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Quantitative immunocytochemistry of pituitary receptors for luteinizing hormone-releasing hormone

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Summary

In Araldite sections of male rat pituitaries, stained after embedding by the unlabeled antibody enzyme method with antisera to native luteinizing hormone-releasing hormone (LH-RH) or LH-RH azo-conjugated to bovine serum albumin, localization is confined mainly to the interior of the large, and to a lesser extent to that of the small, secretion granules of the gonadotrophic cells. Plasma membranes are not demonstrated. Except for weak staining in the granules of corticotrophs, no other pituitary cell is stained. Pretreatment of sections with LH-RH (to dilutions of 4 pg/μl) increases staining intensity in the gonadotrophic granules. Other cells are unaffected. The lesser the gonadotroph staining intensity without pretreatment, the greater the increase (more than 23-fold reactivity). Augmented staining is measurable (P<0.001) to antiserum dilutions of 1∶240 000. Pretreatment with des-Glu-1-LH-RH, porcine corticotropin or rat prolactin has no effect. LH-RH-Gly-10(desamide) inhibits. Rat glycoprotein hormones enhance staining with anti-azo-conjugated LH-RH. With antinative LH-RH these hormones enhance weak staining, but inhibit strong staining. Thick vibrotome sections of male rat or rabbit pituitaries stained before embedding reveal specific localization on plasma membrane and gonadotrophic secretion granules, provided the sections have been pretreated with LH-RH (250 pg/μl). The data show that LH-RH after reaction with receptor is not sterically hindered from binding specific antibodies. Receptor may be found in secretion granules, both in the free state or combined with LH-RH. Plasma membrane receptor, on the other hand, was free under the conditions of the experiments. Immunization with LH-RH elicits not only heteroimmune antibodies specific for LH-RH, but also a group of still ill defined autoimmune antibodies, some of which may conceivably be reactive with glycoprotein hormone α-chains.

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Authors and Affiliations

  1. Immunology Branch, Edgewood Arsenal, Maryland

    Ludwig A. Sternberger & John P. Petrali

  2. Department of Microbiology, Johns Hopkins University, School of Medicine, Baltimore, Maryland, USA

    Ludwig A. Sternberger & John P. Petrali

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  1. Ludwig A. Sternberger
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  2. John P. Petrali
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We would like to thank Drs. Pelletier, Labrie and Dermody for their generous gifts of anti-LH-RH, Dr. Noren of Ferring AB for kindly supplying purified pACTH and Dr. Parlow for his generosity in making available repeated gifts of rat pituitary hormones and their antisera from the NIAMD Rat Pituitary Hormone Distribution Program. Human FSH and LH were donated by the NIAMD Human Pituitary Agency. Dr. N. S. Halmi has kindly agreed to review the manuscript. His valuable comments have been incoperated with deep appreciation.

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Sternberger, L.A., Petrali, J.P. Quantitative immunocytochemistry of pituitary receptors for luteinizing hormone-releasing hormone. Cell Tissue Res. 162, 141–176 (1975). https://doi.org/10.1007/BF00209204

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