TGFp FSH INH INH

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1 (210095) ( genlinwang@hotmail.com) - -(Strept Avidin-Biotin-Peroxidase Complex SABC), 1 32,, 1 inhibin INH α TGFp FSH FSH [1] INHα Sertoli Noguchi 1997 [2] Meunier et al.1988 [3] INH INH INHα INH SD sigma I8641 sigma I2777SABC SA IgG SABC Strept Avidin-Biotin-Peroxidase Complex

2 3,3-3,3 -diamino-benzidine DAB AR1022 (Poly-L-Lysine) ( AR0001 ) SD INH2 g,1 0.5 g 10 6h % 4 m horseradish peroxidase HRP DAB 3 H2O2 10min M ph6.0 20min PBS phosphare buffered saline 0.01M PH min 25 g/ml 37 1hr IgG 37 20min PBS 0.01M PH7.4 2min 3 SABC 37 20min PBS 5min 4 DAB 10min 1min [4], - INH 1-2 -

3 Table1 Immunoreactivity for inhibin- subunit in mature rats Specimen Control The intense of injected Ovariectomized INH / g/ml INH- fragment Hypothalamus - - Pituitary Lympth node Adrenal Heart Liver Spleen Lung Kidney Pancreas result of immunoreactivity- Negative Positive 1 2 ( 80 DAB ( 80 DAB ) ) Fig.1 Mutural rat immunostained for inhibin subunit in the Pituitary of the mature rat. It showed posititve reactivity.(arrow)( 80 DAB stain) Fig.2Mutural rat immunostained for inhibin subunit in the Pituitary of the ovariectomized rat. It showed negative reactivity.( 80 DAB stain) - 3 -

4 3 4 ( 80 DAB 40 DAB ) Fig.3 Mutural rat immunostained for inhibin subunit in the hypothalamus of the mature rat. It showed posititve reactivity.(arrow)( 80 DAB stain). Fig.4 Mutural rat immunostained for inhibin subunit in the liver of the ovariectomized rat. It showed negative reactivity.( 40 DAB stain) 5 6 ( 40 DAB ( 40 DAB ) ) Fig.5 Mutural rat immunostained for inhibin subunit in the kidney of the ovariectomized rat. It showed posititve reactivity.(arrow)( 40 DAB stain) Fig.6 Mutural rat immunostained for inhibin subunit in the pancreas of the ovariectomized rat. It showed posititve reactivity.(arrow) ( 40 DAB stain) - 4 -

5 7 8 ( 80 DAB ) 40 DAB Fig.7 Mutural rat immunostained for inhibin subunit in the lung of the mature rat. It showed posititve reactivity.(arrow)( 40 DAB stain). Fig.8 Mutural rat immunostained for inhibin subunit in the lung of the ovariectomized rat. It showed negative reactivity.( 40 DAB stain). 4 --Strept Avidin-Biotin-Peroxidase Complex SABC INH INH INH Meunier et al 1988 [3] Mckeehan et al. [5] INH Peeters et al (1997) [6] INH INH INH Yamaguchi et al INH ACTH [7] INH INH INH INH INH INH [1]Robertson D M,Cahir N, Findlay J K,Burger H G, et al. The biological and immunological characterization of inhibin A and B forms in human follicular fluid and plasma[j]. J Clin Endorcrinol Metab, 1997,82(3):

6 [2] Noguchi J. Ontogeny of inhibin secretion in the rat testis: Secretion of inhibin-related proteins from fetal leydig cell and of bioactive inhibin from sertdi cells, Journal-of-Endecrinology, 1997, [3] Helene Meunier, Catherine Rivier,Ronald M, et al. Gonadal and extragonadal expression of inhibin α, βa and βb subunits in various tissues predict diverse function [J]. Proc Natl Soci USA,1988, 85: [4] LMA Munro,A Kennedy, et al.the expression of inhibin/activin subunits in the humun adrenal cortex and its tumors. Journal of Endocrinology. 1999,161: [5] Xu J, Mckeehan K, Matsuzaki K, et al. Inhibin antagonizes inhibition of liver cell growth by activin by a dominant negative mechanism.[j]biol Chem,1995;270: [6] Peeters R, Vanmontfort D, Van Isterdael J,R.,.Evidence for the presence of immunoreactive inhibin in extragonadal tissues of ovariectomized ewes.anim Repro Sci.1997,48: [7]Yamaguchi M, Mizunuma H, Miyamoto K,et al..immunoreactive inhibin concentrations in adult men:presence of a circadian rhythm. Journal of Clinical Endocrinology and Metabolism.1991,72: Immunohistochemical localization of inhibin-α subunit in mature rats ZHANG Zhong-xia, LI Lian, XU Lu, YAN Yu-qin, WANG Gen-lin College of Animal Science and Technology,Nanjing Agricultural University, Abstract: Immunohistochemical localization of inhibin α-subunit in normal mature rats was carried out by using Strept Avidin-Biotin-Peroxidase Complex.The results showed that there was positive immunoreactivity of the McAb to inhibinα-subunit in heart,kidney, spleen, lymph,adrenal gland and pancreas cells. It indicated that inhibinα-subunit existed in the above tissues, But no positive sites were seen in hypothalamus, pituitary, lung and liver, that means they have no inhibinα-subunit. Compared with the ovariectomized rats, the hypothalamus and pituitary of the rats injected with inhibin-α fragment 1 32 showed an positive reaction, suggesting that inhibin α-subunit could pass through the blood-brain barrier. Key words: Inhibin, Inhibin-α subunit, immunohistochemical localization