Electron microscope studies on developing oocytes of a coelenterate medusa with special reference to vitellogenesis

In a hydrozoan jellyfish, the female gonad is differentiated from a specialized region of the epidermis near the manubrium. Changes in the oocytes during growth and vitellogenesis are described as observed with electron microscopic and cytochemical techniques. Three major types of yolk are formed; these include lipid, glycogen, and membrane-bound granules consisting of both protein and carbohydrate. The latter first appear evident within vesicular and cisternal elements of the numerous Golgi complexes. The orientation and structural variations noted between the endoplasmic reticulum and forming face of the Golgi complexes suggest that the protein component of the yolk granules may be transferred from the cisternae of the endoplasmic reticulum to the Golgi complex where it is joined to carbohydrate perhaps synthesized by the Golgi complexes. Stages in the release of the precursor yolk material sequestered in cisternal elements of the Golgi complexes are illustrated. The presence of coated and uncoated vesicles in the Golgi regions and their possible role in intracellular transport are described and discussed. The presence and possible method of morphogenesis of vesiculate yolk bodies are also described. What appear to represent invaginations of the oolemma extend into the ooplasm and display a special orientation with respect to lamellae of the rough-surfaced endoplasmic reticulum. Intraooplasmic synthesis appears to constitute the major pathway for protein-carbohydrate yolk deposition.     

An immunogold,cryoultrastructural study of sites of synthesis and storage of chorionic gonadotropin and placental lactogen in human syncytiotrophoblast

Summary The sites of intracellular synthesis and storage of human placental lactogen (hPL) and human chorionic gonadotropin (hCG) are controversial. We have used one of the most sensitive methods, cryoultramicrotomy and immunogold labelling, to localise these hormones at the electron-microscopic level. In both 12-week and term placentas hCG and hPL are present throughout the rough endoplasmic reticulum cisternae, in the Golgi bodies, and in the infrequent small dense granules of the syncytiotrophoblast. Previous assays have shown that hCG is at a higher concentration in early pregnancy and hPL peaks in late pregnancy, and our results corroborate these findings. No significant localisation of either hormone was seen in the cytotrophoblast or villous stroma. The results suggest that both hCG and hPL are synthesised and packaged by the classical secretory pathway, although the level of hormone stored in granules at any one time is small.     

The formation of intracellular crystals in midgut glands of Limnoria lignorum

Certain morphological features of intracellular crystal formation within the midgut glands of Limnoria lignorum (Rathke) have been studied with the electron microscope and cytochemical methods. A correlation has been established between Golgi membranes and formation of the crystals. The Prussian blue reaction reveals quantities of iron localized in the intracellular crystals and in small granular structures seen in the apical region of the cells. These granules can be identified as accumulations of Golgi membranes, with which iron-containing particles are associated. When these membrane configurations are studied with the electron microscope, they can be classified and arranged in an assumed sequence which is thought to represent successive stages in the development of crystals. As the membrane systems become progressively specialized, increasing accumulations of dense granular material appear within their interstices. This material is rich in iron and probably represents the component responsible for the positive Prussian blue reaction. This material also appears to be a precursor substance for iron-containing protein molecules which are synthesized and arranged to make up the crystals. These iron-containing molecules are first deposited in orderly array as double rows of dense particles on certain internal membranes of the specialized Golgi complexes. The membranes later disappear and the particles form definitive crystals by rearrangement into a hexagonal close-packed pattern.     

Ultrastructural localization of human placental lactogen in distinctive granules in human term placenta: comparison with granules containing human chorionic gonadotropin

Human placental lactogen (hPL) is known to originate in the syncytiotrophoblast, as demonstrated by light microscopic peroxidase and immunofluorescent staining. However, ultrastructural localization of hPL has not previously been performed. In these experiments, immunostaining of electron microscopic sections using protein A-gold and avidin-biotin complex techniques was used to study hPL and human chorionic gonadotropin (beta hCG) localization in first trimester and term placentae. HPL was localized in many small (0.12-0.25 micron) granules. In contrast, beta hCG was found in large (0.40-1.2 micron) granule complexes. The results therefore demonstrate that these two hormones are stored in two morphologically distinct types of cytoplasmic granules. Since hPL and hCG have different secretory mechanisms, this methodology will be useful in studying these differing mechanisms in human placenta.     

The Formation of Intracellular Crystals in Midgut Glands of Limnoria lignorum

Certain morphological features of intracellular crystal formation within the midgut glands of Limnoria lignorum (Rathke) have been studied with the electron microscope and cytochemical methods. A correlation has been established between Golgi membranes and formation of the crystals. The Prussian blue reaction reveals quantities of iron localized in the intracellular crystals and in small granular structures seen in the apical region of the cells. These granules can be identified as accumulations of Golgi membranes, with which iron-containing particles are associated. When these membrane configurations are studied with the electron microscope, they can be classified and arranged in an assumed sequence which is thought to represent successive stages in the development of crystals. As the membrane systems become progressively specialized, increasing accumulations of dense granular material appear within their interstices. This material is rich in iron and probably represents the component responsible for the positive Prussian blue reaction. This material also appears to be a precursor substance for iron-containing protein molecules which are synthesized and arranged to make up the crystals. These iron-containing molecules are first deposited in orderly array as double rows of dense particles on certain internal membranes of the specialized Golgi complexes. The membranes later disappear and the particles form definitive crystals by rearrangement into a hexagonal close-packed pattern.     

Fine structure of the formation and fate of the residual bodies of mouse spermatozoa with evidence for the participation of lysosomes

Routine electron microscopy in combination with subcellular localization of acid phosphatase has been employed to study the formation and fate of residual cytoplasmic bodies extruded into the tubular lumen shortly before spermiation. Prior to extrusion the spermatid cytoplasm contains lipid droplets, mitochondria, ribosomes, endoplasmic reticulum, the caudally migrated Golgi apparatus, and numerous multivesicular and multigranular bodies. These membrane-limited bodies and the Golgi zone stain heavily for acid phosphatase. Following extrusion the residual bodies undergo a series of alterations: (1) disruption of multigranular bodies with release of free granules; (2) sequestration of granules, ribosomes, and reticulum inside double-membrane-limited vacuoles derived from Golgi lamellae; (3) appearance of numerous, single-membrane-bound, cytoplasmic vacuoles; (4) fragmentation; (5) peripheral migration toward the tubular wall; and (6) phagocytosis of these migrating fragments by the Sertoli cells. The demonstration of acid phosphatase activity within free granules, the sequestering Golgi lamellae, and both classes of vacuoles suggests that initial residual body degradation occurs through lysosomal cytoplasmic autophagy.     

抗体形成细胞发育的免疫电镜研究

The pre-embedding immunoelectron microscopic method was used to study the development of antibody-producing cells in the guinea pig popliteal lymph nodes of 2, 3, 5, 8 and 10 days after a second challenge with horseradish peroxidase. The results indicated that the antibody activity was located in the perinuclear space, the endoplasmic reticulum and Golgi complex. According to the cellular developmental stages and the characteristics of distribution of the antibody activity, the antibody-producing cells (APC) were divided into four types: (1) Type I cells (lymphocytes) exhibited many positive granules throughout the cytoplasm; (2) Type II cells (proplasmacytes) contained many positive granules and positive short bars, some of them were parallel; (3) Type III cells (proplasmacytes) contained numerous parallel positive lamellae in cytoplasm; (4) the parallel lamellae in cytoplasm of type IV cells (plasmacytes) were arranged into a network-endoplasmic reticulum. According to the kinetic change from granules, short bars to parallel lamellae and the network, the results indicated the developmental course of AFC from lymphocytes, proplasmacytes to plasmacytes.     

The persistence of Golgi complexes during cell division in an insect epidermis

The Golgi complexes of animal cells are said to become vesicular during cell division in order to allow the equal partitioning of organelles between daughter cells (Warren, 1985). However, in the epidermis of fifth stage larval Calpodes ethlius (Lepidoptera, Hesperi idae), cutical deposition is concurrent with cell division in preparation for pupation. We therefore looked at the Golgi complexes of these epidermal cells to see if they maintained their interphase form to allow them to continue to function during cell division. Dividing cells were recognized by changes in the nucleus and nuclear envelope, the form of the cell cortex and cell surface, and by the disposition of microtubules. Epidermal Golgi complexes consist of 3-5 cisternae capped by endoplasmic reticulum with transfer vesicles and rings of GC beads next to the cis face, and secretory vesicles on the trans face. Golgi complexes of dividing cells are structurally indistinguishable from those in interphase, their beads are in the rings characteristic of active GCs, and cuticle continues in uninterrupted lamellae above the apical microvilli. The observations suggest that Golgi complexes in dividing insect cells differ from those of most vertebrates by remaining functional through mitosis.     

The formation of glandular secretion in larval Austramphilina elongata (Amphilinidea)

The ultrastructure of three types of gland cells of embryos and free-swimming larvae of Austramphilina elongata is described. Type I gland cells contain large, more or less round electron-dense granules which are formed by numerous Golgi complexes. Type II gland cells contain thread-like, membrane-bound secretory granules with longitudinally arranged microtubules inside the granules; secretory droplets are produced by Golgi complexes and the microtubules apparently condense in the cytoplasm or in the droplets. Type III gland cells contain irregular-ovoid membrane-bound granules with coiled up microtubules which have an electron-dense core; the granules are formed by secretionderived from Golgi complexes and the microtubules aggregate around and migrate into the secretion; microtubules are at first hollow and the early secretory granules have a central electron-dense region.     

Oogenesis in Xenopus laevis (Daudin). II. The induction and subcellular localization of tyrosinase activity in developing oocytes

Tyrosinase activity is increased at specific stages of development in Xenopus laevis oocytes in mature females by an injection of 1000 units of human chorionic gonadtropin (HCG). Enzyme activity is stimulated slightly in stage II oocytes, greatly (5- to 6-fold) in stage III and early stage IV oocytes, slightly in late stage IV, and not at all in stage V oocytes. Tyrosinase activity has been localized cytochemically in oocytes by the DOPA-reaction. The DOPA-reaction product is found in the distal cisterna of the Golgi complex and in an anastomosing network of smooth-surfaced tubules associated with the Golgi complex. No reaction product is found in the clustered elements of smooth endoplasmic reticulum which gives rise to the premelanosomes. Substantial melanization of the premelanosomes does not occur until the DOPA-positive Golgi complexes move into proximity with the premelanosomes at the oocyte periphery. Biochemical assay of the isolated melanin granules shows that premelanosomes isolated from stage III and IV oocytes contain significant tyrosinase activity. This activity appears to decrease in the later stages of melanization. It is concluded that the metabolic activities leading to the formation of oocyte pigmentation are stimulated by gonadotropins and the degree of response to the stimulation is quantitatively regulated according to parameters typical of the specific stage of oocyte development.     

Immuno-electron microscopy of formation,degradation and exocytosis of the ovulation neurohormone in Lymnaea stagnalis

Summary The cerebral caudodorsal cells (CDC) of the pulmonate snail Lymnaea stagnalis are involved in the control of egg laying and associated behaviour by releasing various peptides. One of these is the ovulation hormone (CDCH). The cellular dynamics of this peptide have been studied using an antiserum raised to a synthetic portion of CDCH comprising the 20–36 amino acid sequence. With the secondary antibody-immunogold technique, specific immunoreactivity was found in all CDC. Rough endoplasmic reticulum and Golgi apparatus showed very little reactivity as did secretory granules that were in the process of being budded off from the Golgi apparatus. However, secretory granules that were being discharged from the Golgi apparatus, were strongly reactive. Secretory granules within lysosomal structures revealed various degrees of immunoreactivity, indicating their graded breakdown. Large electrondense granules, formed by the Golgi apparatus and thought to be involved in intracellular degradation of secretory material, were only slightly reactive. In the axon terminals secretory granules released their contents into the haemolymph by the process of exocytosis. The exteriorized contents were in most cases clearly immunopositive.The possibility has been discussed that CDCH is cleaved from its polypeptide precursor within secretory granules during granule discharge from the Golgi apparatus; subsequently, the mature secretory granules would be transported towards the neurohaemal axon terminals where they release CDCH into the haemolymph. Superfluous secretory material would be degraded by the lysosomal system including the large electron-dense granules.     

Early stages of ciliogenesis in the respiratory epithelium of the nasal cavity of rabbit embryos

Summary Previous studies have shown that ciliogenesis in the epithelial cells of various species exhibits similarities as well as differences. In an attempt to establish whether this process is identical in epithelial cells of a single species, early stages of centriole formation not previously described were encountered. Ciliogenesis was investigated in the respiratory epithelium of the nasal cavity of 18 to 23-day-old rabbit embryos. The appearance of groups of deuterosomes and fibrous granules is followed by the radial formation of procentrioles around the deuterosomes and parent centrioles. The majority of the procentrioles, forming acentriolarly, occur in pairs, with their distal ends facing each other, between the deuterosomes. Subsequent growth of these procentrioles between deuterosomes in a group results in separating the deuterosomes from one another. The deuterosomes, however, still remain interconnected by means of the growing procentrioles. Accordingly, long chains and networks consisting of the above-mentioned structures are formed. After the procentrioles have attained a certain length, the chains and networks split into separate deuterosome-procentriole complexes. During these earlier stages of ciliogenesis fibrous granules are present, however, their function is yet to be determined.     

The Golgi apparatus in the acinar cells of the developing embryonic pancreas: I. Morphology and enzyme cytochemistry

The Golgi apparatus of pancreatic acinar cells of rat embryos was studied during development from day 13 through day 20 of gestation. The morphological and enzyme cytochemical patterns varied characteristically in the course of cell differentiation. A pronounced system of "rigid lamellae" characterized the area near the trans face of the Golgi stacks in the protodifferentiated and early phases of the differentiated states; by contrast, "rigid lamellae" were sparse in the terminal period of gestation. Reaction product of acid phosphatase labeled the "rigid lamellae" in the protodifferentiated state, was extended across the majority of the stacked cisternae in the early differentiated state, but was restricted to the trans side again in the later periods of cell differentiation. The early phase of the differentiated state was characterized by the tight association of the endoplasmic reticulum and Golgi cisternae on the trans side; the close spatial relationship of the two compartments was lessened after production of secretion granules had started. The findings are in line with those of recent studies on the Golgi organization in some other types of cells in different functional states, and they present the embryonic pancreatic tissue as another model for demonstrating the high flexibility of the Golgi complex. In agreement with the patterns previously found in the absorptive cells of the small intestine, the present results show that the close associations of the endoplasmic reticulum and cisternae of the trans Golgi side predominate in the early stages of cell differentiation.     

The Golgi complex in the esophageal mucous glands of the newly hatched chick

The general morphology of the mucous gland cell and the nature of the secretory granule in esophageal glands of the newly hatched chick have been described. Lightly basophilic supporting cells, attached to secretory cells by desmosomes and containing tonofilaments, are located on the basal lamina. Electron microscopic studies showed a morphological polarity of the Golgi complex which suggests that mucous precursors are transported from other sites within the cell to the Golgi complex for further packaging into secretory granules. Finally, acid mucopolysaccharides (AMPS) were specifically stained using the Thorotrast technique and not detected in the rough endoplasmic reticulum, the transitional elements, or in the lamellae at the forming face of the Golgi complex. Conversely, AMPS are found in the vicinity of the mature face of the Golgi complex, and in the secretory granules. The acquisition of cytochemical reactivity for AMPS within the Golgi complex is discussed.     

THE CYTOLOGY OF THE NORMAL PARATHYROID GLANDS OF MAN AND VIRGINIA DEER : A Light and Electron Microscopic Study with Morphologic Evidence of Secretory Activity

The normal parathyroids of six humans and a Virginia deer were studied by light and electron microscopy. The parenchyma of the deer parathyroid is composed of uniform chief cells, which contained 100 to 400 mµ electron-opaque, membrane-limited granules, presumed to be secretory granules, in addition to the usual cytoplasmic organelles. Desmosomes are present between adjacent cells, and rare cilia are observed protruding from the chief cells into the intercellular space. The human parathyroids contain chief cells in two phases—active and inactive—as well as oxyphil cells. Active chief cells have a large Golgi apparatus, sparse glycogen, numerous secretory granules, and rare cilia. Inactive chief cells contain a small Golgi apparatus, abundant glycogen, and few secretory granules. Both forms have the usual cytoplasmic organelles and, between adjacent cells, desmosomes. Oxyphil cell cytoplasm is composed of tightly packed mitochondria and glycogen granules, with rare secretory granules. Cells with cytoplasmic characteristics intermediate between chief and oxyphil cells, possibly representing transitional cells, have been observed. Secretory granules of both man and deer are composed of 100 to 200 A particles and short rods, and the granules develop from prosecretory granules in the Golgi region of the cell. The human secretory granules are smaller and more variable in shape than those of the deer. The granules are iron and chrome alum hematoxylin-positive, argyrophilic, and aldehyde fuchsin-positive, permitting light microscopic identification. They are also found in the capillary endothelial cells of the parathyroid and in its surrounding connective tissue. The secretory granules of the parathyroid cells can thus be followed from their formation in the Golgi apparatus almost to their extrusion into the blood stream.     

The fine structure of the secretory cells of the uterus (shell gland) of the chicken

The secretory processes in the shell gland of laying chickens were the subject of this study. Three cell types contribute secretory material to the forming egg: ciliated and non-ciliated columnar cells of the uterine surface epithelium, and cells of tubular glands in the mucosa. The ciliated cells as well as the non-ciliated cells have microvilli, which undergo changes in form and extent during the secretory cycle. At the final stages of shell formation they resemble stereocilia. It is postulated that the microvilli of both cells are active in the production of the cuticle of the shell. The ciliated cell which has both cilia and microvilli manufactures secretory granules which arise from the Golgi complex in varying amounts throughout the egg laying cycle. Granule production reaches its greatest intensity during the early stages of shell deposition. The ciliated cell probably supplies proteinaceous material to the matrix of the forming egg shell. The non-ciliated cell has only microvilli. Secretory granules, containing an acid mucopolysaccharide, arise from the Golgi complex. Some granules are extruded into the uterine lumen where they supply the egg shell with organic matrix. Others migrate towards the supranuclear zone. Here a number of them disintegrate. This is accompanied by the formation of a large membraneless space, which is termed “vacuoloid.” Subsequently the vacuoloid regresses and during regression an extensive rough endoplasmic reticulum with numerous polyribosomes of spiral configuration appears. It is suggested that material in the vacuoloid originating from the disintegrating granules is resynthesized and utilized for the formation of secretory product. The uterine tubular gland cells have irregular, frondlike microvilli. During egg shell deposition, these microvilli form large blebs and are probably related to the elaboration of a watery, calcium-containing fluid.     

日本沼虾高尔基体在精子发生过程中的变化

用岸民镜技术研究了日本沼虾精子发生过程中生精细胞内高尔基体变化。结果表明：精原细胞内,高尔基体结构典型,分布在核膜附近,许多膜囊通过过连接小管相互连接。初级精母细胞内,高尔基体结构紧凑且更典型,更造近核膜,在反面的分泌活动旺盛,产生大量初级溶酶体;     

ORIGIN OF GRANULES IN POLYMORPHONUCLEAR LEUKOCYTES : Two Types Derived from Opposite Faces of the Golgi Complex in Developing Granulocytes

The origin, nature, and distribution of polymorphonuclear leukocyte (PMN) granules were investigated by examining developing granulocytes from normal rabbit bone marrow which had been fixed in glutaraldehyde and postfixed in OsO₄. Two distinct types of granules, azurophil and specific, were distinguished on the basis of their differences in size, density, and time and mode of origin. Both types are produced by the Golgi complex, but they are formed at different stages of maturation and originate from different faces of the Golgi complex. Azurophil granules are larger (~800 mµ) and more dense. They are formed only during the progranulocyte stage and arise from the proximal or concave face of the Golgi complex by budding and subsequent aggregation of vacuoles with a dense core. Smaller (~500 mµ), less dense specific granules are formed during the myelocyte stage; they arise from the distal or convex face of the Golgi complex by pinching-off and confluence of vesicles which have a finely granular content. Only azurophil granules are found in progranulocytes, but in mature PMN relatively few (10 to 20%) azurophils are seen and most (80 to 90%) of the granules present are of the specific type. The results indicate that inversion of the azurophil/specific granule ratio occurs during the myelocyte stage and is due to: (a) reduction of azurophil granules by multiple mitoses; (b) lack of new azurophil granule formation after the progranulocyte stage; and (c) continuing specific granule production. The findings demonstrate the existence of two distinct granule types in normal rabbit PMN and their separate origins from the Golgi complex. The implications of the observations are discussed in relationship to previous morphological and cytochemical studies on PMN granules and to such questions as the source of primary lysosomes and the concept of polarity within the Golgi complex.     

Subcompartment sugar residues of gastric surface mucous cells studied with labeled lectins.

We examined the intracellular localization of sugar residues of the rat gastric surface mucous cells in relation to the functional polarity of the cell organellae using preembedding method with several lectins. In the surface mucous cells, the nuclear envelope and rough endoplasmic reticulum (rER) and cis cisternae of the Golgi stacks were intensely stained with Maclura pomifera (MPA), which is specific to alpha-Gal and GalNAc residues. In the Golgi apparatus, one or two cis side cisternae were stained with MPA and Dolichos biflorus (DBA) which is specific to terminal alpha-N-acetylgalactosamine residues, while the intermediate lamellae were intensely labeled with Arachis hypogaea (PNA) which is specific to Gal beta 1,3 GalNAc. Cisternae of the trans Golgi region were also stained with MPA, Ricinus communis I (RCA I) which is specific to beta-Gal and Limax flavus (LFA) which is specific to alpha-NeuAc. Immature mucous granules which are contiguous with the trans Golgi lamellae were weakly stained with RCA I, while LFA stained both immature and mature granules. The differences between each lectin's reactivity in the rough endoplasmic reticulum, in each compartment of the Golgi lamellae and in the secretory granules suggest that there are compositional and structural differences between the glycoconjugates in the respective cell organellae, reflecting the various processes of glycosylation in the gastric surface mucous cells.     

Some observations on spermatogenesis in Gelastocoris oculatus (Hemiptera) with the aid of the electron microscope

The nuclear cap in the spermatogonial and early spermatocyte cells of Gelastocoris is an aggregate of closely packed mitochondria with their long axes perpendicular to the nuclear membrane. Eventually in the early growth period, the mitochondria move from the cap and appear to become more or less equally distributed in the cytoplasm where they remain until their fusion in the spermatid to form the nebenkern. The Golgi complex consists of clusters of lamellae and vesicles, the Golgi bodies. Granules form within the vesicles, increase in size, move from their place of origin and become distributed at random in the cytoplasm. They are the pro-acrosomal granules and at the end of the growth period fuse to form the proacrosome, about which Golgi bodies collect. The Golgi bodies, however, never fuse into an acroblast. At one end of the oval-shaped pro-acrosome is a small dark body and a less dense vesicle the future of which is uncertain. The dark body eventually occupies a position at the tip of the acrosome. The pro-acrosome, after moving to the side of the nucleus opposite the nebenkern, differentiates into the acrosome which elongates into a tail-like structure. The nuclear membrane of some spermatocytes may appear wave-like in cross section, with the crest and trough different in appearance. Near the membrane and in the troughs of the waves large clusters of granules are frequently present. Similar clusters may be found elsewhere in the cytoplasm. Presumably they had their origin near the membrane but this is not conclusive. Bodies of indeterminate origin and structure may be present in the cytoplasm. They could be lysosomes but evidence is lacking. In late spermatocytes and in spermatids, a group of ten or twelve granules is present. They are smaller than the pro-acrosomal granules, are always closely associated and pass as a group into the tail. Their significance is unknown. The endoplasmic reticulum is typical of cells in general. There are no granule accumulations within the vesicles as in some secretory cells. Vesicles of various shapes and sizes are present within the centrosphere of the first meiotic division. While their location is similar to that of the centriole, the identity of the vesicles is uncertain.