Biomedical subjects

M I Arenas

Publications and source records attributed to M I Arenas.

28 records · Page 2Linked to original sources

PubMed1997

Immunocytochemical and quantitative study of actin, desmin and vimentin in the peritubular cells of the testes from elderly men.

A quantitative immunohistochemical study using light and electron microscopy was carried out to evaluate the morphological and quantitative distribution of the peritubular cells that immunoreact with actin, vimentin and desmin, alone or in combinations, in normal adult testes and the changes in these cells in elderly men. Seminiferous tubules in ageing testes were classified in three groups according to the degree of lamina propria thickening due to tubular sclerosis: group I, < 8 microns; group II, 8.1-12 microns; and group III, > 12.1 microns. The number of peritubular cells per cross-sectioned tubule increased from group I to group III tubules. However, no significant differences between ageing men and controls were found in the total number of peritubular cells per testis. Most peritubular cells of control testes and of group I and group II tubules displayed immunoreactivity to actin. The peritubular cells in the outermost layers of group III tubules showed no or scanty reaction. The number of actin-immunostained cells per cross-sectioned tubule decreased (P < 0.05) with tubular sclerosis. The total number of these cells per testis was significantly lower (P < 0.05) in elderly men. A narrow band around the seminiferous epithelium immunostained for desmin in control testes and group I tubules. These cells also immunoreacted to actin and vimentin. In group II and, principally, in group III tubules, only isolated peritubular cells were immunostained for desmin. The number of desmin-immunostained cells per cross-sectioned tubule decreased with tubular sclerosis and the total number per testis was also lower in elderly men. Vimentin immunostaining was observed in most peritubular cells in all tubule groups; these cells also immunoreacted to actin. Vimentin and desmin co-localized only in the inner peritubular cell layers. The number of vimentin-immunostained cells per cross-sectioned tubule increased with the degree of tubular sclerosis but the total number of these cells per testis did not differ significantly between control and ageing testes.

Actins↗

PubMed1996

Testicular cell cytoskeleton in the newt, Triturus marmoratus marmoratus, during the annual cycle.

Light and electron microscopy immunohistochemical studies and Western blotting analysis of cytoskeletal proteins have been carried out in the testis of the marbled newt (Triturus marmoratus marmoratus) during the annual testicular cycle. The present findings revealed homologies and differences with regard to those reported in the testes of mammals and other vertebrates. Changes in immunohistochemical expression have also been detected in the course of the annual cycle. Actin and tubulin, which were scanty and diffusely located in spermatogonia and spermatocytes, increased their expression and reorganized during spermiogenesis. Vimentin and keratin, undetected in spermatogonia and spermatocytes, were expressed in differentiating spermatids and spermatozoa. In these cells, actin might be related with the connection of the axial fiber to the undulating membrane and the coordination of movement by both structures, while vimentin might be involved in the maintenance of the spatial relationship between the axoneme and the marginal fiber. During the first stages of spermatogenesis, the cytoplasm of Sertoli cells (follicular cells) showed a diffuse immunoreaction to actin, myosin, and tubulin and no vimentin immunolabeling. In advanced spermiogenesis, the follicular cells showed an intense immunoreaction to actin, myosin, tubulin, and vimentin in the apical projections that surrounded the spermatid heads. These apical cytoskeletal components might be involved in spermatid elongation, since the spermatids display no manchette, and in spermatozoon positioning and grouping. The colocalization of myosin and actin in the follicular cells suggests that actin filaments from contractile bundles and that contraction might be involved in changes in the Sertoli cell shape that accompany germ cell development during spermatogenesis. The interstitial cells immunostained to actin, myosin, tubulin, and vimentin. These cells, together with follicular cells, seemed to form the glandular tissue cells which showed a similar immunophenotype. The cells that surrounded the efferent duct epithelium immunostained to desmin, and they are probably contractile cells involved in sperm evacuation.

Activity Cycles↗

PubMed1996

Immunocytochemical electron microscopic study and Western blot analysis of troponin in striated muscle of the fruit fly Drosophila melanogaster and in several muscle cell types of the earthworm Eisenia foetida.

BACKGROUND: There is little information about troponin in invertebrate muscles, and no previous references to this protein in annelid muscles have been found. The aim of this paper was to study the presence and distribution of troponin in different muscle cell types from the earthworm Eisenia foetida (the muscular body wall, and the inner and outer muscular layer of the pseudoheart). These results were compared with those obtained in the transversely striated muscle of Drosophila melanogaster and in skeletal and smooth muscles of the mouse. METHODS: Immunocytochemical electron microscopic study and Western blot analysis using anti-TnT antibodies were employed in this study. RESULTS: Troponin immunoreaction was detected in the mouse skeletal muscle, the fly flight muscle, and earthworm obliquely striated muscles (body wall musculature and inner muscular layer of the pseudoheart). Immunolabeling for TnT in all these muscle cells appeared in moderate amounts at any point along the sarcomere length, except for the central zone of the A band (H band). This suggests that troponin molecules were located along the thin filaments. The density of immunogold particles was similar in the three muscles, and thus the amount of troponin in each muscle type was proportional to the number and length of actin filaments in each. Troponin was found in neither the mouse smooth muscle nor the outer muscular layer of the earthworm pseudoheart. The latter muscle showed an ultrastructural pattern that was intermediate between obliquely striated and smooth muscle. The estimated molecular weight for TnT in the earthworm was 55 kDa; this is higher than the weight of this protein in the mouse skeletal muscle (40 kDa) but similar to that of the D. melanogaster muscle (52 kDa). CONCLUSIONS: Troponin is present in both types of striated muscle (transversely striated and obliquely striated) of the earthworm with a distribution that is very similar to that observed in the mammalian striated muscle. As in vertebrates, troponin is absent in the smooth muscle of the earthworm. Discrepancies in the classification of some invertebrate muscles are common in the literature, and the use of distinctive markers, such as troponin, may improve our understanding of the nature and properties of many invertebrate muscles showing an ultrastructural pattern that does not resemble any of the classic muscle types.

Animals↗

PubMed1996

Immunocytochemical electron microscopic study and western blot analysis of paramyosin in different invertebrate muscle cell types of the fruit fly Drosophila melanogaster, the earthworm Eisenia foetida, and the snail Helix aspersa.

The presence and distribution pattern of paramyosin have been examined in different invertebrate muscle cell types by means of Western blot analysis and electron microscopy immunogold labelling. The muscles studied were: transversely striated muscle with continuous Z lines (flight muscle from Drosophila melanogaster), transversely striated muscle with discontinuous Z lines (heart muscle from the snail Helix aspersa), obliquely striated body wall muscle from the earthworm Eisenia foetida, and smooth muscles (retractor muscle from the snail and pseudoheart outer muscular layer from the earthworm). Paramyosin-like immunoreactivity was localized in thick filaments of all muscles studied. Immunogold particle density was similar along the whole thick filament length in insect flight muscle but it predominated in filament tips of fusiform thick filaments in both snail heart and earthworm body wall musculature when these filaments were observed in longitudinal sections. In obliquely sectioned thick filaments, immunolabelling was more abundant at the sites where filaments disappeared from the section. These results agree with the notion that paramyosin extended along the whole filament length, but that it can only be immunolabelled when it is not covered by myosin. In all muscles examined, immunolabelling density was lower in cross-sectioned myofilaments than in longitudinally sectioned myofilaments. This suggests that paramyosin does not form a continuous filament. The results of a semiquantitative analysis of paramyosin-like immunoreactivity indicated that it was more abundant in striated than in smooth muscles, and that, within striated muscles, transversely striated muscles contain more paramyosin than obliquely striated muscles.

Animals↗

PubMed1996

Light and electron microscopic immunohistochemical localization of protein gene product 9.5 and ubiquitin immunoreactivities in the human epididymis and vas deferens.

The distribution of protein gene product 9.5 (PGP) and ubiquitin immunoreactivities in the ductuli efferentes, ductus epididymidis, and ductus deferens of humans was studied by Western blot analyses and light and electron microscopic immunocytochemistry. PGP immunoreactivity was intense in the ductuli efferentes and weak in the ductus epididymidis and ductus deferens, while ubiquitin immunoreactivity was intense in the ductuli efferentes and ductus epididymidis and very weak in the ductus deferens. In the ductuli efferentes epithelium, PGP immunolabeling was observed in the cytoplasm of principal cells, whereas ubiquitin immunoreactivity was found in the nucleus and cytoplasm of principal cells and ciliated cells. In the ductus epididymidis epithelium, only scattered cells (mitochondria-rich cells) showed PGP immunoreaction in their cytoplasm, whereas ubiquitin immunostaining was detected in the nucleus and cytoplasm of most epithelial cells, except for the cauda, where ubiquitin immunolabeling was observed only in the nuclei. The ductus deferens showed no immunostaining for PGP, and only nuclear immunoreactivity to ubiquitin. The ultrastructural localization of PGP immunoreactivity was in the apical cytosol and microvilli. In addition to these locations, ubiquitin immunoreactivity was also found in the nucleus of all cell types and cilia of ciliated cells. Although the distribution of PGP and ubiquitin immunoreactivities in humans differs from that reported in rats, it seems that PGP and ubiquitinated proteins are secreted into the epididymal lumen in both species.

Adult↗

PubMed1996

Lectin histochemistry in the human epididymis.

A histochemical study using lectins to identify glycoconjugates present in the efferent ducts and ductus epididymidis of men without testicular or related disease was carried out. The lectins used and the oligosaccharide residues linked were: wheat germ agglutinin (WGA) for beta-N-acetylglucosamine and sialic acid, concanavalin A (ConA) for alpha-mannose, Ulex europaeus agglutinin (UEA-I) for alpha-fucose, Dolichos biflorus agglutinin (DBA) for alpha-N-acetylgalactosamine, soy bean agglutinin (SBA) for beta-N-acetylgalactosamine, and peanut agglutinin (PNA) for beta-galactose. The lectin-binding pattern in the human epididymis presents similarities and differences to those observed in other mammals which also showed differences between species. The present results revealed that regional differences along the human ductus epididymidis were less pronounced than those reported in other mammals. The epithelial cells in the efferent ducts reacted positively to WGA. All along the length of the ductus epididymidis, the principal cells and the luminal content showed staining affinity for WGA and ConA. The epididymal principal cells and luminal content also reacted positively to DBA for alpha-N-acetylgalactosamine but only in the cauda epididymidis. A positive reaction to UEA-I was observed only in the luminal content of the cauda epididymidis. This finding suggests that changes in the oligosaccharide chains of secretions leading to a positive UEA-I reaction occur in the cauda epididymidis. The epididymal principal cells showed positive reactions to SBA and PNA over the apical microvilli but not in the cytoplasm. The reaction was observed in the caput and corpus epididymidis but not in the cauda. Positive reactions to SBA and PNA were also detected in the epididymal fluid and in the cytoplasm of mitochondria-rich cells (a minor population of epididymal epithelial cells). These cells also reacted to other lectins such as WGA, ConA and DBA, which were also detected in the principal cells.

Acetylgalactosamine↗

PubMed1995

Cytoskeleton in Sertoli cells of the mosquito fish (Gambusia affinis holbrooki).

BACKGROUND: There is little information about the distribution of cytoskeletal components in the testes of teleost fish. The aim of this paper was to know the distribution of some major cytoskeletal proteins (tubulin, actin, vimentin, desmin, and cytokeratins) in the Sertoli cells of Gambusia affinis holbrooki and in their efferent duct epithelial cells which are possibly originated from the Sertoli cells. METHODS: Light and electron microscopic immunocytochemical studies and Western blotting analysis were performed in G. affinis testis. RESULTS: Actin immunoreaction was observed in the Sertoli cells at all spermatogenic stages, although the intensity of this reaction varied from one stage to another. Sertoli cells that support spermatogonia or spermatocytes showed a weak immunoreaction which was uniformly distributed throughout the cytoplasm and somewhat more concentrated at the level of the inter-Sertoli specialized junctions. Immunoreaction to actin increased during the first stages of spermiogenesis and was mainly localized beneath the plasma membrane. This immunoreaction was more intense in the basal than in the apical cytoplasm of Sertoli cells. In a more advanced stage of spermiogenesis, actin immunoreaction become stronger in the apical cytoplasm where Sertoli cells displayed cytoplasmic projections around each spermatid. After sperm release, the apical Sertoli cell cytoplasm still showed an intense actin immunoreaction. Intense immunoreaction to actin was also observed in the epithelial cells lining the efferent ducts. Immunoreaction to tubulin was diffuse throughout the Sertoli cell cytoplasm. No immunoreaction to vimentin or desmin was observed in the Sertoli cells during the spermatogenic process. Immunoreaction to both vimentin and desmin was observed in the efferent duct cells. Desmin immunoreaction was also observed in the seminiferous tubule boundary cells, mainly in the sections showing germ cell cysts at the last stages of spermiogenesis and in the peritubular cells that surrounded the efferent duct epithelium. Immunoreaction to cytokeratins was found in the endothelium of testicular blood vessels but not in the Sertoli cells or in the efferent duct epithelium. CONCLUSIONS: Immunoreaction pattern to cytoskeletal proteins in the Sertoli cells of G. affinis differs from that reported in mammalian Sertoli cells. These differences include the distribution of actin filaments and the absence of detectable vimentin immunoreaction in G. affinis Sertoli cells.

Animals↗

PubMed1995

Intermediate filaments in the testis of the teleost mosquito fish Gambusia affinis holbrooki: a light and electron microscope immunocytochemical study and western blotting analysis.

A light and electron microscope immunocytochemical study and Western blotting analysis has been performed on intermediate filaments (vimentin, desmin and cytokeratins) in the testis of the teleost fish Gambusia affinis holbrooki. An immunoreaction to vimentin was observed in the epithelium of the efferent ducts, testicular canal and their surrounding peritubular cells. Positive vimentin immunostaining was also observed in the cells located around seminiferous tubules (boundary cells), Leydig cells, interstitial fibroblasts, chromatophores, and blood vessel endothelial cells. In contrast to mammals, no vimentin immunoreactivity was found in the Sertoli cells. Immunoreactivity to desmin was weak in the epithelial cells of the efferent ducts and testicular canal and intense in the peritubular cells that surrounded these ducts. Desmin immunoreactivity was also observed in the seminiferous tubule boundary cells. The immunoreactivity was weak in the boundary cells that surrounded germ cell cysts containing spermatogonia or spermatocytes and intense in the boundary cells around cysts with elongated or mature spermatids. Immunoreactivity towards cytokeratins was observed only in testicular blood vessels. Cytokeratin immunolabelling was intense in the endothelium and weak in the vascular smooth muscle cells. No cytokeratin immunoreactivity was found in the Sertoli cells, germ cells, interstitial cells or in the efferent duct epithelium. The absence of intermediate filaments in the Sertoli cells, the absence of cytokeratins in the epithelium of the sperm excretory ducts, and the presence of desmin filaments in these epithelial cells are the most important differences with regards to the intermediate filament phenotype in mammalian testes.

Animals↗

PubMedDate not supplied

Identification of N- and O-linked oligosaccharides in human seminal vesicles.

The main oligosaccharide residues and the saccharide linkage in infantile and adult human seminal vesicles were studied by means of lectin histochemistry at light and electron microscopy levels. In adult glands, the epithelial cell cytoplasm and luminal content reacted positively to the following residues: (GlcNAc)n (WGA), Galbeta1,3GalNAc (PNA), GalNAcalpha1,3Gal (SBA), GalNAcalpha1,3GalNAc (HPA), Fucalpha1,2Galbeta1,4GlcNAc (UEA-I), and alphaL-Fuc1,6DGlcNAc-O-Melibiosc (AAA). The presence of intense staining in the luminal content suggest that glycoproteins containing these oligosaccharide moieties are secreted by epithelial cells. Adult epithelial cells also reacted to Neu5Acalpha2,6Gal (SNA), Neu5Acalphaa2,3Galbeta1,4GlcNAc (MAA), Galbeta1,4GlcNAc (DSA), branched mannose chains (ConA), Man1,3Man (GNA), and Fucalpha1,2Galbeta1,4GlcNAcFucalpha1,3GlcNAc (LTA) but reaction to these residues was weak (MAA, DSA, ConA, and LTA) or absent (SNA and GNA) in the gland lumen, which suggests that they belong to intracytoplasmic proteins. The chemical and enzymatic treatments used suggest that the residues recognized by SNA, MAA, PNA, DSA, HPA, and SBA belong to O-linked oligosaccharides; those residues localized by ConA and GNA have an N-glycosidic linkage, and those bound by WGA, LTA, UEA-I, and AAA are linked to both N- and O-oligosaccharides. In prepubertal seminal vesicles, reaction in the epithelial cell cytoplasm was similar to that observed in adults, except for GNA and HPA, which showed a weaker reaction. However, the lumen of prepubertal seminal vesicles showed intense reaction to WGA and SBA only. The chemical and enzymatic treatments suggest that the scanty glycoproteins secreted by the prepubertal glands belong to the mucin-type.

Adult↗

PubMedDate not supplied

Immunoexpressions of p21, Rb, mcl-1 and bad gene products in normal, hyperplastic and carcinomatous human prostates.

A comparative study of the expression of p21, Rb, mcl-1, and bad gene products, which are involved in the control of the cell cycle, was performed in normal, hyperplastic, and carcinomatous human prostates by means of a semiquantitative immunochemical study. This included Western blot, ELISA, and immunohistochemistry procedures. In normal prostates, immunoexpression of the four gene products was scanty or absent. In men with benign prostatic hyperplasia, immunoreactions to the four proteins studied were found in many epithelial cells and some stromal cells. In prostatic carcinoma, the immunostaining pattern was as in hyperplastic prostates but the numbers of both epithelial and stromal cells were higher. Present results indicate that immunoexpression of p21, Rb (both the phosphorylated and dephosphorylated forms), mcl-1, and bad gene products are markedly increased in prostates with proliferative alterations but that these proteins do not discriminate between benignant (hyperplasia) and malignant (adenocarcinoma) prostatic tumours, although immunoexpression is higher in prostatic carcinoma.

Adult↗