An electron microscopical study of epididymal epithelium of rats treated with gonadotropic hormone

The ultrastructural modifications of the epithelial cells of rat corpus epididymis stimulated with gonadotropic hormone were studied. The structural variety of the cells depending on functional conditions becomes more prominent 6 h after the injection of gonadotropic hormone. Light large cells have one or often two nucleus-containing bing nucleoli, in their cytoplasm there are numerous vesicles, a well-developed Golgi apparatus, other organelles and lysosomal bodies. Some other cells are filled with many large vacuoles of different density, dense bodies and vesicles. Cells of another type which are in the majority show an unusually active structure reflecting the function of synthesis. The more prominent nucleolus is associated to clumps of chromatin. Their apical cytoplasm is filled by a structure related to absorption. The whole remaining part of their cytoplasm is covered with a very extensive Golgi apparatus and a very well developed granular endoplasmic reticulum. The extremely enlarged cisternae of this reticulum were found to be very closely applied to the basal cell membrane. There is a flocculent material inside the cisternae. Similar material is observed in the extracellular medium under the basal membrane. The epithelium seems normal 10 h after the injection of hormone, but large light cells make up the majority of them.     

Ultrastructure of the eyes of the grass shrimp,Palaemonetes pugio

Summary The cone cells and corneagenous cells possess extensive networks of smooth tubular endoplasmic reticulum that may be involved in optical reflectance and light-adaptational responses, respectively. The extracellular basal lamina of the basement membrane is confluent with glial cell capillary walls and may prove to be a viaduct for the transmission of hemolymph-borne substances to the retina or of retinal degradation products to the hemolymph. In addition to dense pigment granules, the distal pigment cells are shown for the first time to contain migratory reflecting platelets that are usually polymorphic in light-adapted eyes but are rectangular in dark-adapted eyes. In the latter these plates become aligned against the crystalline cones and presumably contribute to the reflection superposition optics of the grass shrimp. Dark-adapted retinular cells possess well-developed perirhabdomal cisternae, oblong or ovoid mitochondria, generally vesicular rough endoplasmic reticulum, and occasional, spherical, calcium-like intrarhabdomal inclusions. Light-adapted retinular cells possess poorly developed perirhabdomal cisternae, lamelliform rough endoplasmic reticulum, and condensed mitochondria frequently associated with lipid droplets and pigment granules. The cytoplasmic boundaries of the reflecting pigment cells expand into the extracellular spaces between individual ommatidial retinular cells during dark adaptation and recede to the interommatidial extracellular spaces during light adaptation. Cytoplasmic microfilament bundles found only at the bases of partially light-adapted rhabdomeric microvilli may be involved in microvillar shortening.     

The ecdysial gland of the spider crab,Libinia emarginata (L). I. Ultrastructure of the gland in the male

The ecdysial glands of mature male Libinia emarginata are pale, yellowish organs composed of lobes of epithelial cells having oval nuclei which are often eccentric and which have one or two nucleoli containing amorphous granular material and coarse strands. The plasma membrane bordering the basal lamina consists of invaginations containing microtubules which may serve to increase the surface area for metabolic exchange. Masses of smooth endoplasmic reticulum and associated vesicles are scattered throughout the cytoplasm. Two or more vacuoles may coalesce. Larger vesicles lie close to the cell surface. Numerous mitochondria with tubular cristae surround the nucleus and frequently are associated with SER. A few Golgi complexes consisting of flattened sacs, cisternae or vesicles, lipid droplets and free ribosomes were seen. Adjacent plasma membranes may be in close apposition or separated by a space filled with vesicles, granules, or blood or supporting cells. This type of ultrastructure is associated with steroid-secreting cells.     

An ultrastructural study of the clitellar epithelium of the earthworm Metaphire posthuma (vail.)

The ultrastructure of clitellar epithelium of Metuphire posthuma revealed mainly three types of secretory cells. Most prominent among these are the large slender granular cells which contain a large number of secretory granules filling in the entire columncr region of the cell. The secretory granules are 2-4mu in diameter with a limiting membrane and containing numerous tiny vesicles in a matrix of varying electron density. Basolateral rough endoplasmic reticulum and extensive Golgi cisternae were seen interspersed with the secretory granules. The Golgi cisternae in these cells were quite prominent extending all around the secretory granules. The secretory granules of type 2 cells are spheroid bodies with motley appearance due to varying electron density of the matrix. The immature granules contain fibrillar material. Type 3 cells contained electron lucent membrane-bound mucous like secretory granules which are reticulated with filamentous materials. All the three cell types open to the exterior at the cuticular region which is characterised by the presence of numerous microvilli.     

Changes in the structure of Nippostrongylus brasiliensis intestinal cells during development from the free-living to the parasitic stages

The ultrastructure of Nippostrongylus brasiliensis intestinal cells was examined in free-living, feeding second-stage larvae, infective, nonfeeding third-stage larvae, and parasitic, feeding third-stage larvae. The intestinal cells of second-stage larvae were characterized by a well-developed microvillar border, large numbers of ribosomes, Golgi complexes, rough endoplasmic reticulum, and nuclei with prominent nucleoli. The intestinal cells of infective, third-stage larvae had very few microvilli and the cells were extremely narrow. Few ribosomes, Golgi complexes, and little rough endoplasmic reticulum were present. Nuclei did not contain nucleoli. When worms were introduced into an in vitro culture system, development of intestinal cells began. By 36 hr, microvilli were well differentiated and the cysoplasm contained numerous ribosomes and Golgi complexes, and rough endoplasmic reticulum, mitochondria, and nucleoli were prominent. These morphological changes were related to changes in the physiology of Nippostrongylus brasiliensis which occur during development from a free-living to parasitic form.     

Cytological differentiation in the uropygial gland.

Ultrastructural changes were studied in the cells undergoing secretory differentiation in zone I of the tubules of the uropygial gland of White Plymouth Rock chickens. A layer of basal cells and four secretory stages are recognized as the cells migrate from the periphery to the lumen of tubules and progressively elaborate a secretion product. Basal cells, containing rough endoplasmic reticulum and free ribosomes, rest on the basement membrane and are the source from which secretory cells arise. Dilated perinuclear cisternae and the proliferation of smooth endoplasmic reticulum in the form of vesicles, invaginated sacs and cusp-shaped cisternae indicate the onset of lipgenesis in stage I cells. The perinuclear cisternae are more dilated and the endoplasmic reticulum is composed on saccules and cisternae in stage II cells. Stage III cells are characterized by concentric lamellae of endoplasmic reticulum surrounding secretory droplets. Dilated cisternae of endoplasmic reticulum and secretory droplets both contain a reticular substance. The perinuclear cisternae of stage III cells have returned to normal dimensions. Large mature lucent secretory droplets, lined with electron-dense material, fill the cytoplasm ostage IV cells which degenerate and release their secretory product into the tubule lumen. Spherical membrane-bound compartments containing a mottled substance of moderate electron density occur in basal cells and all subsequent secretory stages. These mottled bodies are surrounded by saccules of endoplasmic reticulum in stage II cells and are intimately associated with secretory droplets in stage III cells, but there is no evidence that they give rise to secretory droplets and their role in secretory differentiation is unknown.     

Oligovillous cells of the epidermis: sensory elements of lamprey skin

The epidermis cf Lampetra spp. contains several kinds of differentiated cell; one innervated variety is characterized by bearing a group of large apical microvilli which project from the surface of the skin. In Lampetra planeri such oligovillous cells are numerous under the oral hood of the ammocoete larva, on the papillae fringing the dorsal fin and bordering the gill vents of the adult, and at the tip of the male genital papilla. Elsewhere on the head, body and fins they are present but more scattered, which appears to be the condition also in adult anadromous Lampetra fluviatilis . There are differences in the number and dimensions of the microvilli found on oligovillous cells, but each is supported by a stout core of actin filaments extending a variable distance down into the cytoplasm. Under the apex of the cell there are microtubules and numerous vesicles which are thought to be concerned in the renewal of the membrane on the microvilli. Beside and proximal to the nucleus is a system of channels of rough endoplasmic reticulum, and a stack of membranous cisternae which appears to have been derived from the endoplasmic reticulum. A nerve fibre is associated with the base of the cell which is indented by a spur-like process from the neurite. Typical "synaptic vesicles" are not found in the cell but irregular vesicular or canalicular profiles are associated with the cell membrane adjoining the neurite spur. The space between the cell and neurite membranes contains extracellular material with a characteristic appearance of prickle-like densities on the cell side meeting densities on the neurite membrane. Variations in the cytology of oligovillous cells can be explained in terms of a cycle of development and de-differentiation. Certain cells with vesicles throughout the cytoplasm and with a narrow apex without microvilli are interpreted as degenerate examples. The oligovillous cells are thought to be chemosensory receptors.     

Ultrastructural, histochemical and cytochemical characterization of intestinal epithelial cells in Aplysia depilans (Gastropoda, Opisthobranchia)

To improve the current knowledge about the digestive system in opisthobranchs, light and electron microscopy methods were used to characterize the epithelial cells in the mid‐intestine of Aplysia depilans. This epithelium is mainly formed by columnar cells intermingled with two types of secretory cells, named mucous cells and granular cells. Columnar cells bear microvilli on their apical surface and most of them are ciliated. Mitochondria, multivesicular bodies, lysosomes and lipid droplets are the main components of the cytoplasm in the region above the nucleus of these cells. Peroxisomes are mainly found in middle and basal regions, usually close to mitochondria. Mucous cells are filled with large secretory vesicles containing thin electron‐dense filaments surrounded by electron‐lucent material in which acidic mucopolysaccharides were detected. The basal region includes the nucleus, several Golgi stacks and many dilated rough endoplasmic reticulum cisternae containing tubular structures. The granular cells are characterized by very high amounts of flat rough endoplasmic reticulum cisternae and electron‐dense spherical secretory granules containing glycoproteins. Enteroendocrine cells containing small electron‐dense granules are occasionally present in the basal region of the epithelium. Intraepithelial nerve fibres are abundant and seem to establish contacts with secretory and enteroendocrine cells.     

Cellular organization and peritrophic membrane formation in the cardia (Proventriculus) of Drosophila melanogaster

The peritrophic membrane of Drosophila melanogaster consists of four layers, each associated with a specific region of the folded epithelial lining of the cardia. The epithelium is adapted to produce this multilaminar peritrophic membrane by bringing together several regions of foregut and midgut, each characterized by a distinctively differentiated cell type. The very thin, electron-dense inner layer of the peritrophic membrane originates adjacent to the cuticular surface of the stomadeal valve and so appears to require some contribution by the underlying foregut cells. These foregut cells are characterized by dense concentrations of glycogen, extensive arrays of smooth endoplasmic reticulum, and pleated apical plasma membranes. The second and thickest layer of the peritrophic membrane coalesces from amorphous, periodic acid-Schiff-positive material between the microvilli of midgut cells in the neck of the valve. The third layer of the peritrophic membrane is composed of fine electron-dense granules associated with the tall midgut cells of the outer cardia wall. These columnar cells are characterized by cytoplasm filled with extensive rough endoplasmic reticulum and numerous Golgi bodies and by an apical projection filled with secretory vesicles and covered by microvilli. The fourth, outer layer of the peritrophic membrane originates over the brush border of the cuboidal midgut cells, which connect the cardia with the ventriculus.     

Endocytosis in nonciliated epithelial cells of the ductuli efferentes in the rat

The nonciliated cells lining the ductuli efferentes presented three distinct cytoplasmic regions. The apical region contained, in addition to cisternae of endoplasmic reticulum and mitochondria, two distinct membranous elements. The tubulovesicular system consisted of dilated tubules connected to the apical plasma membrane and subjacent distended vesicular profiles. The apical tubules, not connected to the cell surface, consisted of numerous densely stained tubules of small size which contain a compact, finely granulated material. The supranuclear region, in addition to a Golgi apparatus and ER cisternae, contained dilated vacuoles, pale and dense multivesicular bodies, as well as numerous dense granules identified cytochemically as lysosomes. The basal region contained the nucleus and many lipid droplets. The endocytic activity of these cells was investigated using cationic ferritin (CF) and concanavalin-A-ferritin (Con-A-ferritin) as markers of adsorptive endocytosis; and native ferritin (NF), concanavalin-A-ferritin in the presence of alpha-methyl mannoside, and horseradish peroxidase or albumin bound to colloidal gold for demonstrating fluid-phase endocytosis. These tracers were injected separately into the rete testis, and animals were sacrificed at various time intervals after injection. At 1 min, CF or Con-A-ferritin were seen bound to the apical plasma membrane, to the membrane of microvilli, and to the membrane delimiting elements of the tubulovesicular system. Between 2 and 5 min, these tracers accumulated in the densely stained apical tubules and at 15 min in the dilated vacuoles. Between 30 min and 1 hr, the tracers appeared in multivesicular bodies of progressively increasing density, whereas at 2 hr and later time intervals, many dense lysosomal elements became labeled. The tracers for fluid-phase endocytosis showed a distribution similar to that for CF or Con-A-ferritin except that they did not bind to the apical plasma membrane, microvilli, or membrane delimiting the tubulovesicular system. At no time interval were any of the tracers observed in the abluminal spaces. Thus, the nonciliated epithelial cells of the ductuli efferentes are actively involved in fluid-phase and adsorptive endocytosis, both of which result in the sequestration of endocytosed material within the lysosomal apparatus of the cell.     

Ultrastructure of the nuclear apparatus of the infusorian Loxodes magnus. I. The macronuclei, macronuclear anlagen and the interphasic micronuclei]

The nuclear apparatus of Loxodes magnus Stokes (Holotricha) consists of numerous macronuclei which belong to the diploid type and never divide, and of numerous micronuclei. No nuclear groups exist; individual nuclei often lie in cytoplasmic islets surrounded by large lacunae of the smooth endoplasmic reticulum. Interphasic micronuclei have two-membraned envelopes with numerous pores, usually lined at the cytoplasmic side with a layer of vacuoles, channels, or flattened vesicles of the smooth endoplasmic reticulum. The chromatin of the micronuclei consists of anastomosing threads, 0.1--0.2 mum wide, between which several nucleolus-like bodies of microfibrillar structure occur. Adult macronuclei have a similar nuclear envelope and a similar system of vacuoles, channels, and flattened agranular cisternae outside it. The macronucleus contains a single large composite nucleolus with 3 or 4 fibrillar cores inside the common granular cortex. The fibrillar cores are pierced by channels containing nucleolar organizers in the form of strands of condensed chromatin. The peripheral zone of the macronucleus is filled with decondensed chromatin fibrils and contains a number of small chromocenters and several aggregates of RNP granules. No protein inclusions (spheres) have been observed in Loxodes macronuclei. The macronuclear anlagen, developing in the cycle of every cell division, show progressive decondensation of the chromosomes and formation of several nucleoli, each with its own organizer. Later on, the nucleoli fuse into a single nucleolus. The small chromocentres are the last to form.     

Alterations of the cytoplasmic organization of WIRL cells induced by trifluoperazine

The antipsychotic drug trifluoperazine (TFP) causes a reversible rounding of cells of the rat liver epithelial cell line, WIRL. We have investigated the cytoplasmic organization of these cells after TFP treatment using SEM, TEM and immunofluorescence and have observed significant differences between the control and treated cells. Mitochondria are converted to the condensed configuration with distended cristae and the endoplasmic rcticulum becomes tubular with distended cisternae. Intermediate filaments, visualized with a monoclonal antibody, are aggregated to a cap on the nucleus in an arrangement different from that induced by colcemid.     

Possible relationship between the activity of the adrenal gland and the subcommissural organ in the lizard Lacerta s. sicula raf.

Summary In order to study the possible functional relationship between the adrenal gland and the subcommissural organ (SCO) in the lizard Lacerta s. sicula Raf., ACTH was administered to some specimens of this species in January when both the adrenal gland and the subcommissural organ have a very low activity. In comparison to untreated controls, the adrenals of animals treated with ACTH showed clear signs of stimulation, presenting enlarged blood vessels, very few lipid droplets, numerous polymorphic mitochondria and abundant tubular smooth endoplasmic reticulum. In addition, a distinct increase in secretory material was observed in the subcommissural cells of specimens treated with ACTH. These cells showed large cisternae of the rough endoplasmic reticulum filled with granular material in the basal region, numerous secretory granules of two types in the apical region and a reduced number of microvilli on the free cell surface. These findings, together with the results of preceding studies, lead the authors to the consideration that steroid hormones might play a role in the regulation of the secretory activity of the SCO.     

Nuclear division in the cyst,and white spot nuclei preceding cyst formation,inAcetabularia wettsteinii

Summary Electron microscopy of nuclear division in young cysts ofAcetabularia wettsteinii shows that the dividing nucleus hat two additional cisternae of endoplasmic reticulum immediately outside the nuclear envelope. These additional cisternae are attached to, and apparently formed from a membrane body which develops outside the nucleus in early prophase. The interphase nucleus does not have the additional cisternae. The nucleoli are extruded from the nucleus at anaphase, the nucleolar bodies remaining in the peri-nuclear cytoplasm. The chromosomes have localized centromeres; the stratified ultrastructure characteristic of some chlorophycean and animal kinetochores has not been found inAcetabularia, although the kinetochore appears distinct, projecting from the chromatid, and has attached microtubules. The condensed bodies of the white spot nucleus are discussed.     

The fine structure of the retina of the honey bee drone

Summary The eye of the honey bee drone is composed of approximately 8,000 photoreceptive units or ommatidia, each topped by a crystalline cone and a corneal facet. An ommatidium contains 9 visual or retinula cells whose processes or axons pierce a basement membrane and enter the optic lobe underlying the sensory retina. The visual cells of the ommatidium are of unequal size: six are large and three, small. In the center of the ommatidium, the visual cells bear a brush of microvilli called rhabdomere. The rhabdome is a closed-type one and formed mainly by the rhabdomeres of the six large retinula cells. The rhabdomeric microvilli probably contain the photopigment (rhodopsin), whose modification by light lead to the receptor potential in the retinula cells. The cytoplasm of the retinula cells contains various organelles including pigment granules (ommochromes), and peculiar structures called the subrhabdomeric cisternae. The cisternae, probably composed of agranular endoplasmic reticulum undergo swelling during dark adaptation and appear in frequent connection with Golgi cisternae. Three types of pigment cells are associated with each ommatidium. The crystalline cone is entirely surrounded by two corneal pigment cells. The ommatidium, including its dioptric apparatus and corneal pigment cells, is surrounded by a sleeve of about 30 elongated cells called the outer pigment cells. These extend from the base of the corneal facet to the basement membrane. Near the basement membrane the center of the ommatidium is occupied by a basal pigment cell. Open extracellular channels are present between pigment cells as well as between retinula cells. Tight junctions within the ommatidium are restricted to the contact points between the rhabdomeric microvilli. These results are discussed in view of their functional implications in the drone vision, as well as in view of the data of comparative morphology.This work was supported by a grant from the Fonds National Suisse de la Recherche Scientifique.     

Blastocyst attachment and morphogenesis of ectoplacental cone in mouse

Mouse blastocyst attaches on the antimesometrial side of the uterus through mural trophoblasts. Later the polar trophoblasts begin proliferation, and rapid multiplication towards the mesometrial side of the uterus occurs resulting in the formation of an excrescence designated as ectoplacental cone. The morphogenesis of ectoplacental cone, viewedin utero, initiates on day 6post-coitum when microvilli of the trophoblast and the uterine epithelial cells are lost and as a result of this opposing membranes appear interlocked with each other. Soon following the invasion by surrounding trophoblasts the necrosis of the epithelial cells starts. Mitochondriae of the epithelial cells, at this stage, are shrunken and lack well defined cristae. Several leucocytes are seen at the site and few electron dense structures appear wedged between the trophoblasts and epithelial cells. At places the cell membrane is studded with the basement membrane of the uterine epithelium giving an impression of a bristle coated membrane. By day 7post-coitum the basement membrane has almost disappeared leaving trophoblast cells to develop close contact with stromal cells. Collagen fibres appeared between the trophoblasts and the stromal cells, many large inclusions of high electron density representing engulfed necrotic epithelial cells are discernible. On day 8post-coitum the ectoplacental cone is fully developed. Four types of trophoblast cells can be identified in it: (i) basal cells lying on the base of the cone, are polyhedral and compactly arranged. They have a large nucleus and well developed nucleoli, (ii) central cells forming the middle area of the cone are of two types; one contained several osmiophilic granules enclosing translucent area (eccentric) and a well developed golgi complex around the nucleus, while the other has many heterophagosomes, vacuoles and residual bodies and (iii) peripheral cells contained several pleomorphic structures resembling secondary lysosomes. Minute dense granules and band of microfibrils on the apical region of these cells are seen. Dense granules probably release lytic proteins at the site and microfibrils help in forming cytoplasmic projections.     

Electron microscopic contrast of the cytoskeleton and junctional complexes of intestinal epithelial cells by ethanolic phosphotungstic acid

After glutaraldehyde fixation and treatment with ethanolic phosphotungstic acid (E-PTA) before plastic embedding, sections of rat large intestine showed a characteristic electron contrasting pattern in epithelial cells. The axis of microvilli, terminal web, a thin band below the luminal plasma membrane, centrioles and junctional complexes (tight junctions, adherens junctions, and desmosomes) appeared highly contrasted. In addition to protein components of microfilaments and intermediate filaments, proteins from the junctional complexes could also be implicated in the contrasting reaction with E-PTA. Mitochondrial membranes, chromatin masses, and nucleoli of enterocytes showed considerable electron density, whereas no reaction was found in the glycocalyx and mucin content of goblet cells. The clear visualization of cytoskeleton elements and junctional complexes by E-PTA contrasting represents a simple and valuable method for studies on the normal and pathological organization of these structures in epithelial cells.     

The Eyes of Mesostoma ehrenbergi (Focke, 1836) (Platyhelminthes,Rhabdocoela). Fine Structure and Photoreceptor Membrane Turnover

Abstract Each pigment-cup eye of Mesostoma ehrenbergi consists of two photoreceptor cells, the anterior cell being bilobate. the posterior almost linear, and of a multicellular pigment cup. The nuclei of the photoreceptor cells are located inside the medial region of the brain. Thin cytoplasmic photoreceptor projections provided with neurosecretory-like granules are interposed between the inner surface of the eye cup and the distal extremity of the microvilli. The breakdown and renewal of microvillar membranes was analysed. Membrane turnover is a continuous process. At dusk and during the night abscission of photoreceptive membranes occurs. At dawn the membrane fragments are degraded to granular material, which is then endocytosed into the submicrovillar cytoplasm as coated vesicles. These vesicles form multivesicular bodies. The degradation of multivesicular body content occurs during the following light hours. The dark period is correlated with membrane synthesis for elongation of reticular membranes, which are converted into ellipsoid bodies. The formation of new microvillar membranes occurs at the base of the microvillar border, and involves the fusion with the old microvillar membranes of small vesicles detached from the tubular endoplasmic membranes and from the flattened concentric cisternae of ellipsoid bodies. The correlations with daily cycles of other invertebrates are discussed.     

Early development of the kelp fly,Coelopa frigida (Diptera). II. Morphology of cleavage and blastoderm formation

Cleavage and blastoderm formation in Coelopa frigida are extremely rapid developmental processes. In short (6–7 minutes) successive cell cycles, nuclei multiply and spread out through the egg. The movement seems to be aided by endoplasmic vesicles and cisternae which are in direct contact with the nuclear membrane. The first cells to separate from the egg plasmodium in early superficial cleavage stages are the pole cells. Precursor material from multivesicular bodies forms the pole cell membranes. The primary nuclei from the posterior pole region are removed from the blastoderm by the pole cell segregation. Blastoderm nuclei from the regions adjacent to the posterior pole migrate into the residual periplasm after pole cell segregation has been completed and constitute the blastoderm nuclei in that region of the egg. Nucleoli are not revealed during internal cleavage. They appear in pole cells shortly after their segregation. The generation time of the blastoderm nuclei increases after the twelfth cleavage. Concurrently, nucleoli form in the blastoderm nuclei and permanent cell membranes separate individual blastoderm cells. After blastoderm cells have been separated from each other, they remain in contact with the interior yolk sac by means of cytoplasmic canals. This contact is maintained at least during the early phases of blastokinesis. Observations on nuclear migration and rapid membrane formation are discussed as examples of protein assembly from subunits as an alternative to de novo protein synthesis in early stages of development.     

The ultrastructural differentiation of the endoplasmic reticulum in choroidal epithelial cells of the chick embryo

During the development of the choroidal epithelium in the chick embryo, a substantial concentration of granular endoplasmic reticulum differentiates in the subnuclear cytoplasm of the epithelial cells. The formation of the membranous components of this organelle is preceded by the appearance of a dense, localized population of small, free polyribosomes. Subsequently, numerous membrane-bound vesicles appear in the perinuclear cytoplasm. These primordial ER vesicles measure from 0.1 μ to 0.5 μ or more and they originate from evaginations of the outer nuclear membrane. These vesicles commonly occur in successive rows situated around the margin of the nucleus, and they expand and/or interconnect to form incipient ER tubules. Most vesicles and early tubules are smooth to nearly smooth in appearance. With continued development nuclear evaginations cease, and ER tubules expand in Situ to form an elaborate, laminated system of 7–12 ‘bag-like’ cisternae. Throughout this period of expansive growth, small polyribosomes attach to the developing ER cisternae. As the ER cisternae progressively attain their granular appearance, the number of small, free polyribosomes diminishes. During later stages of development larger polyribosomes appear in association with the subnuclear concentration of ER, and the first accumulations of electron-dense material develop within cisternal spaces.