Superficial Epithelium Research Articles

Changes in c-erbB-2 Immunoexpression in Feline Endometrial Adenocarcinomas.

Human epidermal growth factor receptor type 2 (c-erbB-2), an oncoprotein with potential prognostic marker and therapeutic use, is overexpressed in several human and animal tumours. But information regarding this molecule in feline tumours is scarce. This study aimed to assess the changes in the immunohistochemical expression of c-erbB-2 in feline endometrial adenocarcinomas (FEA) compared to normal endometrium. An immunohistochemistry assay using a specific antibody against c-erbB-2 was performed in FEA samples (n = 34) and in normal endometrium in the follicular (FS; n = 12) and luteal (LS; n = 11) stages. In FEA, the c-erbB-2 immunoexpression was assessed in neoplastic epithelial cells whilst in normal endometria it was individually evaluated in the surface and the superficial and deep glandular epithelia (SE, SGE and DGE, respectively). In FS and in LS, all the epithelia were positive for c-erbB-2; positivity was higher in the SE and the SGE than in DGE. Twenty of the 34 FEA samples (58.8%) were positive for c-erbB-2 immunolabelling. Nevertheless, its expression was higher in all the epithelia in the FS compared to FEA (p ≤ 0.0001) or the LS (p = 0.016). The results presented herein suggest that c-erbB-2 molecule is differently expressed in the feline endometrium through the oestrous cycle and though it may also be involved in feline endometrial carcinogenesis, a question remains unanswered on the importance of additional pathways of epithelial proliferation in the neoplastic changes in feline endometrium.

Histological, surface ultrastructural, and histochemical study of the stomach of red piranha, Pygocentrus nattereri (Kner)

Abstract The structural characterization and function of the stomach in the omnivore Pygocentrus nattereri were described using light and scanning electron microscopy. The sac-like stomach was morphologically divided into the cardiac and pyloric regions. The histological structure of the stomach consisted of four layers of the mucosa, submucosa, muscularis, and serosa. The superficial epithelium of the cardiac stomach was lined with columnar epithelial cells and the glandular epithelium contained numerous gastric glands. Gastric glands were completely absent in the pyloric portion. The mucosal surface of the stomach was a meshwork of various folds, provided with oval or rounded columnar epithelial cells which were densely packed with short, stubby microvilli. The occasional presence of conspicuous gastric pits was surrounded by epithelial cells. The localization and chemical nature of acid and neutral mucins in the various cells of the stomach was studied by employing combined the Alcian Blue-Periodic Acid Schiff (AB-PAS) technique. The deposition of glycogen was detected in the gastric glands as well as in the epithelial lining of the stomach. The utmost reactions for protein and tryptophan were recorded in the gastric glands of mucosa. The cellular organization and histochemical characterization of the stomach are discussed in relation to the feeding and digestion of the fish concerned.

Answer to December 2015 Photo Quiz

Answer: Intestinal spirochetosis. The colon biopsy specimen showed intestinal spirochetosis. The endoscopic appearance in intestinal spirochetosis tends to be nonspecific, ranging from normal to polypoid areas or erythema (1). A basophilic fringe of spirochetes can be seen attached to the luminal side of the colonic epithelium. These organisms are known to attach to the mature microvilli of mature colonic and rectal epithelia. The spirochetes appear as an accentuated brush border-like layer coating the microvilli of the superficial epithelium, often referred to as a false brush border. Typically, the diagnosis is made by a pathologist examining the biopsy specimen. A Warthin-Starry silver stain for spirochetes can highlight the organism (see Fig. 1C and D in the photo quiz). Intestinal spirochetosis in humans is caused predominantly by Brachyspira aalborgi and Brachyspira pilosicoli. This condition is thought to be most prevalent in developing countries (2), among homosexual males (3), and among individuals with human immunodeficiency virus (HIV) infection (3). These organisms are slow-growing fastidious anaerobes and thus are infrequently recovered from stool specimens. While the pathological significance of intestinal spirochetosis is unclear, this condition has been reported to be associated with chronic diarrheal symptoms, especially in HIV-positive individuals. Evidence suggests that invasion of the organism beyond the epithelium may be associated with symptoms that respond to metronidazole (3). Using CLSI guidelines for susceptibility testing of anaerobes, Brooke et al. tested 123 human isolates of Brachyspira pilosicoli, all of which were susceptible to metronidazole, with an MIC90 of 0.25 μg/ml (4). The introduction of PCR-based assays along with immunohistological methods over the past decade may provide the tools to allow us to study the epidemiology and clinical importance of this organism (5). (See page 3711 in this issue [doi:10.1128/JCM.00005-14] for photo quiz case presentation.)

Immunohistochemical characterization of a retinal hamartoma

PurposeTo describe the immunohistochemical features of a retinal hamartoma.MethodsA 38 year old man was referred for preretinal fibrosis on his right eye. There was loss of vision for more than 8 years with metamorphopsia. The left eye was normal. Oct imaging showed an epiretinal fibrosis with extensive macular oedema. A 27Gauge vitrectomy was performed. The lamina limitans interna (ilm) was peeled after staining with Membrane Blue Dual and sent for histopathology.ResultsAfter removal the epiretinal membrane was immediately fixed with PreservCyt® in a ThinPrep® container. The specimen was routinely processed by the Cellient® embedding system. Microscopy showed a folded Pas positive membrane, recognized as ilm with on one side some glial remnants and on the other side a cuboidal non‐pigmented epithelium. The glial remnants stained with glial fibrillar protein (gfap), the epithelium with prekeratin AE1/AE3. CD 34 stain was positive in preretinal capillaries.ConclusionsA strange retinal fibrosis consisted of glial tissue and a row of superficial cuboidal non‐pigmented epithelium cells. This kind of hamartoma with immunohistological description was never investigated or published.

Measurement of In Vivo Three-Dimensional Corneal Cell Density and Size Using Two-Photon Imaging in C57BL/6 Mice

ABSTRACTPurpose: To measure the cell size and cell density in five layers of the central cornea in the widely used inbred C57BL/6 mouse strain using in vivo three-dimensional (3D) two-photon (2PH) imaging.Methods: Corneas were scanned using a 2PH laser scanning fluorescence microscope after staining with plasma membrane stain and Hoechst 33342. Good quality 3D images were selected for the cell density and cell size analysis. Cell density was determined by counting the cell nuclei in a predefined cube of 3D images. Cell size measurements, including cell surface area, cell volume, nuclear surface area and nuclear volume, were automatically quantified using the Imaris software. The cell and nuclear surface-area-to-volume ratio (S:V ratio) and the cell nuclear-cytoplasmic ratio (N:C ratio) were calculated.Results: The highest cell density was observed in the basal epithelium and the lowest in the posterior stroma. The highest cell surface area was found in the anterior stroma, and the highest cell volume was observed in the superficial epithelium. The lowest cell surface area and cell volume were both found in the basal epithelium. The highest S:V ratio was observed in the basal epithelium and the lowest in the superficial epithelium. The highest cell nuclear surface area and volume were both observed in the superficial epithelium and the lowest in the basal epithelium. The highest cell nuclear S:V ratio was observed in the basal epithelium and the lowest in the superficial epithelium. The highest N:C ratio was found in the basal epithelial cells and the lowest in the posterior keratocytes.Conclusions: We are the first to quantify the cell density and size parameters, including cell surface area and volume, cell nuclear surface area and volume, and the S:V ratio, in the five layers of the central cornea. These data provide important cell morphology features for the study of corneal physiology, pathology and disease in mice, particularly in C57BL/6 mice.

Histological and histochemical characterization on stomach of Mystus cavasius (Hamilton), Oreochromis niloticus (Linnaeus) and Gudusia chapra (Hamilton): Comparative study

The histological features and histochemical characterization of the stomach were investigated in Mystus cavasius (Hamilton), Oreochromis niloticus (Linnaeus) and Gudusia chapra (Hamilton) having different feeding habits. Histologically the stomach of all the three fishes was made up of mucosa, submucosa, muscularis and serosa. The mucosa of superficial epithelium consists of a single layer of compactly arranged columnar epithelial cells. Prominent gastric glands are present in M. cavasius whereas in G. chapra the gastric glands are totally absent in the gizzard like stomach. However, in O. niloticus tubular gastric glands are present in the glandular epithelium of caecal like stomach. The distribution and chemical nature of mucopolysaccharides in the aforementioned fishes were studied histochemically by employing Periodic Acid Schiff’s in combination with the Alcian Blue (PAS–AB) technique. Columnar epithelial cells lining the mucosa of the stomach including mucosal border were provided with exclusively neutral mucin which was probably involved in the protective functions against acid and enzymes. The different intensities of reaction of Best Carmine (BC) for glycogen in the epithelial lining and gastric glands of the stomach of the aforesaid three fish species under study were discussed. The intense reaction for protein and tryptophan was noticed in the gastric epithelium and gastric glands of M. cavasius probably due to accumulation of zymogen granules in the gastric glands. On the contrary, moderate reaction for protein and tryptophan was associated with the epithelial cells and gastric glands of O. niloticus and G. chapra. The cytoarchitecture and different degrees of localization of mucopolysaccharides, glycogen, protein and tryptophan in the stomach of M. cavasius, O. niloticus and G. chapra were correlated with the functional significance of the region concerned.

Mo1710 Regulation of Smad7 Translation by microRNA-195 Affects Steroid Resistance of Ulcerative Colitis

G A A b st ra ct s during 24h with agonists and antagonists of the Hh pathway, inflammatory cytokines and bacterial products. Supernatants were analyzed for cytokine production by ELISA, and caspase-3 activity was assayed in epithelial cells isolated from mucosal samples. Chemoattraction of peripheral blood mononuclear cells (PBMC) was analyzed in a transwell system. Primary human fibroblasts were used for analyzing migration and BrDU incorporation. Results: Hh-proteins were basically expressed at the superficial epithelium, and a marked reduction was observed in CD compared to UC (p=0.04), and controls (p=0.001). In the lamina propria, Gli-1 predominantly co-localizes with non-immune cells, such as alphasmooth muscle actin-, vimentin-, and tubulin-positive cells, with lower levels in CD. Levels of Indian and Sonic Hh mRNA, Ptch-1, and the downstream transcription factors Gli-1 and Gli-2 were remarkably lower in CD (p<0.04). In CD colon explants, treatment with Shh peptide resulted in a significant decrease in the levels of TNF-alpha (p=0.04), IL-17 (p= 0.01), and TGF-beta (p=0.03). Caspase-3 activity was higher in CD epithelial cells and significantly decreased after blockade of Hh with anti-Shh antibody, cyclopamine, and Gant61 (p<0.04), in contrast to UC and controls. Levels of Gli-1 mRNA negatively correlated to caspase-3 activity (CC -0.75, p=0.01). PBMC transmigration significantly increased after treatment with Gant61 (p<0.04). Primary fibroblasts incorporated significantly more BrdU (p<0.04), and migrated less (p<0.03) after treatment with Gant61. Conclusion: Loss of Hhsignaling is associated with disruption of the epithelial barrier and pro-inflammatory cytokine production in CD intestinal mucosa. In vitro studies indicate that blocking the Hh pathway induces mononuclear cell chemo-attraction and fibroblast proliferation, but reduces fibroblast migration. Taken together, these data support the notion of a reduced intestinal Hh-signaling underlying the pathogenesis of CD.

In Vivo Confocal Microscopic Changes of the Corneal Epithelium and Stroma in Patients With Herpes Zoster Ophthalmicus

To analyze the density and morphology of corneal epithelial cells and keratocytes by invivo confocal microscopy (IVCM) in patients with herpes zoster ophthalmicus (HZO) as associated with corneal innervation. Prospective, controlled and masked cross-sectional study. setting: Single-center study. Thirty eyes with the diagnosis HZO and their contralateral clinically unaffected eyes, 15 eyes of 15 normal controls. intervention procedures: Invivo confocal microscopy and corneal esthesiometry of the central cornea. Changes in morphology and density of the superficial and basal epithelial cells and stromal keratocytes, and correlation with corneal sensation. The density of superficial epithelial cells in HZO eyes with severe sensation loss (766.5 ± 25.2 cells/mm(2)) was significantly lower than both healthy control eyes (1450.23 ± 150.83 cells/mm(2)) and contralateral unaffected eyes (1974.13 ± 298.24 cells/mm(2)) (P= .003). Superficial epithelial cell size (1162.5μm(2)) was significantly larger in HZO eyes with severe loss of sensation, as compared to contralateral (441.46 ± 298.14) or healthy eyes (407.4± 47.2μm(2); all P < .05). The density of basal epithelial cells, anterior keratocytes, and posterior keratocytes did not show statistical significance between patients, controls, and contralateral unaffected eyes. Changes in superficial epithelial cell density and morphology correlated strongly with corneal sensation. Invivo confocal microscopy reveals profound HZO-induced changes in the superficial epithelium, as demonstrated by increase in cell size, decrease in cell density, and squamous metaplasia. We demonstrate that these changes strongly correlate with changes in corneal innervation in eyes affected by HZO.

Corneal Microstructural Changes in Non-Sjögren Dry Eye Using Confocal Microscopy: Clinical Correlation

Purpose: To investigate the relationship between changes of corneal epithelium and subbasal nerves in non-Sjogren dry eye using in vivo confocal microscope (IVCM) and self-reported clinical symptoms. Methods: The present study included 40 patients with dry eye and 18 healthy control subjects. The dry eye group underwent an evaluation of dry eye symptoms using visual analogue scale (VAS) score and was subdivided into 2 groups; score 0-5 as the low VAS score (LVS) group and score 6 - 10 as the high VAS score (HVS) group. The tear film break-up time, fluorescein staining, Schirmer test and microstructural imaging of epithelium, and subbasal nerve at cornea center with IVCM were performed on both eyes of each patient. Twenty-three normal eyes and 54 eyes of dry eye patients were included in the study. Cell densities and morphological characteristics were analyzed using ImageJ and NeuronJ softwares. Results: Both LVS and HVS groups had decreased cell density of superficial, intermediate, and basal epithelium (p < 0.001). Intermediate epithelial cells were more decreased in the dry eye group with more severe symptoms (p < 0.0001). Subbasal nerve density (p < 0.005) was more decreased and nerve beadings, tortuosity, and reflectivity increased in the HVS group than both LVS and control groups (p < 0.05). Conclusions: The alterations of corneal cellular level in dry eye patients visualized using IVCM are correlated with pathology and clinical symptoms and may be useful objective criteria in diagnosis and monitoring treatment efficacy. J Korean Ophthalmol Soc 2015;56(5):680-686

Endoscopic surveillance of gastric cancers after Helicobacter pylori eradication.

The incidence and mortality of gastric cancer remains high in East Asian countries. Current data suggest that Helicobacter pylori (H. pylori) eradication might be more effective for preventing gastric cancer in young people before they develop atrophic gastritis and intestinal metaplasia. However, the long-term effect of H. pylori eradication on metachronous cancer prevention after endoscopic resection (ER) of early gastric cancer remains controversial, with some discordance between results published for Japanese and Korean studies. The detection ability of synchronous lesions before ER and eradication of H. pylori directly influences these results. After eradication, some gastric cancers are more difficult to diagnose by endoscopy because of morphologic changes that lead to a flat or depressed appearance. Narrow-band imaging with magnifying endoscopy (NBI-ME) is expected to be useful for identifying metachronous cancers. However, some gastric cancers after eradication show a "gastritis-like" appearance under NBI-ME. The gastritis-like appearance correlates with the histological surface differentiation of the cancer tubules and superficial non-neoplastic epithelium atop or interspersed with the cancer. Till date, it remains unclear whether H. pylori eradication could prevent progression of gastric cancer. Until we can establish more useful endoscopic examination methodologies, regular endoscopic surveillance of high-risk groups is expected to be the most beneficial approach for detection.

Mucous membrane pemphigoid as a cause of acute dysphagia - an endoscopic study.

Mucous membrane pemphigoid (MMP, formerly cicatricial pemphigoid) is a chronic autoimmune, subepithelial blistering disease usually of middle aged or elderly patients, more common in women. The prevalence is uncertain and the aetiology is still unknown. Target antigens are epithelial basement membrane proteins BPAG-1, BPAG-2, laminin-331, and laminin-332 [1]. Oral and conjunctival membranes are most frequently affected. Oesophageal involvement is rare but may be complicated with persistent dysphagia [2].

In vivo confocal scanning laser microscopy in patients with primary Sjögren's syndrome: A monocentric experience

Aims. (i) To analyze the in vivo corneal structure and sub-basal plexus nerves in patients with primary Sjögren's syndrome (pSS) and no-SS dry eye by confocal scanning laser microscopy (CSLM) and (ii) to correlate CSLM findings with tear function tests and with patients’ subjective dryness.Methods. Seventeen patients with pSS, 16 no-SS dry eye, and 20 healthy volunteers were included. CSLM parameters taken into consideration included: basal epithelial integrity, corneal thickness, epithelial cellular density, keratocyte activation, and sub-basal plexus morphology. Statistical analysis was carried out using SPSS-13 (Chicago IL, USA).Results. CSLM pachymetric data and the superficial epithelium cell density were significantly lower in pSS versus no-SS dry eye (p < 0.0001); keratocyte activation and sub-basal nerve abnormalities were also more frequent in pSS patients (p < 0.0001). CSLM findings well correlated with both the ocular test results and the patients’ perception of ocular dryness at the baseline and over the follow-up.Conclusion. CSLM might be a useful novel tool in the assessment of the involvement of the lachrymal functional unit in pSS.

Ultrastructural features of eosinophilic oesophagitis: impact of treatment on desmosomes

AimsA growing body of evidence suggests a role for altered epithelial barrier function in the pathophysiology of eosinophilic oesophagitis (EoE), but few have described the epithelial structure during inflammation. The...

The anatomy of the dog soft palate. III. Histological evaluation of the caudal soft palate in brachycephalic neonates.

A thickened and abnormally long soft palate is mostly involved in the pathogenesis of both nasopharyngeal and oropharyngeal narrowing, affecting the respiratory activity in virtually all of the brachycephalic dogs suffering from Brachycephalic Airway Obstructive Syndrome (BAOS). The morphology of the soft palate in adult mesaticephalic and brachycephalic dogs has been previously described. In this article specimens from brachycephalic dog neonates (N=10) dead from one to 3 hr after birth of unrelated conditions were collected and histologically evaluated at three transverse levels to describe the microscopic aspect of the caudal part of the soft palate. The soft palate of neonate brachycephalic dogs was histologically characterized by a musculo-connective axis containing salivary glands and coated by a mucosal layer on both the nasopharyngeal and the oral side. Quali-quantitative features, such as thickened superficial epithelium, broad oedema of the lamina propria, mucous gland hyperplasia and diverse muscular modifications described in adult brachycephalic dogs, were not observed in the soft palate of brachycephalic neonate dogs. The lack of tissue lesions in the soft palate of newborn brachycephalic dogs further supports the hypothesis that inspiratory depression during the inspiratory phase causes chronic vibration and microtrauma, which lead to soft palate alterations in adult brachycephalic dogs starting from the earliest grade of the respiratory syndrome. Overall, this study provides baseline information for the comprehension of the pathogenesis of BAOS.

Clinicopathologic and Diagnostic Features of Esophagitis Dissecans Superficialis

Introduction: Esophagitis dissecans superficialis (EDS) is a desquamative disorder of the esophagus. Little is known about the clinical presentation, predisposing factors, and clinical outcomes of this condition. A cohort of EDS patients were identified and evaluated to further characterize the clinicopathologic features of EDS and to propose diagnostic criteria. Methods: Endoscopy and pathology databases were retrospectively reviewed from 2000-2013 at the Mayo Clinic Rochester to identify potential cases of EDS. Cases were selected based on endoscopic findings and confirmed by re-review of the original pathologic specimens. Clinical, endoscopic, and histologic features of EDS were recorded and evaluated. Results: Forty-one subjects were identified with a mean age of 65.0 years at diagnosis (IQR 52.8-76.1), and 63.4% were female. A large number of patients were taking a psychoactive agent (73.1%) or acid suppressive therapy (58.5%). Endoscopically, strips of sloughed membranes were seen in all patients with a predilection for the distal and/or middle esophagus. Predominant histologic features included parakeratosis (100%) and intraepithelial splitting (86.1%; 25.0% of these showed distinct cysts), while 50.0% or fewer of cases showed partially necrotic long, detached fragments of superficial epithelium. The majority (85.7%) of subjects with endoscopic follow-up showed complete resolution. No complications of stricture or malignancy were noted at a median follow-up of 10.4 months (IQR 1.2-17.2). Conclusion: EDS is a rare entity with a high proportion of individuals with using a psychoactive medication prior to recognition, although the clinical relevance of this association is uncertain. EDS is defined endoscopically by sloughing, superficial sheets of esophageal mucosa and is likely a benign, incidental endoscopic finding without clinical sequelae; however, further studies with prolonged follow-up are needed.

Ascaridia galli induced ulcerative proventriculitis in a poultry bird.

Various possible causes of proventriculitis include virus, bacteria, fungus, protozoans, nematodes, biogenic amines and excessive copper sulphate. In the present case, parasites were found in the lumen of the proventriculus, gizzard and duodenum of a poultry bird. Characteristic features of the parasite were studied and confirmed as Ascaridia galli. An ulcerative proventriculitis evident as denuded superficial epithelium, sub-epithelial hemorrhages, infiltration of the inflammatory cells and fibrosis were seen at histopathology. Proventriculitis caused by A. galli has not been reported till date. Here, we report a case of ulcerative proventriculitis in a poultry bird caused by nematode, A. galli.

Evaluation of the Number of Gastric Mucoid Epitheliocytes and Parietal Cells in Relation to the Amount of Helicobacteria in the Fundic Gland Region of Domestic Dogs (Canis Familiaris)

Abstract This study was done to evaluate the number of gastric mucoid epitheliocytes and parietal cells in relation to the amount of helicobacteria in the fundic gland region of 30 domestic dogs (Canis familiaris) Gastric mucosa samples were taken from the fundic gland region in eight strictly determined places to detect helicobacteria in 10 microscopic visual fields by histological examination, to find out the quantity of helicobacteria at the superficial epithelium, in gastric pits, and in fundic glands by classifying it as absent, mild, moderate, and severe, as well as to evaluate the number of mucoid epitheliocytes of the gastric superficial–pit epithelium and of the parietal cells of fundic glands. Out of all examined 2400 microscopic visual fields, mild and moderate amounts of helicobacteria localizing mainly at the superficial epithelium and in gastric pits were detected in 69.0–77.5% of cases, and only in around 20% of cases they were found in different amounts deeper into the fundic glands. The number of mucoid epitheliocytes (199±20 cells in one visual field) was only slightly larger in places with a severe amount of helicobacteria compared to places with no helicobacteria (195±16 mucoid epitheliocytes in one visual field). Also the number of parietal cells in the fundic glands did not differ markedly among the places with different amounts of helicobacteria; however, in places with a severe amount of helicobacteria, parietal cell count tended to decrease to 76±8 cells in one visual field, but in places free from helicobacteria there were 83±9 parietal cells in one visual field.

Stem Cell-Like Characteristics of Corneal Epithelial Cells

Purpose: The adult corneal epithelium is maintained by a population of limbal stem cells (LSCs), transmembrane protein prominin, regarded as stem cell marker was investigated on mouse corneal tissue, to study weather contains CD133-expressing cells and their distribution. Methods: Enucleated mouse eyes were embedded in OCT and cryosections were performed for mmunohistochemical studies using the avidin-biotin-peroxidase complex (ABC) procedure. Meanwhile, dissected mouse corneas were analyzed by westernblot. Results: In the adult mouse, 13A4 immunoreactivity was detected at the apical side of superficial corneal epithelium, including the limbus region, but not by stroma and endothelium. 115 KDa protein was approved in corneal tissue by Westernblot. Conclusions: The stem cell activity does not occur along the limbus but presumably presented by small portion of corneal epithelial cells which may hold a similar properties of stem cells.

Fibroma odontogénico periférico Reporte de un caso

El Fibroma Odontógenico Periférico es la forma más frecuente del Fibroma Odontógeno y parece proceder del epitelio gingival superficial o de residuos de la lámina dental que permanecen en una localización extraósea. El componente epitelial se asemeja a la lámina dental formada durante las etapas tempranas de la odontogénesis. Se presenta a cualquier edad, aunque tiene predilección por niños y adultos jóvenes; se observa más en mujeres que en hombres.

Effects of vitamin D receptor knockout on cornea epithelium gap junctions.

Gap junctions are present in all corneal cell types and have been shown to have a critical role in cell phenotype determination. Vitamin D has been shown to influence cell differentiation, and recent work demonstrates the presence of vitamin D in the ocular anterior segment. This study measured and compared gap junction diffusion coefficients among different cornea epithelium phenotypes and in keratocytes using a noninvasive technique, fluorescence recovery after photobleaching (FRAP), and examined the influence of vitamin D receptor (VDR) knockout on epithelial gap junction communication in intact corneas. Previous gap junction studies in cornea epithelium and keratocytes were performed using cultured cells or ex vivo invasive techniques. These invasive techniques were unable to measure diffusion coefficients and likely were disruptive to normal cell physiology. Corneas from VDR knockout and control mice were stained with 5(6)-carboxyfluorescein diacetate (CFDA). Gap junction diffusion coefficients of the corneal epithelium phenotypes and of keratocytes, residing in intact corneas, were detected using FRAP. Diffusion coefficients equaled 18.7, 9.8, 5.6, and 4.2 μm(2)/s for superficial squamous cells, middle wing cells, basal cells, and keratocytes, respectively. Corneal thickness, superficial cell size, and the superficial squamous cell diffusion coefficient of 10-week-old VDR knockout mice were significantly lower than those of control mice (P < 0.01). The superficial cell diffusion coefficient of heterozygous mice was significantly lower than control mice (P < 0.05). Our results demonstrate differences in gap junction dye spread among the epithelial cell phenotypes, mirroring the epithelial developmental axis. The VDR knockout influences previously unreported cell-to-cell communication in superficial epithelium.