Resume
Les récepteurs nucléaires des oxystérols LXRx sont impliqués dans l’homéostasie du cholestérol et le métabolisme lipidique. Les souris déficientes pour les 2 isoformes LXRα et LXRβ présentent une perturbation sévère de la structure de l’épithélium des segments 1 et 2 de la tête de l’épididyme ainsi qu’une fragilité des spermatozoïdes. Ces altérations ont pour conséquence d’engendrer une stérilité totale chez les souris mâles âgées de 10 mois.
A ce jour, aucun travail n’a été effectué sur le rôle des LXRs au niveau de l’épididyme. Une lignée cellulaire issue de la tête de l’épididyme murin (cellules B2) a été utilisée pour rechercherin vitro des gènes cibles épididymaires des LXRs. La présence d’une isoforme des LXRs (LXRα) a été mise en évidence par immunocytochimie et la capacité de réponse des cellules B2 à un agoniste synthétique des LXRs (T0901317) a été vérifiée. Ces résultats valident les cellules B2 comme modèle d’étude. Des analyses protéomiques en électrophorèse bidimensionnelle ont été menées sur les cellules B2 traitées au T0901317. Cela a permis d’isoler 8 protéines régulées positivement par les LXRs. Une seule a pu être identifiée: la polyubiquitine, dont l’implication dans l’homéostasie cellulaire du cholestérol a déjà été rapportée.
Abstract
Liver X receptors (LXRs) are involved in cholesterol homeostasis and lipid metabolism.Ixr knock-out mice for the two isoformsIxra andIxrb exhibit severe disruption of the structure of caput epididymidis segment 1 and 2 epithelium and increased sperm fragility. These defects generate infertility in 10-month-old male mice.
The role of LXRs in the epididymis have not yet been investigated. A cell line obtained from mouse caput epididymidis (B2 cells) was used to screen for LXR epididymal target genesin vitro. The presence of one isoform of LXR (LXRα) was detected by immunocytochemistry and the capacity of B2 cells to respond to a synthetic agonist of LXRs (T0901317) was verified. These results validated the use of B2 cells as a model. Bidimensional electrophoresis was performed on B2 cells treated with T0901317. Eight proteins up-regulated by LXRs were isolated. Only one protein has been identified: polyubiquitin, which has already been reported to be involved in cellular cholesterol homeostasis.
Article PDF
References
ABE K., TAKANO H., ITO T.: Ultrastructure of the mouse epididymal duct with special reference to the regional differences of the principal cells. Arch. Histol. Jpn., 1983, 46: 51–68.
ABOU-HAILA A., FAIN-MAUREL M.A.: Regional differences of the proximal part of mouse epididymis: morphological and histochemical characterization. Anat. Rec., 1984, 209: 197–208.
AKMAL K.M., DUFOUR J.M., KIM K.H.: Region-specific localization of retinoic acid receptor-alpha expression in the rat epididymis. Biol. Reprod., 1996, 54: 1111–1119.
ANDERSEN O.M., YEUNG C.H., VORUM H. et al.: Essential role of the apolipoprotein E receptor-2 in sperm development. J. Biol. Chem., 2003, 278: 23989–23995.
APFEL R., BENBROOK D., LERNHARDT E. et al.: A novel orphan receptor specific for a subset of thyroid hormone-responsive elements and its interaction with the retinoid/thyroid hormone receptor subfamily. Mol. Cell Biol., 1994, 14: 7025–7035.
BARONE R.: Anatomie Comparée des mammifères domestiques. Tome 4 Splanchnologie II. Paris, Vigot, 1990.
BRITAN A., LAREYRE J.J., LEFRANCOIS-MARTINEZ A.M. et al.: Spontaneously immortalized epithelial cells from mouse caput epididymidis. Mol. Cell Endocrinol., 2004, 224: 41–53.
COHEN D.J., ROCHWEGER L., ELLERMAN D.A. et al.: Relationship between the association of rat epididymal protein “DE” with spermatozoa and the behavior and function of the protein. Mol. Reprod. Dev., 2000, 56: 180–188.
COOPER T.G.: Role of the epididymis in mediating changes in the male gamete during maturation. Adv. Exp. Med. Biol., 1995, 377: 87–101.
CORNWALL G.A., CAMERON A., LINDBERG I. et al.: The cystatin-related epididymal spermatogenic protein inhibits the serine protease prohormone convertase 2. Endocrinology, 2003, 144: 901–908.
CORNWALL G.A., HANN S.R.: Specialized gene expression in the epididymis. J. Androl., 1995, 16: 379–383.
CORNWALL G.A., LAREYRE J.J., MATUSIK R.J. et al.: Gene expression and epididymal function. In: Robaire B., Hinton B.T. eds. The epididymis: From molecules to clinical practice. New York, Kluwer Academic/Plenum Publishers, 2002: 169–199.
CROSS N.L.: Role of cholesterol in sperm capacitation. Biol. Reprod., 1998, 59: 7–11.
DACHEUX J.L., GATTI J.L., CASTELLA S. et al.: The epididymal protein. In: Hinton B.T., Turner T.T. eds. The third international conference on the epididymis. New York, The Van Doren Company, 2003, 115–122.
DACHEUX J.L., DACHEUX F.: Protein secretion in the epididymis. In: Robaire B., Hinton B.T. eds. The epididymis: From molecules to clinical practice. New York, Kluwer Academic/Plenum Publishers, 2002: 151–168.
DREVET J.R., LAREYRE J.J., SCHWAAB V. et al.: The PEA3 protein of the Ets oncogene family is a putative transcriptional modulator of the mouse epididymis-specific glutathione peroxidase gene gpx5. Mol. Reprod. Dev., 1998, 49: 131–140.
DUFAURE J.P., DREVET J.R.: La régulation de l’expression des gènes dans l’épididyme. Médecine/Science, 1998, 14: 1392–1398.
EDDY E.M., WASHBURN T.F., BUNCH D.O. et al.: Targeted disruption of the estrogen receptor gene in male mice causes alteration of spermatogenesis and infertility. Endocrinology, 1996, 137: 4796–4805.
FRENOUX J.M., VERNET P., VOLLE D.H. et al.: Nuclear oxysterol receptors, LXR, are involved in the maintenance of mouse caput epididymidis structure and sperm post-testicular maturational events. J. Mol. Endocrinol., 2004, 33: 361–375.
FU X., MENKE J.G., CHEN Y. et al.: 27-hydroxycholesterol is an endogenous ligand for liver X receptor in cholesterol-loaded cells. J. Biol. Chem., 2001, 276: 38378–38387.
GARRETT J.E., GARRETT S.H., DOUGLASS J.A.: Spermatozoa-associated factor regulates proenkephalin gene expression in the rat epididymis. Mol. Endocrinol., 1990, 4: 108–118.
GAUDREAULT C., LE GARE C., BERUBE B., SULLIVAN R.: Hamster sperm protein, p26h: a member of the short-chain dehydrogenase/reductase superfamily. Biol. Reprod., 1999, 61: 264–273.
GLOVER T.D., NICANDER L.: Some aspects of structure and function in the mammalian epididymis. J. Reprod. Fertil., Suppl, 1971, 13: 39–50.
HAIDL G., OPPER C.: Changes in lipids and membrane anisotropy in human spermatozoa during epididymal maturation. Hum. Reprod., 1997, 12: 2720–2723.
HALL J.C., HADLEY J., DOMAN T.: Correlation between changes in rat sperm membrane lipids, protein, and the membrane physical state during epididymal maturation. J. Androl., 1991, 12: 76–87.
HALL J.C., KILLIAN G.J.: Changes in rat sperm membrane glycosidase activities and carbohydrate and protein contents associated with epididymal transit. Biol. Reprod., 1987, 36: 709–718.
HAMILTON D.: Anatomy of mamalian male accessory reproductive organs. In: Laming G. ed. Marshall’s Physiology of Reproduction, reproduction in the Male. Edinburg, Churchill Livingston, 1990, Vol 2: 691–746.
HAMPTON R.Y.: Proteolysis and sterol regulation. Ann. Rev. Cell Dev. Biol., 2002, 18: 345–378.
HESS R.A., BUNICK D., LEE K.H. et al.: A role for oestrogens in the male reproductive system. Nature, 1997, 390: 509–512.
HESS R.A., BUNICK D., LUBAHN D.B., ZHOU Q., BOUMA J.: Morphologic changes in efferent ductules and epididymis in estrogen receptor-alpha knockout mice. J. Androl., 2000, 21: 107–121.
HINTON B.T., LAN Z.J., RUDOLPH D.B., LABUS J.C., LYE R.J.: Testicular regulation of epididymal gene expression. J. Reprod. Fertil., Suppl, 1998, 53: 47–57.
HUANG L.S., VOYIAZIAKIS E., CHEN H.L., RUBIN E.M., GORDON J.W.: A novel functional role for apolipoprotein B in male infertility in heterozygous apolipoprotein B knockout mice. Proc. Natl Acad. Sci. USA, 1996, 93: 10903–10907.
JANOWSKI B.A., GROGAN M.J., JONES S.A. et al.: Structural requirements of ligands for the oxysterol liver X receptors LXRalpha and LXRbeta. Proc. Natl Acad. Sci. USA, 1999, 96: 266–271.
KIRCHHOFF C.: Gene expression in the epididymis. Int. Rev. Cytol., 1999, 188: 133–202.
KIRSTEN M.R., GERTRUD U.S., KNUT R.S. et al.: The liver X receptor is essential for maintaining cholesterol homeostasis in the testis. Endocrinology, 2005, 146: 2519–2530.
LAN Z.J., LABUS J.C., HINTON B.T.: Regulation of gammaglutamyl transpeptidase catalytic activity and protein level in the initial segment of the rat epididymis by testicular factors: role of basic fibroblast growth factor. Biol. Reprod., 1998, 58: 197–206.
LAREYRE J.J., CLAESSENS F., ROMBAUTS W., DUFAURE J.P., DREVET J.R.: Characterization of an androgen response element within the promoter of the epididymis-specific murine glutathione peroxidase 5 gene. Mol. Cell Endocrinol., 1997, 129: 33–46.
LAREYRE J.J., MATTEI M.G., KASPER S. et al.: Structure and putative function of a murine epididymal retinoic acid-binding protein (mE-RABP). J. Reprod. Fertil. Suppl, 1998, 53: 59–65.
LEGARE C., BERUBE B., BOUE F. et al.: Hamster sperm antigen P26h is a phosphatidylinositol-anchored protein. Mol. Reprod. Dev., 1999, 52: 225–233.
LEGARE C., GAUDREAULT C., ST-JACQUES S., SULLIVAN R.: P34H sperm protein is preferentially expressed by the human corpus epididymidis. Endocrinology, 1999, 140: 3318–3327.
LOBACCARO J.M., REPA J.J., LU T.T. et al.: Regulation of lipid metabolism by the orphan nuclear receptors. Ann. Endocrinol. (Paris), 2001, 62: 239–247.
LU T.T., REPA J.J., MANGELSDORF D.J.: Orphan nuclear receptors as eLiXiRs and FiXeRs of sterol metabolism. J. Biol. Chem., 2001, 276: 37735–37738.
MERCIER-BODARD C., ALFSEN A., BAULIEU E.E.: Sex steroid binding plasma protein (SBP). Acta Endocrinol (Copenh), Suppl. 1970, 147: 204–224.
MOORE H.D., BEDFORD J.M.: The differential absorptive activity of epithelial cells of the rat epididymus before and after castration. Anat. Rec., 1979, 193: 313–327.
MORTZ E., KROGH T.N., VORUM H., GÖRG A.: Improved silver staining protocols for high sensivity protein identification using matrix-assisted laser desorption/ionization time of flight analysis. Proteomics, 2001, 1: 1359–1363.
NATH D., MAJUMDER G.C.: Maturation-dependent modification of the protein phosphorylation profile of isolated goat sperm plasma membrane. J. Reprod. Fertil., 1999, 115: 29–37.
PEET D.J., TURLEY S.D., MA W. et al.: Cholesterol and bile acid metabolism are impaired in mice lacking the nuclear oxysterol receptor LXR alpha. Cell, 1998, 93: 693–704.
PERRY A.C., JONES R., HALL L.: Isolation and characterization of a rat cDNA clone encoding a secreted superoxide dismutase reveals the epididymis to be a major site of its expression. Biochem. J., 1993, 293: 21–25.
POLLANEN P., COOPER T.G.: Immunology of the testicular excurrent ducts. J. Reprod. Immunol., 1994, 26: 167–216.
RAVID T., DOOLMAN R., AVNER R., HARATS D., ROITELMAN J.: The ubiquitin-proteasome pathway mediates the regulated degradation of mammalian 3-hydroxy-3-methylglutaryl-coenzyme A reductase. J. Biol. Chem., 2000, 275: 35840–35847.
REBECCA L.W., GORDON C., MAYER R.J.: Ubiquitin and ubiquitin-like proteins as multifunctional signals. Nature Reviews, 2005, 6: 599–609.
REPA J.J., MANGELSDORF D.J.: The role of orphan nuclear receptors in the regulation of cholesterol homeostasis. Ann. Rev. Cell Biol., 2000, 16: 59–81.
ROBAIRE B., HERMO L.: Efferents ducts, epididymis, vas deferens: structure, functions, and their regulation. In: Knobil E. ed. The physiology of reproduction. New York, Raven Press, 1988: 999–1080
ROBAIRE B., JERVIS K.M., EZER N.: Cell Dynamics and Death in the Epididymal Epithelium. In: Hinton B.T., Turner T.T. eds. The third international conference on the epididymis. New York, The Van Doren Company, 2003: 35–49.
SHALGI R., SELIGMAN J., KOSOWER NS.: Dynamics of the thiol status of rat spermatozoa during maturation: analysis with the fluorescent labeling agent monobromobimane. Biol. Reprod., 1989, 40: 1037–1045.
SINOWATZ F., VOLGMAYR J.K., GABIUS H.J., FRIESS A.E.: Cytochemical analysis of mammalian sperm membranes. Prog. Histochem. Cytochem., 1989, 19: 1–74.
SORANZO L., DADOUNE J.P., FAIN-MAUREL M.A.: Segmentation of the epididymal duct in mouse: an ultrastructural study. Reprod. Nutr. Dev., 1982, 22: 999–1012.
TEBOUL M., ENMARK E., LI Q., WIKSTROM A.C., PELTOHUIKKO M., GUSTAFSSON J.A.: OR-1, a member of the nuclear receptor superfamily that interacts with the 9-cis-retinoic acid receptor. Proc. Natl. Acad. Sci. USA, 1995, 92: 2096–2100.
THUILLIER P., BAILLIE R., SHA X., CLARKE S.D.: Cytosolic and nuclear distribution of PPARgamma2 in differentiating 3T3-L1 preadipocytes. J. Lipid. Res., 1998, 39: 2329–2338.
TRAMER F., ROCCO F., MICALI F., SANDRI G., PANFILI E.: Role of luminal fluid glycosyltransferases and glycosidases in the modification of rat sperm plasma membrane glycoproteins during epididymal maturation. J. Reprod. Fertil., Suppl. 1998, 53: 85–97.
TULSIANI D.R., SKUDLAREK M.D., HOLLAND M.K.: Orgebin-Crist M.C.: Glycosylation of rat sperm plasma membrane during epididymal maturation. Biol. Reprod., 1993, 48: 417–428.
TURNER T.T., AVERY E.A., SAWCHUK T.J.: Assessment of protein synthesis and secretion by rat seminiferous and epididymal tubules in vivo. Int. J. Androl., 1994, 17: 205–213.
VERNET P., RIGAUDIERE N., GHYSELINCK N., DUFAURE J.P., DREVET J.R.: In vitro expression of a mouse tissue specific glutahione-peroxidase-like protein lacking the selenocysteine can protect stably transfected mammalian cells against oxidative damage. Biochem. Cell Biol., 1996, 74: 125–131.
VISCONTI P.E., NING X., FORNES M.W. et al.: Cholesterol efflux-mediated signal transduction in mammalian sperm: cholesterol release signals an increase in protein tyrosine phosphorylation during mouse sperm capacitation. Dev. Biol., 1999, 214: 429–443.
VOLLE D.H., DÉCHELOTTE P., CUMMINS C.C. et al.: Deficient mice for oxysterol nuclear receptors LXRs show a rapid decrease of fertility associated with testicular destructuration. 2004 (en cours de préparation).
VOLLE D.H., FRENOUX J.M., MOUZAT K. et al.: Rôle des récepteurs nucléaires des oxystérols LXR dans la régulation de l’homéostasie du cholestérol au niveau de l’appareil reproducteur mâle. Andrologie, 2005, 15: 151–159.
WAN Y.J., WANG L., WU T.C.: Detection of retinoic acid receptor mRNA in rat tissues by reverse transcriptase-polymerase chain reaction. J. Mol. Endocrinol., 1992, 9: 291–294.
Author information
Authors and Affiliations
Corresponding author
Additional information
Prix du meilleur DEA-SALF 2004
Rights and permissions
About this article
Cite this article
Kocer, A., Saez, F., Mouzat, K. et al. Criblage de nouveaux gènes cibles des récepteurs nucléairesdes oxystérols LXRs impliqués dans le maintien de l’épithélium épididymaire et la maturation des spermatozoïdes dans l’épididyme. Androl. 16, 11–23 (2006). https://doi.org/10.1007/BF03034827
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF03034827