TSH Cells Research Articles

Immunocytochemical studies on the pituitary pars distalis of the tropical vespertilionid bat, Scotophilus heathi, with reference to the ovarian cycle.

Changes in the anterior pituitary in relation to the ovarian cycle and ovulation in female Scotophilus heathi are described. TSH, LH, FSH and PRL cells have been identified in the pars distalis (PD) by immunocytochemistry using the peroxidase-antiperoxidase technique. LH, FSH and PRL cells exhibited significant variation during the reproductive cycle of female S. heathi. The areal fractions of the PD occupied by LH and FSH cells were significantly (p < 0.05) higher during winter dormancy and the breeding phase as compared to the quiescence phase. The mean areal fraction of LH and FSH cells declined in March, following ovulation. The areal fraction of PD occupied by PRL cells varied from a minimum during winter dormancy to a maximum during pregnancy. Several of the PRL cells exhibited a degranulated appearance during winter dormancy suggesting PRL release. The areal fraction of TSH immunoreactive cells did not show a significant (p > 0.05) variation during the reproductive cycle. No evidence of involution was observed in the PD cells during the period of delayed ovulation.

Immunocytochemical and morphometric study on the changes of TSH, PRL, GH and ACTH cells during the development of Bufo arenarum.

The development and dynamics of thyrotropin (TSH), adrenocorticotropic hormone (ACTH), prolactin (PRL), and growth hormone (GH) cells have been studied using immunocytochemical techniques and rabbit antisera, raised against the relevant human hormone, in the pars distalis of Bufo arenarum larvae at different stages of development. The four types of cells studied were identified in different zones of the pars distalis: TSH cells occurred mainly in the centro-ventral zone, ACTH cells in the rostral and dorsal zones, GH cells in the central and caudal zones, and PRL cells in the anterior two-thirds of the gland. This distribution pattern does not show significant changes with development. Morphometry and stereology were used to evaluate the changes observed in the volume of the pars distalis and the immunoreactive cells during development. The former increased during larval growth and decreased throughout the metamorphic climax. The results obtained on cell number, volume density, and total volume suggest that, during larval growth (pre-prometamorphosis) of B. arenarum, TSH, PRL, GH and ACTH cells show a proliferative period with storage of their hormones; a second period involving hormone release occurs at the metamorphic climax.

Pituitary physiological and ultrastructural changes during aging

Although it is known that both aged human beings and animals exhibit a decrease in pituitary hormone production and release, there is controversy about the true nature of these changes. Whereas some authors postulate an extra pituitary cause, i.e., a dopaminergic failure, others consider that the problem is at the level of the gland itself. CFW mice 2, 6, 12 and 18 months old, were i.p. inoculated withL: -Dopa. The pituitary gland was removed and sectioned, then observed and photographed in an electron microscope. Photomicrographs were scanned into a computer and digital image analysis made to determine secretory granules and organelle kinetics. Normal GH and TSH cells of elder mice responded to stimulation withL: -Dopa in a similar way as did cells of juveniles. The responsiveness rate of those cells to the amine precursor during the first hour of treatment was 38±3.5% and 26±0.3% of the studied cells in young and aged animals, respectively. Fully functional cells, i.e., GH and TSH cells showing 5 to 90% of their cytoplasm occupied by secretory granules (some of them immature), with a developed RER, and with absence of cell damage, were seen to be reduced from 98% in younger to 65.7% in aged animals. In successive steps, cells showed cell desquamation, darkened cytoplasm, differential swollen endoplasmic reticulum without secretory granules, increased number of secondary lysosomes (more than two per cell in a cross-section), differential swollen mitochondria, cytoplasm only containing two to five giant secondary lysosomes and finally, a complete loss of cell architecture. Therefore, GH and TSH cells at the end of their life-span were seen to derive into apoptotic images. In the whole gland, an increasing number of those pathologic images were seen as aging proceeded, thus reducing the number of normal and productive cells. From the results presented here it is proposed that an extrapituitary failure is insufficient to explain the reduced production of GH and TSH, and that the problem evolves also at the level of the gland itself.

Subchronic Toxicity Study in Rats with 1-Methyl-3-propylimidazole-2-thione (PTI): Effects on the Thyroid

Subchronic Toxicity Study in Rats with 1-Methyl-3-propylimidazole-2-thione (PTI): Effects on the Thyroid. Biegel, L. B., Cook, J. C., O'Connor, J. C., Aschiero, M., Arduengo, A. J., III, and Slone, T. W. (1995). Fundam. Appl. Pharmacol. 27, 185-194. A 90-day gavage study was performed to evaluate the subchronic toxicity of 1-methyl-3-propylimidazole-2-thione (PTI) when administered to Crl:CD BR rats. PTI is a chemical catalyst and is structurally similar to the thioureas, which are known to adversely affect the thyroid. Therefore, this study was designed to investigate the effects of PTI on the thyroid. Male and female rats were dosed with 0, 5, 10, 25, or 75 mg PTI/kg/day for 13 weeks. Clinical pathology examinations and pathology examination were performed and the following were measured periodically: serum T 3, T 4, and TSH, hepatic UDP- glucuronyltransferase activity, and cell proliferation of the thyroid and liver. Under the conditions of this study, the overall no-observed-adverse-effect level (NOAEL) for the subchronic effects of PTI in male and female rats was 10 mg PTI/kg/day. The NOAEL was based on the effects on the thyroid gland in male and female rats dosed with 25 and 75 mg PTI/kg/day, as well as the hepatic centrilobular fatty change, increased severity of chronic progressive nephropathy, fatty change in the adrenal medulla, and the substantial reduction in body weight and body weight gain. The primary target organs were the thyroid and liver. Alterations in thyroid hormones (T 3, T 4, and TSH) occurred predominantly at 25 and 75 mg/kg/day. Toxicologically significant alterations in T 3, T 4, and TSH levels, cell proliferation, and UDP-glucuronyltransferase activity occurred in rats dosed with 25 and 75 mg/kg/day, which correlated with organ weight and histopathological effects. Additionally, the effect of PTI on thyroid peroxidase activity, a key step in thyroid hormone synthesis, was evaluated in vitro using microswine thyroid microsomes. PTI was shown to inhibit thyroid peroxidase, with an IC50 of 0.02 M. These data suggest that PTI enhances the excretion of T 4 via induction of glucuronyltransferase and inhibits thyroid hormone synthesis via a direct affect on thyroid peroxidase. Both of these effects contribute to the disruption of the hypothalamic-pituitary-thyroid axis and result in sustained elevation of TSH and the corresponding thyroid hypertrophy and hyperplasia.

Expression of a novel receptor for the calcitonin peptide family and a salmon calcitonin-like peptide in the alpha-thyrotropin thyrotroph cell line.

We have previously shown an increased incidence of alpha-subunit-producing thyrotroph tumors after salmon calcitonin (sCT) injection into rats. However, it is not clear whether the effects of CT are direct or indirect. Our hypothesis was that for sCT to act directly, it must have a binding site on thyrotrophs. The alpha TSH cell line was used as a model for thyrotrophs. Receptor binding studies using alpha TSH membranes revealed a high affinity binding site for sCT [IC50 = 0.97 +/- 0.18 nM (n = 4); Kd = 5.45 +/- 0.43 nM (n = 3); binding capacity = 6.6 pmol/mg protein (n = 3)]. Rat CT did not compete with binding at this site. Receptor screening for other CT peptide family members revealed high specific binding for CT gene-related peptide (CGRP; IC50 = 0.25 +/- 0.08 nM; n = 3) and islet amyloid polypeptide (IC50 = 4.36 +/- 1.1 nM; n = 3). This together with the absence of rat CT binding excluded a conventional CT-binding site, and we propose a site similar to the CGRP subtype III receptor described in the rat nucleus accumbens. Guanosine 5'O-(3-thiotriphosphate) (GTP gamma S) (20 microM), reduced [125I]CGRP binding to 38% of maximal, indicating that this site is G-protein coupled. Immunocytochemically, all of the cells displayed intense sCT-like immunoreactivity, which was totally abolished by preabsorption of the antibody with sCT. The presence of this receptor supports the hypothesis that sCT mediates tumorigenesis via a direct pituitary action and, together with the coexistence of a sCT-like peptide in these cells, provides evidence for a possible autocrine role of this peptide in the control of thyrotroph function.

Expression of a novel receptor for the calcitonin peptide family and a salmon calcitonin-like peptide in the alpha-thyrotropin thyrotroph cell line

We have previously shown an increased incidence of alpha-subunit-producing thyrotroph tumors after salmon calcitonin (sCT) injection into rats. However, it is not clear whether the effects of CT are direct or indirect. Our hypothesis was that for sCT to act directly, it must have a binding site on thyrotrophs. The alpha TSH cell line was used as a model for thyrotrophs. Receptor binding studies using alpha TSH membranes revealed a high affinity binding site for sCT [IC50 = 0.97 +/- 0.18 nM (n = 4); Kd = 5.45 +/- 0.43 nM (n = 3); binding capacity = 6.6 pmol/mg protein (n = 3)]. Rat CT did not compete with binding at this site. Receptor screening for other CT peptide family members revealed high specific binding for CT gene-related peptide (CGRP; IC50 = 0.25 +/- 0.08 nM; n = 3) and islet amyloid polypeptide (IC50 = 4.36 +/- 1.1 nM; n = 3). This together with the absence of rat CT binding excluded a conventional CT-binding site, and we propose a site similar to the CGRP subtype III receptor described in the rat nucleus accumbens. Guanosine 5'O-(3-thiotriphosphate) (GTP gamma S) (20 microM), reduced [125I]CGRP binding to 38% of maximal, indicating that this site is G-protein coupled. Immunocytochemically, all of the cells displayed intense sCT-like immunoreactivity, which was totally abolished by preabsorption of the antibody with sCT. The presence of this receptor supports the hypothesis that sCT mediates tumorigenesis via a direct pituitary action and, together with the coexistence of a sCT-like peptide in these cells, provides evidence for a possible autocrine role of this peptide in the control of thyrotroph function.

Immunocytochemical and Morphometric Study of TSH, PRL, GH, and ACTH Cells in Bufo arenarum Larvae with Inhibited Thyroid Function

Changes in the pituitary cells involved in amphibian metamorphosis were studied in Bufo arenarum tadpoles with inhibited thyroid function. After hatching, larvae were treated for 5 months with potassium perchlorate (KCIO4), a goitrogenic substance known to prevent absorption of iodine from water or food, resulting in impaired thyroid hormone synthesis. Treated larvae continued to grow but halted their development in premetamorphosis, showing hyperplasic thyroid glands with disorganized follicles lacking colloid. Thyrotrop (TSH), lactotrop (PRL), somatotrop (GH), and corticotrop (ACTH) cells were stained immunocytochemically and the changes observed were evaluated morphometrically using an automatic image analyzer. Pars distalis volume increased in treated larvae. Morphometric results showed that, in treated tadpoles, TSH and PRL cell populations and cell volumes increased compared to normal larvae at the same stage. Changes in the GH and ACTH cell morphometry were slight. These results indicate that in Bufo tadpoles, after chronic thyroid hormone withdrawal, TSH, PRL, GH, and ACTH cells are able to develop and that thyroid hormones exert a strong feedback control on the synthesis and storage of TSH and PRL.

Estradiol-induced changes in TSH-like immunoreactivity of pituitary cells in female rats.

Beta-thyrotropin (TSH)-producing cells in the pituitary pars distalis of female rats were studied using rabbit anti-rat beta-thyrotropin (TSH) serum and a peroxidase-antiperoxidase (PAP) immunohistochemical procedure. Animals were neonatally treated with 1 mg estradiol-dipropionate (EDP) and sacrificed at different stages of development up to adulthood. Intact females of the corresponding age served as the controls. Morphometry and stereology were used to evaluate the changes in TSH-cell number and volume densities of the cells and nuclei. All morphometric parameters examined in estradiol-treated animals showed a significant decrease in comparison with immunoreactive TSH cells of age-matched controls. The most prominent EDP-induced changes were evident in peripubertal 38-day-old rats, the number and volumetric densities of both TSH cells and their nuclei being reduced by about 90% compared to intact pituitary. This decrease in the number and volume densities of TSH cells in EDP-treated rats explicitly demonstrated that this hormone, applied neonatally, has an inhibitory effect on TSH-immunoreactive cells up to adulthood, in accordance with our earlier data obtained by light and electron microscopy.

Characterization of Interleukin-2 (IL-2) receptor expression and action of IL-2 and IL-6 on normal anterior pituitary cell growth

The pituitary gland is known to express cytokines and their receptors. Interleukin-2 (IL-2) and IL-2 receptor (IL-2R) transcripts and protein products in corticotrophic cells have been previously described. IL-2R were also observed in PRL and GH-producing cells. The synthesis of IL-1 and IL-6 and their receptors by pituitary cells has also been reported. We recently demonstrated that the cytokines in addition to their regulatory effects on anterior pituitary hormone secretion are involved in the autocrine or paracrine regulation of pituitary growth. In the present study we show in normal rat anterior pituitary cells: (a) expression of IL-2Rα chain mRNA, (b) the co-localization of IL-2Rα chain with TSH, FSH and LH-producing cells, (c) the percentage of co-localization of IL-2R with all types of anterior pituitary hormone producing cells: PRL> > > ACTH> > GH> TSH = FSH = LH. (d) that [(3)H]-thymidine is incorporated into the nucleus of all types of hormoneproducing cells without incorporation into other cell types, following IL-2 and IL-6 stimulation. Our results suggest that IL-2 acts on all types of anterior pituitary hormone-producing cells and, through specific functional receptors on the same or other cells, constitutes, as well as IL-6, an inter or intra-cellular factor involved in the coordinate regulation not only of hormone secretion but also of the proliferation of anterior pituitary hormone-producing cells.

Induction from posterior hypothalamus is essential for the development of the pituitary proopiomelacortin (POMC) cells of the toad (Bufo japonicus).

The role of the posterior hypothalamus in the development of the epithelial hypophysis was studied in Bufo embryos. In animals from which the central part of the neural plate (NP) had been surgically removed at the open neurula stage, the infundibulum did not develop, and the epithelial hypophysis was formed away from the normal site without morphological connection with the brain. Immunoreactive MSH cells and ACTH cells, i.e., the pituitary POMC cells, were not detected in any of the surgically treated animals, while other types of secretory cells (PRL, GH, TSH and GTH cells) were invariably present. In view of the fact that POMC cells originate in the anterior neural ridge, and not in the neural plate, the embryonic brain seems to exert an inductive influence upon the primordial pituitary POMC cells. Since these cells differentiate in a tail graft, isolated from the brain at a later stage (tail-bud stage), the inductive stimuli must be conveyed from/via the posterior hypothalamus to the pituitary anlage between the open neurula and the tail-bud stages.

Histological Changes in the Pituitary, Thyroid Gland and Gonads of the Fourspine Sculpin (Cottus kazika) during Downstream Migration

The Fourspine sculpin (Cottus kazika) is a catadromous fish which is widely distributed in the rivers of Japan. The fish was used to examine the relationship between the migration behavior and hormonal control by studying the histological changes in the pituitary gland, thyroid gland and gonads during its downstream migration. By use of the immonocytochemical and histochemical techniques, 7 types of cells were identified in the pituitary gland namely; immunoreactive (ir)-PRL, GH, TSH, GTH, ACTH, MSH and SL cells. From among the first 4 types of the aforementioned cells, remarkable histological changes were observed in cells containing ir-GTH during the downstream migration. At this time also, the gonads were obsereved to be well developed, while the thyroid glands did not show clear changes morphologically. These results suggest that the gonadotropin regulates gonadal development in the Fourspine sculpin during downstream migration and possibly sex hormones synthesized by the gonads cause the downstream migration of this catadromous fish.

Immunohistochemical and Radioimmunological Assessment of Thyrotrophs in the Pituitary of Aging Rats

Thyrotrophs were studied by quantitative immunohistochemistry in the pituitary gland of young (4 months), old (20 months) and very old (29 months) male rats. An attempt was also made to correlate morphometric parameters with serum levels of thyrotropin (TSH), thyroxine (T4) and triiodothyronine (T3). Cells were immunostained by the peroxidase-antiperoxidase method and hormones were measured in serum by specific radioimmunoassays. There was a marked age-related reduction in TSH cell number, volume density and surface density but a significant increase in TSH cell area and perimeter. Basal serum levels of TSH increased, T4 decreased and T3 remained unchanged with age. There was a highly significant (p < 0.001) negative correlation between serum TSH and T4, but no significant correlation was found between TSH and morphometric parameters. The present results suggest that aged rats possess a reduced but functionally preserved thyrotrophic cell population. The coexistence of high circulating levels of TSH with reduced serum T4 suggests that aging brings about a progressive desensitization of the thyroid to TSH.

Localization of the N-methyl-D-aspartate R1 receptor subunit in specific anterior pituitary hormone cell types of the female rat.

N-methyl-D-aspartate (NMDA), a specific agonist of the NMDA-type glutamate receptor, has been shown to stimulate the release of several anterior pituitary hormones when administered in vivo. The primary site of action of NMDA has been suggested to be at the level of the hypothalamus via the control of hypothalamic releasing factors. However, recent studies have demonstrated that NMDA can exert stimulatory effects directly upon anterior pituitary cells perifused in vitro, suggesting that the anterior pituitary may also be a site of action for glutamate. Hence, the purpose of the present study was to determine whether the NMDA R1 receptor subunit is co-localized in specific hormone-secreting cells of the anterior pituitary. To achieve this aim, immunohistochemical studies were performed using an antibody specific for the NMDA R1 receptor subunit to stain sections from the pituitaries of steroid-treated ovariectomized immature rats. Double-immunohistochemistry was employed to demonstrate co-localization. The results of the studies revealed that moderate immunoreactive NMDA R1 receptor staining was observed in the anterior and intermediate lobe of the pituitary with little staining observed in the posterior lobe. Double-immunohistochemistry using antibodies for the NMDA R1 receptor subunit and specific anterior pituitary hormones revealed that the NMDA R1 receptor subunit is co-localized in many cell types of the anterior pituitary including LH (11.2%), FSH (9%), GH (4.6%), TSH (5.7%) and PRL cells (8%). Only one ACTH cell was found to co-localize the NMDA R1 receptor subunit.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of the Pituitary‐Thyroid Axis in Bilaterally Adrenalectomized or Adrenal Transplanted Rats

The percentage, distribution, shape, intensity of staining and morphometrical parameters of the pituitary TSH immunoreactive cells and the histological features of the thyroid glands, were compared between adult rats with intact adrenals, without adrenals and biadrenalectomized animals with neonatal adrenal grafts. After the removal of the adrenal glands, TSH immunoreactive cells increased in percentage and exhibited a higher complexity of the cellular outline, than that of the intact animals. The nuclear, cytoplasmic and cell areas were significantly increased. However the bigger enhancement of the cytoplasmic area relative to the nuclear area, produced a decrease in the nuclear/cell area ratio. The thyroid glands showed some histological evidences of activation. After the transplantation of neonatal adrenal glands to adult rats, several adrenocortical nodules were present in the lumen of the small bowel segment. These adrenal masses induced a great decrease in the TSH cell area, which coupled with a smaller but significant variation of the nuclear area, led to an increase in the nuclear/cell area ratio relative to that observed in adrenalectomized animals. In addition, the distribution, shape and intensity of the immunoreactive material was similar to that observed in intact animals. In this experimental group, thyroid histology was observed to be similar to that of the intact animals.

Na(+)-I- symport activity is present in membrane vesicles from thyrotropin-deprived non-I(-)-transporting cultured thyroid cells.

The active accumulation of I- in the thyroid gland is mediated by the Na(+)-I- symporter and driven by the Na+ gradient generated by the Na+/K(+)-ATPase. Thyrotropin (TSH) stimulates thyroidal I- accumulation. Rat thyroid-derived FRTL-5 cells require TSH to accumulate I-. TSH withdrawal for over 7 days results in complete loss of Na(+)-I-symport activity in these cells [Weiss, S. J., Philp, N. J. and Grollman, E. F. (1984) Endocrinology 114, 1090-1098]. Surprisingly, membrane vesicles prepared from FRTL-5 cells maintained in TSH-free medium [TSH(-)cells]accumulate I-, suggesting that the absence of Na(+)-I- symport activity in TSH(-) cells cannot be due solely to a decrease in the biosynthesis of either the symporter or a putative activating factor. This finding indicates that the Na(+)-I- symporter is present, probably in an inactive state, in TSH(-) cells despite their lack of Na(+)-I- symport activity. Na(+)-I- symport activity in thyroid membrane vesicles is enhanced when conditions for vesicle preparation favor proteolysis. Subcellular fractionation studies in both TSH(+) and TSH(-) cells show that Na(+)-I- symport activity is mostly associated with fractions enriched in plasma membrane rather than in intracellular membranes, suggesting that the Na(+)-I- symporter may constitutively reside in the plasma membrane and may be activated by TSH.

Re-evaluation of gonadotropin-releasing hormone (GnRH) action on pituitary cell differentiation with special regard to its effect on LH and TSH cell types.

We have previously demonstrated that TRH induces several hypophyseal cell type differentiation. In the present study, we re-evaluated the effect of GnRH on gonadotroph differentiation, and we determined whether gonadotropin is implicated in the appearance of other pituitary cell types. These effects were compared to the influence of TRH on LH and TSH cell differentiation. Pituitary primordia of 11 day-old rat embryos were cultured in a synthetic medium until the equivalent of term, and the LH and TSH cell differentiation was visualized using an immunohistochemical technique. The effect of GnRH on pituitary hormone cells was evaluated by measuring the percentage of immunoreactive area per section. GnRH, when added on the first day of culture, induced LH and TSH cell differentiation in a dose-dependent manner. GnRH was more effective than TRH in inducing gonadotropic phenotype expression. This suggests that GnRH, which evokes LH release, is the most effective factor inducing gonadotroph differentiation in early fetal life. However, GnRH was as effective as TRH in thyrotroph differentiation. Lactotrophs were detected in primordia cultured with GnRH. As lactotroph phenotype differentiation is controlled by the alpha-subunit of glycoproteic hormone, further studies are required to determine whether or not GnRH effect on lactotroph is direct. GnRH did not affect the corticotroph and somatotroph differentiation. These results confirm that hypothalamic factors such as TRH or GnRH are capable of inducing more than one pituitary cell phenotypes.

Species Differences of the Thyroid Protein Kinase C Heterogeneity

Protein kinase C (PKC), the mediator of the phosphoinositide transduction pathway, is a family of at least 11 isozymes and its heterogeneity has been described in many tissues and cells. We studied here the heterogeneity of PKC in thyroid glands from three different species, rat, pig, and dog. By combining immunological and biochemical approaches, we identified in rat thyroids, the PKC alpha, beta II, delta, epsilon, and zeta subspecies, in pig thyroids, the alpha, epsilon, and zeta isozymes, and in dog thyroids, only the alpha and zeta isozymes. The observed species differences of the thyroid gland PKC heterogeneity could be related to the reported species differences in the activation of the phosphoinositide regulatory cascade by TSH and other thyroid cell regulators.

In Situ Hybridization Study of Gonadotropin-Releasing Hormone (GnRH) Receptor mRNA in Female Rat Pituitary Gland during Estrous Cycle and after Ovariectomy.

Expression of rat GnRH receptor mRNA in female rat pituitary was examined by in situ hybridization using 35S-labeled antisense RNA probe. Specific signals were detected in the pars distalis. For detailed analysis of the cellular expression of GnRH receptor mRNA, the pituitaries were double-stained by in situ hybridization of GnRH receptor mRNA and antisera against some pituitary hormones (LH, FSH, TSH and prolactin). The expression of GnRH receptor mRNA was detected in all the immunoreactive gonadotropes, but was not detectable in TSH or prolactin cells. Cellular expression of GnRH receptor mRNA in immunoreactive gonadotropes was compared during the estrous cycle and after ovariectomy. In the pituitary of normal cycling rats, the mean value of the cellular expression of the mRNA in estrus was significantly smaller than those of other phases of the cycle. In ovariectomized rats, the cellular expression was significantly in-creased 4 days after the operation as compared with normal cycling rats, and the elevated level of cellular expression was maintained until 21 days after ovariectomy. These results suggest that GnRH receptor expression in the pituitary is preferentially localized in gonadotropes, and the extent of cellular expression varies during the estrous cycle and after ovariectomy.

Pit-1/ghf-1 transcription factor expression in rodent pituitaries.

The Pit-1/GHF-1 (Pit-1) transcription factor is important for the development of anterior pituitary cells that produce GH and PRL. We examined the expression of Pit-1 mRNA in pituitary tissues from rats and mice. Analysis of pituitaries from normal and GHRH transgenic mice showed that Pit-1 transcripts were readily detected in normal, hyperplastic, and neoplastic pituitaries. A cell line (GHRH-CL1) established from a GhRH transgenic mouse pituitary tumor in our laboratory also expressed Pit-1 mRNA. Normal rat pituitaries and those with estrogen-induced PRL cell hyperplasia expressed Pit-1 mRNA. There was a decrease in Pit-1 mRNA in hyperplastic rat pituitaries concomitant with a decrease In GH mRNA amounts and an increase in PRL mRNA amounts after estrogen treatment. Similarly, analysis of GH3 cells in vitro showed that estrogen and bFGF modulated PRL but not Pit-1 mRNA levels. Pit-1 mRNA was localized by combined in situ hybridization and immunohistochemistry to predominantly GH and PRL cells, although some TSH and LH cells in the rat pituitary also expressed Pit-1 mRNA, indicating wide distribution of the mRNA for this transcription factor in various anterior pituitary cell types. Analysis of cell proliferation in normal rat pituitary and GH3 cells revealed that estrogen and bFGF stimulated cell proliferation in normal pituitaries but inhibited proliferation in GH3 cells, whereas Pit-1 transcripts remained unchanged in both groups of cultured cells. These results indicate that Pit-1 mRNA is readily detected in normal, hyperplastic, and neoplastic rodent pituitaries. Changes in Pit-1 mRNA amounts appear to correlate more closely with changes in GH than PRL mRNA levels in cultured pituitary cells.Endocr Pathol 4:146-154, 1993.

Immunohistochemical study of p53 protein in human and animal pituitary tumors.

In many human cancers, p53 gene mutations are frequently occurring genetic abnormalities, which may be detected by immunohistochemical staining for p53 protein. In the present study, p53 immunoreactivity was investigated in formalin-fixed, paraffin-embedded tissues from human and animal pituitary tumors, using the avidin-biotin-peroxidase complex technique. No p53 was detected in 3 nontumorous human adenohypophyses or in 40 human pituitary tumors including 5 GH cell adenomas, 10 PRL cell adenomas, 2 mixed GH cell-PRL cell adenomas, 2 acidophil stem cell adenomas, 8 ACTH cell adenomas, 1 TSH cell adenoma, 1 FSH/LH cell adenoma, 5 null cell adenomas, 5 oncocytomas, and 1 plurihormonal adenoma. Twenty nontumorous and hyperplastic pituitaries of hGRH transgenic mice and 8 tumors in these transgenic animals were immunonegative for p53. All pituitary tumors found in AVP/SV40 transgenic mice contained p53 immunoreactivity in the nuclei, while the nontumorous adenohypophysis of one such transgenic mouse was negative. It can be concluded that p53 mutations are apparently not involved in the pathogenesis of human pituitary adenomas or of the pituitary tumors which develop in hGRH transgenic mice. However, pituitary tumors in AVP/ SV40 transgenic mice are accompanied by p53 expression.