Immunocytochemical and immunogold localization of two prolactin isoforms in the same pituitary cells and in the same granules in the tilapia (Oreochromis mossambicus).
The tilapia pituitary secretes two forms of prolactin (tPRL) and a single growth hormone (tGH). The tPRLs share only 69% sequence identity and are designated tPRL177 and tPRL188 to indicate the number of amino acid residues in each isoform. Our aim was to develop specific antisera for detection of these three related polypeptides. Ten peptides corresponding to unique epitopes on the tPRLs and two peptides of tGH were synthesized using solid-phase methods, conjugated to carrier proteins, and used as immunogens for antibody production in rabbits. Select antisera for the tPRLs were highly specific, exhibiting only 1% cross-reactivity to the alternate tPRL under noncompetitive ELISA conditions at dilutions used in immunocytochemical analysis. The anti-tGH specifically bound to cells in the proximal pars distalis. Production of both tPRLs by a single cell type was indicated by the binding of both anti-tPRL177 and anti-tPRL188 to the same cells in the rostral pars distalis. Ultrastructural analysis of PRL-producing cells stained sequentially using the two different anti-tPRL antibodies labeled with immunogold of two size classes indicated that both tPRLs appear in the same granules. These findings suggest that the biological significance of two forms of PRL in the adult tilapia is not a function of differential regulation of two different classes of PRL cells or differential release of unique secretory granules.
Specker
J L
JL
Department of Zoology, University of Rhode Island, Kingston 02881-0816.
Kishida
M
M
Huang
L
L
King
D S
DS
Nagahama
Y
Y
Ueda
H
H
Anderson
T R
TR
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D002851:Chromatography, High Pressure Liquid; D003594:Cytoplasmic Granules / Q000737:chemistry*; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D013006:Growth Hormone / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D010902:Pituitary Gland / Q000737:chemistry* / Q000648:ultrastructure; D011388:Prolactin / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D017210:Tilapia / Q000378:metabolism*
D000906:Antibodies; D000939:Epitopes; D010446:Peptide Fragments; D011388:Prolactin; D013006:Growth Hormone