Ultrastructure of the corpus luteum of antarctic seals during pregnancy

Summary The fine structure of granulosa lutein cells from three crabeater seals, Lobodon carcinophagus, and two leopard seals, Hydrurga leptonyx, has been studied from early through mid-pregnancy. Analysis of the arrangement and modifications of the cytoplasmic organelles and inclusions has revealed three types of lutein cells throughout the corpus. Type I cell typically possesses a central nucleus and cytoplasm containing very large amounts of smooth and/or fenestrated endoplasmic cisternae which frequently extend from the juxta-nuclear to the periphery of the cell. Type II cell contains a central or eccentric nucleus, moderate amounts of peripheral, smooth and fenestrated cisternae which often form large and concentric membranous whorls, numerous mitochondria and small lipid droplets. Frequently these cells show polarity in the arrangement of the cytoplasmic organelles and inclusions. Type III cell contains predominant large lipid droplets, many mitochondria, and small amounts of smooth and fenestrated cisternae. In light microscopy the type I cell is evenly granular, while the type III cell is highly vacuolated. Type II cells have both granular and vacuolated conditions. Ultrastructural features of type I and II cells suggest that they probably secrete most of the steroids, whereas the primary role of the type III cells appear to be lipid storage.This research was supported by National Science Foundation, Grant No. 1325 from the Office of Antarctic Biology.     

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 gular gland of the free-tailed bat Tadarida brasiliensis

The gular gland of the bat Tadarida brasiliensis is a specialized sebaceous gland located in the skin of the suprasternal region of adult males. It consists of an aggregation of simple branched tubulo-acinar gland units, the number of which varies seasonally. Each acinus is composed of densely packed sebaceous cells at various stages of differentiation. Acinar basal cells and cells of the epithelium of the ducts can differentiate into sebaceous cells. Two main changes appear in the cytoplasm concurrent with the sebaceous transformation: the differentiation of cytoplasmic organelles and the deposition of lipid material. The appearance of a different type of mitochondrion and the development of large numbers of ribosomes and polyribosomes can be recognized in the cytoplasm at an early stage of differentiation. Concomitant with the deposition of significant numbers of lipid droplets, the cells develop abundant agranular endoplasmic reticulum occurring mainly as scattered tubular cisternae. These at times form whorls surrounding lipid droplets. At later stages, the cisternae of the agranular endoplasmic reticulum often occur in crystalline arrays between secretory oil droplets. The roles of the different cytoplasmic organelles, especially in relation to the production of sebum, are discussed.     

THE FINE STRUCTURE OF TESTICULAR INTERSTITIAL CELLS IN GUINEA PIGS

In guinea pig testes perfused with either glutaraldehyde or osmium tetroxide fixative, the cytoplasm of the interstitial cells contains an exceptionally abundant agranular endoplasmic reticulum. The reticulum in central regions of the cell is a network of interconnected tubules, but in extensive peripheral areas the reticulum is commonly organized into closely packed, flattened cisternae which are fenestrated. Occasional small patches of the granular reticulum occur in the cytoplasm and connect freely with the agranular reticulum. The mitochondria have a dense matrix and contain cristae and some tubules. The Golgi complex is disperse and shows no evidence of secretory material. The cytoplasm also contains lipid droplets. Lipofuscin pigment granules are probably polymorphic residual bodies and contain three components: (1) a dense material which at high magnification shows a 75-A periodicity; (2) a medium-sized lipid droplet; and (3) a cap-like structure. In glutaraldehyde-perfused testis the interstitial cell cytoplasm appears to have the same density from cell to cell, and the agranular reticulum is tubular or cisternal but not in the form of empty vesicles. Thus the "dark" and "light" cells and the vesicular agranular reticulum sometimes encountered in other fixations may be artifacts. Biochemical results from other laboratories, correlated with the present findings, indicate that the membranes of the agranular endoplasmic reticulum in guinea pig interstitial cells are the site of at least two enzymes of androgen biosynthesis, the 17-hydroxylase and the 17-desmolase.     

Ultrastructure and light microscopical study of a Leydig cell tumor of the testis associated with bilateral gynaecomastia

Light and electronmicroscopic study of a Leydig cell testicular tumor in an 18-year-old male is presented. Bilateral gynaecomastia and normal hormonal blood levels were found. Emphasis on the diagnostic value of electronmicroscopy is remarked upon, based on the following ultrastructural characteristics of the cells; 1) Ovoid shaped nuclei with ondulating contours and dispersed and homogeneous chromatin, 2) Rich agranular endoplasmic reticulum with frequent special modifications, such as membranous whorls with a central cytoplasmic mass or lipid droplets, 3) Numerous mitochondria with occasional tubular cristae, 4) Numerous lipid vacuoles. Other structures also identified in this tumor are Reinke crystalloids, cytoplasmic microbodies, myelin figures, gap-type junctional complexes and paracrystalline inclusions of Payer type E, which are less common.     

OBSERVATIONS ON THE FINE STRUCTURE OF LUTEIN CELLS : II. The Effects of Hypophysectomy and Mammotrophic Hormone in the Rat

Corpus luteum formation was induced in 26-day-old rats which were subsequently hypophysectomized and injected with mammotrophic hormone (MH, LTH). Sections of corpora lutea from these animals were examined with the electron microscope and compared with similarly prepared (Caulfield's fixed, Araldite embedded) corpora from normal pregnancy and from controls, the latter consisting of corpora prior to hypophysectomy and corpora from uninjected rats 7 to 14 days after hypophysectomy. Lutein cells from corpora lutea of injected animals and of normal pregnancy are characterized by abundant, tortuous, tubular agranular endoplasmic reticulum and by mitochondria, many of which are disc-shaped with dense matrices and both villiform and lamelliform cristae. The endoplasmic reticulum is most abundant in lutein cells from pregnant animals, in which cells it is in the form of thin, highly tortuous tubules. The form of the lipid droplets seen in cells of stimulated animals varies greatly. Marginal foldings of the lutein cells on the perivascular space were found in all instances. Lutein cells from hypophysectomized animals have a less highly developed agranular endoplasmic reticulum. The mitochondria have irregular outlines and a relatively lucid matrix. The lipid droplets in these cells show less tendency to be extracted, but are not so large or abundant as in the cells of onset controls. Granules believed to contain lipid pigments are common in the lutein cells of these control animals. It is suggested that lutein cells from corpora lutea which are actively secreting progesterone may be readily distinguished from lutein cells from non-active corpora by means of the multiple characteristics enumerated. It is further suggested that mammotrophic hormone has a general effect on the metabolism of lutein cells rather than solely affecting a specific organelle, the abundance or composition of which may be the limiting factor in the production of progesterone.     

OVARIAN STEROID CELLS : II. The Lutein Cell

The lutein cells of the rabbit exhibit fine structural variations during their life-span of 28 to 30 days. A systematic examination of the corpus luteum reveals that cellular distinctions may be recognized during the first, second, and third stages of pregnancy. The agranular endoplasmic reticulum reveals vesicular, tubular, and cisternal profiles after fixation with each of the following fixatives: glutaraldehyde, osmium tetroxide, and permanganate. The osmolality of the fixing solutions was varied with sucrose and recorded with an osmometer in order to determine the effect of osmotic concentration on the intracellular membranous profiles. It was determined that vesicles and short, branched tubules of similar structure are present in the agranular reticulum when the osmolalities are 300 to 800 milliosmols (iso-osmotic considered 300 milliosmols). At 900 milliosmols, the vesicular or tubular lumen is obliterated. Intracellular membrane profiles do not exhibit interconversions due to hyperosmotic fixative solutions. The agranular endoplasmic reticulum is randomly distributed as short tubular profiles during the first third of pregnancy. A continuity between these membranes and irregular, electron-opaque lipid masses is evident. When physiological and histochemical data indicate that the lutein cell may be storing sterol precursors, cytological observations show that the agranular endoplasmic reticulum exists in a more organized pattern within the cytoplasmic matrix. Vesicular and short tubular, circular aggregations as well as whorled cisternal patterns surround the larger, less electron-opaque lipid droplets. Surface views of cisternal agranular endoplasmic reticulum exhibit tubular extensions, accentuating the continuity between these two profiles. During the progress of pregnancy, the lutein cell increases in diameter, and accumulates both lipid inclusions and aggregations of intracellular membranes. The agranular endoplasmic reticulum may be peripherally packed and arranged parallel to the cell surface during later stages. In the postpartum, degenerating lutein cell, large myelin figures are present which form from the agranular endoplasmic reticulum. These cellular events are discussed in relation to lutein cell activity, including both secretion of product and storage of precursors.     

Structural characteristics of the corpus luteum during implantation in the rhesus monkey (Macaca mulatta)

Corpora lutea were obtained from ten pregnant rhesus monkeys during implantation, and the ultrastructure of granulosa and theca lutein cells was characterized. Specimens were individually staged with regard to the extent of implantation and the relationship to the rise in circulating progesterone and estrogen which is characteristic of early pregnancy. Structural changes characteristic of granulosa lutein cells occurring during implantation included: change in form of endoplasmic reticulum from predominantly agranular tubules to predominantly granular cisternae; reduction in size and number of lipid droplets; increase in area occupied by the Golgi and increase in length of the cisternae of the Golgi complex; development of numerous microvillus-lined intracellular spaces; increase in numbers of membrane-bound dense bodies including peroxisomelike bodies, multivesicular bodies within lobopodia, and other lysosomelike bodies; and alterations of mitochondrial cristae. These changes were suggestive of the production of a secretory protein, rapid utilization of existing steroid precursor reserves for the production of progesterone, and a reduction in capability for steroid precursor accumulation and processing by granulosa lutein cells. Structural changes characteristic of theca lutein cells occurring during implantation included an increase in size and number of lipid droplets, an increase in agranular endoplasmic reticulum, and an increase in area occupied by the Golgi complex. These changes were suggestive of an increased capability for steroid precursor accumulation and processing, perhaps for estrogen production, by the theca lutein cells.     

The fine structure of testicular interstitial cells in the adult golden hamster with special reference to seasonal changes

Summary The fine structure of the testicular interstitium was studied in normal adult golden hamsters sacrificed in the reproductive season (spring and summer) and in the winter. The Leydig cells in the reproductively active testes contain abundant endoplasmic reticulum (ER) and numerous mitochondria. The ER occurs in the form of flattened cisternae and tubules, the former prevailing. The cisternae are extremely extensive and are partly granular and partly agranular, their ends being continuous with the tubular reticulum. Mitochondria intervening between the cisternae are closely associated with the agranular portions of the latter. Adjacent to the Golgi complex and continuous with the centrosome a unique filamentous body with a dense laminar core is often observed. In the regressive testes, the Leydig cells show a great reduction of cytoplasmic volume and a remarkable decline of the organelles, especially agranular tubules. The possible functional significance of the tubular and cisternal ER with the associated mitochondria is discussed in relation to the biosynthesis of androgens. Macrophages appear to constitute another important population of the interstitial cell clusters.This study was supported in part by a grant from the National Science Council, the Republic of China     

Fine structure of the human foetal testis

Summary Thirteen male human foetuses ranging in crown-rump length from 29 to 212 mm (ages 8–27 weeks) were studied. Four developmental phases are distinguished. 1. The predifferentiation phase (below 8 weeks): The interstitium contains only undifferentiated mesenchymal cells. 2. The differentiation phase (8–14 weeks): Leydig cells develop and gradually fill the space between the germ cords. 3. The maturity phase (14–18 weeks): The interstitium occupies more than one half of the total area in the testis sections and is filled with mature foetal Leydig cells. 4. The involution phase (18–40 weeks): Most of the Leydig cells gradually degenerate and disappear.The foetal Leydig cells are packed with tubular agranular endoplasmic reticulum (AER). Islets of parallel granular ER membranes and other organelles are embedded in the AER. The mitochondria vary in shape and form, the cristae being mainly tubular. Some mitochondria like organelles contain electron dense inclusions. Dark membrane bound bodies of variable form and resembling the Golgi cisternae are present in most cells. Reinke crystals are never found in the foetal cells. In degenerating Leydig cells the AER appears in vesicular form, membranous whorls are seen in some of them and the cell membrane seems to rupture finally, and cytoplasmic material protrudes outside the cells. The fine structure of the mature foetal Leydig cells is suggested to reflect signs of human chorionic gonadotrophin stimulation.This investigation was supported by the Damon Runyon Memorial Fund (DRG-940) and by the Sigrid Jusélius Foundation.     

The fine structure of the mouse adrenal X zone

Summary Electron microscopically the adrenal X zone was examined in the fourteen SMA female mice aged 40 and 70 days. At these ages, the X zone showed no signs of degeneration. The X zone cell was somewhat smaller than the permanent cortical cell.The mitochondria in the X zone cell were quite bizarre in shape, provided with tubules or cristae. Many intramitochondrial bodies very similar to the cytoplasmic lipid droplets were found in the X zone. A few lipid droplets and globules were also noticed in this zone. The lipid droplets may possibly be formed within the mitochondria.The light and dark cells were differentiated. For the light cells, scant mitochondria and tubular granular endoplasmic reticulum were characteristic in contrast to the abundant mitochondria and multi-lamellated agranular endoplasmic reticulum in the dark cells. The cellular variety in density was discussed with regard to steroid synthesis.The author wishes to express his sincere appreciation to Prof. H. Tauchi, The 2nd Department of Pathology, Nagoya University School of Medicine, for kind advice, to Dr. M. Hoshino for helpful suggestion, and to Mr. J. Aoki for excellent technical assistance.     

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.     

Cytological differentiation of the interrenal tissue of the Japanese quail,Coturnix coturnix

Summary As reported for several other avian species there are clearly distinguishable subcapsular (SCZ) and inner (IZ) zones of interrenal tissue in the Japanese quail. The SCZ contains large columnar cells (type I) with rounded nuclei, polymorphic mitochondria with shelf-like cristae, and relatively small numbers of lipid droplets. The IZ contains two and possibly three types of cells. Type II consists of large columnar cells with moderately dense cytoplasm containing large numbers of lipid droplets and many rounded mitochondria with tubular cristae. Smooth endoplasmic reticulum (SER) and Golgi apparatus are well developed; coated vesicles occur in the Golgi area and at the cell surface. Type-III cells occur in IZ and especially in its more peripheral areas. They are columnar cells with strikingly clear cytoplasm (in comparison with type II) containing mitochondria with plate-like cristae and tubular SER. Type-IV cells are sparsely distributed in IZ and occur rarely in SCZ. Type IV may be a degenerating phase of type III.After adenohypophysectomy or section of portal vessels type-I cells atrophy somewhat with a decrease in lipid droplets; type-II cells, also atrophy with conspicuous increase in size and number of lipid droplets, enlargement of mitochondria, and gradual disappearance of SER; type-III cells decrease in number whereas type-IV cells increase.After injection of ACTH, type-I cells enlarge and their mitochondria, SER and Golgi apparatus become more conspicuous; there is a decrease in lipid droplets in type-II cells and a development of SER, polysomes and Golgi apparatus; there is also a decrease in lipid droplets and a development of SER in type-III cells after injection of 2IU ACTH and an almost complete disappearance of lipid droplets after 4IU ACTH; type-IV cells increase in number.The investigation reported herein was supported by Scientific Research Grants from the Ministry of Education of Japan to Professor Mikami; and by grants from the Japan Society for the Promotion of Science, the National Science Foundation (USA), and the Graduate School Fund of the University of Washington to Professor Farner     

Fine structural changes in Sertoli and Leydig cells during the reproductive cycle of the ground squirrel, Citellus lateralis

During spermatogenesis in sexually mature ground squirrels Leydig and Sertoli cells were morphologically well differentiated. For Leydig cells the most prominent organelles were lipid droplets, mitochondria with tubulo-vesicular cristae and abundant agranular reticulum organized as a mass of anastomosing tubules. These morphological criteria suggest that the Leydig cells were steroidogenically active. Sertoli cells exhibited a topographical distribution of certain organelles with basal regions containing stacks of granular reticulum, and large areas of agranular reticulum. The cytoplasm surrounding maturing germ cells contained numerous microtubules, and an adluminal layer of spermatids at a certain stage of spermiogenesis became enveloped by Sertoli cytoplasm containing an enormous proliferation of agranular reticulum. The presence of these organelles in Sertoli cells suggests that during spermatogenesis they are active in the synthesis of proteins and steroids. In particular the mass of agranular reticulum surrounding late stage spermatids indicates that steroids may be required for spermatid maturation and/or spermiation. By contrast Leydig and Sertoli cells observed during testicular regression, when only spermatogonia remain in the seminiferous tubules, had undergone structural changes. Leydig cells were still numerous and large with abundant agranular reticulum that was now organized as a loose assemblage of single unbranched tubules. Sertoli cells were drastically reduced in both cytoplasmic volume and content of organelles.     

Organelle distribution in chick neuroepithelial cells: effects of colchicine and cytochalasin B

The intracellular distribution of mitochondria, cytoplasmic inclusions and rough endoplasmic reticulum cisternae of chick neuroepithelial cells was investigated at neurulation stages 6, 8, 10 and 12. These neuroepithelial cells were subdivided into three zones: apical, median and basal and the distribution percentages of distribution of these organelles were obtained. Mitochondrial distribution was related to the energy supply that mitochondria provide for apical microfilament contraction. Cytoplasmic inclusions were distributed preferentially in the apical zone of the neuroepithelial cells during the four stages. Rough endoplasmic reticulum cisternae were homogeneously distributed in the three zones at stages 10 and 12, but at stages 6 and 8 there are more elevated percentages of rough endoplasmic reticulum in the apical zones than in the other zones. Experimental treatments with colchicine and cytochalasin B does not modify the patterns of mitochondria and rough endoplasmic reticulum cisternae but alters the distribution of cytoplasmic inclusions. Finally, there is a correlation in the normal neurulating neuroepithelial cells between the distributions of mitochondria and rough endoplasmic reticulum distribution and between the distributions of mitochondria and cytoplasmic inclusions distribution. This relationship is retained in the treated neuroepithelial cells.     

Ultrastructural features of the ventral prostate epithelial cells in the Australian plains rat, Pseudomys australis

The fine structure of epithelial cells of the small ventral prostate of Pseudomys australis males was studied. The cells were sometimes binucleated, exhibited relatively little granular endoplasmic reticulum, generally few secretory granules (probably reflecting a low proteinaceous secretory activity), but had abundant agranular endoplasmic reticulum (AER), similar to that in steroidogenic cells. Some of the mitochondria also showed tubular cristae. Furthermore, most cells had some irregular dense bodies in the cytoplasm which may have developed, by a gradual transformation process, from the membranes of AER, mitochondria and other organelles; they could be the product of degenerative changes in these organelles. These findings indicate a significant difference in the structure of these cells from those present in the ventral prostate of the hopping mouse, Notomys alexis. It is speculated that this gland secretes relatively little protein but perhaps more lipid or cholesterol-derived substance.     

OBSERVATIONS ON THE FINE STRUCTURE OF LUTEIN CELLS

Corpora lutea from the period of delayed implantation and from early postimplantation stages of the armadillo, mink, and rat were fixed in buffered osmium tetroxide-sucrose or potassium permanganate. After rapid dehydration, the portions of the corpora lutea were embedded in either methacrylate or epoxy resin. Examination of the lutein cells by electron microscopy revealed the presence, in the better preserved material, of an extensive development of tubular agranular endoplasmic reticulum. Although the membranes of the endoplasmic reticulum are the most striking feature of the lutein cells of both stages of the three animals examined, very numerous large mitochondria with cristae that exhibit a variety of forms tending toward villiform, and protrusions and foldings of the lutein cell margins on the pericapillary space are also characteristic of these cells. Certain minor differences in the lutein cells of the species examined are also noted. No indications of conversion of mitochondria into lipid, of accumulation of lipid in the Golgi area, or of the protrusion of lutein cells into spaces between the endothelial cells, as suggested by other authors, were noted in these preparations. Some of the difficulties inherent in the visualization of the secretory activity of cells producing steroid hormones are briefly discussed.     

Hormone-induced differentiation of agranular endoplasmic reticulum in the interstitial cells of the mouse testis

Summary The administration of chorionic gonadotrophin to prepuberal mice results in precocious maturation of the testicular interstitial cells. The cytoplasm of the nine-day-old cells is characterized by abundant lipid droplets, large numbers of glycogen particles and mitochondria. By contrast, the membranous organelles are poorly developed.Human chorionic gonadotrophin brings about mobilization of lipid droplets and glycogen particles, and differentiation of large areas of agranular endoplasmic reticulum.The present observations are in agreement with the reports that human chorionic gonadotrophin increases the secretion of testosterone and that the agranular endoplasmic reticulum is the site of storage of steroid and of the enzymes involved in the biosynthesis of androgens.     

THE NORMAL FINE STRUCTURE OF OPOSSUM TESTICULAR INTERSTITIAL CELLS

The interstitial tissue of the opossum testis includes interstitial or Leydig cells, macrophages, and small cells which morphologically resemble mesenchymal cells. The latter are thought to give rise to mature interstitial cells. The most prominent feature of the interstitial cell cytoplasm is an exceedingly abundant agranular endoplasmic reticulum. This reticulum is generally in the form of a meshwork of interconnected tubules about 300 to 450 A in diameter, but occasionally it assumes the form of flattened, fenestrated cisternae resembling those of pancreatic acinar cells, except for the lack of ribonucleoprotein particles on the surface of the membranes. The interstitial cells vary considerably in their cytoplasmic density. The majority are quite light, but some appear extremely dense, and in addition usually have a more irregular cell surface, with numerous small pseudopodia. These differences may well reflect variations in physiological state. Cytoplasmic structures previously interpreted as "crystalloids" consist of long bundles of minute parallel tubules, each about 180 A in diameter, which seem to be local differentiations of the endoplasmic reticulum. The mitochondria are rod-shaped, and contain a moderately complex internal membrane structure, and also occasional large inclusions that are spherical and homogeneous. The prominent juxtanuclear Golgi complex contains closely packed flattened sacs and small vesicles. The results of the present study, coupled with biochemical evidence from other laboratories, make it seem highly probable that the agranular endoplasmic reticulum is involved in the synthesis of the steroid hormones produced by the interstitial cell. This finding therefore constitutes one of the first functions of the agranular reticulum for which there is good morphological and biochemical evidence.     

Ultrastructural analysis of the granulosa — Luteal cell transition in the ovary of the dog

The transition from ovarian granulosa to lutein cell during the estrus cycle of 60 pregnant and non-pregnant beagle bitches was analyzed by light and electron microscopy (both 100 and 1000 KV). Early proestrus was characterized by a gradual rise in serum estrogen levels, hyperplasia of the granulosa cells, the accumulation of follicular fluid, and the development of tortuous intercellular channels. During the second half of proestrus, serum estrogen levels continued to rise, but growth, division, and differentiation of the granulosa cells was minimal. Estrus was marked by the first acceptance of the male and a well-defined LH peak In the subsequent 24 hour period, the granulosa-lutein cells hypertrophy rapidly and develop a large Golgi apparatus, small profiles of granular endoplasmic reticulum, numerous microfilaments, and large gap junctions between the cells. Mitochondria also proliferate, enlarge, and elongate, but retain lamelliform cristae. Luteinization of the cells and progesterone secretion begin just after ovulation which in turn occurs about 24 hours after the LH peak. On the third and fourth day of estrus, numerous small vesicles of agranular endoplasmic reticulum fill the extoplasm and the mitochondria swell up and round off. The vesicles rapidly fuse into whorled and flattened cisternae or anastomosing tubules of agranular endoplasmic reticulum, while the mitochondria develop tubulovesicular cristae. These structures gradually become organized with respect to the basal lamina. The Golgi apparatus is centered over the pole of the nucleus that faces the pericapillary space. Stacked and whorled cisternae of agranular ER develop in the lateral margins and avascular end of the cell while mitochondria and tubular elements of agranular ER predominate in the central medial and most basal portions of the cytoplasm. Microfilaments are ubiquitous and appear to be instrumental in this orientation process. The cell surface develops three distinct regional specializations that coincide with the underlying cellular compartments: interconnecting pleomorphic folds fill the pericapillary space; long tenous microvilli project from the lateral cell surface and form tortuous intercellular channels and canaliculi; and large gap junctions form along the margins of the cell furthest removed from the basal lamina. By the sixth day of estrus, the granulosa-luteal cell transition is nearly complete and serum progesterone levels are on the rise.