Preparation and use of specific antibodies to the β-I and β-II subunits of gonadotropic hormone from Fundulus heteroclitus pituitary

Fundulus heteroclitus naturally spawns with a semilunar periodicity throughout most of the year in its southernmost habitat, an activity that can be maintained in the laboratory. The alpha and two beta subunits comprising F. heteroclitus gonadotropic hormones (GtHs) I and II have been sequenced, and antibodies have been raised against unique peptides found in each of the two beta subunits. On immunoblots of pituitary proteins, each antibody recognizes a single band with a molecular mass of 16-17 kDa, somewhat larger than the deduced sizes (11-13 kDa) of the unglycosylated subunits. Each antibody also recognizes a different subset of pituitary cells in the central (GtH I) and peripheral (GtH II) proximal pars distalis, regions that display the typical tinctorial properties of gonadotrops. The distribution and distinct separation of cells containing GtH beta subunits I and II thus differ from those found for previously described teleost species, most of which are salmonids that engage in a single spawning episode during the year. The availability of these antibodies thus makes F. heteroclitus an inexpensive, easily manipulated model system for studies on the hormonal regulation of fractional spawning common to a large class of commercially important species other than salmonids. (C) 2001 Academic Press.