Identification and stable expression of vitellogenin receptor (VTGR) through vitellogenesis in the European eel

[EN] In teleosts, vitellogenin (Vtg) is a phospholipoglycoprotein synthesized by the liver, released into the blood circulation and incorporated into the oocytes via endocytosis mediated by the Vtg receptor (VTGR) to form the yolk granules. The VTGR is crucial for oocyte growth in egg-laying animals but is also present in non-oviparous vertebrates, such as human. The VTGR belongs to the low-density lipoprotein receptor superfamily (LDLR) and is also named very-low-density lipoprotein receptor (VLDLR). In this study, we identified and phylogenetically positioned the VTGR of a basal teleost, the European eel, Anguilla anguilla. We developed quantitative real-time PCR (qRT-PCR) and investigated the tissue distribution of vtgr transcripts. We compared by qRT-PCR the ovarian expression levels of vtgr in juvenile yellow eels and pre-pubertal silver eels. We also analyzed the regulation of ovarian vtgr expression throughout vitellogenesis in experimentally matured eels. The Vtg plasma level was measured by homologous ELISA experimental maturation. Our in silico search and phylogenetical analysis revealed a single vtgr in the European eel, orthologous to other vertebrate vtgr. The qRT-PCR studies revealed that vtgr is mainly expressed in the ovary and also detected in various other tissues such as brain, pituitary, gill, fat, heart, and testis, suggesting some extra-ovarian functions of VTGR. We showed that vtgr is expressed in ovaries of juvenile yellow eels with no higher expression in pre-pubertal silver eels nor in experimentally matured eels. This suggests that vtgr transcription already occurs during early pre-vitellogenesis of immature eels and is not further activated in vitellogenic oocytes. European eel Vtg plasma level increased throughout experimental maturation in agreement with previous studies. Taken together, these results suggest that vtgr transcript levels may not be a limiting step for the uptake of Vtg by the oocyte in the European eel.M.M., A.G.L. and S.D. were granted with Short Term Scientific Missions by the COST Office (COST Action FA1205: AQUAGAMETE). S.D. was also awarded with a grant from the UPV's School of Doctorate (Accion para la Internacionalizacion de los Programas de Doctorado) in 2017. We thank E. Feunteun and colleagues, MNHN, Dinard, for the ovarian samples from eels of the Fremur, France.Morini, M.; Lafont, AG.; Maugars, G.; Baloche, S.; Dufour, S.; Asturiano, JF.; Pérez Igualada, LM. (2020). Identification and stable expression of vitellogenin receptor (VTGR) through vitellogenesis in the European eel. Animal. 14(6):1213-1222. https://doi.org/10.1017/S1751731119003355S12131222146Abascal, F., Zardoya, R., & Posada, D. (2005). ProtTest: selection of best-fit models of protein evolution. Bioinformatics, 21(9), 2104-2105. doi:10.1093/bioinformatics/bti263Ali, B. R., Silhavy, J. L., Gleeson, M. J., Gleeson, J. G., & Al-Gazali, L. (2012). A missense founder mutation in VLDLR is associated with Dysequilibrium Syndrome without quadrupedal locomotion. BMC Medical Genetics, 13(1). doi:10.1186/1471-2350-13-80Andersen, Ø., Xu, C., Timmerhaus, G., Kirste, K. H., Naeve, I., Mommens, M., & Tveiten, H. (2017). Resolving the complexity of vitellogenins and their receptors in the tetraploid Atlantic salmon (Salmo salar ): Ancient origin of the phosvitin-less VtgC in chondrichthyean fishes. Molecular Reproduction and Development, 84(11), 1191-1202. doi:10.1002/mrd.22881Bidwell, C., & Carlson, D. (1995). Characterization of vitellogenin from white sturgeon, Acipenser transmontanus. Journal of Molecular Evolution, 41(1). doi:10.1007/bf00174046Bujo, H., Hermann, M., Kaderli, M. O., Jacobsen, L., Sugawara, S., Nimpf, J., … Schneider, W. J. (1994). Chicken oocyte growth is mediated by an eight ligand binding repeat member of the LDL receptor family. The EMBO Journal, 13(21), 5165-5175. doi:10.1002/j.1460-2075.1994.tb06847.xBurzawa-Gerard, E., & Dumas-Vidal, A. (1991). Effects of 17β-estradiol and carp gonadotropin on vitellogenesis in normal and hypophysectomized European silver female eel (Anguilla anguilla L.) employing a homologous radioimmunoassay for vitellogenin. General and Comparative Endocrinology, 84(2), 264-276. doi:10.1016/0016-6480(91)90049-cChen, J.-N., López, J. A., Lavoué, S., Miya, M., & Chen, W.-J. (2014). Phylogeny of the Elopomorpha (Teleostei): Evidence from six nuclear and mitochondrial markers. Molecular Phylogenetics and Evolution, 70, 152-161. doi:10.1016/j.ympev.2013.09.002Clelland, E. S., & Kelly, S. P. (2010). Tight junction proteins in zebrafish ovarian follicles: Stage specific mRNA abundance and response to 17β-estradiol, human chorionic gonadotropin, and maturation inducing hormone. General and Comparative Endocrinology, 168(3), 388-400. doi:10.1016/j.ygcen.2010.05.011Damsteegt, E. L., Mizuta, H., Hiramatsu, N., & Lokman, P. M. (2015). How do eggs get fat? Insights into ovarian fatty acid accumulation in the shortfinned eel, Anguilla australis. General and Comparative Endocrinology, 221, 94-100. doi:10.1016/j.ygcen.2014.12.019Dufour S, Burzawa-Gerard E, Le Belle N, Sbaihi M and Vidal B 2003. Reproductive endocrinology of the European eel, Anguilla anguilla. In Eel biology (eds. K Aida, K Tsukamoto and K Yamauchi ), pp 373–383. Springer-Verlag, Tokyo, Japan.Dufour, S., Lopez, E., Le Menn, F., Le Belle, N., Baloche, S., & Fontaine, Y. A. (1988). Stimulation of gonadotropin release and of ovarian development, by the administration of a gonadoliberin agonist and of dopamine antagonists, in female silver eel pretreated with estradiol. General and Comparative Endocrinology, 70(1), 20-30. doi:10.1016/0016-6480(88)90090-1Henkel, C. V., Burgerhout, E., de Wijze, D. L., Dirks, R. P., Minegishi, Y., Jansen, H. J., … van den Thillart, G. E. E. J. M. (2012). Primitive Duplicate Hox Clusters in the European Eel’s Genome. PLoS ONE, 7(2), e32231. doi:10.1371/journal.pone.0032231Henkel, C. V., Dirks, R. P., de Wijze, D. L., Minegishi, Y., Aoyama, J., Jansen, H. J., … van den Thillart, G. E. E. J. M. (2012). First draft genome sequence of the Japanese eel, Anguilla japonica. Gene, 511(2), 195-201. doi:10.1016/j.gene.2012.09.064Herz, J., & Bock, H. H. (2002). Lipoprotein Receptors in the Nervous System. Annual Review of Biochemistry, 71(1), 405-434. doi:10.1146/annurev.biochem.71.110601.135342Hiramatsu, N., Luo, W., Reading, B. J., Sullivan, C. V., Mizuta, H., Ryu, Y.-W., … Hara, A. (2012). Multiple ovarian lipoprotein receptors in teleosts. Fish Physiology and Biochemistry, 39(1), 29-32. doi:10.1007/s10695-012-9612-6Hiramatsu, N., Todo, T., Sullivan, C. V., Schilling, J., Reading, B. J., Matsubara, T., … Hara, A. (2015). Ovarian yolk formation in fishes: Molecular mechanisms underlying formation of lipid droplets and vitellogenin-derived yolk proteins. General and Comparative Endocrinology, 221, 9-15. doi:10.1016/j.ygcen.2015.01.025Hummel, S., Lynn, E. G., Osanger, A., Hirayama, S., Nimpf, J., & Schneider, W. J. (2003). Molecular characterization of the first avian LDL receptor. Journal of Lipid Research, 44(9), 1633-1642. doi:10.1194/jlr.m300014-jlr200Jéhannet P, Kruijt L, Damsteegt EL, Swinkels W, Heinsbroek LTN, Lokman PM and Palstra AP. A mechanistic model for studying the initiation of anguillid vitellogenesis by comparing the European eel (Anguilla anguilla) and the shortfinned eel (A. australis). General and Comparative Endocrinology (in press). https://doi.org/10.1016/j.ygcen.2019.02.018Lafont, A.-G., Rousseau, K., Tomkiewicz, J., & Dufour, S. (2016). Three nuclear and two membrane estrogen receptors in basal teleosts, Anguilla sp.: Identification, evolutionary history and differential expression regulation. General and Comparative Endocrinology, 235, 177-191. doi:10.1016/j.ygcen.2015.11.021Mazzeo, I., Peñaranda, D. S., Gallego, V., Baloche, S., Nourizadeh-Lillabadi, R., Tveiten, H., … Pérez, L. (2014). Temperature modulates the progression of vitellogenesis in the European eel. Aquaculture, 434, 38-47. doi:10.1016/j.aquaculture.2014.07.020Mizuta, H., Luo, W., Ito, Y., Mushirobira, Y., Todo, T., Hara, A., … Hiramatsu, N. (2013). Ovarian expression and localization of a vitellogenin receptor with eight ligand binding repeats in the cutthroat trout (Oncorhynchus clarki). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 166(1), 81-90. doi:10.1016/j.cbpb.2013.07.005Mizuta, H., Mushirobira, Y., Nagata, J., Todo, T., Hara, A., Reading, B. J., … Hiramatsu, N. (2017). Ovarian expression and localization of clathrin (Cltc) components in cutthroat trout, Oncorhynchus clarki: Evidence for Cltc involvement in endocytosis of vitellogenin during oocyte growth. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 212, 24-34. doi:10.1016/j.cbpa.2017.06.021Morini, M., Peñaranda, D. S., Vílchez, M. C., Gallego, V., Nourizadeh-Lillabadi, R., Asturiano, J. F., … Pérez, L. (2015). Transcript levels of the soluble sperm factor protein phospholipase C zeta 1 (PLCζ1) increase through induced spermatogenesis in European eel. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 187, 168-176. doi:10.1016/j.cbpa.2015.05.028Morini, M., Peñaranda, D. S., Vílchez, M. C., Nourizadeh-Lillabadi, R., Lafont, A.-G., Dufour, S., … Pérez, L. (2017). Nuclear and membrane progestin receptors in the European eel: Characterization and expression in vivo through spermatogenesis. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 207, 79-92. doi:10.1016/j.cbpa.2017.02.009Nader, N., Dib, M., Courjaret, R., Hodeify, R., Machaca, R., Graumann, J., & Machaca, K. (2018). VLDL receptor regulates membrane progesterone receptor trafficking and non-genomic signaling. Journal of Cell Science. doi:10.1242/jcs.212522Okabayashi, K., Shoji, H., Nakamura, T., Hashimoto, O., Asashima, M., & Sugino, H. (1996). cDNA Cloning and Expression of theXenopus laevisVitellogenin Receptor. Biochemical and Biophysical Research Communications, 224(2), 406-413. doi:10.1006/bbrc.1996.1040Palstra, A. P., & van den Thillart, G. E. E. J. M. (2010). Swimming physiology of European silver eels (Anguilla anguilla L.): energetic costs and effects on sexual maturation and reproduction. Fish Physiology and Biochemistry, 36(3), 297-322. doi:10.1007/s10695-010-9397-4Pankhurst, N. W. (1982). Relation of visual changes to the onset of sexual maturation in the European eel Anguilla anguilla (L.). Journal of Fish Biology, 21(2), 127-140. doi:10.1111/j.1095-8649.1982.tb03994.xPasquier, J., Lafont, A.-G., Jeng, S.-R., Morini, M., Dirks, R., van den Thillart, G., … Dufour, S. (2012). Multiple Kisspeptin Receptors in Early Osteichthyans Provide New Insights into the Evolution of This Receptor Family. PLoS ONE, 7(11), e48931. doi:10.1371/journal.pone.0048931Perez, L., Aturiano, J. F., Tomas, A., Zegrari, S., Barrera, R., Espinos, F. J., … Jover, M. (2000). Induction of maturation and spermiation in the male European eel: assessment of sperm quality throughout treatment. Journal of Fish Biology, 57(6), 1488-1504. doi:10.1111/j.1095-8649.2000.tb02227.xPérez L, Vílchez MC, Gallego V, Mazzeo I, Peñaranda DS, Weltzien FA, Dufour S and Asturiano JF 2012. Trying to reproduce the European eel (Anguilla anguilla) under captivity: experiments with females. In Proceeding of the Domestication in Finfish Aquaculture, October 2012, Olsztyn, Poland, pp. 11–15.Prat, F., Coward, K., Sumpter, J. P., & Tyler, C. R. (1998). Molecular Characterization and Expression of two Ovarian Lipoprotein Receptors in the Rainbow Trout, Oncorhynchus mykiss 1. Biology of Reproduction, 58(5), 1146-1153. doi:10.1095/biolreprod58.5.1146Reading, B. J., Hiramatsu, N., Schilling, J., Molloy, K. T., Glassbrook, N., Mizuta, H., … Sullivan, C. V. (2014). Lrp13 is a novel vertebrate lipoprotein receptor that binds vitellogenins in teleost fishes. Journal of Lipid Research, 55(11), 2287-2295. doi:10.1194/jlr.m050286Stamatakis, A. (2014). RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics, 30(9), 1312-1313. doi:10.1093/bioinformatics/btu033Takahashi, S., Kawarabayasi, Y., Nakai, T., Sakai, J., & Yamamoto, T. (1992). Rabbit very low density lipoprotein receptor: a low density lipoprotein receptor-like protein with distinct ligand specificity. Proceedings of the National Academy of Sciences, 89(19), 9252-9256. doi:10.1073/pnas.89.19.9252Thompson, J. D., Higgins, D. G., & Gibson, T. J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research, 22(22), 4673-4680. doi:10.1093/nar/22.22.4673Trommsdorff, M., Gotthardt, M., Hiesberger, T., Shelton, J., Stockinger, W., Nimpf, J., … Herz, J. (1999). Reeler/Disabled-like Disruption of Neuronal Migration in Knockout Mice Lacking the VLDL Receptor and ApoE Receptor 2. Cell, 97(6), 689-701. doi:10.1016/s0092-8674(00)80782-5Tyler, C. R., Pottinger, T. G., Coward, K., Prat, F., Beresford, N., & Maddix, S. (1997). Salmonid Follicle-Stimulating Hormone (GtH I) Mediates Vitellogenic Development of Oocytes in the Rainbow Trout, Oncorhynchus mykiss 1. Biology of Reproduction, 57(5), 1238-1244. doi:10.1095/biolreprod57.5.1238Weltzien, F.-A., Pasqualini, C., Vernier, P., & Dufour, S. (2005). A quantitative real-time RT-PCR assay for European eel tyrosine hydroxylase. General and Comparative Endocrinology, 142(1-2), 134-142. doi:10.1016/j.ygcen.2004.12.01

Etude immunocytochimique des systèmes à GnRH dans le cerveau et l'hypophyse d'anguilles européennes normales et traitées à l'hCG

International audienc

Immunocytochemical study of GnRH systemes in the brain and pituitary of normal and hCG treated European Eels.

International audienc

The GnRH systems in the brain and pituitary of normal and hCG treated European silver eels

The distribution of immunoreactive GnRH was studied in the brain and pituitary gland of normal and human chorionic gonadotrophin (hCG) injected silver eels. It was found that the general organization of GnRH systems in this species is similar to that reported in other teleosts. Cell bodies were present in the olfactory bulbs, ventral telencephalon, periventricular hypothalamus and dorsal tegmentum. No positive perikarya could be detected in the preoptic region. Only scarce fibers were observed in the proximal neurohypophysis. Treatment with hCG does not modify the distribution of GnRH but it increases the density of positive structures, in particular at the level of the pituitary. The results are discussed in relation with the present status of knowledge of the mechanisms underlying the blockage of sexual maturation in the European eel at the silver stage

Life + Alosa alosa Rapport Irstea 2 : Actions C1 - D7 Juillet 2012-Juin 2013

[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASARE [Coll_IRSTEA]Irstea, groupement de Bordeaux, étude n°157This report presents works managed by Irstea, Aquarium La Rochelle and Borea Team concerning the set up of an ex situ rearing method for juveniles Allis shads Alosa alosa, and the monitoring of sexual maturation. Fish were moved to a larger tank in September 2012, which probably improve growth. Mortality has been stabilized. Histological analysis of samples collected in June 2012 showed a normal gonadal development in 50% of the samples. A second set of samplings for the monitoring of sexual maturation was carried out in June 2013.Ce rapport présente les travaux conduits par Irstea, l'Aquarium de La Rochelle et l’équipe Borea dans le cadre de la mise au point d’une méthode d’élevage ex situ de juvéniles de grande alose Alosa alosa et du suivi de la maturation sexuelle. Les poissons ont été transférés dans plus grand bac en septembre 2012, ce qui a probablement permis d’améliorer la croissance. La mortalité a été stabilisée. Les analyses histologiques des prélèvements effectués en juin 2012 ont montré un développement normal des gonades dans 50 % des échantillons. Une seconde série de prélèvements a été réalisée en Juin 2013

Life + Alosa alosa Rapport intermédiaire 1 Irstea

[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASARE [Coll_IRSTEA]Irstea, groupement de Bordeaux, étude n°151This report presents works managed by Irstea, Aquarium La Rochelle and Borea Team concerning the set up of an ex situ rearing method for juveniles Allis shads Alosa alosa, and the monitoring of the sexual maturation. Due to incidental loss of the 2008 batch, first samplings carried out in June 2012 were focused on juveniles from the 2011 batch. Considering their age (one year old) the objective of this first sampling was to start histological monitoring of gonads.Ce rapport présente les travaux menés par Irstea, l'Aquarium La Rochelle, et l'équipe Borea dans le cadre de la mise au point d'une méthode d'élevage ex situ de juvéniles de grande alose Alosa alosa, et de suivre l'évolution de la maturation sexuelle dans ces conditions. Du fait de la perte accidentelle du lot 2008, les premiers suivis réalisés en juin 2012 ont été réalisés sur les juvéniles du lot 2011. Considérant leur âge (1 an) l'objectif de cette première intervention avait pour but de commencer le suivi histologique des gonades

Stock captif de grandes aloses à Aquarium La Rochelle - résultats de l'élevage et de la maturation sexuelle

[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASAREInternational audienceThe final objective of this action is to reduce the amount of adults collected for larvae production. In the framework of this action, a batch of allis shad larvae was reared in Aquarium La Rochelle facilities to test the feasibility and to follow the sexual maturation process. Mortality was high during the first 4 months, and the survival rate was stabilized at 22%. At the end of the experiment survival was 12%. From 1 month old larvae, the feeding sequence was made of artificial feed and frozen zooplankton. After a 4 year rearing, mean total length was 35 ± 1 cm, and mean weight was 387 ± 34 g. Two main pathologies were observed: mouth malformation during the first weeks of rearing and renal nodules in 3 years old fish. In June 2015 the first mature fish were observed (males). The first allis shad ex situ rearing from larvae until 4 years old fish has been realized in Aquarium La Rochelle. Some recommendations are listed.Dans l’optique de minimiser les prélèvements de géniteurs pour réaliser des productions de larves destinées au repeuplement, une action C1 Ex situ stock a été développée dans le cadre du projet Life+ Alosa alosa. Dans le cadre de cette action un lot de larves a été élevé pendant 4 ans à l’Aquarium La Rochelle, pour déterminer les particularités de cet élevage et le suivi de la maturation sexuelle. La mortalité a été très forte durant les 4 premiers mois. Le taux de survie s’est stabilisé à 22%. Il est de 12% à la fin des 4 ans d’élevage. A partir de l’âge de 1 mois, la séquence alimentaire a été composée d’un aliment artificiel et de zooplancton congelé. A la fin de l’élevage, les poissons avaient une longueur totale moyenne de 35 ± 1 cm, pour un poids moyen de 387 ± 34 g. Les principales pathologies observées ont concernées les déformations de la bouche et des nodules rénaux sur des poissons de 3 ans. En juin 2015 les premiers individus matures ont été observés (mâles fluents). Ce travail a permis de réaliser le premier élevage ex situ de grande alose, depuis le stade larvaire et jusqu’à l’âge de 4 ans. Des recommandations sont proposées à partir de cette première expérience