Collagen-like Material Research Articles

Benzodiazepine Delorazepam Induces Locomotory Hyperactivity and Alterations in Pedal Mucus Texture in the Freshwater Gastropod Planorbarius corneus

Benzodiazepines, psychotropic drugs, are ubiquitous in the aquatic environment due to over-consumption and inefficient removal by sewage treatment plants. Bioaccumulation with consequent behavioral and physiological effects has been reported in many aquatic species. However, the responses are species-specific and still poorly understood. To improve the knowledge, we exposed the freshwater snail Planorbarius corneus to 1, 5, or 10 µg/L of delorazepam, the most widely consumed benzodiazepine in Italy. Conventional behavioral tests were used to assess the effects on locomotor and feeding behavior. Histological and biochemical analyses were also performed to detect possible changes in the structure and composition of the foot mucus and glands. The results show a paradoxical response with reduced feeding activity and locomotor hyperactivity. Pedal mucus was altered in texture but not in composition, becoming particularly rich in fibrous collagen-like material, and a significant change in the protein composition was highlighted in the foot. In conclusion, exposure to delorazepam induces disinhibited behavior in Planorbarius corneus, potentially increasing the risk of predation, and an increase in mucus protein production, which, together with reduced feeding activity, would severely compromise energy resources.

A Computed Tomographic and Pathological Study of Equine Cheek Teeth Infundibulae Extracted From Asymptomatic Horses. Part 2: MicroCT, Gross, and Histological Findings.

Background: Equine maxillary cheek teeth infundibulae are frequently affected by developmental and acquired disorders, but the imaging, gross, and histological features of normal and abnormal infundibulae remain incompletely understood.Objective: To perform MicroCT, gross examination, and histology on sectioned teeth and compare the imaging and anatomical findings.Study design: Ex vivo original study.Methods: Eight maxillary cheek teeth of different ages and with varying grades of unilateral (n = 5) or bilateral (n = 3) occlusal infundibular caries were extracted from equine heads obtained from an abattoir. The teeth were imaged by MicroCT, then transversely sectioned and grossly and histologically examined, with the imaging and gross and histological anatomical findings examined and compared.Results: Fifteen infundibulae, including two without occlusal caries had subocclusal cemental hypoplasia or caries. One infundibulum without occlusal caries had no subocclusal cemental defects. Histologically, hypoplastic cemental areas consisted of irregularly-shaped, wide central channels, with multiple, large, cylindrical side-branches that extended peripherally to a variable extent. Cementum with extensive, wide, empty channels, and cementum with a more irregular moth-eaten appearance had dark or eroded gross appearance and a low HU on CT. Some infundibulae had cement-free areas that only contained fragments of collagen-like material, especially at their apical aspects (apical cemental hypoplasia). Carious subocclusal areas had connections with the occlusal surface and had disrupted cemental architecture, including of their central vascular channel that, along with their side branches, contained degraded food, and cemental debris.Main Limitations: No clinical histories or accurate ages were available for these eight teeth.Conclusions: Hypoplastic cemental lesions, including at central linear and apical sites, histologically contain areas with multiple wide-branched, cylindrical channels or even areas of total cement hypoplasia visible on gross sections When such cemental defects contact the occlusal surface due to normal wear, food impaction, and caries can ensue.

Fabrication of Dielectrically Sensitive Carbon Nanotube/Polyethylene Nanohybrid Shish-Kebab Structure for Potential Tissue Engineering Applications

Carbon nanotube (CNT)/polymer nanohybrid shish-kebab (NHSK) structure is a unique approach to functionalize CNTs and mimic the hierarchical architecture of native collagen fibrils in the extracellular matrix (ECM), owing to their favorable geometric similarity. This study thus aimed at combining the unique NHSK architecture of CNTs with their favorable dielectric properties to fabricate a collagen-like, dielectrically sensitive and biocompatible material. The NHSK structure was fabricated with CNTs and polyethylene (PE) by means of a solution crystallization technique. Configuration of PE in terms of molecular weight and structure was found to be one of the important factors that determined the morphology of the NHSK. Moreover, the material was exposed to dielectrophoresis (DEP) to guide the deposition of CNT/PE NHSK into the directional pattern. The PE-decorated CNTs exhibited enhanced dielectrophoretic sensitivity and controllable structural alignment, while the neat CNTs did not. Human osteoblast-like MG-63 cells were cultured on the scaffolds to examine their biocompatibility and results indicated that the nanotopography of the kebab crystals on the CNTs facilitated cell adhesion and proliferation while the DEP-patterned NHSK structure further boosted cell attachment and viability, thus suggesting that the CNT/polymer nanohybrid shish kebab structure has potential applications in tissue engineering.

Does the G-spot exist? A review of the current literature.

In 1950, Gräfenberg described a distinct erotogenic zone on the anterior wall of the vagina, which was referred to as the Gräfenberg spot (G-spot) by Addiego, Whipple (a nurse) et al. in 1981. As a result, the G-spot has become a central topic of popular speculation and a basis of a huge business surrounding it. In our opinion, these sexologists have made a hotchpotch of Gräfenberg's thoughts and ideas that were set forth and expounded in his 1950 article: the intraurethral glands are not the corpus spongiosum of the female urethra, and Gräfenberg did not report an orgasm of the intraurethral glands. G-spot amplification is a cosmetic surgery procedure for temporarily increasing the size and sensitivity of the G-spot in which a dermal filler or a collagen-like material is injected into the bladder-vaginal septum. All published scientific data point to the fact that the G-spot does not exist, and the supposed G-spot should not be identified with Gräfenberg's name. Moreover, G-spot amplification is not medically indicated and is an unnecessary and inefficacious medical procedure.

Production of pediocin PA-1 in the methylotrophic yeast Pichia pastoris reveals unexpected inhibition of its biological activity due to the presence of collagen-like material

Expression of the pedA gene from Pediococcus acidilactici, coding for mature bacteriocin Pediocin PA-1, was investigated using the yeast Pichia pastoris to obtain larger quantities of pediocin to support additional studies, including structure–function research. Following various cloning strategies, a KM71H (Mut s) strain was selected. A significant concentration (74 μg/ml) of extracellular recombinant pediocin was obtained but the pediocin showed no biological activity. Supernatant fluids from P. pastoris cultures, harboring or not pedA, inhibited the biological activity of natural pediocin PA-1. The recombinant pediocin appeared as a mixture of three main fractions (7–8, 11, 20 kDa vs. 4.6 kDa for natural pediocin PA-1). The recombinant pediocin was also less hydrophobic and behaved differently when subjected to isoelectric focusing. Strong evidence indicated that some “collagen-like” material was tightly associated, most probably via covalent binding, to the recombinant pediocin. The “collagen-like” material was most probably responsible for the lack of biological activity of the recombinant pediocin and for the differences observed regarding some of the physico-chemical properties. Both the recombinant pediocin and natural pediocin were sensitive to collagenase, suggesting that pediocin PA-1 may possess a somewhat “collagen-like” nature. Interestingly, recombinant pediocin preparations showed the ability to assemble into fibrils.

Identification of seminiferous tubule aberrations and a low incidence of testicular microliths associated with the development of azoospermia

Objective: To evaluate the use of percutaneous testicular sperm aspiration in the assessment of azoospermia and its association with seminiferous tubule microliths. Design: Case report. Setting: Tertiary care fertility center in a university hospital. Patient(s): Male undergoing infertility evaluation. Intervention(s): Testicular biopsy and percutaneous testicular aspiration. Main Outcome Measure(s): Serum hormone analysis, sperm concentration in semen, spermatogenesis in samples from testicular biopsies and aspirations, and microlith composition. Result(s): A patient presented for infertility evaluation with a history of severe oligospermia that progressed to azoospermia. The serum testosterone concentration (357 ng/dL) and LH concentration (9.2 mIU/mL) were normal and the serum FSH concentration (18.3 mIU/mL) was elevated. Testicular biopsy results indicated spermatogenic hypoplasia with limited spermatozoa. Seminiferous tubules obtained by percutaneous testicular aspiration were structurally aberrant, with multiple diverticula. Microliths averaging 120 μm in diameter were observed within and blocking the seminiferous tubules. The microliths were composed of calcium phosphate (hydroxyapatite) in both the core and peripheral regions. Electron microscopy revealed a high degree of collagen-like material within the peripheral zone. Conclusion(s): The presence of seminiferous tubule microliths is associated with the development of azoospermia. In patients with a low incidence of seminiferous tubule microliths and aberrant seminiferous tubule architecture, percutaneous testicular aspiration may provide a diagnostic advantage over testicular biopsy.

Chronic nitric oxide inhibition model six years on.

The discovery in 1987 that endothelium-derived nitric oxide (NO) mediates the vasodilatory effect of certain endothelium-dependent agonists1 2 inaugurated the current huge field of NO biology. It is now recognized that NO plays essential roles in many diverse physiological processes and in some pathophysiologic events. Development of these concepts has been based largely on evidence obtained by limiting NO biosynthesis. This review is centered on the cardiovascular and particularly the renal functional and structural consequences of chronic pharmacologic NO inhibition by l-arginine analogues. We devoted special attention to the mechanisms of hypertension and organ injury that occur under these circumstances, while appreciating the inherent limitations surrounding interpretation of this data. NO is made by the enzymatic action of several widely distributed NO synthases (NOS). In the presence of the substrates l-arginine and oxygen, as well as a number of essential cofactors, NO is produced in response to appropriate stimuli. The constitutively expressed NOS play a major role in the physiological control of vascular tone and kidney function.3 4 Vascular endothelial NOS (eNOS) and brain-type NOS (bNOS) are widely distributed throughout the kidney5 as well as the cardiovascular system and in strategic locations in the peripheral and central nervous system (CNS).6 7 Both eNOS and particularly bNOS are abundant in the kidney, glomeruli, and vasculature as well as in most segments of the tubule,3 5 and NOS activity in medulla is considerably greater than in cortex.8 NO generated within the kidney controls the glomerular filtration rate (GFR), total renal and medullary blood flow, pressure natriuresis, epithelial sodium transport, and production of various vasoactive factors including renin.3 4 5 eNOS is distributed throughout most parts of the arterial and venous circulation, although there is considerable heterogeneity in the extent to which NO controls …

Intracellular collagen-like material in mouse embryonic mesenchymal cells incubated with TNF-alpha and IL-1 beta in organoid culture.

Blastemal cells of embryonic mouse limb buds (day 12) were cultivated in organoid cultures in the presence of the human recombinant cytokines interleukin-1 beta (IL-1 beta) and tumour necrosis factor alpha (TNF-alpha). The effects of both cytokines (applied alone or together) on mesenchymal cells were demonstrated by electron microscopy. Cultures treated with TNF-alpha (alone or in combination with IL-1 beta) showed several mesenchymal cells with numerous irregularly shaped membrane-bordered cavities containing thick bundled tannic-acid-positive fibrillar structures that resembled loosened collagen fibrils, whereas cells exposed to IL-1 beta alone did not exhibit such changes. These findings are discussed in the light of two hypotheses: the phagocytosis of extracellular collagen fibrils, and fibrillogenesis resulting from incongruity of synthesis and secretion rates of procollagen; our results favour the former.

Effects of Axonal Injury on Astrocyte Proliferation and Morphologyin Vitro:Implications for Astrogliosis

Mechanisms inducing gliosis following injury in the central nervous system are poorly understood. We evaluated the effect of axonal injury on astrocyte and Schwann cell proliferation and morphologyin vitro.Purified rat dorsal root ganglion neurons grown on monolayers of rat neonatal cortical astrocytes (N-ASneonatalcultures) or sciatic nerve-derived Schwann cells (N-SC cultures) were mechanically injured. Noninjured cultures served as controls. Cell proliferation near lesions was monitored by autoradiography 1, 2, 4, and 8 days postinjury. Axonal injury caused a significant transient increase in astrocyte proliferation immediately proximal and distal to the lesion. The lesion did not induce marked changes in the intensity of glial fibrillary acidic protein (GFAP) immunoreactivity. However, processes from GFAP-positive cells usually arranged in random fashion in noninjured cultures were aligned perpendicularly to the cut distal to lesions. Ultrastructural analysis in lesioned N-ASneonatalcultures indicated that proximal to the lesion filament-filled astrocytes were intermingled with axons. Distal to the lesion astrocyte processes formed layers, between which an increased amount of collagen-like material appeared with time postlesion. Axons distal to the lesion degenerated by 2 days, coinciding with the early disappearance of neurofilament immunoreactivity. In noninjured and proximally in injured N-SC cultures, Schwann cells extended processes, engulfing some axons. Distal to the lesion, Schwann cells appeared more rounded and neurites remained until 4 days postinjury. Media conditioned by injured or noninjured N-ASneonatalcultures did not affect neuron-induced Schwann cell proliferation. These findings demonstrate that axonal injury and degeneration cause a transient increase in astrocyte proliferation and induce morphological changes in astrocytes consistent with the onset of gliosis.

Blood-borne collagenous debris complexes with plasma fibronectin after thermal injury

Plasma fibronectin augments the clearance of blood-borne foreign and effete complexes by mononuclear phagocytes. The release of a “gelatin- like” ligand into plasma after thermal injury has been reported. We quantified the release of this collagenous debris from thermally injured skin, and its potential interaction with soluble fibronectin in plasma using anesthetized rats. Collagen-like material debris in the plasma was detected by assay of hydroxyproline. Fibronectin was measured by a double antibody enzyme-linked immunosorbent assay (ELISA) technique. Over a 24-hour postburn interval, plasma hydroxyproline increased from 6.7 +/- 0.6 micrograms/mL to a maximum of 19.0 +/- 3.3 micrograms/mL at 60 minutes postburn, and normalized by 6 hours. A direct correlation existed between the magnitude of burn injury and the increase in plasma hydroxyproline. In parallel, plasma fibronectin declined over a 15-minute to 2-hour period postburn, and normalized by 3 to 4 hours with rebound hyperfibronectinemia observed at 24 hours. The elevation in total plasma hydroxyproline was not due to an increase in plasma Clq (zero time, 26.2 +/- 1.4 micrograms/mL; 60 minutes, 23.9 +/- 1.1 micrograms/mL). Tracer studies with 125I-fibronectin showed that the acute decline of plasma fibronectin was due to its uptake by the liver and binding to sites of tissue injury. Total hydroxyproline in extracts of burn skin, used as an index of soluble collagenous material, rose from 15 +/- 3.3 micrograms/g skin at zero time to 129.3 +/- 43.7 micrograms/g skin by 5 minutes postburn, with a decline to 38 +/- 22 micrograms/g skin by 24 hours. The formation of circulating fibronectin-gelatin complexes in vivo was documented by cross- immunoelectrophoresis coupled with autoradiography using 125I-gelatin as a model ligand. Thus, collagenous tissue debris from burned skin may enter the plasma after thermal injury and directly complexes with soluble fibronectin before hepatic phagocytic clearance.

Blood-Borne Collagenous Debris Complexes with Plasma Fibronectin After Thermal Injury

Plasma fibronectin augments the clearance of blood-borne foreign and effete complexes by mononuclear phagocytes. The release of a "gelatin-like" ligand into plasma after thermal injury has been reported. We quantified the release of this collagenous debris from thermally injured skin, and its potential interaction with soluble fibronectin in plasma using anesthetized rats. Collagen-like material debris in the plasma was detected by assay of hydroxyproline. Fibronectin was measured by a double antibody enzyme-linked immunosorbent assay (ELISA) technique. Over a 24-hour postburn interval, plasma hydroxyproline increased from 6.7 +/- 0.6 micrograms/mL to a maximum of 19.0 +/- 3.3 micrograms/mL at 60 minutes postburn, and normalized by 6 hours. A direct correlation existed between the magnitude of burn injury and the increase in plasma hydroxyproline. In parallel, plasma fibronectin declined over a 15-minute to 2-hour period postburn, and normalized by 3 to 4 hours with rebound hyperfibronectinemia observed at 24 hours. The elevation in total plasma hydroxyproline was not due to an increase in plasma Clq (zero time, 26.2 +/- 1.4 micrograms/mL; 60 minutes, 23.9 +/- 1.1 micrograms/mL). Tracer studies with 125I-fibronectin showed that the acute decline of plasma fibronectin was due to its uptake by the liver and binding to sites of tissue injury. Total hydroxyproline in extracts of burn skin, used as an index of soluble collagenous material, rose from 15 +/- 3.3 micrograms/g skin at zero time to 129.3 +/- 43.7 micrograms/g skin by 5 minutes postburn, with a decline to 38 +/- 22 micrograms/g skin by 24 hours. The formation of circulating fibronectin-gelatin complexes in vivo was documented by cross-immunoelectrophoresis coupled with autoradiography using 125I-gelatin as a model ligand. Thus, collagenous tissue debris from burned skin may enter the plasma after thermal injury and directly complexes with soluble fibronectin before hepatic phagocytic clearance.

Oviductal morphology and egg shelling in the oviparous lizards Crotaphytus collaris and Eumeces obsoletus.

Morphological changes occurring in the oviduct and epithelial cells of the lizards Crotaphytus collaris and Eumeces obsoletus during the natural reproductive cycle were examined and quantified. Additionally, development of the eggshell at different stages of gravidity was described. The anterior uterus of each species has a distinct glandular type which differs between species: in E. obsoletus, the glands are tubular and in C. collaris, branched saccular. The branched saccular glands in the anterior uterus of C. collaris produce collagen-like material that forms the fibers of the shell membranes. However, fibers from the eggshell of E. obsoletus did not stain for collagen. The shell of both species is composed of a multilayered inner boundary covered externally by fibers of varying thickness. Initial layers are composed of thick fibers all lying along the same general axis. Outer layers of fibers are progressively thinner and an external surface layer composed of glycosaminoglycans (GAGs) is also present. In C. collaris, calcium, which is deposited in relatively small amounts on the shell surface, appears to be secreted by the epithelium of the anterior uterus. The nonciliated secretory epithelial cells covering the villi-like folds of the posterior infundibulum secrete GAGs. Epithelial cell height of the infundibular villi is greatest during early gravidity. A functional relationship may exist between luteal activity and oviductal secretory activity because the activity of the glandular epithelium varied as gravidity progressed.

Collagen in the egg shell membranes of the hen

Collagen-like proteins have been found in the egg shell membranes of the hen. Materials similar to types I and V collagens were detected in each of the two layers of this membrane, the thick outer membrane and the thin inner membrane. Collagen was extracted by acid-pepsin digestion and isolated by differential salt precipitation. Identification of type-specific collagen-like material was established by coelectrophoresis on SDS-polyacrylamide gels using known collagen standards. These bands were susceptible to digestion by bacterial collagenase. From differential staining of the gels it was estimated that the ratio of collagen types I:V was approximately 100:1. Further confirmation of these biochemical results was obtained with immunofluorescence microscopy using type-specific antisera against chicken types I and V collagen with the indirect sandwich technique. Both the inner and outer shell membranes contained the two types of collagen. Within each membrane, the large, coarse 2.5-μm fibers contained predominantly type I collagen-like material, while type V collagen was mainly associated with the delicate narrower fibers of approximately 0.6-μm diameter. These tended to be concentrated in the inner membrane. At the electron microscopic level, both types of fibers were coated with glycoproteins that stained positively with ruthenium red. The deposition of these collagen-like substances by the hen oviduct on to the surface of the developing egg is an additional example of interstitial-type collagen synthesis and secretion by epithelial rather than by measenchymal cells.

Collagen in the pterobranch coenecium and the problem of graptolite affinities

Amino-acid analysis of the extracellular skeleton (coenecium) of two pterobranchs (Hemichordata) Rhabdopleura normani and Cephalodiscus (C.) hodgsoni shows that both contain considerable quantities of collagenous material with relatively high hydroxyproline and low hydroxylysine levels. The appearance of the fibrous material in the skeleton of pterobranchs, although collagenous, differs from standard EM characteristics of collagen. The identification of the collagenous nature of the pterobranch skeleton with the presumed presence of collagen-like material in the periderm of fossil graptolites, taken in conjunction with other data, supports the hypothesis that both groups may be closely related phylogenetically.

Light microscopical studies of the acute toxic effects of N-nitrosodimethylamine on the marine mussel, Mytilus edulis

Mussels, Mytilus edulis, exposed to N-nitrosodimethylamine (DMN), 100 mg DMN/liter sea water, exhibited the following tissue responses: Congestion of the larger vessels in the Leydig cell tissue and infiltration of the tissue with numerous hemocytes, well-demarcated lesions composed of hemocytes in the Leydig cell tissue, degeneration of the epithelial lining of the ducts of the digestive diverticula, and congestion of the branchial vessels with numerous hemocytes accompanied by distortion of the cilia and the ciliated columnar epithelium of the gills. Furthermore, deposition of collagen-like material in the walls of the vessels was observed. DMN also induced spawning prior to the normal spawning time.

Nature of the autofluorescent material in cerebrocortical necrosis.

A distinctive autofluorescence under ultraviolet light has been observed in the cerebral cortex of young ruminants suffering from cerebrocortical necrosis. The fluorescence is intimately connected with structural elements of the brain and cannot be extracted into solvents by normal procedures. The work presented here shows it to be associated with a collagen-like material.

Structural response of adult rat sertoli cells to peritubular fibroblasts in vitro

Sertoli cells were harvested from sexually mature rats and maintained in vitro for up to ten days (Sertoli cell-enriched cultures) or co-cultured with rat peritubular fibroblasts. Cultures were examined by differential light microscopy and by scanning or transmission electron microscopy. The presence of peritubular fibroblasts in co-culture greatly enhanced plating efficiency and viability of adult Sertoli cells. Sertoli cell aggregates preferentially adhered to peritubular cells, and by ten days had flattened and spread across these cells. Sertoli cells retained their characteristic ultrastructural features. Early in co-culture a collagen-like extracellular material was seen bridging Sertoli and peritubular cells, and its appearance was coincident with the presence of swollen rough endoplasmic reticulum in peritubular cells. Interperitubular cell spaces became engorged with a fibrillar material morphologically similar to the basal lamina of the seminiferous tubule wall. In the absence of peritubular cells, in culture medium "conditioned" by prior incubation with peritubular cells but not containing them, or when cultured with other fibroblastic cells, plating efficiency was low; Sertoli cells never flattened and cell ultrastructure progressively degenerated. The results indicated that peritubular cells support adult Sertoli cells in culture, possibly via extracellular material derived from peritubular cells, and suggest that Sertoli cells have an inductive effect on peritubular cell secretory activity.

Electron microscopic and cytochemical studies of rat aorta. Intracellular vesicles containing elastin- and collagen-like material

Small, rounded vesicles with a dense core of amorphous material were observed in all cell types in the young rat aorta, that is, endothelial cells, smooth muscle cells and fibroblasts. They were particularly numerous in the Golgi complex but were also found in the cell periphery. The content of the vesicles had staining characteristics identical to those of elastin. Material of the same type was also found in cisternae on the maturing side of the dictyosomes and in vesicles budding from them. Reaction product for thiamine pyrophosphatase was present in both these structures, indicating that the Golgi complex is responsible for the formation of the dense-cored vesicles. This was further supported by the absence of reaction product for acid phosphatase in the cisternae and in the vesicles. Moreover, no uptake of exogenous markers was noted in the latter. On the basis of these findings it is suggested that the dense-cored vesicles have a secretory function and contain precursors of elastin. Elongated vesicles or profiles containing collagen fibrils were observed in smooth muscle cells and fibroblasts. In the cell periphery, these vesicles were often found to communicate with the extracellular space. Further inside the cells, they showed a close spatial relationship to the Golgi complex. Neither thiamine pyrophosphatase nor acid phosphatase activity was demonstrated in the elongated vesicles. Like the plasma membrane, their limiting membrane was positively stained for alkaline phosphatase. On the basis of these findings and the absence of uptake of exogenous markers in them, it is suggested that the elongated vesicles represent a means for collagen secretion in the growing aortic wall. The Golgi complex is believed to be involved in the transfer of collagen to these vesicles.

Ultrastructural alterations during the development of retinopathy in sucrose-fed and streptozotocin-diabetic rats

Normal and streptozotocin-diabetic rats were maintained for 6–11 months on diets in which the sole source of carbohydrate was either 68% corn starch or sucrose. Perpendicular cross-sections of 15 capillaries from the retinal outer plexiform layer from each animal were examined qualitatively and quantitatively. The retinopathy produced by sucrose feeding to normal animals although identical by light microscopy to that produced by streptozotocin was not so ultrastructurally. Ramifications of the basement membrane, nodules and deposition of collagen-like material within the basement membrane were observed but no uniform thickening took place. Similar pathological changes were present in the diabetic capillaries except that a significant increase in the thickness of the basement membrane occurred. This thickening was not related to either blood glucose levels or the severity of the retinopathy indicating that the development of retinopathy is not related to basement membrane thickening, but rather to the deposition of material within the membrane.

Interaction between mesenchymal cells and the posterior iris epithelium in chicken embryos.

The iris anlage of 3--10 day old chicken embryos was studied by both light and electron microscopy. Serial semithin sections showed that some of the mesenchymal cells overlying the eye cup moved into the primitive eye cavity by the 3rd day of incubation. On the 4th day some of these cells came into close contact with the basement membrane of the dorsal iris epithelium. The bases of the epithelial cells were flat at this stage. Towards the 10th day they formed cytoplasmic processes which did not penetrate the basement membrane. Fine mesenchymal cytoplasmic processes and a large number of extracellular fibrils developed in the epithelial--mesenchymal interface. The fine mesenchymal processes came into close contact with the basement membrane of the posterior iris epithelium but did not penetrate it. Collagen-like material was observed within the cisternae of the rough ER of the mesenchymal cells at certain stages of development. Both, the mesenchymal cells and the collagen fibrils adjacent to the posterior iris layer disappeared by the 10th day when the entire iris epithelium was completely pigmented. The possible origin of the collagen fibrils and the differentiation of the posterior iris epithelium are discussed.