultrastructural immunolocalization in rat testis during ontogenesis

Summary The synthesis of one of the main glycoproteins of the basement membrane, the laminin, was demonstrated by ultrastructural immunolocalization during rat foetal (16th day to 20th day of gestation) and postnatal development of the testis. The lamina densa, part of seminiferous tubular basement membrane, is labeled uniformly at all studied stages. The lamina lucida is not well defined before the postnatal stages, at which times discrete immunostaining extends from the lamina densa to the adjacent seminiferous epithelial cells (spermatogonia and Sertoli cells). The extracellular matrix around the peritubular cells is not labeled before birth. Intracellular immunostaining was detected as early as the 16th day of gestation in both Sertoli cells and cells around the seminiferous tubules which will transform later into peritubular cells. It was located in rough endoplasmic reticulum (RER) cisternae and secretory vesicles. After 18–20 days of postnatal life, the immunostaining faints progressively. Some positive material is seen in the RER of the gonocytes at all studied stages.Sertoli cells and peritubular cells are the main producing cells of laminin after the 16th of gestation. The laminin secreted by gonocytes may play an important role in adhesion of gonocytes to the lamina densa and adjacent Sertoli cells before their transition from basal compartment to adluminal compartment.     

Laminin ultrastructural immunolocalization in rat testis during ontogenesis

The synthesis of one of the main glycoproteins of the basement membrane, the laminin, was demonstrated by ultrastructural immunolocalization during rat foetal (16th day to 20th day of gestation) and postnatal development of the testis. The lamina densa, part of seminiferous tubular basement membrane, is labeled uniformly at all studied stages. The lamina lucida is not well defined before the postnatal stages, at which times discrete immunostaining extends from the lamina densa to the adjacent seminiferous epithelial cells (spermatogonia and Sertoli cells). The extracellular matrix around the peritubular cells is not labeled before birth. Intracellular immunostaining was detected as early as the 16th day of gestation in both Sertoli cells and cells around the seminiferous tubules which will transform later into peritubular cells. It was located in rough endoplasmic reticulum (RER) cisternae and secretory vesicles. After 18-20 days of postnatal life, the immunostaining faints progressively. Some positive material is seen in the RER of the gonocytes at all studied stages. Sertoli cells and peritubular cells are the main producing cells of laminin after the 16th of gestation. The laminin secreted by gonocytes may play an important role in adhesion of gonocytes to the lamina densa and adjacent Sertoli cells before their transition from basal compartment to adluminal compartment.     

Differentiation of the golden hamster oviduct epithelial cells during postnatal development: an electron microscopic study

The ultrastructural changes in the process of differentiation of the epithelial cells of the golden hamster oviduct during postnatal development were investigated by means of electron microscopy. In the epithelium of the ampulla of the neonatal oviducts, no differentiated ciliated cells or secretory cells were identified. In these undifferentiated cells, free ribosomes were well developed, but rough endoplasmic reticulum (RER) and the Golgi apparatus were undeveloped. Cells undergoing ciliogenesis were first identified at 3.5 days after birth, and some ciliated cells appeared at 4.5 days. In the nonciliated cells, marked changes in the organelles were observed at this time. Subsequently, some nonciliated cells containing well-developed RER and Golgi apparatus were observed at 9.5 to 10.5 days after birth, and a few mature secretory cells were observed at 10.5 days. An increase in secretory granules occurred in the secretory cells at 12.5-15.5 days after birth. Many fully mature ciliated and secretory cells were observed at 15.5 days after birth. After 20.5 days after birth, the epithelium was identical with that of the adult golden hamster. Quantitative data indicated that the differentiation of ciliated cells began earlier and took place over a more extended period of time than did that of the secretory cells in the golden hamster oviduct during postnatal development.     

Orderly arrangement of ribosomes in the embryogenic callus tissue of Corylus avellana L

Cultured callus tissue of hazel (Corylus avellana L.), which has the potency of somatic embryogenesis, was used for the study of cell ultrastructure in the course of callus growth and embryoid formation. The meristematic cells of this tissue exhibit a specific organization of rough endoplasmic reticulum (RER), stacked into extensive parallel sheets. The membranes of the aggregated RER are associated with orderly arrays of bound ribosomes. The high regularity of the alignment of the attached ribosomes seems to be influenced by the distance between the two neighbouring membranes in the RER aggregate. The RER aggregates with orderly attached ribosomes are more frequently found in callus cells and in early embryogenesis than in the advanced stages of embryo development.     

The Ultrastructure of Circulating Hemolymph Cells of the Marine Snail Cerithidea californica (Gastropoda: Prosobranchiata)

Two morphologically distinct blood cell-types, the granulocyte and hyalinocyte, are found in the hemolymph circulation of the marine prosobranch Cerithidea californica. Granulocytes, measuring 12.7 µ (9.0–15.0 µ) in diameter, possess well-defined ectoplasmic and endoplasmic regions of the cytoplasm, granules of moderate to heavy electron density, tubular rough endoplasmic reticulum (RER), short vesicles of smooth endoplasmic reticulum (SER), and a large cytoplasm to nucleus ratio. Two morphological variants of this cell-type are distinguished depending upon the presence or absence of dense granules or RER. Hyalinocytes, measuring 5.3 µ (4.0–8.0 µ) in diameter, are distinguished from gran ulocytes by possessing a smaller cytoplasm to nucleus ratio and a general lack of dense cytoplasmic granules and SER.     

Polyribosomal attachment to rat liver and hepatoma endoplasmic reticulum in vitro. A method for its study

A system for study and measurement of the attachment in vitro of exogenous polyribosomes to membranes has been presented. Its main features are use of low temperature, post-microsomal supernatant, pyrophosphate and citric acid to remove ribosomes from the surface of rough endoplasmic reticulum, and a method for quantitative separation of unattached from membrane-associated polyribosomes. The following were found. (1) Rough endoplasmic reticulum, from which ribosomes had been removed by treatment with pyrophosphate and citrate, bound over 50% of added polyribosomes, whereas the untreated (or control) rough and smooth endoplasmic reticulum and the smooth endoplasmic reticulum treated with pyrophosphate-citrate did not bind polyribosomes. (2) The polyribosome-binding capacity of rough endoplasmic reticulum stripped of its ribosomes decayed upon storage of the membranes at 0-4 degrees C. The half-life of this decay was about 6 days whereas that of the polyribosome-binding capacity of hepatoma stripped rough endoplasmic reticulum was about 1.5 days. (3) Preparations of stripped rough endoplasmic reticulum after reassociation with polyribosomes in vitro were quite similar to preparations of native rough endoplasmic reticulum as viewed with the electron microscope. Evidence is presented to support the contention that association of polyribosomes with membranes was the result of polyribosomal reattachment to the membranes rather than trapping of the polyribosomes between vesicles of the membranes.     

The ultrastructure of Blastocystis galli from chickens

The ultrastructure stages of Blastocystis galli were studied in chicken's intestine and in laboratory cultures. There were found morphological structures: surface coat (cell from chickens' intestine showed a very thick surface coat); cell membrane--there were some small electron-opaque deepening "pockets" on the membrane; inner membrane; endoplasmic reticulum with attached ribosomes, which present in the cytoplasm; all cells contained numerous of small vacuoles and large glycogen inclusions in cytoplasm; mitochondria with tubular cristae; nucleus with granules condensed chromatin; central vacuole; Golgi complex was represented by number of plates grouped in a pite; the cyst-like forms were surrounded by multilayered wall.     

A quantitative morphological study of human Leydig cells from birth to adulthood

Summary Human testicular specimens were obtained from biopsies and autopsies covering the period from birth to adulthood. The number of testosterone-containing Leydig cells was determined using the peroxidase-anti-peroxidase method. This number decreased markedly from 3–6 months of age to the end of the first year of life and, up to 6 years of age, only a small number of testosterone-containing cells was found. From 6 years onwards the number of Leydig cells progressively increased. Ultrastructural examination revealed four types of Leydig cells: (1) fetal-type Leydig cells (from birth to 1 year of age) with round nuclei, abundant smooth endoplasmic reticulum and mitochondria with tubular cristae; (2) infantile-type Leydig cells (from birth to 8–10 years of age), showing a multilobated nucleus, moderately abundant smooth endoplasmic reticulum, some lipid droplets and mitochondria with parallel cristae; (3) prepubertal, partially differentiated Leydig cells (from 6 years of age onwards) with regularly-outlined round nuclei, abundant smooth endoplasmic reticulum, mitochondria with tubular cristae, and some lipid droplets and lipofuscin granules; and (4) mature adult Leydig cells (from 8–10 years of age onwards). The ultrastructure of the infantile-type Leydig cells and the lack of delay between the disappearance of the fetal-type Leydig cells and the appearance of infantile-type Leydig cells suggest that fetal-type Leydig cells give rise to the infantile-type Leydig cells. Before puberty, myofibroblast-like precursor cells differentiate into the prepubertal, partially differentiated Leydig cells, which complete their differentiation into the adult Leydig cells.This work was supported by grants from the Comisión Asesora de Investigation Científica y Técnica, and the Fondo de Investigaciones Sanitarias de la Seguridad Social, Madrid, Spain