Histology and histopathology Vol.28, nº11 (2013)
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- PublicationOpen AccessDifferential expression and localization of transient receptor potential vanilloid 1 in rabbit and human eyes(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Martínez-García, M. Carmen; Martínez, Tamara; Pañeda, Covadonga; Gallego, Patricia; Jimenez, Ana I.; Merayo, JesusIntroduction: The superfamily of transient receptor potential (TRP) cation channels is involved in nociception. Members of this family, such as the vanilloid receptor type 1 (TRPV1) channel, are activated by a wide range of stimuli including heat (>43°C), low pH (<6.5), hypoxia, and hypertonicity. Here we report TRPV1 expression in rabbit and human eyes. Material and methods: We analyzed the expression of TRPV1 mRNA by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and protein by immunohistochemistry in eyes of New Zealand White rabbits and humans. Results: In rabbit and human eyes, TRPV1 protein was present in all layers of the corneal epithelium, but only in the basal layer of the conjunctiva. It was also in the ciliary and lens epithelia of both species as well as in the secretory cells of the rabbit lacrimal gland. The retinal pigment epithelium was positive for this protein in both species. TRPV1 was also present in rabbit Müller cells, where it had a similar pattern of expression to vimentin intermediate filaments. Analysis by qRT-PCR showed that TRPV1 mRNA was found in all of the structures where the protein was present. The highest level was in the lens and the lowest in the retina. Conclusion: TRPV1 is expressed in cells that are particularly active in Ca2+ exchange as well as in cells with significant water transport activity. Because TRPV1 is a Ca2+ channel, it probably functions in the regulation of both water and Ca2+ movements in ocular tissues.
- PublicationOpen AccessExpression of tricellulin in epithelial cells and non-epithelial cells(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Kojima, Takashi; Ninomiya, Takafumi; Konno, Takumi; Kohno, Takayuki; Taniguchi, Masahiko; Sawada, NorimasaTricellulin is the first molecular component of tricellular tight junctions at tricellular contacts where three epithelial cells meet, and it is required for the their formation and maintenance of the epithelial barrier. Tricellulin binds other tight junction proteins, and its expression and distribution are affected by the bicellular tight junction protein occludin and lipolysis-stimulated lipoprotein receptor (LSR) which is expressed at tricellular contacts. Tricellulin is also detected in endothelial cells, neurons, microglia and astrocytes. Here, we focused tricellulin expression in various types of epithelial cells, nasal epithelial cells, pancreatic duct epithelial cells cells and hepatocytes, and non-epithelial cells, dendritic cells and Schwann cells, compared to expression of the bicellular tight junction protein occludin and LSR, and discuss the regulation and the role of tricellulin in cellular specificity.
- PublicationOpen AccessSignaling molecules and pathways involved in MSC tumor tropism(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Ho, Ivy AW; Lam, Paula YPHuman bone marrow is a reservoir containing cells with different self-renewal capabilities, such as mesenchymal stem cells (MSC) and hematopoeitic stem cells (HSC). MSC in particular have been increasingly used in preclinical and clinical treatment of tissue regenerative disorder. Understanding the molecular mechanisms underlying MSC homing and mobilization is critical to the design of rational cell therapy approaches. In this review, we will discuss the key molecular mechanisms that govern the homing of MSC to bone marrow, the mobilization of MSC to tumors and injured sites via circulation, and strategies that enhance MSC migration.
- PublicationOpen AccessSPARC is up-regulated during skeletal muscle regeneration and inhibits myoblast differentiation(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Petersson, Stine Juhl; Jørgensen, Louise Helskov; Andersen, Ditte C.; Nørgaard, Rikke C.; Jensen3, Charlotte Harken; Schrøder, Henrik DaaSkeletal muscle repair is mediated primarily by the muscle stem cell, the satellite cell. Several factors, including extracellular matrix, are known to regulate satellite cell function and regeneration. One factor, the matricellular Secreted Protein Acidic and Rich in Cysteine (SPARC) is highly up-regulated during skeletal muscle disease, but its function remains elusive. In the present study, we demonstrate a prominent yet transient increase in SPARC mRNA and protein content during skeletal muscle regeneration that correlates with the expression profile of specific muscle factors like MyoD, Myf5, Myf6, Myogenin, NCAM, CD34, and M-Cadherin, all known to be implicated in satellite cell activation/proliferation following muscle damage. This up regulation was detected in more cell types. Ectopic expression of SPARC in the muscle progenitor cell line C2C12 was performed to mimic the high levels of SPARC seen in muscle disease. SPARC overexpression almost completely abolished myogenic differentiation in these cultures as determined by substantially reduced levels of myogenic factors (Pax7, Myf5, Myod, Mef2a, Myogenin, and Myostatin) and a lack of multinucleated myotubes. These results demonstrate that there is a delicate temporal regulation of SPARC to which more sources in the micro environment contribute, and that disturbances in this, such as extensive up regulation, may have an adverse effect on muscle regeneration.
- PublicationOpen AccessSimultaneous phenotypic and genetic characterization of single circulating tumor cells from colon cancer patients(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Campos, María; Luque, Rafael; Jiménez, Juan; Martínez, Rafael; Warleta, Fernando; Sánchez-Quesada, Cristina; Delgado-Rodríguez, Miguel; Calvo, Alfonso; Gaforio, José J.Since circulating tumor cells (CTCs) have metastatic potential, their genetic and phenotypic characteristics could provide crucial information to establish the most effective therapy. We assessed the clinical utility of a methodology that allows the simultaneous analysis of CTC phenotype and genotype in colon cancer patients and, in addition, whether this methodology could provide complementary information to that obtained by the primary tumor biopsy. Thirty-three non-metastatic (stages 0-III) colon cancer patients and 9 healthy donor samples were evaluated. All peripheral blood samples (10 ml) were analyzed by cytokeratin immunomagnetic enrichment. Eight samples were analyzed by immunocytochemistry and 25 samples were analyzed by FICTION technique for simultaneous cytokeratin expression and chromosome 17 and ERBB2 gene status. A further study was carried out in one patient who showed CTC heterogeneity in chromosomal abnormalities. We analyzed HER2 protein expression on CTCs and FISH and HER2 protein expression in primary tumor of this patient. Our results show that 9.09% of patients had cytokeratin-positive CTCs (CK+/CTCs in peripheral blood). One of the patients showed heterogeneity in chromosomal 17 abnormalities and two different CK expression patterns on CTCs: one CK+/CTCs and one CK-/CTCs. Furthermore, 63.33% of these CTCs overexpressed HER2 protein while the primary tumor of this patient was diploid and did not express HER2 protein. We describe a methodology that allows the simultaneous genetic and phenotypic analysis of CTCs in colon cancer patients, which may provide essential information to select patients who might benefit from specific therapy.
- PublicationOpen AccessThe vertebrae of prematurely aging mice as a skeletal model of involutional osteoporosis(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Portal Núñez, Sergio; Cruces, Julia; Lozano, Daniel; Ardura, Juan Antonio; Villanueva-Peñacarrillo, María L.; Fuente, Mónica de la; Esbrit, PedroOxidative stress in bone increases with age, which leads to bone frailty and a high fracture risk. Animal models show that early changes in trabecular structure occur in age-related osteopenia. These models might be valuable to assess the contribution of oxidative stress in age-related bone loss. Premature aging mice (PAM) have previously been characterized as a model of premature immunological and neurological senescence. PAM long bones (mainly consisting of cortical bone) display features of aging bone. Thus, we aimed to evaluate the vertebrae, representing a unique poorly loaded type of trabecular bone in mice, in PAM and no PAM (NPAM) controls. PAM showed an anxious behaviour, based on physical activity evaluation. These mice had decreased bone mineral density (0.078 mg/cm2 in NPAM vs 0.070 g/cm2 in PAM; p<0.05); a decreased number of osteocytes per bone field (404±36 in NPAM vs 320±27 in PAM; p<0.01); and downregulation of various osteoblastic genes and low eroded surface/bone surface, 4.2±0.5 in NPAM vs 1.9±0.2 in PAM; p<0.01). This was associated with increased expression of oxidative stress markers, Foxo1 and GADD45, in PAM vertebrae. Mesenchymal progenitors in the bone marrow of PAM have a poor mineralization capacity (assessed by the number of mineralized nodules and suface), and showed a lower response to an osteogenic input - represented by parathormone-related protein-, compared to NPAM. Collectively, these results indicate that PAM vertebrae show osteopenia related to diminished bone formation and remodeling. Our findings further support the validity of PAM as a suitable model for involutional osteoporosis and its treatment.
- PublicationOpen AccessDistribution of zinc and zinc transporters in the mouse ovarian follicles and corpus luteum(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Zhong, Man-Li; Guo, Chuang; Chi, Zhi-Hong; Shan, Zhong-Yan; Teng, Wei-Ping; Wang, Zhan-YouZinc is essential for female reproduction and it plays a role in sexual development, ovulation, menstruation and estrous cycles. Zinc deficiency may lead to female reproductive system dysfunction. The present study aimed to investigate the expression and distribution patterns of free zinc and the members of zinc transporter (ZnT) family, with zinc autometallographic (AMG), immunohistochemistry and real-time PCR, to explore the relationship of zinc homeostasis in the development and function of the ovary in the mouse. Our data revealed that the free zinc ions and ZnTs are predominantly distributed in the mouse ovarian follicles and corpus luteum. Specifically, AMG staining presented in various stages of the ovarian follicles and corpus luteum. ZnT1-9 mRNA was variously expressed, whereas ZnT10 mRNA was almost undetectable in the ovary. Moreover, the immunoreactivity of all the tested ZnTs, except for ZnT10, was detected with various intensity in the mouse primordial follicles, primary follicles, secondary follicles and antral follicles. In the corpus luteum, the immunoreactivity of ZnT1-5, 7, 8, 10, was abundantly observed in the granular and theca lutein cells and interstitial cells. Collectively, our results suggest that ZnT family proteins are differently distributed and might exert different biological functions in controlling cellular zinc levels, which regulate ovarian development and function in the mouse ovary.
- PublicationOpen AccessRole of PRDM16 in the activation of brown fat programming. Relevance to the development of obesity(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Becerril, Sara; Gómez-Ambrosi, Javier; Martín, Marina; Moncada, Rafael; Sesma, Pilar; Burrell, María A.; Frühbeck, GemaFrom a histological and functional point of view, two types of adipose tissue can be identified. As opposed to the mainly unilocular white adipocytes, brown adipocytes possess plenty of small multilocular lipid droplets and dissipate energy as heat. Moreover, two distinct types of brown adipose cells exist. In vivo fate mapping experiments of brown adipose tissue (BAT) precursors suggest that classical brown adipocytes and skeletal myoblasts originate from a common mesenchymal, myogenic factor 5 (Myf5)-positive precursor cell. In addition to the classical brown adipocytes, thermogenic brown-like adipocytes (brite/beige cells) may appear within white adipose tissue (WAT) depots, sharing many of the morphological and functional features of brown adipocytes, but arising from a Myf5-negative lineage. In humans, the conversion of white fat cells into brite adipocytes could be a strategy to increase energy expenditure. The zinc finger transcription factor Prdm16 controls the bidirectional fate decision between brown adipocytes and myoblasts. Prdm16 determines the brown fat-like programme and thermogenesis in both brown and white adipose tissues. Moreover, the expression of this transcriptional regulator is strongly correlated with beige cell-selective genes. From a therapeutical point of view, the potential of inducing BAT or the transdifferentiation of WAT into beige cells by enhancing Prdm16 expression, as well as the identification of mechanisms of Prdm16 function and regulation represent potentially exciting new approaches for treatment or prevention of obesity and related diseases.
- PublicationOpen AccessLower expression of ER-α36 is associated with the development of endometrial hyperplasia in PCOS patients(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Lin, Sheng-Li; Tu, Bin-Bin; Du, Xiao-Guo; Yan, Li-Ying; Qiao, JieObjective: To explore the expression change of ER-α36 in endometria of PCOS patients. Methods: Proliferative endometria were collected and divided into three groups: CE group (n=30), proliferative endometria from control women; PCOSE group (n=30), proliferative endometria from PCOS patients; HPCOSE group (n=15), hyperplastic endometria from PCOS patients. The cellular localization of ER-α36 and ER-α66 was analyzed by immunohistochemistry, and expression of ER-α36 and ER-α66 was determined by immunohistochemistry and Real-Time Quantitative PCR. The correlation between serum hormone concentration and expression of ER-α36 and ER-α66 was analyzed. Results: ER-α36 was localized on the cell surface, cytoplasm and nucleus of glandular epithelial cells of both CE and PCOSE groups, mainly on the cell surface, and ER-α66 was localized on the nucleus. The protein and mRNA expression of ER-α36 was decreased from CE, PCOSE to HPCOSE group, while expression of ERα66 showed no obvious difference among groups. The expression ratio of ER-α36 to ER-α66 of CE, PCOSE and HPCOSE group showed a decreasing tendency. The expression of ER-α36 and its expressive ratio to ER-α66 was negatively correlated with serum concentration of LH, LH/FSH, testosterone and androstenedione. Conclusions: Lower expression of ER-α36 is associated with the development of endometrial hyperplasia in PCOS patients.
- PublicationOpen AccessApocrine carcinoma of the breast: A comprehensive review(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Vranic, Semir; Schmitt, Fernando; Sapino2, Anna; Costa, José Luis; Reddy, Sandeep; Castro, Michael; Gatalica5, ZoranApocrine carcinoma of the breast is a rare, special type of breast carcinoma showing distinct morphologic, immunohistochemical and molecular genetic features. Apocrine epithelium has a characteristic steroid receptor profile that is estrogen receptor and progesterone receptor negative and androgen receptor positive. This combination of morphologic and immunohistochemical characteristics is essential for the proper recognition of the apocrine carcinomas. Strictly defined, apocrine carcinomas express either Her-2/neu or EGFR, which along with androgen receptor positivity make patients with the apocrine carcinoma eligible for targeted therapies.
- PublicationOpen AccessSerum amyloid A-positive hepatocellular neoplasms in the resected livers from 3 patients with alcoholic cirrhosis(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Sasaki, Motoko; Kondo, Fukuo; Sawai, Yoshiyuki; Imai, Yasuharu; Kadowak, Susumu; Sano, Keiji; Fukusato, Toshio; Matsui, Osamu; Nakanuma, YasuniTwelve hepatocellular nodules were characterized in the resected livers from 3 patients (2 men and a woman) with alcoholic cirrhosis. Imaging techniques suggested that the nodules were hypervascular and may be hepatocellular carcinoma. Five nodules (4-31 mm in diameter) were serum amyloid A-positive hepatocellular neoplasm, which shares features with inflammatory hepatocellular adenoma. The remaining 7 nodules (5-8 mm) were focal nodular hyperplasia-like nodules showing focal or no immunostaining for serum amyloid A. The serum amyloid A-positive hepatocellular neoplasms showed increased cellular density, inflammatory infiltrate, sinusoidal dilatation, and ductular reaction to various degrees. These histologic features tended to be less extensive in focal nodular hyperplasia-like nodules. Three of 4 serum amyloid A-positive hepatocellular neoplasms showed slight hypointensity in the hepatobiliary phase on the magnetic resonance (MR) imaging with gadolinium ethoxybenzyl diethylenetriaminepentaacetic acid (Gd-EOB-DTPA) enhancement. In contrast, 3 focal nodular hyperplasia-like nodules showed iso-intensity in the hepatobiliary phase. This study further confirms characteristics of serum amyloid A-positive hepatocellular neoplasm arising in alcoholic cirrhosis that share features with inflammatory hepatocellular adenomas. Serum amyloid A-positive hepatocellular neoplasms sometimes co-exist with focal nodular hyperplasia-like nodules and may show different findings on Gd-EOB-enhanced MR imaging.
- PublicationOpen AccessDifferential expression of Yes-associated protein and phosphorylated Yes-associated protein is correlated with expression of Ki-67 and phospho-ERK in colorectal adenocarcinoma(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Kim, Dong-Hoon; Kim, Seok-Hyung; Lee, Ok-Jun; Huang, Song-Mei; Kwon, Ju-Lee; Kim, Jin Man; Kim, Ji-Yeon; Seong, In Ock; Song, Kyu Sang; Kim, Kyung HeeYes-associated protein (YAP) is a transcriptional co-activator and functions as a nuclear downstream effector of the Hippo pathway. Differential expression of YAP and phosphorylated Yes-associated protein (pYAP), which are involved in the expression of Ki-67 and phosphorylated extracellular signal-regulated kinase (pERK) in colorectal adenocarcinoma (CRAC), is not clear. Herein, we hypothesized that nuclear expression of YAP could predict cell proliferation and poor prognosis, while cytoplasmic expression of pYAP would show a reverse correlation with cell proliferation. Paraffin-embedded samples from 144 CRAC patients were studied using immunohistochemistry for YAP, pYAP, Ki-67 and pERK. Frozen samples from 20 CRAC patients were examined for YAP mRNA in tumor and non-tumor tissues, using quantitative real-time PCR. High nuclear YAP expression coincided with high Ki-67 expression (P=0.002). The high nuclear YAP expression group tended to display a poor overall and disease-free survival (P=0.089 and P=0.089, respectively), but YAP mRNA levels in the 20 CRAC tissues were not significantly different in comparison with the 20 nontumor tissues (P=0.929). We observed an inverse correlation between high cytoplasmic pYAP expression and high Ki-67 expression (P=0.001). Nuclear pERK expression was positively correlated with nuclear YAP expression, but negatively correlated with cytoplasmic pYAP expression (P=0.017 and P=0.020, respectively). Activated nuclear YAP and inactivated cytoplasmic pYAP in CRAC showed a positive correlation with Ki67 and nuclear pERK expression, suggesting that the expression of YAP and pYAP is a possible predictor of tumor cell proliferation and prognosis in CRAC.
- PublicationOpen AccessClinicopathologic characteristics of STAT1 positive/interleukin-8 negative subgroup in triple negative breast cancer defined by surrogate immunohistochemistry(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Kim, Sewha; Kim, Do Hee; Jung, Woo-Hee; Koo, Ja SeungBackground: The aim of this study was to define immune-related triple negative breast cancer (TNBC) using immunohistochemistry for STAT1, CD20, CD3, IL-8, and IFN-γ and to assess its clinicopathologic characteristics. Material and methods: Tissues from 133 cases of TNBC were used for a tissue microarray. Expression of STAT1, CD20, CD3, IL-8, and IFN-γ were evaluated by immunohistochemical staining of the tissue microarrays. Immune-related type was defined as TNBC which was positive for STAT1 and negative for IL-8. A separate assessment of IL-8 and STAT1 status in tumor and stroma compartment was used to further classify immune-related type into tumor-based and stroma-based immune-related TNBC. Results: Stroma-based, immune-related TNBC showed a significantly smaller central acellular zone (p=0.043), more lymphocytic infiltration (p<0.001), higher CD20 index (p=0.001), and higher CD3 index (p=0.018) than stroma-based, non-immune-related TNBC. IL-8 was independently associated with shorter disease-free survival (Hazard ratio: 3.804, 95% CI: 1.234-11.729, p=0.020) and shorter overall survival (Hazard ratio: 3.434, 95% CI: 1.132-10.414, p=0.029). Conclusions: Immune-related proteins such as STAT1, IFN-γ, IL-8, and CD20 were variably expressed in TNBCs. Stroma-based, immune-related TNBC (when positive for stromal STAT1 and negative for stromal IL-8) showed significantly higher lymphocytic infiltration including both CD3 positive T cell and CD20 positive B cell.