Ultrastructural changes in the oviductal mucosa throughout the sexual cycle in Bufo arenarum

In Bufo arenarum the oviduct exhibits conspicuous changes throughout the sexual cycle. In the present study, we analyzed the optical and ultrastructural characteristics of the oviductal pars convoluta mucosa, the portion responsible for jelly secretion, during both the preovulatory and postovulatory periods. Secretory epithelial cells, ciliated cells, basal cells, and glandular cells are described. Secretory epithelial cells are characterized by the presence of secretory granules, the size, shape and electron density of which vary markedly. Their contents are mainly released by exocytosis into the oviductal lumen. Moreover, in the preovulatory period, apocrine, and holocrine secretion processes frequently occur. During the postovulatory period, these cells exhibit a marked diminution of secretory granules. Ciliated cells show a typical ultrastructural organization. Basal cells are distinguished in the lower part of the epithelium by their heterochromatic nuclei and electron‐lucent cytoplasm. These cells, to the best of our knowledge, are reported for the first time in Amphibia. Glandular cells exhibit oval, round, or polyhedric granules, most of them with one or more cores. Our results indicate that the contents of epithelial and glandular secretory cells are partially secreted during the preovulatory period. Additional secretion occurs during the transit of the oocytes. J. Morphol. 239:61–73, 1999. © 1999 Wiley‐Liss, Inc.     

Complex secretory products in the oviduct of the plethodontid salamander Bolitoglossa dofleini (Amphibia, Urodela)

The ultrastructure of the secretory granules in the cells of the subdivisions of the oviduct in the neotropical plethodontid salamander Bolitoglossa dofleini was studied by transmission electron microscopy. In addition, we applied the cationic dye Cuprolinic Blue (CB) at different electrolyte concentrations to demonstrate proteoglycans, and the pyrogallol red-copper (PR-C) method to stain proteins at the ultrastructural level. The entire oviduct is lined by a simple epithelium that contains ciliated and microvillous cells in the first subdivision, the aglandular pars recta; microvillous cells show a moderate secretory activity. The following pars convoluta is differentiated into five glandular subdivisions and the aglandular “uterine portion”. Especially in the glandular parts, the epithelium is arranged in longitudinal folds. At their crests ciliated and microvillous cells similar to those in the pars recta occur. Gland cells are crowded with secretory granules that differ in their structural complexity (with and without electron-dense spheres or masses; elaborated, homogeneous or granular matrix; spherical; distorted) along the various subdivisions. Further, as suggested by the CB-technique, the cranial subdivisions contain large amounts of sulphated proteoglycans that decrease in the caudal direction. Carboxylated proteglycans appear to be present in all subdivisions examined. Electron-dense spheres of secretory granules are largely free of CB-precipitates, but stain more or less intensely with PR-C. The ultrastructure of the pars recta, and especially the “uterine portion” indicates transporting capability. The epithelial cells of the “uterus” have coated pits and a considerable amount of lysosome-like bodies.     

Oviductal Localization of the Cortical Granule Lectin Ligand Involved in the Block to Polyspermy of Xenopus Laevis

The fertilization layer of Xenopus laevis is formed upon egg activation by the binding of the cortical granule lectin (CGL) to its ligand in the egg extracellular matrix. Using Western blotting methods with biotinylated CGL as a probe, oviductal tissue extracts were examined to determine the site of origin of the CGL ligand. Three glycoprotein ligands of M_rs= > 250,000, 160,000, and 90,000 (reduced samples) were localized to the pars convoluta oviduct immediately posterior to the pars recta oviduct. The binding of CGL to these glycoproteins was inhibited in the presence of 200 mM galactose, but not with 200 mM mannose indicating a specific lectin interaction. The M_rs= > 250,000 and 90,000 glycoproteins were linked by disulfide bonds. In addition, these ligands were secreted from a more anterior region of the pars convoluta oviduct than the M_r=160,000 ligand. No binding of CGL was detected to pars recta secretory granule lysate components. The highest molecular weight CGL ligand seen in the pars convoluta corresponded to the CGL ligand in isolated fertilization envelopes. Thus, the CGL ligand involved in the formation of the fertilization layer is a product of the pars convoluta oviduct.     

The effect of estrogen and progesterone on structural changes in the uterine glandular epithelium of the ovariectomized sheep.

The development of epithelial cells of the uterine glands of ovariectomized sheep in response to estradiol-17 beta (E) and progesterone (P) was studied using light and electron microscopy. Animals that had been ovariectomized for six weeks were placed in one of three experimental treatment groups. Group I animals (untreated controls) received no steroid treatment. Group II animals (E alone) received one 4-cm E implant (E approximately 5-10 pg/ml) and their uteri were removed after 2, 4, or 6 days. Group III animals (E-primed, P-treated) received an E implant (E approximately 5-10 pg/ml) for 6 days and then were treated with six 13-cm P implants (P approximately 1.5-3 ng/ml), in the continued presence of E, for 2, 4 or 6 days. Six weeks after ovariectomy the epithelial cells of the uterine glands were low cuboidal and morphologically appeared to be synthetically inactive. Following 2 days of E treatment the epithelial cells had significantly increased in cell height, and protein-synthesizing organelles were well developed. Maturation of the secretory apparatus continued throughout E treatment. The Golgi complex and rough endoplasmic reticulum (RER) were abundant. Lysosomal-like granules and granules of varying electron density were present in the cytoplasm. The chronic administration of P to E-primed animals did not result in any further increase in cell height. Elongated mitochondria, a cup-shaped Golgi apparatus, extensive apical microvilli, and irregularly shaped membranous profiles in the supranuclear cytoplasm characterized these uterine epithelial cells. Lysosomal-like granules, small vesicles, and scattered patches of glycogen were seen in the cytoplasm. These data show that the uterine epithelial cells of the ovariectomized sheep undergo morphological alterations in protein-synthesizing organelles and apical specializations that depend on the presence of E and P.     

Light and electron microscopic study of the pars intermedia of the lizard,Klauberina riversiana

Summary Two cell types can be distinguished in the pars intermedia of Klauberina: (1) Glandular cells, which form a single-layered columnar epithelium on the vascular septum which divides the pars nervosa from the pars intermedia. (2) Marginal cells which form a flattened epithelium over the glandular cells and line the hypophysial cleft. Occasional projections from the marginal cells extend between the glandular cells to contact the basement membrane of the vascular septum, and occasional projections of the vascular septum extend across the glandular epithelium to reach the marginal epithelium. Both cell types are AF negative. The granules of the glandular cells are strongly PAS positive, and acidophilic in response to Mallory's trichrome stain. In electron micrographs, the glandular cells contain large quantities of secretory granules. In one class of cells, they range from 2,000 to 2,500 Å in diameter, in the other, from 4,000 to 5,000 Å. Electron-dense granules 1,000 to 1,500 Å in diameter occur in the cytoplasm of the marginal cells in the region of contact with the vascular septum. Hence more than one active principle may be produced by the pars intermedia.No nerve endings of any kind are present in the pars intermedia. Therefore, synaptic contact of neurons with the secretory cells seems not to be necessary for the regulation of their secretory activity as appears to be the case in other vertebrate groups. It is suggested that regulatory factors are secreted in the pars nervosa and transported to the pars intermedia via the vascular septum.Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina.This investigation was supported in part by a Public Health Service fellowship 1 FZ HD 32, 949-01 REP from the National Institute of Child Health and Human Development. The authors wish to thank Professor H. Heller for his encouragement and kind cooperation during this study and Professor H. D. Dellmann for providing the facilities of his department. They are indebted to the officers and men of the Naval Ordinance Test Station, Pasadena and San Clemente Island, California, for their help in obtaining animals for this investigation.     

Oviduct histochemistry and site of synthesis of a 29.7 kDa jelly coat glycoprotein in the anuran Lepidobatrachus laevis

The oviduct of the anuran Lepidobatrachus laevis contains three morphological regions, each of which contains a histochemically distinct luminal mucosa. In the pars recta, the most anterior portion of the oviduct, there are periodic acid-Schiff base (PAS)-positive simple glands and epithelia. In the pars convoluta, there are alcian blue-positive, combined alcian blue- and PAS-positive and PAS-positive gland types. The most posterior region, the pars uterina, contains alcian blue-positive and alcian blue-negative epithelial cells. Previous work has shown that solubilized egg jelly contains a major 29.7 kDa glycoprotein subunit that was detected in oviduct tissue extracts from the pars convoluta in the present study. Rabbit antisera to the 29.7 kDa egg jelly glycoprotein of L. laevis reacted with the major pars convoluta glycoprotein and there were no immunoreactive components in the pars uterina. The slight immunoreactivity detected at 29.0–37.0 kDa in pars recta extracts is not believed to be the jelly molecule, based on low immunoreactivity and subunit molecular weight measurements. We conclude that the synthesis of the 29.7 kDa egg jelly glycoprotein is restricted to the pars convoluta region of the oviduct.     

Fine structure of the ventral and dorsal lobes of the prostate in the young adult Syrian hamster, Mesocricetus auratus

The normal ventral and dorsal prostatic lobes of the young adult Syrian hamster were examined at the light and electron microscopic levels. Each lobe is composed of branched tubular secretory units separated from each other by loose interacinar connective tissue and draining into the urethra. The lumen of each acinus is lined by a simple epithelium composed of columnar secretory cells with occasional small basal cells. The epithelial layer, with the thin underlying lamina propria, forms a mucosa that is often highly folded. The whole acinus is bounded by a thick muscular stroma. In each of the ventral lobes, there are three main ducts, each one formed of tubular branched tributary secretory units. The walls of the secretory acini are moderately folded. Microvilli dominate the lumenal surface of the secretory epithelial cells. The Golgi complex is very extensive and shows dilated cisternae and secretory vesicles and vacuoles of various sizes. Membrane-bounded secretory granules populate the Golgi and apical areas and are released into the acinar lumen by exocytosis. The rough endoplasmic reticulum is dispersed throughout the cytoplasm, except in the region of the Golgi apparatus. In each of the dorsal lobes, there are several main tubular ducts that open into the urethra. Both proximal (ductal) and distal portions of the glandular tree are secretory in nature. Microvilli and cytoplasmic bulges and blebs dominate the lumenal surface of the secretory cells. The cells are also characterized by highly dilated cisternae of rough endoplasmic reticulum. The secretory cells show heterogeneity in the degree of dilation and distribution of rough endoplasmic reticulum, and this heterogeneity may reflect location in the glandular tree.(ABSTRACT TRUNCATED AT 250 WORDS)     

Scanning electron microscopy of the oviduct of Salamandra salamandra (L.) (Amphibia, Urodela)

The oviduct of non-pregnant females of the ovoviviparous salamander, Salamandra salamandra, was examined using SEM-techniques. In the luminal epithelium polygonal ciliated cells were found along the entire surface of the oviduct, except the uterus, and non-ciliated cells with a varying number of short or long microvilli. The ciliated cells occur in the most anterior portion of the oviduct, the pars recta; they are sparsely distributed in the p. convoluta I, but abundant in the p. convoluta II and III. Non-ciliated cells comprise several small gland cells, restricted to the p. convoluta I, II, III, and undifferentiated cells both provided with microvilli, but difficult to be discerned from their surface appearance. The p. convoluta I, II, III is characterized by three types of secretory cells forming tubular glands, each type confined to a given zone. The secretory cells have slender microvilli at their surfaces. In freeze-cracked glands details of their secretory products can be visualized. The findings are compared to previously published TEM-investigations and discussed with regard to some functions of the oviduct during reproduction.     

Effect of different photoperiods on the ultrastructure of the specific secretory cells and α-subunit mRNA level in the chicken pars tuberalis

The effect of different photoperiods on the specific secretory cells of the pars tuberalis was examined in male chicks. Animals were placed in one of three different photoperiod regimens: (1) normal control (light:dark = 12 h:12 h), (2) continuous light (L:D = 24 h:0), and (3) extended darkness (L:D = 1 h:23 h). The levels of common alpha-subunit mRNA in the pars tuberalis were examined by Northern blot analysis and compared with those in the pars distalis. In chicks exposed to continuous light for 1 week, alpha-subunit mRNA level in the pars tuberalis was decreased, although the level in the pars distalis was increased. Exposure to continuous light for 30 days also induced a decrease in alpha-subunit mRNA level in the pars tuberalis. On the other hand, in chicks exposed to extended darkness for 1 week, the alpha-subunit mRNA level of the pars tuberalis was markedly increased. In situ hybridization with digoxigenin-labeled common alpha-subunit cRNA probe also showed that the hybridization signals for alpha-subunit mRNA in the pars tuberalis cells become weak under continuous light for 30 days but they are very intense under extended darkness. Thus, the synthesis of alpha-subunits in the chick pars tuberalis was inhibited by continuous light but stimulated by extended darkness. These results were confirmed by semiquantitative electron-microscopic analyses. After exposure to continuous light for 30 days, many pars tuberalis (PT)-specific cells were filled with enlarged secretory granules, showing the reduction of secretory activity. On the contrary, extended darkness for 30 days induced hypertrophy of the PT-specific cells; the areas of cytoplasm and nucleus were significantly increased. In addition, secretory granules became small in size and exocytotic features were more frequent. Mitochondria and lysosomes were also increased in number. Thus, the synthetic and secretory activities of the PT-specific cells were increased under extended darkness. The data indicate that the specific cells of the pars tuberalis are responsive to photoperiodic changes in the chick.     

Morphological changes in the oviductal endometrium during the reproductive cycle of the tortoise,Gopherus polyphemus

The oviducts of 24 tortoises (Gopherus polyphemus) were examined using histological techniques and scanning electron microscopy to determine endometrial morphology. Measurements of endometrial characteristics (epithelial cell height, cilia length, thickness of endometrial glandular layer, and glandular diameter) in the uterus and tube (tuba uterina) were obtained to determine changes during the reproductive cycle. Epithelial cell height increases in both the uterus and the tube during vitellogenesis and remains hypertrophied during gravidity. Cilia length increases in the uterus during late vitellogenesis and gravidity, but the length of tubal cilia does not change during the reproductive cycle. The ratio of secretory to ciliated epithelial cells in the oviduct increases from quiescence to gravidity. The thickness of the glandular endometrial layer increases in both the uterus and tube during vitellogenesis. In the uterus, the glandular layer decreases in thickness during gravidity. The diameter of the uterine glands increases throughout vitellogenesis and gravidity; however, following ovulation glandular cells become depleted of secretory granules and cell height diminishes. The diameter of the tubal glands is unchanged during the reproductive cycle. Oviductal hypertrophy during vitellogenesis coincides with elevated circulating estradiol, whereas during gravidity progesterone concentrations peak (Taylor, '82, PhD Dissertation, University of Florida, Gainesville) and may induce secretion of albumen and eggshell components.     

The amylase-secreting cells of the stomach of the scallop, Pecten maximus: ultrastructural, immunohistochemical and immunocytochemical characterizations

(1) alpha-amylase was extracted and purified from the stomach/digestive gland complex of the scallop Pecten maximus and an anti-serum was induced against the purified amylase by rabbit immunization. (2) The anti scallop amylase was used to localize the amylase-secreting cells in the stomach of Pecten maximus by immunofluorescence and immunogold labelling. The amylase-secreting cells are glandular cells particularly numerous in the main sorting area of the stomach. Their secretory granules were found strongly positive for anti-amylase. Three types of glandular cells were observed, actually corresponding to the three stages of the glandular-cell activity, synthesis, secretion and excretion. (3) The synthesizing cell shows the characteristic features of a protein-synthesizing cell: a conspicuous nucleolus and abundant granular endoplasmic reticulum. In the secretory cell, the secretory granules are formed by the Golgi apparatus and accumulate in the apical part of the cell. The secretory cell is filled with two types of secretory granules which are released in the stomach lumen by apocrine excretion. (4) The present study brings the first demonstration of the synthesis and extracellular release of amylase by glandular cells of the stomach epithelium of a bivalve.     

Disorganization of stroma alters epithelial differentiation of the glandular stomach in adult mice

Summary The response of adult epithelium in contact with heterologous mesenchymes/stromas was studied in three digestive organs (forestomach, glandular stomach, and duodenum). After various tissues were implanted beneath the epithelial layer of adult mice, the epithelial differentiation was examined after sacrifice of animals at intervals up to 24 weeks. In the forestomach and duodenum, the epithelial differentiation was not affected at all by the tissue implantation. In the glandular stomach, in contrast, epithelial cells exhibited altered differentiation in which chief and parietal cells disappeared and were replaced by columnar epithelial cells with PAS-positive granules. These epithelial cells often formed immature villi. Such differentiation-altered columnar epithelium (DACE) was induced by implanting any type of tissue and even by sham operation, indicating that it was induced by disorganization of the tissue-implanted stroma. The size of DACE was significantly influenced by the stage of implanted tissue; 14.5-day fetal mesenchyme induced the largest DACE, and was followed by 16.5-day fetal mesenchyme, adult stroma, and sham operation. These results suggest the importance of stromal organization in maintaining epithelial differentiation in the glandular stomach.     

Ultrastructure of the pars intermedia of the adult sheep hypophysis

Summary Light microscopy of coronal sections of the sheep pars intermedia revealed a compact, incompletely lobulated V-shaped region about 15–20 cells thick, situated between the pars distalis and the pars nervosa. A prominent hypophysial cleft and follicles containing a colloid-like substance were seen.Using electron microscopy, five cell types could be distinguished: pars intermedia glandular cells, pars distalis-like glandular cells, interstitial cells, follicular cells and cleft lining cells. The polyhedral to pear-shaped pars intermedia glandular cells predominated. They contained dense-cored, membrane-bound granules near the Golgi complex, and larger, irregular vesicles with finely granular contents of varying electron density throughout the remaining cytoplasm; exocytotic release of granules was occasionally observed. Smaller numbers of cells resembling those seen in the pars distalis were scattered throughout the pars intermedia. Interstitial cells usually possessed elongated cytoplasmic processes which extended between the glandular cells, and were characterized by deeply indented nuclei, elaborate junctional complexes and an absence of cytoplasmic granules. Cells lining the follicles resembled the interstitial cells. The major cells bordering the hypophysial cleft were triangular in section and bore irregular microvilli on their free surface. The pars intermedia appeared to be less vascular than the remainder of the hypophysis and only occasional fenestrated capillaries were seen. Nerve profiles were rare.     

Abnormal ion content, hydration and granule expansion of the secretory granules from cystic fibrosis airway glandular cells

The absence or decreased expression of cystic fibrosis transmembrane conductance regulator (CFTR) induces increased Na(+) absorption and hyperabsorption of the airway surface liquid (ASL) resulting in a dehydrated and hyperviscous ASL. Although the implication of abnormal airway submucosal gland function has been suggested, the ion and water content in the Cystic Fibrosis (CF) glandular secretory granules, before exocytosis, is unknown. We analyzed, in non-CF and CF human airway glandular cell lines (MM-39 and KM4, respectively), the ion content in the secretory granules by electron probe X-ray microanalysis and the water content by quantitative dark field imaging on freeze-dried cryosections. We demonstrated that the ion content (Na(+), Mg(2+), P, S and Cl(-)) is significantly higher and the water content significantly lower in secretory granules from the CF cell line compared to the non-CF cell line. Using videomicroscopy, we observed that the secretory granule expansion was deficient in CF glandular cells. Transfection of CF cells with CFTR cDNA or inhibition of non-CF cells with CFTR(inh)-172, respectively restored or decreased the water content and granule expansion, in parallel with changes in ion content. We hypothesize that the decreased water and increased ion content in glandular secretory granules may contribute to the dehydration and increased viscosity of the ASL in CF.     

Renal transport of neutral amino acids. Cation-dependent uptake of L-alanine by luminal-membrane vesicles.

The characteristics of L-alanine transport in luminal-membrane vesicles isolated either from whole cortex or from pars convoluta or pars recta of rabbit proximal tubules were studied by a rapid filtration technique and by a spectrophotometric method. Uptake of L-alanine by vesicles from whole cortex was mediated by both Na+-dependent and Na+-independent, but electrogenic, processes. The nature, mechanism and tubular localization of the transport systems were studied by the use of vesicles derived from pars convoluta and pars recta. In vesicles from pars recta transport of L-alanine was strictly dependent on Na+ and occurred via a dual transport system, namely a high-affinity (half-saturation 0.14 mM) and a low-affinity system (half-saturation 9.6 mM). The cation-dependent but Na+-unspecific transport system for L-alanine was exclusively localized to the pars convoluta, which also contained an Na+-preferring system of intermediate affinity (half saturation 2.1 mM). A closer examination of the mechanism of transport of L-alanine in vesicles from pars convoluta revealed that an H+ gradient (extravesicular greater than intravesicular) can drive the transport of L-alanine into the vesicles both in the presence and in the absence of Na+. The physiological importance of various L-alanine transporters is briefly discussed.     

Cytodifferentiation of the adenohypophysis of the domestic fowl

Summary In the chick embryo the first membrane-bound secretory granules occur in the cytoplasm of occasional cells in the cephalic lobe of pars distalis at the 7th day of incubation. On the 8th day most of the cells in both the cephalic and caudal lobes contain secretory granules that are variable in size, form and density.On the 9th day at least two types of glandular cells are distinguishable in the cephalic and in the caudal lobes; however, these cells are not comparable with those of the adult gland. Differentiation of acidophils and basophils occurs, apparently simultaneously, in 11-day embryos.The cells of the cephalic and caudal lobes are morphologically distinct from their first appearance. Thus it is concluded that these two lobes develop independently and differently from an early stage of ontogenesis.The secretory granules are formed in the Golgi area of the hypophysial cells after the 8th day of incubation. However, secretory material may be synthesized also by a process not involving the Golgi apparatus.Nerve fibers containing granules first appear in the superficial layer of the median eminence on the 8th embryonic day and by the 12th day three types of granules and two types of clear vesicles are identifiable.The investigation reported herein was supported by grant from Japan-U.S. Cooperative Science Program of Japan Association for Science Promotion to Professor Mikami and by U.S.-Japan Cooperative Science Program Grant No. GF-33334 to Professor Farner.     

Structural and ultrastructural features of boar seminal vesicles

The morphological features of boar seminal vesicles were examined by light and transmission microscopy. Boar seminal vesicles consist of glandular tissue arranged in multiple lobules containing a system of ramified secretory tubules. The secretory tubules are composed of a mucosa formed by an epithelium and an underlying lamina propria and, are surrounded by a muscular layer. The epithelium is made up of columnar cells and occasional basal cells. Mast cells are frequently found among epithelial cells. Three types of columnar cells, considered different stages of the secretory cell cycle, are present: principal cells, clear cells and dense cells. Principal cells are functionally differentiated cells characterised by abundant mitochondria, great development of the rough endoplasmic reticulum and presence of secretory granules in their cytoplasm. The apical surface of many principal cells shows apical blebs filled with PAS-positive material. No acid mucosubstances are detected. Microvilli cover the apical surface except in the apical blebs. Dense cells, arranged between principal cells, are also functional differentiated cells but with signs of cellular degeneration. Clear cells are an initial differentiated stage of columnar cells and are characterised by the presence of a poorly developed rough endoplasmic reticulum and by the absence of secretory granules. Proliferating cells are present among columnar cells. Basal cells contain scarce cytoplasm, few organelles and no secretory granules. The lack of mitotic activity in these cells suggests that they do not act as precursors of columnar cells.     

Influence of neurosecretion on the activity of median eminence and pars intermedia in hereditary nephrogenic diabetes insipidus mice with bilateral supraoptic lesions

Summary The extremely hypertrophied hypothalamo-hypophysial neurosecretory system of the hereditary nephrogenic diabetes insipidus (DI Os/+) mice, shows distinctly hypertrophied median eminence (ME) indicated by the hypertrophied ependymal cells, increased number of Herring bodies and a great number of neurosecretory fibers loaded with secretion converged towards blood capillaries and towards pars tuberalis. The other distinct feature of the DI Os/+ mice is the marked hypertrophy and hyperplasia of the pars intermedia (PI) infiltrated with many beaded neurosecretory fibers. The majority of the PI glandular cells are of the large light type rather than the small type, the latter being comparatively numerous in normal mice. Very often these large light glandular cells and the marginal cuboidal epithelial cells of the hypophysial cleft contain or are surrounded by AF⁺ granules and/or colloid.When the SON is destroyed bilaterally by means of electrolytic lesions, loss of NSM from the ME, infundibulum and the pars nervosa, is accompanied by the disappearance of AF⁺ granules in the PI, and the shrinkage of all the above mentioned parts. The changes also occur in the three glandular cell types. The large light glandular cells, decrease in number as well as in size, while the granularity of the dark cells increases. The marginal cuboidal cells of the hypophysial cleft and the large light glandular cells loose most of the AF⁺ material from the cells as well as from the surroundings. Wherever the AF⁺ material does not disappear in the PI, the PI cells remain much big, as in the controls. The colloidal secretion on the margin of the PI and from the cleft may be the secretion of PI glandular cells, as a result of the stimulation of the neurosecretory fibers in the PI of DI Os/+ mice. This colloid disappears when the neurosecretory fibers are destroyed from the PI as a result of lesioning. Therefore, it suggests that this secretory material may belong to SON, or possibly to some other secretory cells residing in the anterior hypothalamic region. Or more probable explanation could be that with the disappearance of supraoptic neurons and/or anterior hypothalamic neurons due to electryolytic lesions, the secretory activity of the PI cells and its AF⁺ material, might have been hampered due to lack of stimulation of these fibers in PI.Therefore, from the previous work and from the present one, it is suggested that the NSM may contain some substance which stimulates the glandular cells of the PI and this substance may be either vasopressin and/or a type of CRF. However, it is concluded that the activated neurosecretory system plays an important role in the hypertrophy of the ME and PI and certain changes in PI cell types. The impairment of catecholamine fibers and their lack of influence on synthesis and/or release in PI parenchyma cells is discussed.This study was supported by MRC of Canada Grant No. MA-3759. A preliminary report on a part of this work appeared (Naik, 1970b) in Proc. Can. Fed. Biol. 13, 147.     

Localization and development of chromogranin A and luteinizing hormone immunoreactivities in the secretory-specific cells of the hypophyseal pars tuberalis of the chicken

 The pars tuberalis mainly consists of the secretory cells specific to this portion of the pituitary. We examined the localization and development of luteinizing hormone (LH) and chromogranin A in the chicken pars tuberalis by immunohistochemistry. The vast majority of the chicken pars tuberalis was occupied by cells immunoreactive for both LH and chromogranin A. Furthermore, immunoblot analysis of chicken pars tuberalis extracts with LH antiserum demonstrated that two bands, the large α-subunit and small β-subunit of the LH molecule, were expressed in this tissue as well as in the pars distalis. A band for chromogranin A was also detected in pars tuberalis extracts with chromogranin A antiserum. In contrast to the cells of mammalian species that contain only a few small secretory granules, the specific cells of the chicken pars tuberalis were characterized by the presence of many secretory granules ranging from 90 to 400 nm in diameter. Postembedding immunogold labeling showed that gold particles representing immunoreactivity for LH were densely located on all secretory granules of the secretory-specific cells. Many secretory granules, especially the large ones, of the cells were also loaded with immunogold particles for chromogranin A. Double immunogold labeling confirmed that LH and chromogranin A were colocalized on the same secretory granules. During embryonic development, the primordium of the pars tuberalis was first detected at 8 days of incubation as a small group of cells containing LH- and chromogranin-immunoreactive cells. In the pars distalis, the onset of LH and chromogranin expression occurred earlier, at 6 days of incubation. At 10 days of incubation, the pars tuberalis primordium became large cell masses consisting of LH- and chromogranin-immunoreactive cells, which were located close to the median eminence. Subsequently, the primordium extended along the median eminence progressively with age. At 14 days of incubation, it reached to the rostral end and surrounded the median eminence as slender cell cords. These results indicate that specific cells of the chicken pars tuberalis synthesize a glycoprotein hormone related to the LH molecule, which is stored in the secretory granules together with chromogranin A. The pars tuberalis may be involved in the regulation of gonadal function in a different way from that of the pars distalis. Accepted: 26 August 1997