Studies on the regulation of growth hormone release from the proximal pars distalis of male tilapia, Oreochromis mossambicus, in vitro

The in vitro effects of several factors, including cortisol, somatostatin (SRIF), and medium osmotic pressure, on growth hormone (GH) release from the tilapia pituitary were examined in relation to fish size. Spontaneous GH release from the proximal pars distalis (PPD) of approximately 60-g fish was significantly less than that from tissue of fish weighing either approximately 120 or approximately 280 g when incubated in 340 m phi smolal medium. While GH content of the PPD cultures (tissue + medium measured by densitometry) increased consistently with fish size, GH concentration (per microgram of tissue protein) was variable, being highest in 120-g fish and lowest in 280-g fish. Moreover, GH concentration was not related to GH release. Fish size also appeared to be important in the responsiveness of GH cells to stimulation by cortisol (Nishioka et al., 1985) and by increased osmotic pressure. In cultures of PPD from approximately 60-g fish, in which spontaneous release was relatively low, cortisol and increased medium osmotic pressure significantly enhanced release. Cortisol and hyperosmotic medium were without significant effect, however, on GH release from PPD of approximately 120-g fish, which showed high spontaneous release. In contrast, SRIF, a potent inhibitor of GH secretion, was effective in lowering GH release regardless of fish size. Nevertheless, SRIF was apparently more effective in inhibiting GH release from tissue of 60-g fish than from tissue of 120-g fish. Our data suggest that GH secretion may be augmented when smaller tilapia (approximately 60 g) are transferred to seawater, a situation in which blood cortisol and osmotic pressure would presumably be elevated.     

Somatostatin and altered medium osmotic pressure elicit rapid changes in prolactin release from the rostral pars distalis of the tilapia, Oreochromis mossambicus, in vitro

Prolactin (PRL) cells in the rostral pars distalis of the tilapia Oreochromis mossambicus respond to somatostatin (SRIF) and reduced medium osmotic pressure within 10-20 min of exposure during perifusion incubation. Pieces of rostral pars distalis tissue were removed from freshwater-adapted tilapia and were preincubated in [3H]leucine in static culture (355 m phi smolal) for 48 hr. Following preincubation, they were placed in the perifusion apparatus and baseline release was established for 3 hr in hyperosmotic medium (355 m phi smolal). Exposure to hyposmotic medium (280 m phi smolal) resulted in a rapid and steep rise in the release of [3H]PRL, which remained elevated for more than 2 hr. When SRIF was added simultaneously with hyposmotic medium, the rise in PRL release normally initiated by reduced osmotic pressure was prevented. Somatostatin also quickly reduced release that had been previously elevated by exposure to hyposmotic medium. The time course of these changes suggests that SRIF and altered osmotic pressure act on PRL secretion in at least partial independence of effects which they may have on PRL synthesis in the tilapia pituitary.     

Prolactin and growth hormone secretion during long-term incubation of the pituitary pars distalis of mature chum salmon, Oncorhynchus keta

The effects of hypotonicity of the medium on prolactin (PRL) and growth hormone (GH) release from the pars distalis (PD) of mature chum salmon pituitary were examined during culture for 4 days. Large amounts of PRL and GH were released during the first 6 hr and secretion decreased rapidly thereafter. The amounts of GH released as well as the residual content in PD were 10 times greater than those of PRL. There was no difference in PRL or GH release between the PD incubated in isotonic medium (325 mOsm) and those in hypotonic medium (250 mOsm) even during the first 6 hr. Female PD secreted more PRL than male PD during the first 6 hr, whereas a sex difference was not observed in GH release. Considerable amounts of both PRL and GH remained in the PD at the end of the culture period. The amount of PRL released during 4 days of culture from female PD (37-39%) was greater than that from male PD (23-26%), and the amount of GH released was less in females (28-32%) than that in males (53-54%). Release of PRL and GH appears to be mainly under stimulatory hypothalamic control in the mature chum salmon.     

Novel effect of vasoactive intestinal polypeptide and peptide histidine isoleucine: inhibition of in vitro secretion of prolactin in the tilapia, Oreochromis mossambicus

The effects of vasoactive intestinal polypeptide (VIP) and peptide histidine isoleucine (PHI) on the in vitro secretion of two prolactins (PRL) from the rostral pars distalis (RPD) and of growth hormone (GH) from the proximal pars distalis (PPD) of the pituitary of the tilapia (Oreochromis mossambicus) were studied. RPDs were incubated for 20 hr in hypoosmotic (280-300 mOsm) or hyperosmotic (340-350 mOsm) Krebs-Ringer bicarbonate medium with added peptide concentrations of 0 (control), 0.3, 3.0, 30, and 300 nM; similarly, PPDs were incubated with the same peptide concentrations in isoosmotic (325 mOsm) medium supplemented with cortisol. PRL and GH in the tissue and secreted into the medium were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by soft laser densitometry of the protein band(s). Neither VIP nor PHI has a detectable effect on the secretion of GH. Secretion of the two PRLs is significantly inhibited by VIP and PHI in both hyperosmotic and hypoosmotic medium. In hyperosmotic medium, 300 nM VIP inhibits secretion of both PRLs by 47%, whereas in hypoosmotic medium, 300 nM VIP inhibits their secretion by 27%. PHI inhibits their secretion by ca. 65% in hyperosmotic medium and by 40% in hypoosmotic medium. There is preliminary immunocytochemical evidence for some VIP-like immunoreactivity (IR), but no conclusive indication of PHI-like IR, in the hypothalamo-hypophysial area. The inhibitory actions of VIP and PHI on PRL secretion in tilapia are in contrast to the known stimulatory actions of VIP and PHI on PRL secretion in tetrapods.     

In vitro effects of somatostatin and urotensin II on prolactin and growth hormone secretion in tilapia, Oreochromis mossambicus

The control of release of two recently characterized forms of prolactin (PRL) of molecular mass 24 and 20 kDa was investigated. The rostral pars distalis of male tilapia was incubated singly in a hypotonic modified Krebs-Ringer bicarbonate medium in order to stimulate PRL release; for comparison, the proximal pars distalis containing growth hormone (GH) cells was incubated in isotonic medium with or without 1 microgram/ml cortisol to stimulate GH release. The release of both PRLs and GH into the medium was measured by sodium dodecyl sulfate (SDS)--polyacrylamide gel electrophoresis followed by densitometry. Both somatostatin and synthetic (Gillichthys) urotensin II, a partial somatostatin homolog and analog from the teleost caudal neurosecretory system, significantly inhibited the release of both PRLs. Somatostatin significantly inhibited GH release, but urotensin II had no significant effect.     

The loss of 45Ca2+ associated with prolactin release from the tilapia (Oreochromis mossambicus) rostral pars distalis

The relationship between tritium 3H-labeled prolactin (PRL) release and the loss of tissue-associated 45Ca2+ was examined in the tilapia rostral pars distalis (RPD) using perifusion incubation under conditions which inhibit or stimulate PRL release. Depolarizing [K+] (56 mM) and hyposmotic medium (280 mOsmolal) increased both the release of [3H]PRL and the loss of 45Ca2+. The responses to high [K+] were faster and shorter in duration than those produced by reduced osmotic pressure. The depletion of Ca2+ from the incubation medium with 2 mM EGTA suppressed the [3H]PRL response evoked by high [K+] or reduced osmotic pressure. Exposing the tissues to Ca(2+)-depleted medium in the absence of high [K+] or reduced osmotic pressure produced a sharp, but brief, increase in 45Ca2+ loss. Cobalt (10(-3) M), a competitive inhibitor of calcium-mediated processes, inhibited the [3H]PRL response to hyposmotic medium and to high [K+]. Cobalt also diminished the increased loss of 45Ca2+ evoked by exposure to reduced osmotic pressure, but was ineffective in altering responses to high [K+]. Methoxyverapamil (D600; 10(-5) M), a blocker of certain voltage-sensitive Ca2+ channels, did not alter either the [3H]PRL or the 45Ca2+ responses to high [K+] and reduced osmotic pressure. Taken together with our earlier studies, the present findings suggest that exposure to high [K+] or hyposmotic medium produces rapid changes in the Ca2+ metabolism of the tilapia RPD that are linked to the stimulation of PRL secretion. Nevertheless, the increased 45Ca2+ loss, but not [3H]PRL release, upon exposure to Ca(2+)-depleted media suggests that Ca2+ loss may not always reflect intracellular events that lead to PRL release.     

Colocalization of GnRH binding sites with gonadotropin-, somatotropin-, somatolactin-, and prolactin-expressing pituitary cells of the pejerrey, Odontesthes bonariensis, in vitro.

Previous studies in the pejerrey, Odontesthes bonariensis, have demonstrated that fibers with immunoreactivity to gonadotropin-releasing hormone (ir-GnRH) reach all areas of the pituitary gland, the rostral pars distalis (RPD), the proximal pars distalis (PPD), and the pars intemedia (PI). A close association was shown between ir-GnRH fibers and gonadotropin (GtH)-, growth hormone (GH)-, somatolactin (SL)-, and prolactin (PRL)-expressing cells. The presence of only one GnRH variant, suspected to be a novel form, has been shown in pituitary extracts of this fish. In addition, GnRH may stimulate GtHs, GH, SL, and PRL levels in different fish species. The objective of the present study was to seek GnRH receptors and therefore colocalization with GtHs, GH, SL, and PRL cells in O. bonariensis using a pituitary primary cell culture system. GnRH binding sites were revealed by autoradiography of an iodinated superactive GnRH agonist ([(125)I]GnRH-A) and pituitary cells were identified by immunocytochemistry using piscine antisera. Following autoradiography, silver grains representing specific [(125)I]GnRH-A binding were associated with anti GtH, GH, SL, and PRL positive cells. These results demonstrate the presence of GnRH binding sites on these cells. It is suggested that GnRH may play a wide role in the neuroendocrine control of different pituitary hormones in addition to the GtHs.     

Effects of GnRH-associated peptide and its component peptides on prolactin secretion from the tilapia pituitary in vitro

The rostral pars distalis (RPD), containing mainly prolactin (PRL)-secreting cells, of the pituitary from immature and mature tilapia was incubated for 16 hr at 27 degrees in hypoosmotic medium (300 mOsm/kg) in the presence (10(-8) and 10(-11) M) or absence of the human GnRH-associated peptide (GAP) molecule, a potent PRL-inhibiting factor in mammals (Nikolics et al., Nature (London) 316, 511, 1985), and of a series of its component peptides. The release of the two forms of PRL in tilapia into the medium was measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by densitometry. The variability inherent in this method was normalized by calculating PRL release as the percentage of the total hormone present in both tissue and medium. Newly synthesized PRL was detected by incorporation of [35S]methionine, introduced into the culture medium, by the PRL molecules. In immature tilapia, GAP inhibited the release of total PRL while stimulating the release of newly synthesized large PRL. Among the GAP fragments tested, 28-36 was the fragment that most significantly affected PRL secretion. Both concentrations of fragment 28-36 stimulated the release of newly synthesized PRL from immature rostral pars distalis (RPDs). This stimulation appears to be dependent on the osmotic pressure of the medium since this fragment did not affect PRL secretion in hyperosmotic medium (340 mOsm/kg). Fragment 38-49 inhibited total PRL release from mature RPDs. Fragment 51-66 stimulated the release of total PRL from mature RPDs. Examination of tissue and medium values in densitometric units after incubation with fragments 28-36 and 51-66 indicated that while the tissue content of PRL was decreased, the medium content of PRL was not affected. This suggests that fragments 28-36 and 51-66, in opposition to the situation found when the data are expressed as percentage release of PRL, may not stimulate PRL release but may instead decrease the tissue content of PRL. These results suggest that the entire human GAP molecule, as well as some of its fragments, may have direct effects on the PRL cells in the tilapia pituitary.     

Immunocytochemical study of growth hormone, prolactin, and thyroid-stimulating hormone in the adenohypophysis of the sea lamprey, Petromyzon marinus L., during its upstream migration

Immunoreactivity to growth hormone (GH), prolactin (PRL), and thyroid-stimulating hormone (TSH) was found in the adenohypophysis of the sea lamprey, Petromyzon marinus L., during its upstream migration, by immunoperoxidase techniques with antisera to mammalian pituitary hormones. Cross-reactivity to GH and PRL was found in two different cell populations in the proximal pars distalis. Specific immunostaining for GH and PRL was absent in other parts of the lamprey pituitary. Immunoreactive TSH cells were located only in the rostral pars distalis, and corresponded in shape and size to the large basophils in this region of the lamprey pituitary. These results suggest that mammalian-like GH, PRL, and TSH are produced in the pituitary of the lamprey.     

In vitro effects of thyrotropin-releasing hormone and somatostatin on prolactin and growth hormone release by the pituitary of Poecilia latipinna. I. An electrophoretic study

Pituitary glands were removed from Poecilia latipinna which had been maintained in one-third seawater and were incubated for 18 hr in media of either 300 mosmol/kg (OP300) or 340 mosmol/kg (OP340) osmotic pressure for measurement of both total and newly synthesised prolactin (PRL) and growth hormone (GH) release. Thyrotropin-releasing hormone (TRH) at 100 ng/ml increased release of total and newly synthesised PRL into OP340, but not into OP300, medium. Conversely, 300 ng/ml of somatotropin-release-inhibiting factor (SRIF) inhibited total and newly synthesised PRL release into OP300, but not OP340, medium. At 1000 ng/ml, SRIF inhibited total PRL release into both media, but newly synthesised PRL release was reduced significantly only in OP300 medium. The release of GH was unaffected by 100 ng/ml TRH in OP300 medium, but both total and newly synthesised GH release were enhanced by this dose in OP340 medium. SRIF at 300 ng/ml reduced total GH release into OP300 medium, whereas the release of newly synthesised GH was inhibited in OP340 medium. At 1000 ng/ml, SRIF inhibited total GH release into both media, but release of the newly synthesised hormone was not significantly altered. These results suggest that TRH can stimulate and SRIF inhibit both PRL and GH release by Poecilia pituitaries, but that these effects may be modulated by plasma osmotic pressure.     

Effects of osmotic pressure and calcium ion on prolactin release in vitro from the rostral pars distalis of the tilapia Sarotherodon mossambicus

As in previous studies, prolactin (PRL) release from the rostral pars distalis of the euryhaline teleost Sarotherodon mossambicus in vitro was inversely related to the osmotic pressure of the incubation medium. In contrast, PRL release was independent of Ca²⁺ concentration when this divalent ion was included in the medium. Release was completely blocked, however, when calcium was excluded. While it appears unlikely that variations in plasma Ca²⁺ within the physiological range can act directly at the pituitary to alter prolactin secretion, our findings are consistent with the notion that calcium is an essential ion mediating PRL release.     

Effects of somatostatin and urotensin II on tilapia pituitary prolactin release and interactions between somatostatin, osmotic pressure Ca++, and adenosine 3',5'-monophosphate in prolactin release in vitro

Both somatostatin (SRIF) and urotensin II, a dodecapeptide from the teleost caudal neurosecretory system, inhibit PRL release from the organ-cultured rostral pars distalis of the tilapia, Sarotherodon mossambicus, in a dose-related manner. The inhibitory action of SRIF on PRL release was completely prevented by the presence of the calcium ionophore A23187. PRL release was also blocked when Ca++ was excluded from the incubation medium, even in the presence of the ionophore. Both dibutyryl cAMP (dbcAMP) and the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, alone or in combination, stimulated PRL release during incubation in high osmotic pressure medium. The effect of dbcAMP appeared to be dose related. Together, dbcAMP and 3-isobutyl-1-methylxanthine were also effective in preventing the inhibition of PRL release by SRIF. These results are consistent with the notion that Ca++, and possibly cAMP, may be important mediators of PRL secretion, and it is likely that SRIF may inhibit PRL release by blocking a Ca++- or cAMP-mediated mechanism.     

Ontogeny of adenohypophyseal cells in the pituitary of the American shad (Alosa sapidissima)

The distribution and ontogeny of adenohypophyseal cells have been studied in the pituitary gland of embryos, larvae, and juveniles of the clupeid American shad (Alosa sapidissima) using immunocytochemical techniques. In juvenile specimens, adenohypophysis was composed of rostral pars distalis (RPD), formed by cavities lined by prolactin (PRL), adrenocorticotropic hormone (ACTH), and gonadotropic hormone (GTH) cells; proximal pars distalis (PPD), containing growth hormone (GH), GTH, and putative thyroid stimulating hormone (TSH) cells; and pars intermedia (PI) with somatolactin (SL) and melanophore stimulating hormone (MSH) cells. At 3 days post-fertilization (3 days pre-hatching) the pituitary of embryos consisted of an oval mass of cells, close to the ventral margin of the diencephalon, divided in rostral and caudal regions. At this time PRL and ACTH cells appeared in the rostral region of the adenohypophysis, while SL cells were observed in the caudal region where MSH cells showed reactivity 1 day before hatching. At variance, GH cells showed a weak immunoreactivity in the rostral portion at hatching that increased 2 days latter. GTH cells also showed weak immunoreactivity in the rostral region of the adenohypophysis at hatching time. Two days later GTH cells were located in the rostral and central regions of the adenohypophysis. At hatching, the neurohypophysis was very small and no nerve processes were seen to penetrate the adenohypophysis tissue. After hatching, the pituitary gland elongated and in 7 days old larvae, the RPD showed a small lumen surrounded by a palisade of PRL, ACTH, and GHT cells; the PPD showed GH and GTH cells while the PI contained SL and MSH cells. The adenohypophysis and neural lobe increased in size with development and, in 42 days old larvae, they were similar to those of juvenile specimens.     

Immunocytochemical identification of the prolactin- and growth hormone-secreting cells in the teleost pituitary with antisera to tilapia prolactin and growth hormone

Antisera raised to highly purified tilapia (Sarotherodon mossambicus) prolactin (PRL) and growth hormone (GH) were used to locate PRL and GH cells in the adenohypophysis of seven species of teleosts by immunoenzymological methods using horseradish peroxidase. The only pituitary cells in the tilapia that immunologically reacted with anti-tilapia PRL were the PRL cells located in the rostral pars distalis (RPD). In the pituitary glands of two species of salmonids and two species of marine fishes, the tilapia PRL antibodies bound to both PRL and GH cells; no prolactin-immunoreactive cells were identified in the goldfish and eel pituitaries. Antiserum to tilapia GH reacted specifically with the GH cells located in the proximal pars distalis of all teleosts examined in this study, but not with any presumed PRL cells in the RPD. These results indicate that tilapia PRL antisera are specific for tilapia PRL cells but not necessarily for these cells in other teleost species. Tilapia GH antisera showed no species specificity among the teleosts tested.     

Ontogenesis of the three parts of the fetal rat adenohypophysis. A detailed immunohistochemical analysis

The sequential changes in the histological pattern of anterior pituitary cytodifferentiation of the rat are described. The first labeled cells were ACTH positive and were detected in the pars tuberalis on postconceptual day 13. On day 14 ACTH cells also appeared in the ventral periphery of the pars distalis. On fetal day 15 the pars tuberalis anlage was characterized by numerous well-stained ACTH cells and by some weakly labeled FSH-beta, LH-beta, TSH-beta, GH and PRL cells while the pars distalis showed only ACTH positivity. On day 16 of gestation the ACTH cells were equally distributed throughout the whole pars distalis, while LH-beta, FSH-beta, TSH-beta, PRL and GH immunoreactive cells were localized either in the ventral region of the pars distalis only or were evenly distributed throughout the pars distalis. The present immunocytochemical data suggest that in the pars distalis the hypophyseal cell differentiation follows a clear rostrocaudal, ventrodorsal direction and that the time sequence of the functional differentiation of the adenohypophysis is pars tuberalis, pars distalis and pars intermedia.     

Immunohistochemical localization of gonadotropin releasing hormones in the brain and pituitary gland of the Nile perch,Lates niloticus (Teleostei,Centropomidae)

In the present study we investigated the distribution of gonadotropin-releasing hormones (GnRH) in the brain of Lates niloticus and their association with different pituitary cell types using immunohistochemical techniques. We found immunoreactive (ir) chicken GnRH-II (cGnRH-II) and mammalian GnRH (mGnRH) as the main components of the GnRH-ir system within the brain of the Nile perch. The results indicate that mGnRH and cGnRH are localized in different neurons: mGnRH-ir perikaria were observed in the preoptic region particularly in the organum vasculosum laminae terminalis (OVLT) and in the nucleus lateralis tuberis pars posterior (NLTP) of the mediobasal hypothalamus. These cell bodies are located along a continuum of ir-fibers that could be traced from the olfactory nerve to the pituitary. mGnRH-ir fibers were detected in many parts of the brain (olfactory bulbs, ventral telencephalon, hypothalamus, and mesencephalon) and in the pituitary. cGnRH-ir cell bodies are restricted to the optic tract, but few scattered fibers could be detected in different parts of the brain. The pituitary exhibited very few cGnRH-II ir fibers, contrasting with an extensive mGnRH innervation. Moreover, mGnRH-ir fibers were targeting the three areas of the pituitary gland: rostral pars distalis (RPD), proximal pars distalis (PPD), and pars intermedia (PI). Double immunolabeling studies showed GnRH-ir fibers in close proximity with prolactin (PRL)- and adrenocorticotropic hormone (ACTH)-producing cells in the RPD, growth hormone (GH)-producing cells in the PPD, gonadotropins (GTHs)-producing cells in the PPD in the external border of the PI, and with somatolactin (SL)- and alpha-melanocyte stimulating hormone (alpha-MSH)-producing cells in the PI. Our results showed direct morphological evidence for a close association of GnRH-ir fibers with the different adenohypophysial cell types. These results suggest a multiple role of GnRH in the regulation of various pituitary hormones' release.     

Immunocytochemical identification and localization of the different cell types in the pituitary of the seabass (Dicentrarchus labrax)

Antisera raised against chum salmon prolactin (PRL), trout growth hormone (GH), mammalian adrenocorticotropic hormone (ACTH), thyroid-stimulating hormone (TSH), luteinizing hormone (LH), and alpha-melanophore-stimulating hormone (alpha-MSH) were used to localize PRL, GH, ACTH, gonadotropic, TSH, and MSH cells in the hypophysis of the teleost Dicentrarchus labrax using the unlabeled peroxidase anti-peroxidase method. In the rostral pars distalis, ACTH cells stained very intensively with anti-ACTH; so did the MSH cells in the pars intermedia. The prolactin cells stained very specifically with anti-prolactin without staining the growth hormone cells. In the proximal pars distalis anti-GH, anti-TSH beta, and anti-LH stained selectively the corresponding cells; with these antisera no cross-reaction with any other cell type was observed. Anti-alpha-MSH only stained cells in the pars intermedia. Some cells in the pars intermedia did not react at all; these could correspond to the PAS-positive cells. A characteristic feature was positive staining with anti-LH in some cell groups encircling the pars intermedia, indicating the fact that in the seabass some cells of the proximal pars distalis surround the pars intermedia.     

Chronological appearance of the different hypophysial hormones in the pituitary of sea bass larvae (Dicentrarchus labrax) during their early development: an immunocytochemical demonstration

Antisera raised against chum salmon prolactin (PRL), rainbow trout growth hormone (GH), mammalian adrenocorticotropic hormone (ACTH), thyroid-stimulating hormone (TSH), and luteinizing hormone (LH) were used to study the chronological appearance of immunoreactivity for PRL, GH, ACTH, TSH, LH, and melanocyte-stimulating hormone (MSH) in the pituitary of sea bass larvae (Dicentrarchus labrax) during the first 26 days after hatching. The anti-ACTH gives positive immunostaining in the ACTH cells as well as in the MSH cells; however, the two cell types can easily be distinguished by their different localization in the pituitary: ACTH in the rostral pars distalis, MSH in the pars intermedia. The first day after hatching cells immunoreactive for TSH, GH and ACTH could already be noticed, ACTH reacted strong in the pars intermedia but very weak in the rostral pars distalis. Cells immunopositive for PRL became visible between Days 9 and 15. With anti-LH, no positive reaction could be obtained during the first 26 days after hatching.     

Estradiol-17 beta and thyrotropin-releasing hormone stimulate prolactin release from the pituitary gland of a teleost fish in vitro

The effects of estradiol-17 beta (E2) and thyrotropin-releasing hormone (TRH) on prolactin (PRL) release were investigated using the organ-cultured rostral pars distalis (RPD) of the tilapia, Oreochromis mossambicus. Spontaneous PRL release into hyperosmotic medium increased in a dose-related manner following E2 pretreatment in vitro. In addition, TRH stimulated a dose-related increase in PRL release from E2-preincubated RPD's, but had no effect on tissues not previously exposed to E2. The maximal PRL response, nearly three times control levels, occurred at 50 nM TRH. Higher doses of TRH were less effective in stimulating PRL release. These findings indicate that TRH may be an important hypothalamic prolactin-releasing factor in the tilapia. Furthermore, the marked potentiation of the action of TRH on PRL release following exposure to E2 suggest that there may be a shift in the control of PRL secretion with changes in the reproductive state of the tilapia.