| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Articles |
a Research and Development, N.V. Organon, 5340 BH Oss, The Netherlands
Two IFMAs for rFSH were developed with mMAbs against the recombinant human (rec h)FSHß subunit (FSH56A) attached to the wall and two different rabbit polyclonal antibodies (PAbs) against the
An IFMA for rLH was set up with mMAb (hCG77A) reacting with rLHß as capture and rabbit PAb to rec rLH
In conclusion, two IFMAs for rFSH and one for rLH have been developed with high sensitivity and specificity for intact gonadotropins. The LH pattern during the estrous cycle was comparable between IFMA, RIA, and FIA, although the overall level in the IFMA was much lower, as were HYPEX levels. The FSH pattern differed only on proestrus Day 2 in the IFMA from that of RIA/FIA, showing a peak level with RIA/FIA and a basal level with the IFMA. This implies that in RIA/FIA measurements, proteins other than intact FSH and LH interfere with the analysis at proestrus Day 2 for FSH and in HYPEX, cycling, and OVX rats for LH.
The aim of this study was to develop time-resolved immunofluorometric assays (TR-IFMA) for measuring rat (r)FSH and rLH. The advantages of these IFMAs are higher sensitivity due to lower background values, higher specificity as only intact molecules of FSH and LH can be measured, and a very long shelf life of the nonradioactive biotin antigens compared with radiolabeled iodine antigens. For rFSH, IFMAs are lacking, while for rLH, if present, the resources for antibodies are scarce or the mouse monoclonal antibodies (mMAbs) against LH
are inactive with FSH. Thus specific antibodies need to be obtained. With the final TR-IFMAs, rFSH and rLH levels were assessed during the estrous cycle and compared with those obtained with the more classical RIAs and fluoroimmunoassays (FIAs). subunit of rec hFSH (R93–2705) or recombinant rat (rec r)LH (R95–2715) conjugated with biotin as signal antibody. With both IFMAs, rFSH holo-molecules can be measured. Rat FSH standards could be assessed between 0.02 and 10 ng/ml with a detection limit of 0.05 and 0.24 ng/ml in buffer and serum, respectively. These detection limits in four IFMAs were 8- to 16-fold lower than those in RIAs and FIAs. This detection level allowed the measurement of FSH levels in serum of hypophysectomized (HYPEX) rats at 0.18 ng/ml. In serum of cycling rats, the FSH levels of the IFMA were 2-fold lower than those of the FIA, while in ovariectomized (OVX) rats the IFMA levels were comparable. A peak level of FSH was found during proestrus of Day 2 and gestation with both RIA and FIA, but with IFMAs at gestation only. (R95–2712) as signal antibody. Rat LH standard could be assessed between 0.001 and 10 ng/ml with a detection limit of 0.012 and 0.1 ng/ml in buffer and serum, respectively, which was 8-fold lower than that in RIA/FIA. In serum of HYPEX rats, LH was undetectable (< 0.04 ng/ml), whereas a high background level of 2.5 ng/ml was measured in the FIA. In serum of cycling rats, only a very low LH level of 0.14 ng/ml was measured, which strongly deviated from the level of 3.46 ng/ml with an FIA. The load of LH in serum of OVX rats was 2.91 ng/ml, which was 12-fold lower than that for the FIA. The peak level of LH was detected on proestrus Day 2 with RIA, FIA, and IFMA.
1 Correspondence: W.G.E.J. Schoonen (RE 2218), Lead Discovery Unit, N.V. Organon, Molenstraat 110, P.O.Box 20, 5340 BH Oss, The Netherlands. FAX: 31 412 662542; w.schoonen{at}organon.oss.akzonobel.nl
This article has been cited by other articles:
|
|
 |
|
  M. H. Abel, P. J. Baker, H. M. Charlton, A. Monteiro, G. Verhoeven, K. De Gendt, F. Guillou, and P. J. O'Shaughnessy Spermatogenesis and Sertoli Cell Activity in Mice Lacking Sertoli Cell Receptors for Follicle-Stimulating Hormone and Androgen Endocrinology, July 1, 2008; 149(7): 3279 - 3285. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Makanji, K. L. Walton, M. C. Wilce, K. L. Chan, D. M. Robertson, and C. A. Harrison Suppression of Inhibin A Biological Activity by Alterations in the Binding Site for Betaglycan J. Biol. Chem., June 13, 2008; 283(24): 16743 - 16751. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Mittag, W. Oehr, H. Heuer, T. Hamalainen, B. Brachvogel, E. Poschl, and K. Bauer Expression and thyroid hormone regulation of annexins in the anterior pituitary J. Endocrinol., December 1, 2007; 195(3): 385 - 392. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Saloniemi, T. Lamminen, K. Huhtinen, M. Welsh, P. Saunders, H. Kujari, and M. Poutanen Activation of Androgens by Hydroxysteroid (17{beta}) Dehydrogenase 1 in Vivo as a Cause of Prenatal Masculinization and Ovarian Benign Serous Cystadenomas Mol. Endocrinol., November 1, 2007; 21(11): 2627 - 2636. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. J Teerds, E. Rijntjes, M. B Veldhuizen-Tsoerkan, F. F G Rommerts, and M. de Boer-Brouwer The development of rat Leydig cell progenitors in vitro: how essential is luteinising hormone? J. Endocrinol., September 1, 2007; 194(3): 579 - 593. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. A. Walters, C. M. Allan, M. Jimenez, P. R. Lim, R. A. Davey, J. D. Zajac, P. Illingworth, and D. J. Handelsman Female Mice Haploinsufficient for an Inactivated Androgen Receptor (AR) Exhibit Age-Dependent Defects That Resemble the AR Null Phenotype of Dysfunctional Late Follicle Development, Ovulation, and Fertility Endocrinology, August 1, 2007; 148(8): 3674 - 3684. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Zhongyi, P. Rantakari, T. Lamminen, J. Toppari, and M. Poutanen Transgenic Male Mice Expressing Human Hydroxysteroid Dehydrogenase 2 Indicate a Role for the Enzyme Independent of Its Action on Sex Steroids Endocrinology, August 1, 2007; 148(8): 3827 - 3836. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Makanji, C. A. Harrison, P. G. Stanton, R. Krishna, and D. M. Robertson Inhibin A and B in Vitro Bioactivities Are Modified by Their Degree of Glycosylation and Their Affinities to Betaglycan Endocrinology, May 1, 2007; 148(5): 2309 - 2316. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Shetty, C. C. Y. Weng, K. L. Porter, Z. Zhang, P. Pakarinen, T. R. Kumar, and M. L. Meistrich Spermatogonial Differentiation in Juvenile Spermatogonial Depletion (jsd) Mice with Androgen Receptor or Follicle-Stimulating Hormone Mutations Endocrinology, July 1, 2006; 147(7): 3563 - 3570. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. A. Harrison, K. L. Chan, and D. M. Robertson Activin-A Binds Follistatin and Type II Receptors through Overlapping Binding Sites: Generation of Mutants with Isolated Binding Activities Endocrinology, June 1, 2006; 147(6): 2744 - 2753. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. M. Walden, T. D. Butters, R. A. Dwek, F. M. Platt, and A. C. van der Spoel Long-term non-hormonal male contraception in mice using N-butyldeoxynojirimycin Hum. Reprod., May 1, 2006; 21(5): 1309 - 1315. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Shetty, C. C. Y. Weng, S. J. Meachem, O. U. Bolden-Tiller, Z. Zhang, P. Pakarinen, I. Huhtaniemi, and M. L. Meistrich Both Testosterone and Follicle-Stimulating Hormone Independently Inhibit Spermatogonial Differentiation in Irradiated Rats Endocrinology, January 1, 2006; 147(1): 472 - 482. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Ledent, I. Demeestere, D. Blum, J. Petermans, T. Hamalainen, G. Smits, and G. Vassart Premature ovarian aging in mice deficient for Gpr3 PNAS, June 21, 2005; 102(25): 8922 - 8926. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H Santti, L Mikkonen, A Anand, S Hirvonen-Santti, J Toppari, M Panhuysen, F Vauti, M Perera, G Corte, W Wurst, et al. Disruption of the murine PIASx gene results in reduced testis weight J. Mol. Endocrinol., June 1, 2005; 34(3): 645 - 654. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. M. Robertson, G. U. Schuster, K. R. Steffensen, O. Hovatta, S. Meaney, K. Hultenby, L. C. Johansson, K. Svechnikov, O. Soder, and J.-A. Gustafsson The Liver X Receptor-{beta} Is Essential for Maintaining Cholesterol Homeostasis in the Testis Endocrinology, June 1, 2005; 146(6): 2519 - 2530. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Pakarainen, F.-P. Zhang, S. Makela, M. Poutanen, and I. Huhtaniemi Testosterone Replacement Therapy Induces Spermatogenesis and Partially Restores Fertility in Luteinizing Hormone Receptor Knockout Mice Endocrinology, February 1, 2005; 146(2): 596 - 606. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Jimenez, J.A. Spaliviero, A.J. Grootenhuis, J. Verhagen, C.M. Allan, and D.J. Handelsman Validation of an Ultrasensitive and Specific Immunofluorometric Assay for Mouse Follicle-Stimulating Hormone Biol Reprod, January 1, 2005; 72(1): 78 - 85. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Shetty, C. C. Y. Weng, O. U. Bolden-Tiller, I. Huhtaniemi, D. J. Handelsman, and M. L. Meistrich Effects of Medroxyprogesterone and Estradiol on the Recovery of Spermatogenesis in Irradiated Rats Endocrinology, October 1, 2004; 145(10): 4461 - 4469. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Zaleska, A. Waclawik, G. Bodek, A. Zezula-Szpyra, X. Li, T. Janowski, W. H. Hansel, N. A. Rahman, and A. J. Ziecik Growth Repression in Diethylstilbestrol/Dimethylbenz[a]anthracene-Induced Rat Mammary Gland Tumor Using Hecate-CG{beta} Conjugate Experimental Biology and Medicine, April 1, 2004; 229(4): 335 - 344. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. M. Allan, A. Garcia, J. Spaliviero, F.-P. Zhang, M. Jimenez, I. Huhtaniemi, and D. J. Handelsman Complete Sertoli Cell Proliferation Induced by Follicle-Stimulating Hormone (FSH) Independently of Luteinizing Hormone Activity: Evidence from Genetic Models of Isolated FSH Action Endocrinology, April 1, 2004; 145(4): 1587 - 1593. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Shetty and C. C. Y. Weng Cryptorchidism Rescues Spermatogonial Differentiation in Juvenile Spermatogonial Depletion (Jsd) Mice Endocrinology, January 1, 2004; 145(1): 126 - 133. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. L. Meistrich, G. Wilson, K. L. Porter, I. Huhtaniemi, G. Shetty, and G. A. Shuttlesworth Restoration of Spermatogenesis in Dibromochloropropane (DBCP)-Treated Rats by Hormone Suppression Toxicol. Sci., December 1, 2003; 76(2): 418 - 426. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F.-P. Zhang, T. Pakarainen, M. Poutanen, J. Toppari, and I. Huhtaniemi The low gonadotropin-independent constitutive production of testicular testosterone is sufficient to maintain spermatogenesis PNAS, November 11, 2003; 100(23): 13692 - 13697. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. B. Rulli, P. Ahtiainen, S. Makela, J. Toppari, M. Poutanen, and I. Huhtaniemi Elevated Steroidogenesis, Defective Reproductive Organs, and Infertility in Transgenic Male Mice Overexpressing Human Chorionic Gonadotropin Endocrinology, November 1, 2003; 144(11): 4980 - 4990. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Perlman, B. van den Hazel, J. Christiansen, S. Gram-Nielsen, C. B. Jeppesen, K. V. Andersen, T. Halkier, S. Okkels, and H. T. Schambye Glycosylation of an N-Terminal Extension Prolongs the Half-Life and Increases the in Vivo Activity of Follicle Stimulating Hormone J. Clin. Endocrinol. Metab., July 1, 2003; 88(7): 3227 - 3235. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Lue, A. P. Sinha Hikim, C. Wang, A. Leung, and R. S. Swerdloff Functional Role of Inducible Nitric Oxide Synthase in the Induction of Male Germ Cell Apoptosis, Regulation of Sperm Number, and Determination of Testes Size: Evidence from Null Mutant Mice Endocrinology, July 1, 2003; 144(7): 3092 - 3100. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Nellemann, M. Dalgaard, H. R. Lam, and A. M. Vinggaard The Combined Effects of Vinclozolin and Procymidone Do Not Deviate from Expected Additivity in Vitro and in Vivo Toxicol. Sci., February 1, 2003; 71(2): 251 - 262. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. C. van der Spoel, M. Jeyakumar, T. D. Butters, H. M. Charlton, H. D. Moore, R. A. Dwek, and F. M. Platt Reversible infertility in male mice after oral administration of alkylated imino sugars: A nonhormonal approach to male contraception PNAS, December 24, 2002; 99(26): 17173 - 17178. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Haywood, N. Tymchenko, J. Spaliviero, A. Koch, M. Jimenez, J. Gromoll, M. Simoni, V. Nordhoff, D. J. Handelsman, and C. M. Allan An Activated Human Follicle-Stimulating Hormone (FSH) Receptor Stimulates FSH-Like Activity in Gonadotropin-Deficient Transgenic Mice Mol. Endocrinol., November 1, 2002; 16(11): 2582 - 2591. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. M. Vinggaard, C. Nellemann, M. Dalgaard, E. B. Jorgensen, and H. R. Andersen Antiandrogenic Effects in Vitro and in Vivo of the Fungicide Prochloraz Toxicol. Sci., October 1, 2002; 69(2): 344 - 353. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. B. Rulli, A. Kuorelahti, O. Karaer, L. J. Pelliniemi, M. Poutanen, and I. Huhtaniemi Reproductive Disturbances, Pituitary Lactotrope Adenomas, and Mammary Gland Tumors in Transgenic Female Mice Producing High Levels of Human Chorionic Gonadotropin Endocrinology, October 1, 2002; 143(10): 4084 - 4095. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Gabant, L. Forrester, J. Nichols, T. Van Reeth, C. De Mees, B. Pajack, A. Watt, J. Smitz, H. Alexandre, C. Szpirer, et al. Alpha-fetoprotein, the major fetal serum protein, is not essential for embryonic development but is required for female fertility PNAS, October 1, 2002; 99(20): 12865 - 12870. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Shetty, G. Wilson, M. P. Hardy, E. Niu, I. Huhtaniemi, and M. L. Meistrich Inhibition of Recovery of Spermatogenesis in Irradiated Rats by Different Androgens Endocrinology, September 1, 2002; 143(9): 3385 - 3396. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Shetty, G. Wilson, I. Huhtaniemi, H. Boettger-Tong, and M. L. Meistrich Testosterone Inhibits Spermatogonial Differentiation in Juvenile Spermatogonial Depletion Mice Endocrinology, July 1, 2001; 142(7): 2789 - 2795. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Li, E. Nokkala, W. Yan, T. Streng, N. Saarinen, A. Warri, I. Huhtaniemi, R. Santti, S. Makela, and M. Poutanen Altered Structure and Function of Reproductive Organs in Transgenic Male Mice Overexpressing Human Aromatase Endocrinology, June 1, 2001; 142(6): 2435 - 2442. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F.-P. Zhang, M. Poutanen, J. Wilbertz, and I. Huhtaniemi Normal Prenatal but Arrested Postnatal Sexual Development of Luteinizing Hormone Receptor Knockout (LuRKO) Mice Mol. Endocrinol., January 1, 2001; 15(1): 172 - 183. [Abstract] [Full Text]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
