Immunocytochemical localization of luteinizing hormone and follicle-stimulating hormone in the equine pituitary.

Gonadotropin-specific primary antisera and gold-conjugated secondary antibodies were used to immunocytochemically localize gonadotropins in the anterior pituitary of intact pony mares. Electron microscopy was then used to characterize the ultrastructure and immunoreactive staining characteristics of equine gonadotropes. Cells containing LH were morphologically indistinguishable from those containing FSH. Gonadotropes were relatively large and commonly had eccentric nuclei. The rough endoplasmic reticulum was well developed and dilated. Secretory granules were present in two morphologically distinct forms. Large polymorphic granules were generally located in perinuclear cytoplasmic areas, whereas small and uniformly shaped granules were in the peripheral cytoplasm, close to the cell membrane. Double-labeling revealed cells with granules that stained for both LH and FSH as well as cells that stained for either LH or FSH. Gonadotropes constituted 15 to 32% of all pituitary cells in the anterior pituitaries from the three mares included in this study. Cells that stained for only LH constituted 2 to 16% of all pituitary cells, cells that stained for only FSH ranged from 1 to 4.5%, and cells staining for both hormones constituted 6.2 to 24% of the pituitary cells. These results indicate that there are in fact three distinct subclasses of gonadotropes in the equine anterior pituitary based on immunocytochemical staining, which is similar to the situation described for several other mammalian species.

Immunocytochemical localization of prolactin and growth hormone in the equine pituitary.

The ultrastructural and immunoreactive staining characteristics of cells containing prolactin (lactotropes) and growth hormone (GH; somatotropes) in the anterior pituitaries of gonadally intact pony mares were studied at the electron microscopic level. Lactotropes included two morphological subsets: Type I cells were larger and contained large, dense, polymorphic granules that were scattered throughout the cytoplasm; Type II cells were smaller and contained small, dense, polymorphic granules that were predominantly found in peripheral areas of the cytoplasm. Lactotropes constituted 5 to 16% of the total number of cells in the pituitary. Somatotropes were medium-sized cells containing uniform, large, dense secretory granules. The somatotropes contained the largest secretory granules in the pituitary and represented 11 to 26% of the total number of cells. Type I lactotropes and somatotropes were readily distinguishable without immunocytochemical staining. Double-labeling of pituitary sections allowed for characterization of cells that contained both hormones (mammosomatotropes). These cells were morphologically indistinguishable from Type I lactotropes and constituted 6.5 to 16.5% of the total number of cells. Results from this study demonstrated that there are two cell populations that contain only prolactin (Type I and II lactotropes) and one cell population that contains only GH (somatotropes) in the equine pituitary, and an additional subset of cells that contains GH and prolactin in the same secretory granules.

Growth hormone and prolactin concentrations in plasma of horses: sex differences and the effects of acute exercise and administration of growth hormone-releasing hormone.

Three experiments were conducted to determine 1) the relationship between prolactin and growth hormone (GH) secretion in mares and the response to GH-releasing hormone (GHRH), 2) whether plasma GH and prolactin concentrations differed among mares, stallions, and geldings, and 3) whether sexual differences existed after administration of GHRH and acute exercise. In Exp. 1, 10-min blood samples were collected from 12 mares for 8 h, and GHRH (0, 45, 90, or 180 micrograms) was administered at 6 h. In Exp. 2, 15-min blood samples were collected for 4 h from 10 mares, stallions, and geldings. In Exp. 3, eight horses of each sexual status were administered GHRH at 0900; later that day, each horse was exercised for 5 min. Blood samples were collected every 10 min around each event. In Exp. 1, prolactin concentrations decreased (P < .01) over the 8-h period, and there was an average of 2.9 +/- .5 episodes of increased secretion during that time; there was no correlation between these episodes and those in GH secretion. Prolactin concentrations were not affected (P > .1) by GHRH. In Exp. 2, average concentrations of GH were 2.4, 8.6, and 8.5 ng/mL for mares, stallions, and geldings, respectively; males differed from females (P < .05). Stallions and geldings had more (P < .05) peaks in GH concentrations and greater (P < .05) amplitude of peaks than mares. In contrast, prolactin concentrations were greater (P < .02) in mares and stallions than in geldings. In Exp. 3, GH response to GHRH was greater (P < .03) in stallions than in mares or geldings.(ABSTRACT TRUNCATED AT 250 WORDS)

Growth hormone in mares and stallions: pulsatile secretion, response to growth hormone-releasing hormone, and effects of exercise, sexual stimulation, and pharmacological agents.

Short-term patterns of growth hormone (GH) secretion and factors affecting it were studied in mares and stallions. In Exp. 1, hourly blood samples were collected from three mares and three stallions in summer and winter. Although GH concentrations varied in a pulsatile manner in all horses, there was no effect of sex or season (P greater than .1) on plasma GH concentrations and no indication of a diurnal pattern of GH secretion. In Exp. 2, 10-min blood samples were drawn for 8 h from 12 mares; after 6 h, porcine GH-releasing hormone (GHRH) was administered i.v. at 0, 45, 90, or 180 micrograms/mare (three mares per dose). Pulsatile secretion of GH occurred in all mares and averaged 2.4 +/- .3 peaks/6 h; amplitudes were variable and ranged from 2.6 to 74.4 ng/mL. Eight of nine mares responded within 20 min to GHRH injection, but there was no difference (P greater than .1) among the three doses tested. In Exp. 3, plasma GH concentrations in stallions increased (P less than .05) 8- to 10-fold after 5 min of acute physical exercise or exposure to an estrual mare. Restraint via a twitch (5 min) and epinephrine administration (3 mg i.v.) also increased (P less than .05) plasma GH concentrations by approximately fourfold. In Exp. 4 and 5, administration of either .4, 2, or 10 mg of thyrotropin-releasing hormone (TRH) or 100 or 500 mg of sulpiride (a dopamine receptor antagonist) increased (P less than .01) plasma prolactin concentrations but had no effect (P greater than .1) on GH concentrations during the same period of time.(ABSTRACT TRUNCATED AT 250 WORDS)

Plasma concentrations of cortisol, prolactin, luteinizing hormone, and follicle-stimulating hormone in stallions after physical exercise and injection of secretagogue before and after sulpiride treatment in winter.

Ten lighthorse stallions were used to determine 1) whether prolactin (PRL) and cortisol responses previously observed after acute exercise in summer would occur in winter when PRL secretion is normally low, 2) whether subsequent treatment with a dopamine receptor antagonist, sulpiride, for 14 d would increase PRL secretion and response to thyrotropin-releasing hormone (TRH) and exercise, and 3) whether secretion of LH, FSH, and cortisol would be affected by sulpiride treatment. On January 11, blood samples were drawn from all stallions before and after a 5-min period of strenuous running. On January 12, blood samples were drawn before and after an i.v. injection of GnRH plus TRH. From January 13 through 26, five stallions were injected s.c. daily with 500 mg of sulpiride; the remaining five stallions received vehicle. The exercise and secretagogue regimens were repeated on January 27 and 28, respectively. Before sulpiride injection, concentrations of both cortisol and PRL increased (P less than .05) 40 to 80% in response to exercise; concentrations of LH and FSH also increased (P less than .05) approximately 5 to 10%. Sulpiride treatment resulted in (P less than .05) a six- to eightfold increase in daily PRL secretion. The PRL response to TRH increased (P less than .05) fourfold in stallions treated with sulpiride but was unchanged in control stallions. Sulpiride treatment did not affect (P greater than .05) the LH or FSH response to exogenous GnRH.(ABSTRACT TRUNCATED AT 250 WORDS)

Function of ovine corpora lutea after administration of luteinizing hormone-releasing hormone.

Ewes were treated with an agonistic analog of luteinizing hormone-releasing hormone (LH-RH) during the luteal phase (d 10) of the estrous cycle. Function of natural and hormonally-induced corpora lutea (CL) was evaluated by measurements of progesterone in sera or luteal tissue. Synthesis and secretion of progesterone by natural CL were not chronically altered by LH-RH. Likewise, there was no in vitro effect of LH-RH on luteal function. When natural CL were surgically removed, newly formed CL functioned at a defective level. Hysterectomy shortly after ovulation did not significantly influence such luteal activity. Induction of ovulation by LH-RH during the follicular phase (d 16) in uterus-intact ewes was followed by normal profiles of luteal secretion of progesterone; serum concentrations of progesterone in animals that were hysterectomized increased in association with development of the CL, but then plateaued at a subnormal level. There were no differences in patterns of secretion of luteinizing hormone in response to LH-RH due to stage of the estrous cycle. Follicles stimulated to ovulate during the luteal phase contained low numbers of steroidogenically-deficient granulosal-lutein cells. These results indicate that: ovine CL are not sensitive to exogenous LH-RH; luteal dysfunction is a consequence of ovulation during the luteal phase, and the etiology of this abnormality appears to be linked with the developmental status of the ovulatory follicle; and CL that are formed from ovulation of a matured follicle begin to develop normally, but then function at a defective rate in the absence of the uterus.