Immunostaining For Neuron-specific Enolase Research Articles

Rosette-forming glioneuronal tumor of the fourth ventricle

Objective To explore the clinicopathological features of rosette-forming glioneuronal tumor (RGNT) of the fourth ventricle. Methods The clinical manifestations, neuroimaging, histopathological and immunohistochemical features were analysed in one case of RGNT of the fourth ventricle, and related literatures were reviewed. Results A 24-year-old female presented with progressive dizziness under no obvious predisposing causes and dyskinesia such as stumbling. MRI revealed expansion of the fourth ventricle, and a mass with long T 1 WI and T 2 WI signal and clear boundary could be seen within the fourth ventricle. The border of tumor showed slight enhancement. At surgery, it was observed that the solitary tumor arised from the fourth ventricle and appeared well demarcated with rhomboid fossa. The tumor was blocking the aqueduct of sylvius before it was removed. Microscopically, the tumor exhibited both neuronic and astrocytic components. In the neuronic components, neurocytes formed neurocytic rosettes and perivascular pseudorosettes. At the center of the neurocytic rosettes, there was an eosinophilic core and some region consisted of microcysts. While the astrocytic components of the tumor revealed typical pilocytic astrocytoma structure. The center of neuronic rosettes and perivascular pseudorosettes displayed strong positive staining with synaptophysin (Syn) and oligodendrocytes transcription factor-2 (Olig-2). The astrocytic components showed positive immunostaining of glial fibrillary acidic protein (GFAP). There were focal and partial positive immunostaining of neuron-specific enolase (NSE) in both components of the tumor. The Ki-67 labeling index was 1.50%-2.00% in two components. Conclusions Rosette-forming glioneuronal tumor of the fourth ventricle is an unusual neuronal and mixed neuronal-glial tumors. The imaging examination showed solid or mixed solid-cystic mass at the fourth ventricle with well demarcated border. The lesion has two characteristic components and the distinctive immunostaining of GFAP and Syn expression will help the differential diagnosis. DOI: 10.3969/j.issn.1672-6731.2015.11.013

Carcinoma neuroendócrino de pequenas células

O presente estudo reporta o caso de uma mulher de 63 anos da qual a única informação clínica era a suspeita de um sarcoma da cérvix. Simultaneamente à colpocitologia, foram enviadas biópsias do colo e do endométrio para diagnóstico. A visualização da amostra citológica revelou vários agregados de número variável de células monótonas, com tamanho pequeno, formato redondo e citoplasma escasso, num fundo com diátese. Os núcleos apresentavam moldagem, hipercromasia, cromatina “sal-e-pimenta” e ausência de nucléolos. O aspeto microscópico das biópsias foi concordante com os achados citológicos, tendo sido igualmente identificados focos glanduliformes com características atípicas. A neoplasia mostrou expressão imunohistoquímica dos antigénios enolase neurónio-específica (neuron specific enolase, NSE), sinaptofisina e citoqueratina (clones AE1/AE3), e uma elevada atividade proliferativa demonstrada pela imunorreactividade para o marcador nuclear Ki67/Mib1. Os achados citológicos, histológicos e imunohistoquímicos foram consistentes com o diagnóstico de carcinoma neuroendócrino de pequenas células. Dos tumores cervicais, esta neoplasia maligna é das mais raras, mostrando um comportamento muito agressivo, com prognóstico muito pobre, em que as terapêuticas existentes são pouco consensuais quanto à sua eficácia. A sua etiologia ainda é estudada, podendo estar relacionada com a infeção pelo Vírus do Papiloma Humano.

Late effects of enriched environment (EE) plus multimodal early onset stimulation (MEOS) after traumatic brain injury in rats: Ongoing improvement of neuromotor function despite sustained volume of the CNS lesion

Recently we showed that the combination between MEOS and EE applied to rats for 7–15 days after traumatic brain injury (TBI) was associated with reduced CNS lesion volume and enhanced reversal of neuromotor dysfunction. In a continuation of this work, we tested whether these effects persisted for longer post-operative periods, e.g. 30 days post-injury (dpi). Rats were subjected to lateral fluid percussion (LFP) or to sham injury. After LFP, one third of the animals (injured and sham) was placed under conditions of standard housing (SH), one third was kept in EE-only, and one third received EE + MEOS. Standardized composite neuroscore (NS) for neurological functions and computerized analysis of the vibrissal motor performance were used to assess post-traumatic neuromotor deficits. These were followed by evaluation of the cortical lesion volume (CLV) after immunostaining for neuron-specific enolase, caspase 3 active, and GFAP. Finally, the volume of cortical lesion containing regeneration-associated proteins (CLV-RAP) was determined in sections stained for GAP-43, MAP2, and neuronal class III β-tubulin. We found (i) no differences in the vibrissal motor performance; (ii) EE + MEOS rats performed significantly better than SH rats in NS; (iii) EE-only and EE + MEOS animals, but not SH rats, showed better recovery at 30 dpi than at 15 dpi; (iv) no differences among all groups in CLV (larger than that at 15 dpi) and CLV-RAP, despite a clear tendency to reduction in the EE-only and EE + MEOS rats. We conclude that EE + MEOS retards, but cannot prevent the increase of lesion volume. This retardation is sufficient for a continuous restoration of neurological functions.

Glutamate induces the expression and release of tumor necrosis factor-α in cultured hypothalamic cells

Tumor necrosis factor-α (TNFα) affects several CNS functions such as regulation of sleep, body temperature, and feeding during pathology. There is also evidence for TNFα involvement in physiological sleep regulation, e.g., TNFα induces sleep and brain levels of TNFα increase during prolonged wakefulness. The immediate cause of enhanced TNFα production in brain is unknown. We investigated whether glutamate could signal TNFα production because glutamate is a neurotransmitter associated with cell activation and wakefulness. We used primary cultures of fetal rat hypothalamic cells to examine the expression and release of TNFα. Immunostaining for neuron specific enolase revealed that the cultures were 50–60% neuronal and 40–50% non-neuronal cells. TNFα was detected in both the media and cells under basal conditions. Stimulation of the cells with 1 mM glutamate for 2 h produced an increase in media content of TNFα, whereas cell content was elevated at earlier time points. Using trypan blue exclusion and MTT assays, there was no evidence of cell toxicity with this stimulation protocol. Immunocytochemical staining revealed that TNFα was expressed by ∼25% of the neurons and ∼75% of the glial cell in the culture. Stimulation of the cultures with glutamate did not increase the percentage of cells expressing TNFα. We conclude that TNFα is constitutively expressed and released by healthy cultures of hypothalamic cells and that activation of the cells with a non-toxic challenge of glutamate increases TNFα production. These findings support the hypothesis that TNFα can participate in normal physiological regulation of sleep and feeding.

Extracellular ATP activates chloride and taurine conductances in cultured hippocampal neurons.

We investigated regulation by extracellular ATP of channels important for volume regulation of rat hippocampal neurons. Cultures made from fetuses at the eighteenth gestational day were predominantly neuronal after 10-20 days in vitro, as indicated by immunostaining for neuron specific enolase. Neurons recorded with whole-cell patch clamp showed inward currents when membrane voltages were driven to values greater than -50 mV. Chloride conductance increased with 10 microM-100 microM extracellular ATP in a dose-dependent fashion. Similarly, an increase in taurine conductance was observed with 50 microM ATP. These currents were inhibited by the anion channel and purinergic receptor antagonists niflumic acid and suramin, respectively. The chloride conductance response to 10 microM ATP was increased over eight-fold in hypoosmotic medium (250 mOsm); however, chloride conductance in 0 mM ATP was not altered by this osmolality. Thus anion and osmolyte conducting channels activated via purinergic receptors may mediate volume regulation of hippocampal neurons.

Altered expression of immune‐related antigens by neuronophages does not improve neuronal survival after severe lesion of the facial nerve in rats

Injection of Fluoro-Gold (FG) into the whiskerpad muscles of rats yields a permanent retrograde labeling of motoneurons in the facial nucleus. Following subsequent resection of 10 mm of the facial nerve, one-third of the facial motoneurons die and the microglia phagocytize the dead FG-labeled neurons, take up FG, and get labeled in vivo. The resulting identification of all FG-labeled cells allows long-term comparative investigations on the behavior of neuronophages. In this study, we used two groups of rats to test whether the quantified expression of five immune-related antigens by neuronophages was related to quantified decline in neuron number (counts after immunostaining for neuron-specific enolase) 3 to 224 days after resection of the facial nerve. Rats of the first group received standard food and those of the second group, pellets containing 1,000 ppm of the calcium channel blocker nimodipine. Image analysis of the number of FG-containing cells and the number and projection area of immunopositive neuronophages in serial sections for each antigen showed that nimodipine significantly attenuated the immunostaining for CR3, MHC class I, and class II antigens (monoclonal antibodies [MAbs] OX-42, OX-18, and OX-6); enhanced the expression of monocyte–macrophage-specific antigen (MAb ED1); and did not change the expression of rat macrophage differentiation antigen (MAb ED2). The altered expressions, however, had no effect on the loss of motoneurons in the lesioned facial nucleus. We conclude that the degree of expression of immune-related antigens by neuronophages has no influence on the delayed neuronal cell death induced by permanent target deprivation. GLIA 24:155–171, 1998. © 1998 Wiley-Liss, Inc.

Altered expression of immune-related antigens by neuronophages does not improve neuronal survival after severe lesion of the facial nerve in rats

Injection of Fluoro-Gold (FG) into the whiskerpad muscles of rats yields a permanent retrograde labeling of motoneurons in the facial nucleus. Following subsequent resection of 10 mm of the facial nerve, one-third of the facial motoneurons die and the microglia phagocytize the dead FG-labeled neurons, take up FG, and get labeled in vivo. The resulting identification of all FG-labeled cells allows long-term comparative investigations on the behavior of neuronophages. In this study, we used two groups of rats to test whether the quantified expression of five immune-related antigens by neuronophages was related to quantified decline in neuron number (counts after immunostaining for neuron-specific enolase) 3 to 224 days after resection of the facial nerve. Rats of the first group received standard food and those of the second group, pellets containing 1,000 ppm of the calcium channel blocker nimodipine. Image analysis of the number of FG-containing cells and the number and projection area of immunopositive neuronophages in serial sections for each antigen showed that nimodipine significantly attenuated the immunostaining for CR3, MHC class I, and class II antigens (monoclonal antibodies [MAbs] OX-42, OX-18, and OX-6); enhanced the expression of monocyte-macrophage-specific antigen (MAb ED1); and did not change the expression of rat macrophage differentiation antigen (MAb ED2). The altered expressions, however, had no effect on the loss of motoneurons in the lesioned facial nucleus. We conclude that the degree of expression of immune-related antigens by neuronophages has no influence on the delayed neuronal cell death induced by permanent target deprivation.

Neuron-specific enolase and chromogranin A as markers of neuroendocrine tumours.

Circulating neuron-specific enolase (NSE) and chromogranin A (CgA) were measured in 128 patients with neuroendocrine tumours (NET) to compare their sensitivity and specificity, to investigate factors associated with elevated serum levels and to determine the usefulness of these markers in the follow-up of NET patients. NSE (Cispack NSE, Cis Bio International, Gif-sur-Yvette, France; normal <12.5 microg l(-1)), and chromogranin A (CgA-Riact, Cis Bio International, normal <100 microg l(-1)) were measured in 128 patients without renal insufficiency. There were 99 patients with gastroenteropancreatic (GEP) NET, 19 with medullary thyroid carcinoma and ten with phaeochromocytoma. Fifty-three patients with non-NET were studied as controls. Serum NSE and CgA levels were elevated in 48 (38%) and 76 (59%) of the 128 NET patients respectively. In all groups of NET patients, CgA proved to be more sensitive than NSE. NSE and CgA had a specificity of 73% and 68% respectively. Immunostaining for NSE was positive in three out of eight controls with elevated CgA levels, whereas immunostaining for CgA and synaptophysin was negative in all cases. Elevated CgA levels were significantly associated with two independent parameters, namely the presence of other secretions (P = 0.0001) and a heavy tumour burden (P = 0.001). Elevated NSE levels were exclusively associated with poor tumour differentiation (P = 0.01). Among six patients with NET followed for 11-37 months, CgA appeared to be a better marker of tumour evolution than NSE. We suggest that CgA ought to be the only general marker screened in NET patients.

Fetal pulmonary immunoreactive adrenocorticotropin: molecular weight and cellular localization

The hypothalamus–pituitary–adrenal axis of the sheep fetus plays a critical role in fetal development, responsiveness to stress, and initiation of parturition. We have recently reported that the fetal lung contains and secretes significant amounts of immunoreactive adrenocorticotropin (iACTH). The present study was designed to identify the molecular weight profile and the cellular location of iACTH in this tissue. iACTH extracted from fetal lung was immunoprecipitated, electrophoresed, and immunoblotted. Pulmonary iACTH was found in several molecular forms. The largest peptides appeared as doublets, and had molecular weights similar to POMC (32, 33 kD). Smaller peptides appeared in molecular weights (17, 24, and 27 kD) which were not consistent with the post-translational processing of POMC in fetal pituitary, but which were consistent with known processing of POMC by chromaffin granule aspartyl protease. None of the molecular forms of iACTH were glycosylated. Immunohistochemistry revealed that the iACTH was contained within bronchial epithelium and within groups of cells within the parenchyma of the lung. Both of these types of cells are consistent with pulmonary neuroendocrine cells. The distribution of neuroendocrine cells and apparent concordance with the iACTH-positive cells was confirmed by immunostaining for neuron specific enolase, a marker for neuroendocrine cells within the lung. We conclude that the lung contains unprocessed and partially processed POMC within cells known to contain neuropeptides. We speculate that secretion of the POMC-related peptides from these cells is physiologically important in the late-gestation fetus.

Temporospatial Relationships between Macroglia and Microglia during in vitro Differentiation of Murine Stem Cells

Embryonic stem (ES) cells of the permanent line BLC6 derived from a 129/Sv Gat mouse blastocyst were differentiated as spheroid aggregates (embryoid bodies, EBs) in the presence of retionic acid. After 2 days in suspension, EBs were plated on gelatine-coated glass coverslips and cultivated for 5, 9, and 16 days post plating (DPP) in normal medium. In this study we investigated whether the well-known retinoic acid-induced differentiation of ES cells into neurons (identified by immunostaining for neuron-specific enolase and synaptophysin) was accompanied by cells expressing astroglial (GFAP), oligodendroglial (O4), and microglial (5C6, galectin-3) markers. Whereas differentiation of neurons was closely related to their centrifugal migration towards the periphery of the EBs, the maturation of neuroglia followed a strict time-dependent manner. At 5 DPP, only neurons but no cells expressing glia-specific markers, were observed. At 9 DPP, GFAP-positive and O4-positive macroglial cells appeared. At 16 DPP, microglial cells (5C6-positive and galectin-3-positive) occurred. The established dynamic of relationships between neuronal and nonneuronal cells shows that the model of EBs is similar to the sequence differentiation of the nervous tissue. Thus, enabling in vivo observation of neurons, astrocytes, oligodendrocytes, and microglia, the model of EBs provides a basis for further investigations on the relationships between neurons and neuroglia under various experimental conditions.

Sclerosing Hemangioma: A Case With Neuroendocrine Differentiation

The sclerosing hemangioma is a rare benign tumor whose histogenesis continues to be controversial. Since the initial description by Liebow and Hubbell in 1956, theories on the cell of origin have included endothelial, alveolar pneumocyte, mesothelial, and epithelial. The tumor is generally solitary, with a female predominance. The patient, a 51-year-old woman, presented with a left hilar mass, grossly, a solid 3 to 4 cm nodule. Histologically, classic features of sclerosing hemangioma were found with solid areas composed of polygonal cells. Also noted were foamy histiocytes and regions of dense sclerosis. Immunohistochemical studies were performed. High and low molecular weight cytokeratin and carcinoembryonic antigen were nonconclusive. Ultrastructural analysis revealed tumor cells that were uniform with few cytoplasmic organelles and cell membranes closely interlocked that were connected by small adhering-type junctions. Thus, the cells were not found to be typically epithelial, endothelial, nor mesothelial; however, a few intracytoplasmic dense core granules were noted, suggesting neuroendocrine origin. Subsequent immunostaining for neuron specific enolase was positive. Vasoactive intestinal peptide, substance P, calcitonin gene related peptide, and bombesin were detected in the cytoplasm of different cell populations with varying intensities, which supported the ultrastructural analysis findings of neuroendocrine features. The findings suggest that sclerosing hemangioma may be of neuroendocrine origin and is capable of producing a number of bioactive peptides.

Progressive reorganization of the myenteric plexus during one year following reanastomosis of the ileum of the guinea pig

The enteric nervous system appears to play a pivotal role in the functional recovery of the gastrointestinal tract after partial resection and reanastomosis, but the structural changes following surgery are not fully understood. The present study was designed to clarify the processes of myenteric plexus regeneration up to one year after transection and reanastomosis of the ileum of the guinea pig. The following techniques were used: nicotinamide adenine dinucleotide (NADH) diaphorase histochemistry, immunostaining of neuron-specific enolase (NSE) in whole-mount preparations, and transmission electron microscopy. Two months after transection and reanastomosis, myenteric ganglion cells with NADH diaphorase reactions were scarce in the center of the lesion, and were less numerous in adjacent areas (3 mm in width) than in the control ileum. In the areas adjacent to the lesion, a few large extraganglionic neurons that did not completely compensate for the loss of ganglion neurons were observed. The remaining ileum showed no changes in NADH diaphorase staining pattern at this stage. Two to 12 months after transection and reanastomosis, ectopic large neurons gradually increased in number not only in the areas adjacent to the lesion but also in part of the remaining ileum, up to 10 cm from the lesion. Concomitantly, large ganglion neurons decreased in number in these areas. In other ileal regions (more than 10 cm distant from the site of transection), no obvious changes in NADH diaphorase staining were noted throughout the observation period. The outgrowth of NSE-containing nerve fibers from the severed stumps was seen two weeks after transection. Six weeks later, numerous bundles of fine nerve fibers with NSE were shown to interconnect the oral and anal cut ends of the myenteric plexus, but they exhibited no subsequent alterations. Transmission electron microscopy revealed that regenerating nerve fiber bundles appeared initially among irregularly arranged smooth muscle cells eight weeks after the operation, as expected from light-microscopic observations. These findings suggest that myenteric ganglion cell bodies, unlike myenteric nerve fibers, require a longer term of reconstruction than previously believed after transection and reanastomosis of the ileum of the guinea pig.

Malignant Retroperitoneal Paraganglioma in a Horse

A large primary retroperitoneal sublumbar neoplasm in a horse, with disseminated neoplastic foci in the brain, lung, kidney and spleen is described. The diagnosis was based on light microscopical studies and positive immunostaining for neuron-specific enolase. Because of the location of the primary tumour mass, the aortico-sympathetic ganglion (organ of Zukerkandl) is proposed as the origin.

Diagnosis and treatment of bronchial carcinoid tumors: clinical and pathological review of 120 operated patients

Clinical and pathological review is presented of 120 patients operated on for bronchial carcinoid tumors between 1976 and 1986. The usual oncologic features were analyzed. The Grimelius reaction, immunohistochemical staining for neuron specific enolase (NSE), serotonin and chromogranin, and DNA-analysis by flow cytometry were performed in these tumors and in small cell lung cancers (SCLC). In our experience the usual oncologic criteria--atypia, tumor-size, localization, history and regional lymph node metastasis--fail to give clear information for the prognosis. The Grimelius reaction has no significant differential diagnostic importance. Immunostaining for NSE can aid in distinguishing between neuroendocrine and non-neuroendocrine pulmonary tumors. The carcinoids and SCLCs could be differentiated by immunostaining for chromogranin and by flow cytometry but none of these methods are suitable for differential diagnosis within the carcinoid group. Resection by thoracotomy is the only treatment of choice: it can provide an excellent result (the 5-year survival rate is above 90%) with a low hospital mortality (0.8%). Parenchyma-sparing resections are to be encouraged.

Immunohistochemical study of NSE in small cell lung cancer (SCLS) combined with serum assay.

Background: Neuron specific enolase (NSE) is a neuronal form of the glycolytic enzyme enolase which was first found in extracts of brain tissue, and later in a variety of APUD cells and neurons of the diffuse endocrine system. SCLC shares many APUD properties with normal neuroendocrine cells. NSE immunostaining and serum NSE measurement may be a useful marker of neuroendocrine differentiation in lung tumors and diagnosis of small cell carcinoma. Methods: NSE immunohistochemical staining was done and at the same time serum NSE levels were measured in 22 small cell lung cancer and 21 non small cell lung cancer which were confirmed histologically. Results: 1) NSE immunoreactivity was detected in 9 of the 18 (50%) small cell lung cancer, in 5 of the 16 non small cell lung cancer. 2) Whereas the mean value in non-small cell lung cancer group was , the mean level of serum NSE in small cell lung cancer increased up to . In small cell lung cancer patients, mean value of limited disease group was , while mean value of extended disease group was showing statistically significant difference. If serum levels above 20 ng/ml were tentatively defined as positive, 16 of 22 (73%) patients with SCLC had positive serum NSE level, but only one patient with NSCLC did. There was no correlation between serum NSE level and immunoreactivity of NSE. Conclusion: These studies indicate that serum NSE measurement may be a useful marker for the diagnosis and disease extent and NSE immunostaining can be used to demonstrate the neuroendocrine components of lung tumor.

Immunohistochemical positivity for neuron-specific enolase and Leu-7 in malignant mesotheliomas

The discovery of immunostaining for neuron-specific enolase (NSE) and Leu-7 in a small cell mesothelioma prompted us to study some putative immunohistochemical markers of neuroendocrine differentiation in malignant mesotheliomas and to examine any diagnostically important immunohistochemical distinctions or similarities between malignant mesothelioma and other histologically similar lung tumours. Most mesotheliomas were positive for NSE (96 per cent) and Leu-7 (70 per cent) and positivity for these two markers was also found in small cell carcinomas (NSE 25 per cent, Leu-7 81 per cent) and adenocarcinomas (NSE 28 per cent, Leu-7 28 per cent) but carcinosarcomas were positive for only NSE (44 per cent). Chromogranin A positivity was found only in occasional small cell carcinomas (6 per cent) and adenocarcinomas (6 per cent). No tumour was positive for bombesin. The high incidence of NSE and Leu-7 positivity in mesotheliomas is an important original observation because it guards against the unjustified exclusion of mesothelioma from a differential diagnosis on the basis of positivity for these two markers.

Histological classification of nesidioblastosis: Efficacy of immunohistochemical study of neuron-specific enolase

Pancreatic specimens of six patients with nesidioblastosis were studied by light microscopy and immunohistochemistry in order to classify the changes of the endocrine pancreas and relate them to the clinical outcome. Neuron-specific enolase immunostaining was useful in demonstrating the distribution of whole pancreatic endocrine cells. The six cases with nesidioblastosis were classified into two types; four cases of focal type (head 1, body 1, tail 2), and two cases of diffuse type. The histological classification show a correlation to the clinical outcome. The patients with a focal body or tail type showed normal blood sugar levels immediately after operation. On the other hand the patients with both diffuse type and a focal head type needed medical treatment within a few months after operation. Therefore, these cases should be followed with great care postoperatively. A partial 80% to 90% pancreatectomy might be sufficient for each type except for the focal head type. When the intraoperative examination (both macroscopic and microscopic) is suggestive of focal head type, a pancreatectomy of 95% or more is called for.

Cytogenetic and Molecular Evaluation of Clinically Aggressive Esthesioneuroblastoma

We report a 16-year-old boy with esthesioneuroblastoma that presented with a unilateral tumor extending to the maxillary sinus and periorbital region. Despite initial therapy with gross resection, 5,682 cGy to the tumor bed and chemotherapy, the patient subsequently had a rapid local recurrence with distant metastases. Immunocytochemical, ultrastructural, cytogenetic, and molecular techniques were performed to determine if this tumor was biologically similar to childhood neuroblastoma. Urinary excretion of vanillylmandelic acid (VMA) and homovanillic acid (HVA) were markedly elevated. Chromogranin and neuron specific enolase immunostaining of tumor cells was positive, as seen in neuroblastoma. Electron microscopic studies showed cells that were closely packed and connected by occasional cell junctions. The cell cytoplasm contained moderate amounts of filaments and microtubules. Numerous electron dense granules were observed; however, these granules lacked distinct nucleoids and generally reacted strongly for acid phosphatase, indicating a lysosomal rather than a secretory function. Tumor cells contained near-pseudotetraploid chromosomes, with all chromosomes represented at least three times, and chromosome 5 was present in multiples of eight. Clonal structural abnormalities included 2q+ and 5q+ and multiple double minutes. Northern blot analysis revealed both c-myc and N-myc expression; however, N-myc amplification was not demonstrated, and c-myc expression appeared increased, unlike cases of rapidly progressive neuroblastoma. These results suggest that despite biologic similarities to neuroblastoma in catecholamine excretion and some ultrastructural features, molecular genetic abnormalities differ in this comparatively aggressive case of estesioneuroblastoma.

Immunohistochemical localization of synaptophysin (p38) in the pineal gland of the Mongolian gerbil (Meriones unguiculatus)

Synaptophysin (protein p38), a major integral membrane glycoprotein of small presynaptic vesicles, was localized immunohistochemically in semithin sections of the superficial pineal gland of the Mongolian gerbil (Meriones unguiculatus). Synaptophysin immunoreactivity could be detected in all pinealocytes, which were visualized with antibodies directed against neuron-specific enolase (NSE) in adjacent sections. No p38 immunoreactivity was discernible in the interstitial glial cells, which showed a heterogeneous pattern of immunostaining for the intermediate filament proteins glial fibrillary acidic protein (GFAP) and vimentin. Pinealocytes exhibited considerable intercellular differences in the densities of immunostaining. The various degrees of synaptophysin immunoreactivities in pinealocytes were not correlated with the densities of NSE immunostaining. Nerve terminals and varicosities displayed stronger immunoreactivities than pinealocytes. They were particularly numerous in the perivascular spaces. It is not clear whether this distribution indicates an innervation of pineal capillaries in addition to the functionally important innervation of pinealocytes. Several highly p38-positive dots of variable size were a conspicuous feature throughout the gland. By the consecutive semithin-thin section technique, they could be identified as processes of pinealocytes, filled with accumulations of small clear vesicles. Obviously, these vesicles represent the major site of synaptophysin immunoreactivity in pinealocytes. In the gerbil, similar vesicles have been ascribed a role in the secretory activity of the gland, and/or in the transport of calcium. The intercellular differences in the degrees of p38 immunostaining may, therefore, reflect different states of a specific cellular activity. The presence of synaptophysin in pinealocytes of the normal pineal, including the deep portions of the gland, emphasizes the paraneuronal character of these cells.

Endocrine Cell Hyperplasia of the Uterine Cervix: A Precursor of Neuroendocrine Carcinoma of the Cervix?

A 33-year-old woman who presented with vaginal bleeding was diagnosed to have neuroendocrine small cell carcinoma based on cervical smear and biopsy. Hysterectomy was performed, and a tumor measuring 5.5 X 2 mm was found at the squamocolumnar junction of the uterine cervix. In the immediate vicinity of the tumor, there was proliferation of cytologically benign endocrine cells in the normal endocervical glands and in the glands showing intraepithelial glandular neoplasia. Both the hyperplastic endocrine cells and the invasive tumor cells showed argyrophilia and immunostaining for neuron-specific enolase, neurofilament, and chromogranin. The topographical relationship suggests that endocrine cell hyperplasia may represent a precursor of neuroendocrine carcinoma of the cervix.