Prostaglandin protection of human isolated gastric glands against indomethacin and ethanol injury. Evidence for direct cellular action of prostaglandin.

Isolated human gastric glands from surgical specimens were preincubated in an oxygenated medium with placebo or 16,16 dimethyl prostaglandin E2 (dmPGE2) and incubated at 37 degrees C in either medium alone, medium containing 4.43 mM indomethacin or medium containing 8% ethanol. We assessed the viability of gland cells with fast green exclusion, release of lactate dehydrogenase (LDH) into the medium, and ultrastructural damage by scanning and transmission electron microscopy. Both indomethacin and ethanol significantly reduced the viability of placebo-pretreated glands, increased LDH release into the medium, and produced prominent ultrastructural damage. DmPGE2 significantly reduced both indomethacin and ethanol-induced injury, increased the number of viable cells, reduced LDH release, and diminished the extent of ultrastructural damage. These studies indicate that PG protection of gastric mucosal cells has a direct cellular action that is not limited to replacement of depleted endogenous PGs. PG protection in our experiments did not depend on PG's previously described systemic actions, such as protection of the microvessels, preservation of the mucosal blood flow, or stimulation of bicarbonate and mucus secretion.

Prouroguanylin and proguanylin: purification from colon, structure, and modulation of bioactivity by proteases.

Uroguanylin and guanylin are peptides isolated from urine and intestinal mucosa, which regulate cyclic GMP production in enterocytes by activating an apical membrane, receptor-guanylate cyclase. This study extended our previous findings, which showed that colonic mucosa of opossums contained uroguanylin and guanylin peptides, by purifying prouroguanylin and proguanylin from this tissue. Prouroguanylin and proguanylin coeluted from Sephadex G-75 gelfiltration columns with a similar molecular size between 6 and 12 kDa. Mass spectrometry indicated that proguanylin (approximately 8.7 kDa) had a 10% lower molecular mass than prouroguanylin (approximately 9.7 kDa). Isoelectric focusing separated prouroguanylin (pI approximately 4.5) from proguanylin (pI approximately 7.5). N-terminal sequence analysis of reverse phrase-HPLC purified prohormones revealed 13 amino acids in opossum proguanylin that shared 77-85% identity with human and rat proguanylin, but only 23% identity with opossum prouroguanylin. The N-terminal 19 residues obtained for opossum prouroguanylin shared 32-42% identity with rat and human proguanylin. Prouroguanylin and proguanylin were both inactive and required pretreatment with proteases to elicit cyclic GMP responses in T84 cells. V8 protease treatment of proguanylin liberated a bioactive, 16-amino acid form of guanylin. Chymotrypsin treatment activated prouroguanylin, but inactivated the bioactive peptide domain within proguanylin. In summary, colonic mucosa contains the bioactive peptide and inactive prohormone forms of uroguanylin and guanylin. Thus, after proteolytic processing of prouroguanylin and proguanylin, bioactive uroguanylin and guanylin could both function to regulate guanylate cyclase activity by autocrine and/or paracrine actions on enterocytes. Also, these peptide hormones are implicated in an intestinal-renal axis for the endocrine regulation of salt and water homeostasis.

Signaling pathways for guanylin and uroguanylin in the digestive, renal, central nervous, reproductive, and lymphoid systems.

Guanylin and uroguanylin are peptides that stimulate membrane guanylate cyclases (GC) and regulate intestinal and renal function via cGMP. Complementary DNAs were isolated encoding opossum preproguanylin and a 279-amino acid portion of a receptor-guanylate cyclase expressed in opossum kidney (OK) cells (GC-OK). The tissue expression of messenger RNA transcripts for these signaling molecules were then compared. Northern and/or reverse transcription-PCR assays revealed that guanylin, uroguanylin, and GC-OK messenger RNAs are expressed in tissues within the digestive, renal, central nervous, reproductive, and lymphoid organ systems. Receptor autoradiography localized the receptors for uroguanylin and guanylin to renal proximal tubules and seminiferous tubules of testis. Synthetic guanylin and uroguanylin peptides activated the receptor-GCs in opossum kidney cortex and in cultured OK cells eliciting increased intracellular cGMP. Expression of agonist and receptor-GC signaling molecules provides a pathway for paracrine and/or autocrine regulation of cellular functions via cGMP in the digestive, renal, central nervous, reproductive, and lymphoid/immune organ systems. Uroguanylin also links the intestine and kidney in a potential endocrine axis that activates tubular receptor-GCs and influences renal function.

Lymphoguanylin: cloning and characterization of a unique member of the guanylin peptide family.

Guanylin and uroguanylin are small peptides containing two disulfide bonds that activate membrane guanylate cyclase-receptors in the intestine, kidney and other epithelia. Hybridization assays with a uroguanylin complementary DNA (cDNA) detected uroguanylin-like messenger RNAs (mRNAs) in the opossum spleen and testis, but these transcripts are larger than uroguanylin mRNAs. RT of RNA from spleen to produce cDNAs for amplification in the PCR followed by cloning and sequencing revealed a novel lymphoid-derived cDNA containing an open reading frame encoding a 109-amino acid polypeptide. This protein shares 84% and 40% of its residues with preprouroguanylin and preproguanylin, respectively. A 15-amino acid, uroguanylin-like peptide occurs at the COOH-terminus of the precursor polypeptide. However, this peptide is unique in having only three cysteine residues. We named the gene and its peptide product lymphoguanylin because the source of the first cDNA isolated was spleen and its mRNA is expressed in all of the lymphoid tissues tested. A 15-amino acid form of lymphoguanylin containing a single disulfide bond was synthesized that activates the guanylate cyclase receptors of human T84 intestinal and opossum kidney (OK) cells, although with less potency than uroguanylin and guanylin. Northern and/or RT-PCR assays detected lymphoguanylin mRNA transcripts in many tissues and organs of opossums, including those within the lymphoid/immune, cardiovascular/renal, reproductive, and central nervous organ systems. Lymphoguanylin joins guanylin and uroguanylin in a growing family of peptide agonists that activate transmembrane guanylate cyclase receptors, thus influencing target cell function via the intracellular second messenger, cGMP.

Purification and properties of parathyroid hormone-related peptide isolated from milk.

The occurrence and properties of PTH-related peptide (PTH-RP) in milk was investigated. PTH-RP was purified to homogeneity from human and bovine milk using heat and acid to precipitate milk proteins followed by ion exchange chromatography and reverse-phase HPLC. The peak of PTH-RP from HPLC was detected using a sensitive bone cell bioassay. A single band of peptide was detected on silver-stained polyacrylamide gels, which migrated as a 20-21-kDa macromolecule. PTH-RP isolated from either human or bovine milk had similar electrophoretic mobilities on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The partially purified bovine PTH-RP stimulated cAMP production in UMR106-01 and OK cell lines and elicited a concentration-dependent inhibition of sodium-dependent phosphate transport in OK cells. Incubation of milk extracts with an anti-PTH antiserum did not affect their bioactivity, whereas an antihuman PTH-RP 1-34 antiserum markedly reduced the cAMP response of UMR106-01 cells to the immunoabsorbed milk extracts. A PTH antagonist, norleu PTH 3-34, blocked the stimulation of cAMP production in UMR106-01 cells treated with milk extracts. PTH-RP immunoreactivity and bioactivity occurred in milk extracts of diverse animals from both eutherian and metatherian (marsupial) species. Porcine colostrum also had immunoreactive PTH-RP, although the levels were lower than the immunoreactive PTH-RP concentrations observed in milk samples collected at 7 and 14 days of lactation. Thus, a 20-21-KDa PTH-RP is secreted into milk where it could play a role in the development of suckling, newborn animals.

Brunner's glands: a structural, histochemical and pathological profile.

Brunner's glands are unique to mammalian species and in eutherians are confined primarily to the submucosa of the proximal duodenum. In the majority of species examined, they begin at the gastrointestinal junction and extend for variable distances distally in the wall of the proximal small intestine. Ducts of individual glands empty either directly into the intestinal lumen or unite with overlying intestinal glands (crypts of Lieberkühn) dependent on the species. Secretory units of Brunner's glands consist of epithelial tubules that show frequent distal branchings. The secretory units, with the exception of those found in rabbits and horses, consist primarily of a mucin producing cell type. However, other cell types normally associated with the overlying intestinal epithelium may be encountered scattered within the secretory units reflecting the developmental origin of these glands. Secretion from Brunner's glands contributes to a layer of mucus that forms a slippery, viscoelastic gel that lubricates the mucosal lining of the proximal intestinal tract. The unique capacity of this mucus layer to protect delicate underlying epithelial surfaces is due primarily to the gel-forming properties of its glycoprotein molecules. Mucin glycoproteins produced by Brunner's glands consist primarily but not exclusively of O-linked oligosaccharides attached to the central protein core of the glycoprotein molecule. Human Brunner's glands produce class III mucin glycoproteins and are thought to be the product of mucin gene MUC6 which is assigned to chromosome 11 (11p15-11p15.5 chromosome region). In addition to mucin glycoproteins and a limited amount of bicarbonate, numerous additional factors (epidermal growth factor, trefoil peptides, bactericidal factors, proteinase inhibitors, and surface-active lipids) have been identified within the secretory product of Brunner's glands. These factors, incorporated into the mucus layer, guard against the degradation of this protective barrier and underlying mucosa by gastric acid, pancreatic enzymes, and other surface active agents associated with this region. Yet other factors produced by Brunner's glands function to provide active and passive immunological defense mechanisms, promote cellular proliferation and differentiation, as well as contribute factors that elevate the pH of luminal contents of this region by promoting secretion of the intestinal mucosa, pancreatic secretion and gall bladder contraction. Additional insights concerning the role of Brunner's glands in the mammalian gastrointestinal tract as well as their possible evolution in this class of vertebrates have been gained from a basic understanding of their pathobiology.

Quality of gastric ulcer healing: histological and ultrastructural assessment.

It has long been assumed that the mucosa in areas of grossly 'healed' gastric or duodenal ulcers returns to normal, either spontaneously or after treatment. This assumption is based almost entirely upon visual, superficial examination by endoscopy. Few, if any, histological and ultrastructural studies examined the deeper mucosa in the areas of grossly healed ulcers. In several experimental studies, we analysed the development, evolution, and healing of acetic acid-induced gastric ulcers in rats and assessed the histological and ultrastructural features (structure and cellular composition) of the gastric mucosa in areas of grossly healed ulcers. The gastric mucosa of grossly 'healed' ulcers showed re-epithelialization of the mucosal surface at every study interval (2 weeks, 2, 3, and 4 months), but the subepithelial mucosa displayed prominent abnormalities. Two patterns of scarring were distinguished: (a) the mucosa in the area of healed ulcer was thinner (25-45% thinner than normal mucosa) with increased connective tissue and poor differentiation and/or degenerative changes in the glandular cells; and (b) the mucosa displayed a marked dilation of gastric glands with poor differentiation of the glandular cells and a reduction in the supportive microvascular network. It is theorized that these abnormalities could interfere with oxygenation, nutrient supply, and mucosal resistance and defence; therefore, they could be a basis for ulcer recurrence. These observations indicate that the quality of mucosal structural restoration rather than the speed of ulcer healing is the most important factor in determining risk of ulcer recurrence. The clinical relevance of these findings is supported by a preliminary study in which marked histological abnormalities were found in the subepithelial mucosa in patients with 'healed' duodenal ulcers.

The portal hypertensive gastric mucosa: histologic, ultrastructural, and functional analysis after aspirin-induced damage.

We assessed macroscopic, histologic, ultrastructural, and functional features of aspirin-induced gastric mucosal injury in portal hypertensive and sham-operated rats. Portal hypertension was produced by staged portal vein ligation. Four hours after intragastric acidified aspirin administration, intraluminal pH in portal hypertensive rats was 6.6 +/- 0.2 and 4.3 +/- 0.5 in sham-operated controls (p less than 0.01). Gross mucosal damage was significantly greater in portal hypertensive rats compared with controls (18 +/- 2 versus 7 +/- 1% of total mucosal area). Histologic deep necrosis involved 22 +/- 2% of mucosal section lengths in portal hypertensive rats compared with 7 +/- 1% in sham-operated rats (p less than 0.01). In portal hypertensive rats, histologic and ultrastructural evaluation demonstrated capillary endothelial abnormalities, arterialization of submucosal veins, and markedly greater severity of microvascular damage than in sham-operated controls. Neutralized aspirin (pH, 7.0) did not produce any significant damage detectable grossly, histologically, or by transmission electron microscopy in portal hypertensive rats. We conclude that acid-dependent aspirin-induced gastric mucosal damage is significantly increased in portal hypertension.

A scanning electron microscopic study of the opossum nasal cavity prior to and shortly after birth.

The nasal cavities of opossums prior to and shortly after birth were examined by scanning electron microscopy. Numerous morphologically mature olfactory receptor neurons are observed in the dorso-rostral-most extent of the olfactory epithelium positioned adjacent to the opening of the nares in all prenatal stages and newborn animals examined. The remainder of the olfactory epithelium, occupying a more dorso-caudal position within the nasal cavity, is undifferentiated, and lacks morphologically mature receptor neurons. A short transition zone of stratified squamous epithelium lies between the epithelium lining the nares and olfactory epithelium. It forms an abrupt junction with the latter. The remainder of the nasal cavity in this group of animals is lined by a non-ciliated pseudostratified type (undifferentiated respiratory) of epithelium. By the end of the second postnatal week the morphologically mature olfactory epithelium is no longer observed in the vestibular area of the nasal cavity, which is lined by stratified squamous epithelium at this time. Mature receptor neurons are now observed within the olfactory epithelium lining the roof of the nasal cavity and covering the turbinates. The greater part of the nasal cavity is lined by a ciliated respiratory epithelium. It is proposed that the precocious differentiation of mature olfactory receptor neurons within the rostral-most extent of the olfactory epithelium just prior to birth is important in guiding the newborn young to the pouch.

Morphological observations on the harderian gland of the North American opossum (Didelphis virginiana).

The Harderian gland of the North American opossum (Didelphis virginiana) is large and well developed, despite the absence of a nictitating membrane in the adult of this species. The elongate glands are surrounded by a delicate connective tissue capsule from which thin septae extend, subdividing the gland into numerous lobules. The secretory units of the opossum Harderian gland are drained by a well defined but not extensive intralobular and interlobular duct system. Most of the secretory end pieces consist of tubulo-alveolar units with widely dilated lumina filled with secretory product. Numerous intact lipid vesicles suspended within an amorphous material constitute the luminal contents. Cells lining the tubulo-alveolar secretory end-pieces are usually columnar in shape, and characterized by numerous lipid-containing secretory vesicles and aggregations of poly-tubular complexes 40-60 nm in diameter. In addition, these cells contain numerous large irregularly shaped mitochondria, whose matrix is of considerable electron density. Intralobular and interlobular ducts are lined by electron-lucent epithelial cells that lack both the lipid-containing vesicles and the large mitochondria, although typical smaller mitochondria are found scattered within the cytoplasm. Both secretory end-pieces and ductal elements are invested by an abundance of myoepithelial cells. A second, smaller serous type of secretory unit may occur near the centre of some Harderian gland lobules. In these units secretory tubules and acini are compactly arranged surrounding a narrow lumen. Serous cells are pyramidal in shape and the cytoplasm is characterized by numerous electron-dense secretory granules and scattered profiles of rough endoplasmic reticulum.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of effects of uroguanylin, guanylin, and Escherichia coli heat-stable enterotoxin STa in mouse intestine and kidney: evidence that uroguanylin is an intestinal natriuretic hormone.

BACKGROUND: Uroguanylin and guanylin are intestinal peptides that activate a receptor-guanylate cyclase, which is also a receptor for Escherichia coli heat-stable enterotoxin (STa). These peptides may have a role in the body's regulation of fluid and electrolytes. METHODS: STa, bioactive guanylin, and bioactive uroguanylin were evaluated for effects in: 1) the suckling mouse intestinal fluid secretion assay; 2) an in vitro suckling mouse intestinal loop assay; 3) an intestinal receptor autoradiography assay; 4) a control or agonist-stimulated assay for cGMP response in T84 cells; and 5) an in vivo renal function assay in mice. RESULTS: In vivo, orally administered uroguanylin and STa but not guanylin, stimulated intestinal fluid secretion. All three peptides activated intestinal guanylate cyclase and had common intestinal receptors. In vitro, after pretreatment with chymotrypsin, only uroguanylin and STa retained agoinst activity. Chymostatin preserved guanylin activity. STa and uroguanylin induced diuresis, natriuresis, and kaliuresis. Guanylin was less potent than uroguanylin and STa. CONCLUSIONS: The results suggest that the endogenous intestinal peptides, uroguanylin and guanylin, regulate water and electrolyte homeostasis both through local effects on intestinal epithelia and endocrine effects on the kidney.

Milk composition in the North American opossum (Didelphis virginiana).

The composition of milk samples collected from captive opossums (Didelphis virginiana) was determined at various intervals during lactation. The milk solids increased from 9% at week one to a maximum of 34% at 11 weeks post-partum. There were changes in the relative proportions of protein, lipid and carbohydrate at different stages of lactation. Lipid represented the greatest fraction of the solids except for a period at mid-lactation when there was a peak in protein concentration. The concentrations of sodium, potassium and magnesium were relatively constant, 41 +/- 4, 35 +/- 11 and 9.2 +/- 1.6 mmol respectively, although calcium increased from 13 +/- 5 mmol at week one to a peak of 112 +/- 35 mmol at 9 weeks.

Guanylin peptides: cyclic GMP signaling mechanisms.

Guanylate cyclases (GC) serve in two different signaling pathways involving cytosolic and membrane enzymes. Membrane GCs are receptors for guanylin and atriopeptin peptides, two families of cGMP-regulating peptides. Three subclasses of guanylin peptides contain one intramolecular disulfide (lymphoguanylin), two disulfides (guanylin and uroguanylin) and three disulfides (E. coli stable toxin, ST). The peptides activate membrane receptor-GCs and regulate intestinal Cl- and HCO3- secretion via cGMP in target enterocytes. Uroguanylin and ST also elicit diuretic and natriuretic responses in the kidney. GC-C is an intestinal receptor-GC for guanylin and uroguanylin, but GC-C may not be involved in renal cGMP pathways. A novel receptor-GC expressed in the opossum kidney (OK-GC) has been identified by molecular cloning. OK-GC cDNAs encode receptor-GCs in renal tubules that are activated by guanylins. Lymphoguanylin is highly expressed in the kidney and heart where it may influence cGMP pathways. Guanylin and uroguanylin are highly expressed in intestinal mucosa to regulate intestinal salt and water transport via paracrine actions on GC-C. Uroguanylin and guanylin are also secreted from intestinal mucosa into plasma where uroguanylin serves as an intestinal natriuretic hormone to influence body Na+ homeostasis by endocrine mechanisms. Thus, guanylin peptides control salt and water transport in the kidney and intestine mediated by cGMP via membrane receptors with intrinsic guanylate cyclase activity.

Morphological, developmental and immunohistochemical observations on the opossum pituitary with emphasis on the pars intermedia.

Development of the pituitary in Didelphis follows the general pattern of that described for most mammalian species. The dorsal region of a multichambered pituitary vesicle, which forms from Rathke's pouch, comes to lie adjacent to the presumptive infundibulum by the 10 1/2 d of gestation. The epithelial wall of this vesicle consists of spindle-shaped cells. The dorsal wall of the upper chamber of the pituitary vesicle forms the pars intermedia; the ventral wall of this chamber gives rise to cells of the pars distalis. Corticotropes, somatotropes, and lactotropes are seen in the presumptive adenohypophysis of the 11 1/2 d embryo. Gonadotropes and thyrotropes appear about 1 d later, shortly after birth. By the 2 postnatal week, the adult distribution of all 5 cell types within the pars distalis appears to have been established. The wall bounding the pituitary cleft in the adult represents an epithelial continuum limited by a basal lamina and corresponds to the upper chamber of the original pituitary vesicle. Ultrastructurally, the limiting walls of the pituitary cleft consists of stellate (marginal) cells, large, bulbous cells, and granulated cells. The latter correspond to the various endocrine cell types normally associated with the pars distalis. Non-granular folliculo-stellate cells also are observed within the epithelial cords of the pars distalis.

Immunohistochemical evidence for the presence of oxytocin in the opossum corpus luteum.

Corpora lutea from opossums late in pregnancy were examined by immunohistochemistry for the presence of oxytocin. Oxytocin-immunoreactivity was observed in all corpora lutea examined but not elsewhere in ovarian tissue. The immunoreactive staining observed was confined primarily to the perinuclear cytoplasm of reactive luteal cells. Not all luteal cells showed oxytocin-immunoreactivity. The immunohistochemical localization of oxytocin in the pregnant opossum corpus luteum demonstrates for the first time this peptide in a metatherian ovary. Its presence in this primitive species suggests that oxytocin has a fundamental role in the physiology of the mammalian ovary.

Brain growth and neocortical development in the opossum.

General brain growth and differentiation of the neocortex have been studied in the marsupial, Didelphis virginiana from the 10.5 day embryo through adulthood. Didelphis is born after a short gestation period of about 12.5 days, at a time when the telencephalic wall consists only of two layers and is considered to be at an embryonic stage of development. The cortical plate does not appear until late in the first postnatal week, thus neocortical development is totally a postnatal phenomenon in Didelphis as has been shown in other marsupial species examined to date. The general pattern of development and the establishment of the six-layered adult neocortex in Didelphis is similar to that described in eutherian mammals. Signs of cortical lamination can be seen as early as postnatal day 35 and the cytoarchitecture of a typical mammalian neocortex is well defined by postnatal day 60 in Didelphis prior to the onset of weaning.

Immunohistochemical localization of relaxin in the reproductive system of the female opossum (Didelphis virginiana).

Relaxin-immunoreactivity was demonstrated in the cytoplasm in the luteal cells from pregnant and lactating opossums. Immunoreactivity for relaxin was not demonstrated elsewhere in the ovary, in the reproductive tract or in the placenta. The corpus luteum is thought to be the primary source of relaxin in Didelphis and in this regard is similar to several eutherian mammals including man.

Morphometric analysis of endometrial periglandular fibrosis in mares.

OBJECTIVES: To develop an objective, quantifiable assay for endometrial periglandular fibrosis (EPF) and correlate assay results with histologic and ultrastructural changes in equine endometrial biopsy specimens. SAMPLE POPULATION: Endometrial biopsy specimens from 70 mares from 3 to 27 years old in estrus. PROCEDURE: In a double-blinded study design, endometrial biopsy specimens were graded histologically (modified Kenney classification) for EPF and inflammation. Endometrial periglandular collagen volume fraction (%EPCVF) was determined by light microscopic image analysis of picrosirius red-stained sections. Specimens from selected mares were examined by transmission electron microscopy. RESULTS: %EPCVF values varied significantly among the 4 modified Kenney EPF categories (I, IIA, IIB, and III) and increased with increasing age of mares. Morphologically, EPF consisted of concentric layers of transformed fibroblasts with myofibroblastic features and deposition of fibrillar collagen around unaltered glandular basal laminae. CONCLUSIONS AND CLINICAL RELEVANCE: %EPCVF correlates well with morphologic changes in endometrial biopsy specimens. Determination of %EPCVF could be useful in evaluation and clinical management of subfertile mares and in investigations of the pathogenesis of EPF.

Ultrastructure of the platypus and echidna mandibular glands.

The secretory units of the platypus and echidna mandibular glands consist of a single serous cell type. Secretory granules within the cells of the platypus mandibular gland stained intensely with the periodic acid-Schiff staining procedure but failed to stain with Alcian Blue, suggesting the granules contained neutral glycoproteins. Secretory granules within the mandibular glands of the echidna failed to stain with the methods used indicating little if any glycoprotein was associated with the secretory granules. Ultrastructurally, secretory granules of the platypus mandibular gland were electron dense with a central core of less electron-dense material and were membrane bound. In contrast, those of the echidna presented a lamellated appearance and also were limited by a membrane. These secretory granules appeared to form as a result of concentric layering of lamellae within cisternae of the Golgi membranes. The intralobular ductal system of the platypus was more extensively developed than that of the echidna. The striated ducts of both species were characterized by elaborate infoldings of the basolateral plasmalemma and an abundance of associated mitochondria.

Microscopy of the koala mandibular (submandibular) glands.

Koala mandibular (submandibular) glands are compound tubuloacinar glands, the secretory units of which consist only of serous cells. Intercellular canaliculi occur between the serous cells, which are continuous with a minute lumen that courses through the centre of each secretory unit. Intercalated ducts are abundant and join striated ducts, the latter being characterized by elaborate basolateral infoldings of the plasmalemma. Secretory granules within the serous cells fail to stain with either the PAS or Alcian Blue (pH 2.5) staining procedures. Ultrastructurally, the secretory granules are membrane bound, and consist of a homogeneous electron lucent material with a fine filamentous texture. The granules tend to coalesce into irregular shaped complexes of secretory material. Discharge of secretory material into the canalicular lumen is a common observation.