The regulation of aromatase and androgen receptor expression during gonad development in male and female European eel

[EN] This research investigated the regulation of aromatase and androgen receptor gene expression in the brain–pituitary– gonad (BPG) axis of male and female European eels (Anguilla anguilla) during induced sexual maturation. Complete A. anguilla aromatase (aa-cyp19a1) and partial androgen receptor a and b (aa-ara and aa-arb) sequences were isolated, and qPCR assays were validated and used for quantification of transcript levels for these three genes. Expression levels of the genes varied with sex, tissue and stage of maturation. aa-arb was expressed at higher levels than aa-ara in the pituitary and gonad in both sexes, suggesting aa-arb is the physiologically most important androgen receptor in these tissues. In the female brain, a decrease in aa-ara and an increase in aa-cyp19a1 were observed at the vitellogenic stage. In contrast, a progressive increase in all three genes was observed in the pituitary and ovaries throughout gonadal development, with aa-arb and aa-cyp19a1 reaching significantly higher levels at the vitellogenic stage. In the male pituitary, a decrease in aa-arb and an increase in aa-cyp19a1 were observed at the beginning of spermatogenesis, and thereafter remained low and high, respectively. In the testis, the transcript levels of androgen receptors and aa-cyp19a1 were higher during the early stages of spermatogenesis and decreased thereafter. These sex-dependent differences in the regulation of the expression of aa-ara, aa-arb and cyp19a1 are discussed in relation to the role of androgens and their potential aromatization in the European eel during gonadal maturationThis work was funded by the European Community's 7th Framework Programme under the Theme 2 'Food, Agriculture and Fisheries, and Biotechnology', grant agreement no 245257 (PRO-EEL). D. S. P. received a postdoc grant from UPV (CEI-01-10), a mobility grant from UPV (PAID-00-11) and has also been supported by a contract cofinanced by MICINN and UPV (PTA2011-4948-I). V. G. and I. M. received predoctoral grants from the Spanish Ministry of Science and Innovation (MICINN) and Generalitat Valenciana, respectively. F.-A. W. received funding from the Norwegian School of Veterinary Science. The fish farm Valenciana de Acuicultura, S. A. supplied the male eels used in the experiments. The English revision was carried out by Professor Lucy Robertson (Lucy Robertson Writing Services, Norway).Peñaranda, D.; Mazzeo, I.; Gallego Albiach, V.; Hildahl, J.; Nourizadeh-Lillabadi, R.; Pérez Igualada, LM.; Weltzien, FA.... (2014). The regulation of aromatase and androgen receptor expression during gonad development in male and female European eel. Reproduction in Domestic Animals. 49(3):512-521. https://doi.org/10.1111/rda.12321512521493Aroua, S., Weltzien, F.-A., Belle, N. L., & Dufour, S. 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Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology, 109(3), 219-245. doi:10.1016/0742-8413(94)00063-gChang, X. T., Kobayashi, T., Kajiura, H., Nakamura, M., & Nagahama, Y. (1997). Isolation and characterization of the cDNA encoding the tilapia (Oreochromis niloticus) cytochrome P450 aromatase (P450arom): changes in P450arom mRNA, protein and enzyme activity in ovarian follicles during oogenesis. Journal of Molecular Endocrinology, 18(1), 57-66. doi:10.1677/jme.0.0180057Choi, J. Y., Park, J. G., Jeong, H. B., Lee, Y. D., Takemura, A., & Kim, S. J. (2005). Molecular cloning of cytochrome P450 aromatases in the protogynous wrasse, Halichoeres tenuispinis. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 141(1), 49-59. doi:10.1016/j.cbpc.2005.01.009Diotel, N., Page, Y. L., Mouriec, K., Tong, S.-K., Pellegrini, E., Vaillant, C., … Kah, O. (2010). 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General and Comparative Endocrinology, 68(1), 91-103. doi:10.1016/0016-6480(87)90064-5Kishida, M., & Callard, G. V. (2001). Distinct Cytochrome P450 Aromatase Isoforms in Zebrafish (Danio rerio) Brain and Ovary Are Differentially Programmed and Estrogen Regulated during Early Development**This research was supported by grants from the National Science Foundation (IBN-96-05053) and the NIH (P42 ES-07381). The nucleotide sequences reported in this paper have been submitted to the GenBank/EMBL Data Bank with accession numbers AF226619 and AF226620. Endocrinology, 142(2), 740-750. doi:10.1210/endo.142.2.7928Kwon, J. Y., McAndrew, B. J., & Penman, D. J. (2001). Cloning of brain aromatase gene and expression of brain and ovarian aromatase genes during sexual differentiation in genetic male and female Nile tilapiaOreochromis niloticus. Molecular Reproduction and Development, 59(4), 359-370. doi:10.1002/mrd.1042Levavi-Sivan, B., Biran, J., & Fireman, E. (2006). Sex Steroids Are Involved in the Regulation of Gonadotropin-Releasing Hormone and Dopamine D2 Receptors in Female Tilapia Pituitary1. Biology of Reproduction, 75(4), 642-650. doi:10.1095/biolreprod.106.051540Lin, C.-J., Wu, G.-C., Lee, M.-F., Lau, E.-L., Dufour, S., & Chang, C.-F. (2010). Regulation of two forms of gonadotropin-releasing hormone receptor gene expression in the protandrous black porgy fish, Acanthopagrus schlegeli. Molecular and Cellular Endocrinology, 323(2), 137-146. doi:10.1016/j.mce.2010.04.003Liu, X., Su, H., Zhu, P., Zhang, Y., Huang, J., & Lin, H. (2009). Molecular cloning, characterization and expression pattern of androgen receptor in Spinibarbus denticulatus. General and Comparative Endocrinology, 160(1), 93-101. doi:10.1016/j.ygcen.2008.10.026Lokman, P. M., George, K. A. N., & Young, G. (2003). Effects of steroid and peptide hormones on in vitro growth of previtellogenic oocytes from eel, Anguilla australis. Fish Physiology and Biochemistry, 28(1-4), 283-285. doi:10.1023/b:fish.0000030556.34592.41Lokman, P. M., George, K. A. N., Divers, S. L., Algie, M., & Young, G. (2007). 11-Ketotestosterone and IGF-I increase the size of previtellogenic oocytes from shortfinned eel, Anguilla australis, in vitro. Reproduction, 133(5), 955-967. doi:10.1530/rep-06-0229MATSUBARA, H., KAZETO, Y., IJIRI, S., HIRAI, T., ADACHI, S., & YAMAUCHI, K. (2003). Changes in mRNA levels of ovarian steroidogenic enzymes during artificial maturation of Japanese eel Anguilla japonica. Fisheries Science, 69(5), 979-988. doi:10.1046/j.1444-2906.2003.00716.xMatsubara, M., Lokman, P. M., Senaha, A., Kazeto, Y., Ijiri, S., Kambegawa, A., … Yamauchi, K. (2003). Synthesis and possible function of 11-ketotestosterone during oogenesis in eel (Anguilla spp.). Fish Physiology and Biochemistry, 28(1-4), 353-354. doi:10.1023/b:fish.0000030585.22093.7aMazzeo, I., Peñaranda, D. S., Gallego, V., Hildahl, J., Nourizadeh-Lillabadi, R., Asturiano, J. F., … Weltzien, F.-A. (2012). Variations in the gene expression of zona pellucida proteins, zpb and zpc, in female European eel (Anguilla anguilla) during induced sexual maturation. General and Comparative Endocrinology, 178(2), 338-346. doi:10.1016/j.ygcen.2012.06.003Miura, T., Yamauchi, K., Takahashi, H., & Nagahama, Y. (1991). Hormonal induction of all stages of spermatogenesis in vitro in the male Japanese eel (Anguilla japonica). Proceedings of the National Academy of Sciences, 88(13), 5774-5778. doi:10.1073/pnas.88.13.5774Miura, T., Miura, C., Ohta, T., Nader, M. R., Todo, T., & Yamauchi, K. (1999). Estradiol-17β Stimulates the Renewal of Spermatogonial Stem Cells in Males. Biochemical and Biophysical Research Communications, 264(1), 230-234. doi:10.1006/bbrc.1999.1494Montero, M., Le Belle, N., King, J. A., Millar, R. P., & Dufour, S. (1995). 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Rosen's (M,R) system in Unified Modelling Language

Robert Rosen's (M,R) system is an abstract biological network architecture that is allegedly non-computable on a Turing machine. If (M,R) is truly non-computable, there are serious implications for the modelling of large biological networks in computer software. A body of work has now accumulated addressing Rosen's claim concerning (M,R) by attempting to instantiate it in various software systems. However, a conclusive refutation has remained elusive, principally since none of the attempts to date have unambiguously avoided the critique that they have altered the properties of (M,R) in the coding process, producing merely approximate simulations of (M,R) rather than true computational models. In this paper, we use the Unified Modelling Language (UML), a diagrammatic notation standard, to express (M,R) as a system of objects having attributes, functions and relations. We believe that this instantiates (M,R) in such a way than none of the original properties of the system are corrupted in the process. Crucially, we demonstrate that (M,R) as classically represented in the relational biology literature is implicitly a UML communication diagram. Furthermore, since UML is formally compatible with object-oriented computing languages, instantiation of (M,R) in UML strongly implies its computability in object-oriented coding languages

Foundations in Wisconsin: A Directory [32nd ed. 2013]

The 2013 edition of Foundations in Wisconsin marks the 32nd production of the print directory and the 13th year of the online version. The directory is designed as a research tool for grantseekers interested in locating information on private, corporate, and community foundations registered in Wisconsin. Each entry in this new edition has been updated or reviewed to provide the most current information available. Most of the data was drawn from IRS 990-PF tax returns filed by the foundations. Additional information was obtained from surveys, foundation websites, annual reports, and newsletters.https://epublications.marquette.edu/lib_fiw/1011/thumbnail.jp

Foundations in Wisconsin: A Directory [28th ed. 2009]

The 2009 edition of Foundations in Wisconsin marks the 28th production of the print directory and the 9th year of the online version. The directory is designed as a research tool for grantseekers interested in locating information on private, corporate, and community foundations registered in Wisconsin. Each entry in this new edition has been updated or reviewed to provide the most current information available. Most of the data was drawn from IRS 990-PF tax returns filed by the foundations. However, additional information was obtained from surveys, foundation Web sites, annual reports, and newsletters. Wisconsin foundations have continued to grow in key areas even with the economic downturn. Active grantmaking foundations now number 1,286, with 54 new foundations identified. Total grants increased to an all-time high of $507 million, a 7$6.8 billion.https://epublications.marquette.edu/lib_fiw/1000/thumbnail.jp

Foundations in Wisconsin: A Directory [35th ed. 2016]

The 2016 release of Foundations in Wisconsin marks the 35th edition of the print directory and the 16th edition of the online version. The directory is designed as a research tool for grantseekers interested in locating information on private, corporate, and community foundations registered in Wisconsin. Each entry in this new edition has been updated or reviewed to provide the most current information available. Most of the data was drawn from IRS 990-PF tax returns filed by the foundations. Additional information was obtained from surveys, foundation websites, and annual reports. This edition paints a very positive picture of financial growth for Wisconsin foundations. Both grant and asset totals have risen to all-time highs. Of particular note, total grants broke the $600 million barrier, increasing by 8$623 million. Additionally, 58 new foundations have been identified this year. (See page 269 for the complete list.) The following table illustrates the 10-year financial pattern as documented in Foundations in Wisconsin.https://epublications.marquette.edu/lib_fiw/1014/thumbnail.jp

Overexpression of the nitroreductase NfsB in an E. coli strain as a whole-cell biocatalyst for the production of chlorinated analogues of the natural herbicide DIBOA

Los ácidos benzohidroxámicos, como el DIBOA (2,4-dihidroxi-2 H)-1,4-benzoxazin-3(4 H)-ona), son productos vegetales que presentan interesantes propiedades herbicidas, fungicidas y bactericidas. Una alternativa viable a su purificación a partir de fuentes naturales es la síntesis de compuestos análogos como el D-DIBOA (2-deoxi-DIBOA) y sus derivados clorados. Su síntesis química se ha simplificado en dos pasos. Sin embargo, el segundo paso es una reacción exotérmica e implica la liberación de hidrógeno, lo que hace que esta metodología sea cara y difícil de ampliar. En el presente estudio se estudia la posibilidad de producir los derivados clorados de las benzoxazinonas mediante un proceso biocatalítico in vivo que utiliza la capacidad de la cepa de E. coli nfsB-/pBAD-NfsB modificada para catalizar la síntesis de 6-Cl- D-DIBOA y 8-Cl- D-DIBOA a partir de sus respectivos precursores (PCs). Los resultados muestran que esta cepa es capaz de crecer en medios que contienen estos compuestos y producir el producto de biotransformación con unos rendimientos del 59,3% y el 46,7%. respectivamente. Además, la cepa es capaz de procesarel precursor (PC) no purificados del primer paso químico para obtener rendimientos similares a los obtenidos a partir del PC purificado. Se estudió la cinética de la reacción in in vitro con la nitrorreductasa NfsB recombinante purificada para caracterizar la catálisis y evaluar los efectos que varios componentes del precursor no purificado tiene durante el proceso de síntesis. Los resultados revelaron que la cinética es la de una enzima alostérica. El efecto inhibitorio del sustrato en el primer paso de la síntesis química, se encuentra también en algunas etapas previas a la purificación del precursor

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