Granular Cell Differentiation Research Articles

The Association Between Runx Signaling and Craniofacial Development and Disease.

The Runx family genes (Runx1, Runx2, Runx3, and Cbfb) are important transcriptional regulators in the development of various tissues. We herein highlight the roles of the Runx family genes in morphogenesis in the craniofacial regions and in the pathogenesis of congenital morphological problems in these regions. A recent analysis using conditional Runx mutant animals and a human genetic study identified the novel roles of Runx genes in the development of the tooth, salivary glands, and the palate. In an animal study, Runx1/Cbfb signaling was found to regulate the Lgr5 expression and maintain the stem cells in the dental epithelium in the growing incisors. Aberrant Runx1/Cbfb signaling induced male-specific involution of the convoluted granular cell differentiation of the submandibular gland. In palatogenesis, Runx1/Cbfb signaling regulated the Tgfb3 expression in the fusing palatal epithelium through Stat3 activation. The combination of a human genetic study and a phenotype analysis of mutant animals revealed the various roles of Runx genes in the development of the tooth, palate, and salivary glands. Runx genes have functional redundancy in various tissues, which still hinder the roles of Runx genes in morphogenesis. Future studies may reveal the novel roles of Runx signaling.

Spinal Cord Compression in a Horse due to a Granular Cell Variant of Neurofibroma

A neurofibroma of granular cell subtype is described in a 7-year-old horse. The horse had a 3-month history of ataxia affecting the forelimbs and hindlimbs, suggesting a C1-C6 neuroanatomical localization. Post-mortem examination revealed an intradural mass arising from the right sixth cervical spinal nerve and compressing the spinal cord. Histologically, the mass was composed largely of wavy spindle cells (a mixture of Schwann cells, perineurial cells and fibroblasts) intimately associated with ropy collagen fibres. Approximately 25% of the spindle cells were swollen and contained densely-packed, eosinophilic and periodic acid-Schiff-positive cytoplasmic granules. Immunohistochemistry for S100 and glial fibrillary acidic protein antigens labelled a proportion of neoplastic cells, while the cytoplasmic granules were positive for S100 and neuron specific enolase. This is the first report of a neurofibroma with granular cell differentiation in an animal. Granular cell differentiation in other peripheral nerve sheath tumours of animals is briefly discussed.

Granular cell differentiation: A review of the published work.

Since the initial description of the granular cell tumor in 1926, numerous other neoplasms, both benign and malignant, have been described to exhibit granular cell change. In most cases, diagnosis remains straightforward via recognition of retained histopathological morphology of the archetypal tumor, despite the presence of focal granular appearance. However, tumors with granular cell differentiation can present a diagnostic challenge either by mimicking alternative diagnoses, or by failing to exhibit architectural clues of the primary entity, thus requiring an immunohistochemical work-up. In light of this, it is important to be aware of the various entities that have been reported to exhibit granular cell morphology. In this review such tumors are discussed along with pertinent clinical and histopathological features.

Marsupialization of unicystic ameloblastoma: A conservative approach for aggressive odontogenic tumors

Unicystic ameloblastoma (UA) is known as a distinct entity which has a less aggressive behavior when compared with conventional ameloblastoma. In this report, we have presented two cases of UAs, (of which one case showed a more aggressive behavior with mural invasion into the adjacent tissues and granular cell differentiation), both of which were successfully managed with enucleation following marsupialization. We aim to highlight how this method can be used for the successful management of such cases, rather than following more aggressive approaches. In both the cases, marsupialization was done for the UA lesions initially and follow-ups were maintained. When the tumor size had regressed on radiographic follow up, an enucleation procedure with ostectomy of the margins was carried out. Special importance was also given to the endodontic treatment of the teeth involved in the area of the lesion. The patients were free of the condition and did not show any signs of recurrence on radiographic follow-ups even after 30 months of the final procedure. Granular variant of UA is quite rare and had been considered to be more aggressive. Marsupialization of UA is an alternative treatment option of resection even for more aggressive variants, as long as the histological behavior of the lesion was carefully evaluated and strict radiographic follow-up is maintained.

Ancient Cytokines, the Role of Astakines as Hematopoietic Growth Factors

Hematopoiesis is the process by which hemocytes mature and subsequently enter the circulation. Vertebrate prokineticins (PKs) are known to take part in this process, as are the invertebrate prokineticin domain proteins, astakines. In Pacifastacus leniusculus, astakine 1 is essential for the release of new hemocytes into the open circulatory system of these animals. In addition to astakine 1, we have now cloned a homologue of astakine 1 with an insert of 13 amino acids, named as astakine 2. Both crustacean astakines lack the N-terminal AVIT motif, which is present in vertebrate PKs, and hence receptor binding differs from that of vertebrate PKs. We have found astakine-like sequences in 19 different invertebrate species, and the sequences show that some motifs are conserved among invertebrate groups. Previously we showed that astakine 1 is directly involved in hematopoiesis, and now we show that astakine 1 and astakine 2 have different roles in hemocyte lineage differentiation. Astakine 1 can stimulate proliferation of hematopoietic tissue (Hpt) cells (precursor of hemocytes) as well as specifically induce differentiation of Hpt cells along the semigranular cell lineage, whereas astakine 2 plays a role in granular cell differentiation. Moreover, we discuss the impact of the putative structures of different astakines in comparison with the vertebrate prokineticins.

Giant dermatofibroma with granular cell changes: side-effect of bee-venom acupuncture?

Dermatofibroma (DF) is a common benign fibrohistiocytic tumour with a predilection for the legs in middle-aged women. Giant DF, a rare clinical variant of DF, is characterized by its unusually large size. Granular cell change is typical of granular cell tumour, but can be observed in diverse cell lineages. Traumatic factors may be involved in the pathogenesis of giant DF and cellular granularity. We describe a 49-year-old Korean man with a giant DF showing granular cell differentiation, which may have been caused in part by multiple treatments with bee-venom acupuncture.

Expression of cortactin and survivin in renal cell carcinoma associated with tumor aggressiveness

We tested the hypothesis that the expression of cortactin and survivin in renal cell carcinomas (RCCs) correlates with more advanced stages of the disease. Immunohistochemical analysis of cortactin and survivin expression (scored on a scale of 0-400) was performed in 124 renal cell carcinomas including clear cell renal cell carcinoma (CRCC), papillary RCC (PRCC), CRCC with sarcomatoid differentiation (SRCC), chromophobe RCC (ChRCC), and CRCC with granular cell differentiation (GRCC). Higher cortactin scores in CRCC were significantly correlated with higher T (p = 0.021) and N stages (p = 0.036), and nuclear grade (p = 0.012). Higher cortactin immunostaining scores were associated with higher mortality (p = 0.035). In addition, the survivin scores were significantly higher in the more aggressive GRCC and SRCC than in CRCC, suggesting a significant role of survivin expression in transformation of tumor cells to a more malignant phenotype. Higher expression of cortactin and survivin significantly correlated with advanced clinicopathological stage. Our findings support the potential targeting of survivin and cortactin for the development of novel therapeutic strategies for renal cell carcinoma.

Intramedullary astrocytoma with granular cell differentiation

Granular cell astrocytomas are uncommon tumors of the central nervous system (CNS) of which no cases have been documented in the spinal cord. This variant of glioma should not be confused with benign granular cell tumor which, although rare, has been well characterized in the spinal cord. We describe here the clinical, pathological, and radiological features of such an astrocytoma arising within the spinal cord at the dorsal level. A 48-year-old female was seen after about 1 year of dorsal pain and gradual spastic paraparesis. Magnetic resonance imaging (MRI) studies showed a 2-cm contrast-enhanced mass in the spinal cord at T6-T7, which had the appearance of an astrocytoma. At surgery, the tumor was found to be infiltrating a posterior column with no dural attachment. It was debulked and dissected. The histological diagnosis was astrocytoma with granular cell differentiation. In addition to documenting a unique example of intramedullary granular cell astrocytoma, we review the literature to investigate differences from other tumors with granular changes described in the spinal cord.

Increasing expression of fascin in renal cell carcinoma associated with clinicopathological parameters of aggressiveness.

To determine whether higher expression of fascin, an actin-bundling protein associated with motility, in conventional renal cell carcinoma (RCC) is associated with more advanced stages of the disease. Immunohistochemical analysis of fascin expression was performed in tissue microarrays of 108 RCCs including 55 clear cell RCCs (CRCCs), 39 CRCCs with granular cell differentiation (GRCCs), 8 CRCCs with sarcomatoid differentiation (SRCCs) and 6 metastatic RCCs. The expression of fascin was undetectable in normal renal tubules of all control cases. However, among the 108 RCC cases, fascin immunoreactivity was seen on the cell membrane and cytoplasm. The average immunostaining score for fascin was 128/400 in grade I, 170/400 in grade II, 207/400 in grade III, and 323/400 in grade IV RCC. The average immunostaining score of fascin was 187/400 for stage T1, 205/400 for stage T2, 288/400 for stage T3, and 355/400 for stage T4 cases of RCCs. Higher fascin scores in RCC were significantly correlated with higher T and N stages and nuclear grade. In addition, the fascin scores in GRCC (368+/-19) and SRCC (263+/-21) were significantly higher than in CRCC (95+/-18). Our findings demonstrate for the first time that increased expression of fascin is associated with clinicopathological parameters of aggressiveness in patients with RCC. Fascin may be a novel biomarker for diagnosis and treatment of RCC.

Expression of Serine Protease Matriptase in Renal Cell Carcinoma: Correlation of Tissue Microarray Immunohistochemical Expression Analysis Results with Clinicopathological Parameters

Serine protease matriptase (matriptase) cleaves and activates proteins implicated in the progression of cancer and represents a potential therapeutic target. Immunohistochemical analysis of matriptase was performed in tissue microarrays of 168 renal cell carcinomas (RCCs). All subtypes of RCC showed significant immunohistochemical expression of matriptase. In contrast, no expression occurred in areas of RCC with sarcomatous differentiation (SRCC) and in normal collecting tubules. The matriptase scores were significantly higher in papillary RCC (341+/-28) and clear cell RCC with granular cell differentiation (GRCC; 324+/-27) than in other histologic subtypes of RCC. In GRCC, matriptase scores were correlated with TNM staging and nuclear grading. Matriptase was overexpressed in all subtypes of RCC, and matriptase scores could distinguish between conventional clear cell RCC, GRCC, and SRCC.

Protein kinase C isozymes regulate proliferation and high cell density-mediated differentiation in HaCaT keratinocytes.

Protein kinase C (PKC) isoforms play pivotal roles in the regulation of differentiation of normal human epidermal keratinocytes (NHEK). In this study, we investigated the participation of the PKC system in the proliferation and high cell density-induced differentiation of the human immortalized keratinocyte line HaCaT. HaCaT keratinocytes possessed a characteristic PKC isoform pattern (PKC alpha, beta, gamma, delta, epsilon, eta, theta, zeta), which altered during proliferation and differentiation. The GF109203X compound, a selective PKC inhibitor, suppressed the expressions of the lat (granular cell) differentiation markers involucrin (INV) and filaggrin (FIL), and the terminal marker keratinocyte-specific transglutaminase-1 (TG), but did not affect the level of the early (spinous cell) marker keratin 10 (K10) and cellular proliferation. Phorbol 12-myristate 13-acetate (PMA), an activator of PKC, inhibited proliferation, elevated intracellular calcium concentration, decreased the expression of K10, and increased the expressions of INV, FIL, and TG. These data indicate that the endogenous activation of PKC regulates the expressions of the late differentiation markers, and that the exogenous activation of PKC by PMA results in the induction of terminal differentiation. Because the cellular effects of PMA were accompanied by differential down-regulations of the sensitive PKC isoforms in proliferating and differentiating cultures, our findings argue for the differential roles of the existing PKC isoforms in the regulation of cellular proliferation and high cell density-induced differentiation of HaCaT cells.

Type I bone morphogenetic protein receptors are expressed on cerebellar granular neurons and a constitutively active form of the type IA receptor induces cerebellar abnormalities

Cerebellar granular neurons are the most abundant neuronal type in the CNS. Their number and experimental accessibility have made these neurons a valuable model for investigating mechanisms of cell proliferation and differentiation in the CNS. Proliferation of granular neurons is regulated, at least in part, by the secreted protein Sonic Hedgehog, whereas induction and differentiation both appear to be controlled by bone morphogenetic protein (BMP) signaling. Given the role of BMPs in granular cell differentiation, we postulated that BMP receptors (BMPRs) would be expressed on cerebellar granular neurons and that signaling through these receptors is required for normal differentiation. We found that both BMPRIA and BMPRIB are expressed on granular neuron precursors and on mature granular neurons in the developing cerebellum. To determine if these receptors are sufficient for granular cell induction and/or differentiation in vivo, we introduced a constitutively active BMPRIA construct into the developing cerebellum. The resulting cerebelli showed a simplified folial pattern and ectopic collections of small cells located deep in the cerebellar white matter. Phenotypic analysis demonstrated that the ectopic cells are granular neurons. From these data we suggest that signaling through the type I BMPRs occurs during normal cerebellar development and ectopic activation of this pathway affects normal granular neuron development. Furthermore, the similarity of the cerebellar anomalies arising from perturbed BMPR signaling to human cerebellar malformations suggests that dysregulation of BMP signaling may play a pathogenic role in some human cerebellar abnormalities.

Combined dermatofibroma: co-existence of two or more variant patterns in a single lesion.

Based on a series of 25 cases, we define and characterize combined dermatofibroma, a tumour comprising two or more variant patterns of dermatofibroma in a single lesion. Dermatofibroma may present with a wide variety of architectural, cellular or stromal peculiarities. Architectural peculiarities include deep penetration, atrophy, collarette formation, fascicular to plexiform architecture, massive haemorrhage, prominent haemangiopericytoma-like vascularity and palisading; cellular peculiarities the presence of epithelioid cells, clear cells, granular cells, prominent myofibroblastic differentiation and atypical giant cells ('monster cells'); or stromal peculiarities such as prominent sclerosis, mucin, haemosiderin and cholesterotic deposits. In combined dermatofibromas two or more of these features are seen in complex or inhomogenous combination such as the silhouette of a deep penetrating dermatofibroma with an 'ordinary' storiform pattern in the upper and granular cell differentiation in the lower part of the lesion; or a dermatofibroma with ordinary features in the upper, prominent sclerosis in the middle and clear cells in the lower portion of the lesion; or the characteristic epidermal collarette and cells of epithelial cell histiocytoma with a plexiform ('neurothekeoma-like') architecture surrounded by a myxoid stroma with spindle-shaped to stellate cells. Clinically, these lesions preferentially occur on the lower extremities of young to middle-aged females, frequently with the diagnosis of a fibrohistiocytic lesion. Apart from one recurrence follow-up was uneventful in all other cases. Immunohistochemically, lesions are consistently positive with KiM1p, variably positive for factor XIIIa, smooth muscle specific actin and with KP1 (CD68), NK1C3 and E9. Recognition of combined dermatofibroma allows the histopathologist to apply a confident benign label to unusual lesions which might otherwise elude diagnosis, or tempt description of 'new' entities and to avoid a misdiagnosis of malignancy.

Increased induction of Ca 2+-mediated differentiation by gamma ray is mediated by endogeneous activation of the protein kinase C signaling pathways in mouse epidermal cells

Purpose: The aim of this study was to determine whether γ-rays can affect Ca 2+-induced differentiation in normal and neoplastic mouse epidermal cells. Methods and Materials: After γ-ray irradiation, primary and v-ras Ha transformed mouse keratinocytes were cultured for 48 h in 0.12 mM Ca 2+-containing media, and cellular translocation from cytosolic to particulated fraction of each PKC isozyme and expressions of differentiation markers were examined. Results: Morphological difference was seen at 48 h after irradiation in both Ca 2+-shifted normal and v-ras Ha transformed cells; v-ras Ha cells were more resistant to the radiation than normal cells. Radiation potentiated granular cell-differentiation marker expressions (filaggrin, loricrin, and SPR-1) in both normal and v-ras Ha transformed cells. In the case of spinous cell markers, the expression of keratins K1 and K10, which are usually blocked in v-ras Ha cells was increased after irradiation. However, there was no change of K8 expression level, which can be seen only after v-ras Ha transfection. Cellular fractionation and immunoblot analysis with antibodies against PKCα, δ, ϵ, η, and ξ revealed that PKCα was responsible for the differentiation marker expression. Conclusions: These findings suggest that PKCα is an important component of the signaling pathway regulating radiation-induced differentiation in both normal and neoplastic epidermal cells.

Low Frequency of Loss and Heterozygosity At the Nevoid Basal Cell Carcinoma Locus and Other Selected Loci in Appendageal Tumors

Previous studies of loss heterozygosity (LOH) have revealed distinct patterns of allelic loss in some skin tumors. In basal cell carcinomas (BCCs) loss of heterozygosity is virtually restricted to chromosome 9, whereas in squamous cell carcinomas (SCCs) and actinic keratoses loss is more widespread involving chromosomes 3, 9, 13, and 17. Because there are histological similarities between BCCs and some appendageal tumors, and because lines of evidence suggest that BCCs are appendageal in origin, we carried out a limited allelotype in 41 appendageal tumors. The overall frequency of allelic loss was low (4 out of 247 informative loci; 1.6%). LOH was seen in a proliferating trichilemmal cyst (17p), a sebaceous epithelioma (17q), an eccrine porocarcinoma (17q), a trichoepithelioma (9q), and in two basal cell carcinomas showing eccrine or granular cell differentiation that were originally misdiagnosed (9Q). The pattern of loss in this mixed group of appendageal tumors shows differences from both BCCs and SCCs, and further emphasizes the unique genetic profile and behavior of BCCs. The finding of 9q loss in BCCs with eccrine or granular cell differentiation shows that 9q loss occurs in differential histological subtypes of BCCs.

Effective treatment for acute alkali injury to the esophagus using weak-acid neutralization therapy: an ex-vivo study.

1) To evaluate whether neutralization therapy with weak acid is effective in reducing observed histopathologic esophageal tissue injury secondary to liquid alkali, 2) to quantify the temperature change of the neutralizing agent, and 3) to determine the effect of interval to therapy on injury severity. Harvested Sprague-Dawley rat esophagi were catheterized and placed in an oxygenated saline bath (37 degrees C) for 60 minutes and then fixed in 10% formalin. Nine groups (n = 10) were perfused with 50% sodium hydroxide (NaOH). Six of the groups were treated by neutralization with cooled orange juice (OJ) or cola that was maintained between 2 degrees C and 4 degrees C. This was performed at 0, 5, or 30 minutes after injury. In addition, two positive control groups were exposed to OJ or cola at time 0 and were not exposed to strong alkali. A third control group was exposed to strong alkali but was not administered any subsequent treatment. The temperature of the neutralizing agent was recorded prior to instillation and after exiting the esophagus. Blinded pathologic scoring of 0 (no injury) to 3 (severe) was recorded performed for six histopathologic categories: epithelial cell viability, cornified epithelial cell differentiation, granular cell differentiation, epithelial cell nuclei, muscle cells, and muscle cell nuclei. Comparisons were made among treatment times using the Kruskal-Wallis test and linear trend analysis. For each histopathologic category and each treatment mode, the Kruskal-Wallis test showed significant differences between the groups (p < 0.002) over time. Trend analyses showed more severe injury with delayed neutralization therapy (p < 0.05) for each treatment mode and histopathologic category. Early neutralization therapy with OJ or cola reduces acute esophageal alkali injury. Additional in-vivo study is needed before neutralization therapy is adopted for clinical use.

Therapeutic Effects of Water and Milk for Acute Alkali Injury of the Esophagus

Study background: Alkali ingestions cause progressive and devastating injury to the esophagus by liquefaction necrosis. However, the therapeutic efficacy of water or milk dilution for alkali-induced esophageal injury has not been determined. This study used our previously reported model of alkali-induced esophageal injury to evaluate the effectiveness of water and milk dilution. Hypothesis: Early dilution with water or milk is efficacious in decreasing esophageal damage from alkali exposure. Methods: The esophagi of 75 Sprague-Dawley rats were harvested, and each end was cannulated with a 20-gauge catheter. Specimens were maintained in an oxygenated saline solution (at 37º C) during a 60-minute experimental period and then fixed immediately in 10% Formalin solution for histologic examination. Esophagi from six experimental groups (total of 60) were perfused with 50% NaOH solution at time 0. Water or milk dilution was performed immediately at 0 minutes, 5 minutes after injury, and 30 minutes after injury. Blinded pathologic examination was performed using a score of 0 (no injury), 1 (minimal), 2 (moderate), or 3 (severe) for the following six histologic categories: epithelial viability, comified epithelial cell differentiation, granular cell differentiation, epithelial cell nuclei, muscle cells, and muscle cell nuclei. Results: Positive and negative controls showed expected outcomes. Significant progressions of injury over time were seen for every histologic category for both water and milk dilution. The injury scores for the milk-treated group at 0 minutes were less than or equal to the injury score for the water-treated group for all categories. However, these differences were significant only for the cornified epithelial cells. Conclusion: Early dilution therapy with water or milk reduces acute alkali injury of the esophagus and supports use of these forms of emergency treatment. [Homan CS, Maitra SR, Lane BP, Thode HC, Sable M: Therapeutic effects of water and milk for acute alkali injury of the esophagus. Ann Emerg Med July 1994;24:14-20.]

Effective treatment of acute alkali injury of the rat esophagus with early saline dilution therapy

Controversy persists regarding the appropriate treatment of acute alkali injury to the esophagus. The current study establishes a controlled model of alkali esophageal injury and examines the efficacy of saline dilution therapy. Early saline dilution therapy effectively reduces esophageal injury resulting from acute alkali exposure. The esophagi were harvested from 60 Sprague-Dawley rats. Each end was cannulated with a 20-gauge catheter. Specimens were maintained in an oxygen-perfused saline bath (37 C) during a 60-minute experimental period and then fixed immediately in 10% formalin solution for histologic examination. Three experimental groups (A, B, and C) were perfused with 50% NaOH solution at time zero. Treatment with saline perfusion was performed immediately in group A, five minutes after injury in group B, and 30 minutes after injury in group C. The positive control group D was perfused with saline at time zero. A negative control, group E, was perfused with 50% NaOH at time zero. This group did not receive subsequent treatment with saline. Pathologic examination was performed in a blinded fashion using a score of 0 to 3 (0, no injury; 1, minimal; 2, moderate; 3, severe) for seven histologic criteria: epithelial viability, extent of injury, cornified epithelial cell differentiation, granular cell differentiation, epithelial cell nuclei, muscle cells, and muscle cell nuclei. The positive control group demonstrated scores of zero. Nonparametric analysis showed a significant difference among treatment groups for each injury category. Trend analysis revealed a significant progression of injury for each category associated with time to treatment. Discriminant analysis indicated that the muscle cells category was the most useful category with which to distinguish injury among groups. In our model, saline lavage decreased objective evidence of esophageal injury after a severe alkaline exposure, and early therapy enhanced this beneficial effect.

Evolution of the granular cells of the juxtaglomerular apparatus during postnatal development in the albino rat. Effect of age, body weight, renal weight and sex.

The postnatal development of the juxtaglomerular apparatus was studied by fluorescent microscopy in a series of 275 albino Wistar rats aged from 2 to 90 days. After treatment of kidney sections with thioflavine T, the appearance and evolution of secretory function in the terminal segment of the afferent glomerular arteriole were assessed in terms of the granular cell index (GCI) and the juxtaglomerular cell granulation index (JGI). Variations in these indices were analyzed in function of age, body weight, renal weight and sex of the animals. It emerges quite clearly from these studies that granular cell differentiation and maturation takes place in four successive stages: (a) a growth and differentiation phase lasting from the 2nd day to the beginning of the 5th week of extra-uterine life; (b) a critical phase around day 30; (c) a declining phase from day 30 to day 45, and (d) a stabilization phase from day 45 onwards. Until day 30 this evolutionary pattern is paralleled by rising body weight in animals of both sexes; afterwards it becomes independent of the parameters studied, in the males as well as in the females. The pattern of kidney weight parallels that of growth in body size and weight and does not constitute a more specific variable than body weight in the study of the developing granules in the juxtaglomerular apparatus.