Immunocytochemical localization of S-100 protein in interstitial cells of the monkey Macaca irus pineal gland

Pineal interstitial cells of the monkey Macaca irus were shown to react with an anti-human S-100 protein antibody, using the indirect immunoperoxidase technique on sections of paraplast-embedded pineal glands. Immunoreactivity was seen in the cytoplasm of the cells, stellate in shape and intermingled with pinealocytes; the latter did not stain with the antiserum against S-100 protein. Immunoreactivity was also present in the nuclei, as was reported in various other cell types immunostained with anti-S-100 protein antibodies. The present results support the view that interstitial cells of the monkey Macaca irus pineal gland may be of astroglial origin.     

Immunohistochemical localization of intermediate filament and S-100 proteins in several non-endocrine cells of the human pituitary gland

The presence and distribution of glial fibrillary acidic protein, vimentin, neurofilament protein, cytokeratins No. 8 (52 Kd), No. 18 (45 Kd) and No. 19 (40 Kd) and S-100 protein in pituicytes, folliculo-stellate cells, the epithelium of the Rathke's cysts and squamous cell nests of the pars tuberalis were investigated immunohistochemically by the peroxidase-antiperoxidase (PAP) method in eleven normal human pituitary glands. An identical immunostaining pattern was expressed by both folliculo-stellate cells and pituicytes. In both cell types the immunostaining for glial fibrillary acidic protein (GFAP), S-100 protein and vimentin was strongly positive. These results indicate the probable glial origin of the folliculo-stellate cell, and enlarge the group of glial cell types expressing vimentin. The co-expression of cytokeratins No. 8 and 19, both characteristic for simple epithelia, and S-100 protein was evident in the epithelial cells lining the Rathke's cysts and the squamous cell nests of the pars tuberalis. Furthermore, some epithelial cells of the Rathke's cysts co-expressed cytokeratins, S-100 protein and GFAP, a fact seldom reported and only in relation to rare neoplasms. The cytokeratin No. 18, characteristic for glandular epithelia, was not clearly demonstrated. Finally, the neurofilament protein was detected only in axons of the neurohypophysis; no immunopositive cells could be found throughout the adenohypophysis. Similarities in the antigenic patterns of these cell populations and the possible relation with their origin and nature are discussed.     

Analysis of S-100 protein positive folliculo-stellate cells in rat pituitary tissues.

The effects of diethylstilbestrol (DES) treatment and withdrawal on the folliculo-stellate (FS) cells in hyperplastic pituitaries were examined in Fischer 344 (F344) and Wistar Furth (W/F) rats by immunochemistry and electron microscopy. The presence of S-100 protein positive cells was examined by immunostaining in spontaneous and in transplantable rat pituitary tumors. The pituitaries of F344 rats showed a fivefold greater increase in weight in response to 5 weeks of DES treatment compared to pituitaries from W/F rats. S-100 protein (S-100) positive cells were significantly increased in both strains with hyperplastic pituitaries (P less than 0.05) 2 days after DES withdrawal. Ultrastructural studies revealed increased phagocytic activity in FS cells. S-100 positive cells were absent in both MtT/W15 and MtT/F4 transplantable tumors even after treatment with DES. Some spontaneous pituitary tumors in aged female rats had S-100 positive cells within the tumors at the periphery of the tumor nodules (3 of 12 cases), while most of these neoplasms did not contain S-100 positive cells. Incubation of normal dissociated pituitary cells with S-100 protein increased PRL secretion in vitro. The effectiveness of S-100 protein in increasing PRL secretion in vitro was less than thyrotropin releasing hormone (TRH). These results indicate that S-100 positive cells are present in normal and hyperplastic pituitaries, but not in spontaneous or in transplantable rat pituitary tumors and also suggest that the FS cells may exert a paracrine type regulation on PRL secretion.     

Intercellular communication within the rat anterior pituitary gland: X. Immunohistocytochemistry of S-100 and connexin 43 of folliculo-stellate cells in the rat anterior pituitary gland

Since Rinehart and Farquhar reported the presence of agranulated cells in the anterior pituitary gland in 1953, the functions of the folliculo-stellate cell remain to be clarified. Intercellular junctions have been described in the monkey, rat, and teleost anterior pituitary glands, indicating the existence of cell-to-cell communication within the organ. We pointed to their possible role in the rapid dissemination of information through a complex interconnecting system of follicles involving gap junctions. The gap junctional/folliculo-stellate cellular network was essential in the maturation and regulation of the pituitary gland system such as the hypothalamic-pituitary-gonadal axis. It has been was shown that a network participated in the conduction of electrophysiological information over a long distance using the ion Ca(++), which propagates to other folliculo-stellate cells by signaling through gap junctions. Sixty-day-old male rats were used in this study for light microscopic immunohistochemistry of S-100 protein, type I collagen, and connexin 43, and for electron microscopy to observe the morphological relationships between the cellular networks of folliculo-stellate cells and granulated pituitary cells. Clusters of anti-S-100 protein-positive cells were clearly observed in a region of the hypophysis tentatively named the transition zone. Anti-S-100 protein-positive cells and their cytoplasmic processes were also present in the anterior lobe and assembled together to form follicular lumina. Type I collagen was clearly shown outlining the incomplete lobular or ductule-like structure making cell cords in the anterior pituitary gland. Numerous microvilli were present within the follicular lumen while around the lumina, junctional specializations including gap junctions were positive for the connexin 43 protein. A nonuniform distribution of the connexin 43-positive sites were observed. Small or dot-shaped positive sites were noted where two clusters of cells were connected; the cells were identified as S-100 cells. Double immunohistochemical staining of the connexin 43 and growth hormone (GH) or connexin 43 and luteinizing hormone (LH) was also performed, demonstrating no direct relationship between the connexin 43 and either the GH or LH cells. These findings indicate that there are two kinds of messages necessary for the hormone release in the pituitary gland. One is via the portal vein system, the other is through the gap junction-mediated networks of folliculo-stellate cells. The granulated cells directly associate with cell membrane of folliculo-stellate cells are able to discharge secretory granules through communication via gap junctions, while those granulated cells that are more distant from the folliculo-stellate cells are only able to discharge hormones via the pituitary hormone-releasing hormone from the portal vein system.     

Immunocytochemical localization of a galanin-like peptide (GALP) in pituicytes of the rat posterior pituitary gland.

A galanin-like peptide (GALP) was recently isolated as a ligand of GalR2, a galanin receptor subtype. The GALP mRNA is expressed in the arcuate nucleus of the hypothalamus and the posterior pituitary (PP). In this study, we demonstrated the localization of GALP-immunoreactive (-ir) cells in the rat PP. In normal conditions, a few GALP-ir cells were detected in the PP, and these cells increased on dehydration for 4 days. The GALP-immunopositive reaction was dramatically enhanced by the intraperitoneal injection of colchicine. For the identification of GALP-ir cells in the PP, we performed electron microscopic observation, and also double immunocytochemical staining for GALP and S-100 protein. Both studies clearly indicated that the GALP-ir cells in the PP are pituicytes.     

Immunocytochemical localization of a developmentally regulated,isoproterenol-inducible protein (LM protein) in rat submandibular gland

Administration of the β-adrenergic drug isoproterenol (IPR) produces hyperplastic and hypertrophic enlargements of the submandibular gland of the rat and induces the synthesis of specific proteins in this organ. One of these proteins, the LM (large mobile) protein, was demonstrated immunocytochemically in the submandibular glands of developing untreated and IPR-treated rats. Immunoreactive LM protein was absent in the glands of 20-day-old fetuses and 1- and 2-day-old rats. It was localized in the proacinar and immature acinar cells in the glands of 6- to 21-day-old animals, but it was undetectable at 28 days of age. In the glands of adult rats, secretory granules of the granular convoluted tubule cells showed immunostaining for the LM protein which was also present in trace amounts in the acinar cells. Daily administration of IPR for 5 days to newborn or 8- or 15-day-old rats caused an apparent acceleration of proacinar/acinar cell differentiation, and consequently it increased the frequency of cells immunostained for the LM protein as well as the amount of immunoreactive material in these cells. Thus, the expression of LM protein in the submandibular gland is developmentally regulated, and it is restricted to the stage of differentiation of proacinar cells from terminal tubule cells. IPR is capable of inducing this protein in fully differentiated acinar cells in 3-week-old or older animals.     

Immunohistochemical demonstration of S-100 protein in salivary gland neoplasms

A peroxidase-antiperoxidase technique for S-100 protein has been applied to 68 salivary glands. These included 17 pleomorphic adenomas, seven adenoid cystic carcinomas, 23 adenolymphomas and a number of other neoplasms. In addition, five specimens of myoepithelial sialadenitis ('benign lymphoepithelial lesion') and five normal parotid glands were included. Consistent results were obtained, myoepithelial cells and cells in myxoid and chondroid areas in pleomorphic adenomas staining intensely. In adenoid cystic carcinoma, on the other hand, there was no staining. The adenolymphomas possessed intensely S-100 protein-positive cells in the interfollicular lymphoid areas; these were probably interdigitating reticulum cells. In addition, branching structures, probably corresponding to Langerhans' cells, were observed in the epithelium of adenolymphomas.     

水通道蛋白4在大鼠脑垂体中的表达

目的：研究水通道蛋白4(AQP4)及其mRNA在脑垂体中的表达,探讨其在脑垂体激素分泌过程中的作用。方法：应用免疫组织化学和原位杂交技术,观察成年Wistar大鼠脑垂体中AQP4及其mRNA的正常分布。结果：AQP4及其mRNA在成年大鼠神经垂体的垂体细胞上表达呈阳性,分布在毛细血管窦周围的垂体细胞表达尤为强烈。腺垂体的所有细胞均有AQP4的表达,胞质中AQP4 mRNA表达呈阳性。中间叶所有细胞AQP4及其mRNA的表达呈弱阳性,其中滤泡星形细胞表达较内分泌细胞强烈。结论：AQP4广泛分布于脑垂体的各种组织细胞表面,可能在垂体激素的正常分泌过程中起重要的调节作用。     

S-100 protein immunopositivity in human nontumorous hypophyses and pituitary adenomas

The presence, distribution, and morphological appearance of S-100 protein-immunoreactive cells in the human hypophysis were studied by immunocytochemistry. One hundred and twelve nonadenomatous pituitaries from fetuses to adults and pituitaries affected by several lesions including metastases, acute infarcts, and lymphocytic hypophysitis, as well as 115 pituitary adenomas were examined.S-100 protein immunoreactivity was detected in neurohy-pophyseal pituicytes and stellate cells of the pars distalis from 5 months following birth. In adults, S-100 protein-immunopositive cells displayed a preferential topographical association with growth hormone-, follicle-stimulating hormone-, luteinizing hormone-, and alpha-sub-unit-immunoreactive cells and with capillary walls. Colloid-containing follicles were mainly lined by hormone-containing cells, although scattered S-100 protein-immunoreactive processes or cell bodies were also observed forming their walls.No major changes in S-100 protein-immunoreactive cells were observed in the pituitary parenchyma bordering metastatic, inflammatory, necrotic, or adenomatous tissues. Eighteen of 115 pituitary adenomas contained a variable number of S-100 protein-immunoreactive cells. No preferential association of these cells with any type of pituitary adenoma was found.We propose that S-100 protein expression in the nontumorous adenohypophysis and pituitary adenomas may constitute a dynamic process and that S-100 protein-positive cells may constitute a heterogeneous cell population composed of pure, fully differentiated stellate cells and of transdifferentiated follicular cells.     

Neurospecific S-100 protein in synaptosomes of the rat cerebral cortex

A nonspecific S-100 protein was found in the composition of low-molecular-weight acid proteins from synaptosomes of the rat cerebral cortex by capillary microdisc electrophoresis in 15% polyacrylamide gel with 0.1% sodium dodecysulfate and with the aid of a highly purified marker protein. The S-100 protein accounted for 15–20% of the lowmolecular-weight acid synaptosomal proteins.Research Institute of Experimental Medicine, Academy of Medical Sciences of the USSR, Leningrad. (Presented by Academician of the Academy of Medical Sciences of the USSR V. S. Il'in). Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 2, pp. 164–166, February, 1976.     

Ultrastructural localization of exogenous silver in the anterior pituitary gland of the rat

Silver accumulations in the anterior pituitary of argyric rats were demonstrated using a histochemical method that visualizes minute traces of the metal. Silver was localized intra- and extracellularly throughout the anterior pituitary. Intracellular deposits were found within the lysosomes of somatotrophs and gonadotrophs. Extracellularly, the grains were located in basal laminae of portal veins and sinusoidal capillaries and in the membrane separating the anterior pituitary and part intermedia. The amount of deposited silver was dependent upon the dose of silver administered. Increasing the dose of silver lactate from 10 to 30 mg resulted in increased deposition, whereas a further increase to 60 mg did not alter the amount of silver deposited.     

Distinctive localization of N- and E-cadherins in rat anterior pituitary gland

In the rat anterior pituitary gland, folliculo-stellate cells aggregate preferably to form pseudofollicles, and each type of hormone-producing cell shows adhesive affinity with particular types of heterologous hormone-producing cells. Distribution of cadherin types in the rat anterior pituitary was examined immunohistochemically to clarify the unique cell arrangements caused by homologous and heterologous affinities among cells. N- and E-cadherins were detected continuously along cell membranes, while P-cadherin was not. N- and E-cadherins showed distinct isolation in localization, with N-cadherins localized in hormone-producing cells of distal and intermediate lobes in various amounts, and E-cadherins limited to folliculo-stellate cells and marginal layer cells facing the residual lumen of Rathke's pouch. A similar distribution of cadherins was observed in cell clusters of primary cultured anterior pituitary cells. These findings suggest that differential expression of cell adhesion molecules may be partially responsible for localization of hormone-producing cells and folliculo-stellate cells.     

Immunocytochemical study of human lymphoid tissues with monoclonal antibodies against S-100 protein subunits

Summary The present study concerns the immunocytochemical localization of S-100 protein and subunits in the cells of human lymphoreticular tissue and their related tumours. The subunit is mainly localized in dendritic cells, most likely the dendritic reticulum cells (DRCs) located within the germinal centers, while the subunit is mainly localized in the interdigitating reticulum cells (IRCs) in the paracortical area and in Histiocytosis X cells. No immunoreactivity for either subunit was found in the majority of normal lymphocytes, macrophages, malignant lymphoma cells, or xanthoma cells.The DRCs and IRCs are generally considered to show different distribution in the lymphoid tissues and demonstrate some difference in their immunocytochemical and enzyme-histochemical features. It is suggested that S-100 subunits can be used as useful markers for these two types of dendritic cells and investigation of these subunits may provide more information for the study of human lymphoreticular system.     

垂体腺瘤中垂体激素与S-100蛋白免疫组化双重染色观察

目的：探讨滤泡星状细胞在垂体腺瘤分类中的意义及滤泡星状细胞与内分泌细胞之间的关系。方法：应用免疫组化双重染色方法,对４２ 例人重体腺瘤的垂体激素与 Ｓ１００ 蛋白表达进行对照观察。结果：垂体腺瘤组织中的滤泡星状细胞有两种情况,一种为腺瘤组织中可见散在分布的滤泡星状细胞,并可见１ 个瘤细胞既有 Ｓ１００ 蛋白表达,又含激素分泌颗粒;另一种为滤泡星状细胞构成了腺瘤的一种主要的细胞成分。结论：滤泡星状细胞与内分泌细胞的功能密切相关,可能在调整内分泌细胞的产生和激素释放方面起一定的作用;滤泡星状细胞腺瘤应作为垂体无功能腺瘤的一个单独类型。     

Electroencephalographic effects of intracentral injection of antibodies against brain-specific S-100 antigen

The effects of intracentral injection of antibodies against brain-specific S-100 antigen (S-100 protein) were studied by an electroencephalographic method with estimation of the power of the EEG rhythms by analog computer. Intracerebral injections of antibodies against S-100 protein caused an increase in power of the principal EEG rhythms in the hippocampus, caudate nucleus, and mesencephalic reticular formation, followed by the development of epileptiform activity in these same structures.Laboratory of Neurophysiological Mechanisms of Adaptation, Institute of Clinical and Experimental Medicine, Academy of Medical Sciences of the USSR, Siberian Branch, Novosibirsk. (Presented by Academician of the Academy of Medical Sciences of the USSR V. P. Kaznacheev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 8, pp. 158–160, August, 1977.     

Immunocytochemical studies on the pituitary gland and spontaneous pituitary tumors of Sprague-Dawley rats

The review of our studies indicates a predictive value of the pituitary gland and spontaneous pituitary tumors of Sprague-Dawley rats for detecting and evaluating the effects and interactions of estrogens and dopamine agonists on the pathogenesis, functional morphology and therapy of human pituitary tumors, such as prolactinomas, gonadotroph adenomas and null cell adenomas. The functional classification of spontaneous pituitary tumors in old female Sprague-Dawley rats reveals a diversity of hormone content and morphologic appearance. They seem to be useful models of the human disease.     

Immunocytochemical localization of angiotensinogen in the rat brain

The distribution of angiotensinogen-like immunoreactivity in the rat brain was investigated using specific antisera against pure rat plasma angiotensinogen in conjunction with the sensitive streptavidin-biotin peroxidase method. Angiotensinogen antisera were shown by radioimmunoassay and Western blotting to recognize angiotensinogen from both rat plasma and cerebrospinal fluid, and to cross-react with des-AI-angiotensinogen (100%) but not with angiotensin I and II, tetradecapeptide, luteinizing hormone-releasing hormone, rat albumin and angiotensinogen from eight other species. Angiotensinogen-like immunoreactivity was detected throughout the rat brain in both neuroglia and neurons. The highest concentration of neuroglial angiotensinogen-like immunoreactivity was in the hypothalamus and preoptic areas, with moderate to heavy concentrations in the mesencephalon and myelencephalon. The cerebellum demonstrated neuroglial staining in the granular layer and fibre tracts. Very little neuroglial staining was noted in the cerebral cortex or olfactory bulbs. Neuronal immunostaining was observed throughout the globus pallidus and the caudate putamen, in various parts of the thalamus and the supraoptic nucleus of the hypothalamus. In the midbrain moderate immunostaining was observed in periaquaductal central gray, the deep mesencephalic nucleus, the inferior colliculus and in scattered cells in the anterior mesencephalon. In the medulla, neuronal staining was localized to the vestibular nuclei and to other cell bodies mainly in the dorsolateral regions. In the cerebellum, staining was noted mainly in the deeper cerebellar nuclei and in the Purkinje cells. Immunostaining in the cerebral cortex was localized to the cingulate cortex and the primary olfactory cortex. Light staining was present in the endopiriform cortex and in scattered neurons adjacent to the external capsule. In the olfactory bulbs light neuronal staining was mainly associated with the mitral cell layer. The widespread distribution of angiotensinogen-like immunoreactivity supports the view that it is synthesized in the central nervous system and forms part of a brain renin-angiotensin system. In addition, its presence at sites other than those normally associated with the control of blood pressure and fluid and electrolyte homeostasis suggests that its involvement may not be limited to these regulatory functions.     

Immunocytochemical localization of the sigma(1) receptor in the adult rat central nervous system

In order to characterize the localization of the sigma(1) receptor in the adult rat central nervous system, a polyclonal antibody was raised against a 20 amino acid peptide, corresponding to the fragment 143-162 of the cloned sigma(1) receptor protein. Throughout the rostrocaudal regions of the central nervous system extending from the olfactory bulb to the spinal cord, intense to moderate immunostaining was found to be associated with: (i) ependymocytes bordering the entire ventricular system, and (ii) neuron-like structures located within the parenchyma. Double fluorescence studies confirmed that, throughout the parenchyma, sigma(1) receptor-immunostaining was essentially associated with neuronal structures immunostained for the neuronal marker betaIII-tubulin. In all rats examined, high levels of immunostaining were always associated with neurons located within specific regions including the granular layer of the olfactory bulb, various hypothalamic nuclei, the septum, the central gray, motor nuclei of the hindbrain and the dorsal horn of the spinal cord. In contrast, only faint immunostaining was associated with neurons located in the caudate-putamen and the cerebellum. Electron microscope studies indicated that sigma(1) receptor immunostaining was mostly associated with neuronal perikarya and dendrites, where it was localized to the limiting plasma membrane, the membrane of mitochondria and of some cisternae of the endoplasmic reticulum. At the level of synaptic contacts, intense immunostaining was associated with postsynaptic structures including the postsynaptic thickening and some polymorphous vesicles, whereas the presynaptic axons were devoid of immunostaining.These data indicate that the sigma(1) receptor antibody prepared here, represents a promising tool for further investigating the role of sigma(1) receptors.