Cytochemical characterization of pituitary target cells for biotinylated gonadotropin releasing hormone |
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Authors: | G V Childs Z Naor E Hazum R Tibolt K N Westlund M B Hancock |
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Affiliation: | 1. Department of Anatomy, The University of Texas Medical Branch, Galveston, TX 77550, USA;2. Department of Hormone Research, The Weizmann Institute of Science, Rehovot, 76-100, Israel |
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Abstract: | These studies describe the application of new cytochemical stains that co-localize a biotin-labeled gonadotropin releasing hormone (GnRH) analog and FSH or LH in the same field or cell. Pituitary monolayer cells were stimulated with the [D-Lys6] GnRH analog or the same analog labeled with biotin. Biotinylated [D-Lys6] GnRH exhibited a higher affinity and was 7-10 X more potent than unlabeled [D-Lys6] GnRH. The avidin-biotin peroxidase complex technique (ABC) was applied to localize the biotinylated GnRH on the cells with the use of a dense black peroxidase substrate. Specificity tests showed that the stain could be eliminated by competition with unlabeled [D-Lys6] GnRH. The GnRH stain was followed by immunocytochemical stains for LH beta, FSH beta or 25-39ACTH with a different peroxidase substrate (amber or orange-red). Stain for GnRH was found on the surfaces of 16% of the cells and 60-90% of the GnRH stained cells also stained for one of the gonadotropins. Most (90-100%) of the gonadotropes showed stain for GnRH. Our studies demonstrate that a potent biotinylated GnRH analog binds cells that can be identified specifically as gonadotropes. |
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Keywords: | GnRH and GnRH analogs LH FSH Gonadotropes Receptor affinity cytochemistry Immunocytochemistry Avidin-biotin Peroxidase Tissue culture Female Rats |
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