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Journal of Histochemistry and Cytochemistry, Vol. 45, 985-990, Copyright © 1997 by The Histochemical Society, Inc.

ARTICLE

Production and Characterization of Specific Anti-peptide Antiserum Against Free {alpha}-subunit of Rat Pituitary Glycoprotein Hormones

Shigeyasu Tanakaa, Shingo Kurabuchic, Hiroshi Mochidaa, Hiroaki Hayashid, and Katsumi Wakabayashib
a Department of Cell Biology, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Japan
b Biosignal Research Center, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Japan
c Department of Histology, The Nippon Dental University School of Dentistry at Tokyo, Chiyoda-ku, Tokyo, Japan
d Gunma Prefectural College of Health Science, Maebashi, Japan

Correspondence to: Shigeyasu Tanaka, Dept. of Cell Biology, Inst. for Molecular and Cellular Regulation, Gunma Univ., Showa-machi 3-39-15, Maebashi 371, Japan.

To obtain an antibody specific for the {alpha}-subunit of rat pituitary glycoprotein hormones, we synthesized a peptide corresponding to the sequence 37-53 (ST-7: Phe-Ser-Arg-Ala-Tyr-Pro-Thr-Pro-Ala-Arg-Ser-Lys-Lys-Thr-Met-Leu-Val) of the rat {alpha}-subunit. The polyclonal antiserum against this peptide was generated in rabbits. This region is hydrophilic and highly conserved among several mammalian species. Noncompetitive binding tests showed that the ST-7 antiserum had specific affinity for the rat free {alpha}-subunit, but not for rat intact LH, FSH, and TSH. The ST-7 antiserum immunostained two types of cells in the rat anterior pituitary, i.e., gonadotrophs and thyrotrophs. This was also the case in mouse, cattle, sheep, and pig, which have an identical sequence of ST-7 in their {alpha}-subunit. The pituitary cells of horse (Arg substituted for Lys as residue 48 of the rat {alpha}-subunit), human, and eel (Leu for Ala at residue 45), chicken (Met for Ala at residue 45), and bullfrog (Tyr for Phe at residue 37 and Met for Ala at residue 45) were not stained with the ST-7 antiserum. This study indicated that the ST-7 antiserum is sequence-specific for the {alpha}-subunit and is therefore useful for immunohistochemical studies on the secretory pathway of the free {alpha}-subunit. (J Histochem Cytochem 45:985-990, 1997)

Key Words: {alpha}-subunit, pituitary glycoprotein hormone, anti-peptide antiserum, region-specific antiserum, immunocytochemistry, pituitary gland, rat


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