Specific alterations of the basement membrane and stroma antigens in Human pituitary tumours in comparison with the normal anterior pituitary. An immunocytochemical study

Summary Our report is the first immunocytochemical study of the principal elements of the basement membrane (BM) and connective tissue in normal and adenomatous human anterior pituitaries. In normal tissues, both the parenchymatous BM limiting the endocrine cell cords and the endothelial BM around the capillaries were continuous and were stained with anti-laminin (LM), anti-type IV collagen (CIV) and anti-fibronectin (FN) antisera. Antiserum to type I collagen (CI) stained the connective tissue only. The same antigens were investigated in 23 human pituitary adenomas, 6 of them having been diagnosed as locally invasive by the radiologist and the neurosurgeon. In all cases a lack of cordai structure was observed and the parenchymatous BM was completely absent (9 cases) or fragmented (14 cases). No correlation could be established between the extent of parenchymatous BM alterations and the invasive behaviour of the tumour. In contrast, a continuous endothelial BM was observed around the blood vessels in all cases and its presence was confirmed in double immunofluorescence experiments using anti-von Willebrand factor and anti-LM or anti-CIV antisera. Anti-FN and CI also stained the wall of the vessels. The tumours showed arterial development, in addition to the capillaries found in normal tissue. The present results favour the hypothesis of a decreased synthesis of parenchymatous BM by human adenomatous pituitary cells in comparison with normal cells and show that these tumours are the site of an active arterial neovascularization.     

Grading of pituitary adenomas in acromegaly

Summary In a series of 284 adenomas from cases of acromegaly we studied major morphological variables using light microscopical techniques and compared them with immunocytochemical and clinical results.Using our semiquantitative estimations many inter-relationships were observed. We established the density of secretory granules, nuclear pleomorphism and the rate of occurrence of multinuclear tumour cells, as essential features of tumour differentiation. Mitotic activity and invasive growth patterns did not reveal clear dependences.Immunocytochemical analysis of 105 cases showed growth hormone (GH) in nearly all adenomas (98%), prolactin in 68%, and LH in 40%. The other hormones (ACTH, FSH, and TSH) were present at a much lower rate. Monohormonal GH-adenomas were found in only 29% of our cases.Many different combinations of hormone content could be demonstrated without any relationship to morphological or clinical data. From the linear correlations and advanced method of semiquantitative evaluation, the granular density of the tumour cells is the most useful variable for subclassification and grading of pituitary adenomas in acromegaly.This publication contains results from the doctorthesis submitted by M. Riedel (Hamburg 1984)     

Culture of normal human anterior pituitary cell monolayers

Summary A detailed procedure is described for the culture of functional normal human anterior pituitary cell monolayers for periods up to 2 months. The tissue is dispersed using a double digestion method using collagenase and hyaluronidase followed by trypsin. Attachment of the cells is accomplished in 5 to 9 d. Verification of the presence of functional cells is demonstrated by release of prolactin and luteinizing hormone in response to appropriate stimuli.     

Occurrence of both growth hormone- and prolactin-immunoreactive material in the cells of human somatotropic pituitary adenomas containing Mammotropic elements

Summary With the use of immunoperoxidase staining, both growth hormone- and prolactin-immunoreactivity were demonstrated in the cells of 6 pituitary adenomas removed because of frank or suspected acromegaly. By double immunostaining of individual sections or comparison of adjacent immunostained sections, partial to almost complete identity of the cell populations containing growth hormone and prolactin was found in 5 of the tumors.     

Morphological and biochemical relationships in 31 human pituitary adenomas with acromegaly

Summary In 22 pure GH cell adenomas and 9 mixed GH cell-prolactin cell adenomas with acromegaly, we compare the morphological and functional data (secretory activity and granular appearance) with GH levels (radioimmunoassays) in the blood and in the tumor. According to morphological criteria, the secretory activity is marked in 13 cases (Group I), mild in 9 cases (Group II), and weak in 9 others (Group III). The mean values of the plasma GH levels in the 3 groups (80 ng/ml±22; 26.5 ng/ml±2; and 16.89 ng/ml±2 respectively) are significantly different. In 17 densely granulated adenomas and 14 sparsely granulated adenomas, the plasma GH values were very variable. The mean levels of these 2 groups (49.76 ng/ml±22 and 41.8 ng/ml±7.8 respectively) are not significantly different. The GH concentrations in the tumor were also very variable (358 to 78,900 ng/mg). Their highly significant relationship with the granular appearance is an indirect proof of the granular localisation of GH. We distinguish between 4 functional aspects of the GH cell adenoma which define the different levels of synthesis, storage, and excretion. The secretory activity of the GH adenomatous cell varies with the adenomas and differs from that of the normal cell.     

Human pituitary gonadotroph adenomas: Relationship between gonadotropin immunoreactivity and clinicopathologic findings

Clinically nonfunctioning pituitary adenomas are generally seen in middle-aged and older patients, and most of them may be gonadotropin-immunoreactive adenomas, that is, gonadotroph adenomas. Our aim was to clarify the relationships between the gonadotropin immunoreactivity, patient age, sex, and microscopic features in 68 gonadotroph adenomas with special reference to either gonadotropin-immunonegative or intensively immunopositive adenomas. There were 68 patients with gonadotroph adenomas (mean age 54.7 yr) in the study, including 39 men (mean age, 52.8 yr) and 29 women (mean age, 57.4 yr). The adenomas were diagnosed on the basis of immunoreactivity for gonadotropins (β-subunit of follicle-stimulating hormone: β-FSH; β-subunit of luteinizing hormone: β-LH; and the α-subunit of the pituitary glycoprotein hormone: α-SU) by the avidin-biotin peroxidase complex (ABC) method or by the characteristic histological feature of a perivascular or pseudorosette pattern, that is, the cells aligned polarity directed toward the capillaries. Fifty-four adenomas (79%) were positive for one or more gonadotropin subunits and β-FSH was the most common subunit encountered (47/68, 69%). In men β-FSH immunoreactivity was similar among all age groups, whereas in women, it was significantly less frequent in patients who were 50 yr or older, compared to younger patients. Gonadotropin-immunonegative adenomas were seen in 4 men (mean age, 46.8 yr) and 10 women (mean age, 61.5 yr). Among the 22 women aged 50 or over, β-FSH was negative in 12 tumors (55%), whereas in men of the same age group, it was negative in 3 of 26 tumors (12%). The reason for this reduced frequency is not clear, but the postmenopausal state and associated changes in the systemic endocrine state may play a role. Adenomas that were intensively positive for β-FSH showed an unusual morphology other than the characteristic perivascular pattern, regardless of the patients' age and sex; the tumor cells had abundant vacuolated cytoplasms and were arranged in a sheet-like pattern. Electron microscopically, these cells with abundant cytoplasm had well-developed Golgi complexes, suggesting an enhanced activity of gonadotropin synthesis, and these adenomas seem to be endocrinologically, if not clinically, functioning. The results indicate that gonadotroph adenomas may vary from functioning adenomas with intense immunoreactivity and unusual histology to immunonegative and less functioning adenomas, which are more frequent in women 50 yr or older.     

Immunohistochemical analysis of tumor angiogenic factors in human pituitary adenomas

Microvessel density (MVD) has been studied in a number of neoplasias, and apparently, there is a relationship between angiogenesis and tumor progression, response to treatment, and outcome. In pituitary adenoma, the association between MVD and vascular endothelial growth factor (VEGF) with tumor behavior has been described, but correlation with other angiogenic factors such as fetal liver kinase 1 (Flk-1) or proliferative markers is unknown. We investigated MVD, VEGF, and its receptor Flk-1 expression in 60 human pituitary adenomas: 13 growth hormone cell adenomas, 7 prolactin cell adenomas, 5 corticotroph cell adenomas, 2 thyrotroph cell adenomas, and 33 nonfunctioning adenomas (30 gonadotroph cell adenomas and 3 null cell adenomas). We performed immunohistochemistry for CD34, Ki-67, VEGF, and Flk-1. To evaluate MVD, we used 2 methods: the number of vessels per square millimeter and the Chalkley method. Immunohistochemistry results were correlated, as well as with clinicopathologic factors. Adenomas with higher MVD were thyrotroph cell adenomas (299.9 +/- 87.5), and those with lower MVD were prolactin cell adenomas (168.6 +/- 63.3; P = .45, analysis of variance). We found a trend toward higher MVD in the adenomas of older patients (P = .142), but no difference was found regarding sex, extrasellar extension, or Ki-67 (P > .05). However, extrasellar extension was nearly significant when the Chalkley method score was high (P = .056). Low expression of VEGF was seen predominantly in prolactin cell adenomas, and high in nonfunctioning adenomas, or in cases of older patients (P < or = .032). Flk-1 score correlated with VEGF (P = .006). High expression was observed in nonfunctioning adenomas, cases presenting at older ages, and with extrasellar extension (P < or = .022). Our study shows that VEGF and Flk-1 are widely expressed in pituitary adenomas, predominantly in nonfunctioning adenomas and those presenting at older ages. Moreover, Flk-1 is associated with a more aggressive phenotype, and it may have potential therapeutic interest.     

Correlation between PCNA expression and AgNOR dots in pituitary adenomas

Nucleolar organizer regions are segments of DNA associated with argyrophilic proteins (AgNORs). Our previous findings showed that the number, the area, and the intranuclear localization of AgNOR dots differ according to tumor aggressiveness and to the hormone-immunopositivity of pituitary adenomas. Proliferating cell nuclear antigen (PCNA) is a nuclear protein, whose expression is correlated with cell proliferation. The aim of the present paper was to examine PCNA-labeling indexes in pituitary adenomas and to correlate them with AgNOR dots in various immunohistochemical types of the tumors. Histological slides from 32 pituitary tumors and one normal pituitary were silver-stained and analyzed with a computerized system for microscopic image analysis. We found that the percentage of PCNA-positive cells did not differ significantly among examined groups of monohormonal adenomas. However, tumors immunopositive for α-subunit (α-SU) showed a significantly higher (p<0.05) PCNA index than adenomas immunonegative for that unit. PCNA index in recurrent tumors was significantly higher than in primary adenomas. There was a moderate positive correlation between the PCNA index and the mean area of AgNOR dots and a similar correlation between the PCNA index and the area of the biggest dot in the nucleus. The obtained results reveal that the PCNA indexes and estimated parameters of AgNOR dots differ according to tumor aggressiveness.     

垂体腺瘤免疫微环境及免疫治疗的研究进展

垂体腺瘤(PAs)是第二常见的原发性中枢神经系统肿瘤,多数可通过手术、药物或放射治疗治愈。但是,部分PAs经过传统治疗后仍反复复发并侵袭性生长,甚至发生远端转移,缩短了患者生存期并降低了生活质量。近年来,关于脑肿瘤免疫微环境的研究逐渐兴起,尤其在胶质瘤领域,为免疫治疗提供了潜在有效靶点,为提高患者生存创造了条件。但是,既往研究仅对PAs微环境中免疫细胞、免疫分子及细胞因子进行了初步分析,对动物和人PAs的免疫治疗研究刚刚起步。     

Chromogranin a processing in human pituitary adenomas and carcinomas: analysis with region-specific antibodies

The expression of various chromogranin A (CgA) peptide fragments was examined with region-specific antisera in benign and malignant pituitary tumors. Analysis of the proconvertases responsible for proteolytic processing of CgA, prohormone convertase 1/3 (PC1/3), and PC2 was also performed. Adenomas were studied using tissue microarrays, and a larger tissue section of a subset of the prolactin (PRL) adenomas was used to compare to the tissue microarray analysis. Carcinomas were analyzed using large tissue sections. There were differences in CgA proteolytic products detected between the functional (PRL, adrenocorticotropic hormone [ACTH], and growth hormone tumors and the nonfunctional (gonadotroph and null cell) tumors, with the former group, expressing lower levels of many peptides. These differences were most notable in the PRL adenomas and carcinomas in which the region-specific antisera against vasostatin I and vasostatin II detected these fragments in the lowest percentage of tumors and/or had the weakest immunoreactivity. The CgA peptide fragment detected by CgA 176–195 (chromacin) antiserum was expressed by the highest percentage of most functional and nonfunctional benign and malignant pituitary tumors. ACTH carcinomas (n+3) were more strongly immunoreactive compared to the ACTH adenomas. These results show that there is differential expression of CgA peptide fragments and PC1/3 among different types of pituitary tumors and that ACTH pituitary carcinomas have higher levels of immunoreactive CgA peptide fragments compared to ACTH adenomas. This study also shows the utility of tissue microarrays in the analysis of a large group of tumors with region-specific antisera.     

Expression of vascular endothelial growth factor in normal pituitary cells and pituitary adenomas producing adrenocorticotropic hormone

Vascular endothelial growth factor (VEGF) induces endothelial cell proliferation and an increase in capillary permeability. Because the anterior pituitary gland and pituitary adenomas are highly vascular, expression of VEGF was examined immunohistochemically. Some normal pituitary cells stained positively for VEGF, and restaining for ACTH, prolactin, TSH, LH, FSH, and S-100 protein after VEGF staining revealed that almost all cells staining positively for ACTH also stained for VEGF. Only adenomas staining positively for ACTH stained for VEGF. These results suggest that VEGF is produced by normal pituitary cells and adenomas producing ACTH.     

Histologic and immunohistochemical study of clinically non-functioning pituitary adenomas

Seventy-five clinically non-functioning pituitary adenomas were characterized in terms of their histologic and immunohistochemical features. Fourteen adenomas (18.7%) were positive for hormones other than gonadotropins. These included two somatotroph adenomas, three lactotroph adenomas, four thyrotroph adenomas, four corticotroph adenomas and one unusual plurihormonal adenoma. Fifty-five adenomas (73.3%) were exclusively positive for one or more gonadotropin subunits (β-follicle stimulating hormone, p-luteinizing hormone, and α-subunit of glycoprotein hormones), but negative for other hormones such as growth hormone and β-thyrotropin. Histologically, a papillary arrangement along the capillary was most characteristically observed in the gonadotropin-positive adenomas. Five of six adenomas negative for any pituitary hormones exhibited the typical papillary structure. Thus, approximately 80% of clinically non-functioning adenomas constituted a single tumor group that shared the common histologic features of gonadotroph adenomas. These findings suggest that nearly all tumor types of clinically non-functioning adenomas can be diagnosed solely by light microscopy in combination with immunohistochemistry, and that the vast majority of them may be gonadotroph adenomas.     

Oncocytomas and null cell adenomas of the human pituitary: Morphometric and in vitro functional comparison

Summary In this study, light microscopic and ultrastructural morphometric features of oncocytomas and null cell adenomas were compared and the morphometric data were correlated with in vitro endocrine activity. All tumours were unassociated with clinical or biochemical evidence of hormone excess and were diagnosed as oncocytomas or null cell adenomas, using histology, immunohistochemistry and electron microscopy. In oncocytomas, when compared with null cell adenomas, light microscopic morphometry revealed that total cell areas were significantly larger and nuclear cytoplasmic ratios were smaller due to an increase in cytoplasmic areas. Ultrastructural morphometry disclosed an abundance of mitochondria in oncocytomas. Absolute volumes of cytoplasmic organelles per cell were not reduced in oncocytomas compared with those of null cell adenomas. These results indicate that accumulating mitochondria do not replace other cytoplasmic organelles, and furthermore that the functional potential of oncocytomas is not lost. In vitro study demonstrated the production of pituitary hormones, primarily gonadotropins in oncocytomas and null cell adenomas. It can be concluded that oncocytomas, which represent the final stage of oncocytic transformation, have a close relationship with null cell adenomas based on morphometric comparison as well as in vitro studies.     

Increased expression of HMGA1 correlates with tumour invasiveness and proliferation in human pituitary adenomas

Wang E L, Qian Z R, Rahman M M, Yoshimoto K, Yamada S, Kudo E & Sano T
(2010) Histopathology 56, 501–509 Increased expression of HMGA1 correlates with tumour invasiveness and proliferation in human pituitary adenomas Aims: High‐mobility group A1 (HMGA1) is highly expressed in various benign and malignant tumours. The development of pituitary adenoma in Hmga1 transgenic mice has been reported. However, no studies have investigated HMGA1 expression and its clinical significance in human pituitary adenomas. Methods and results: Immunohistochemical expression of HMGA1 was analysed with respect to various clinicopathological factors in 95 pituitary adenomas. Nuclear expression of HMGA1 was observed in 62% of pituitary adenomas, whereas normal adenohypophysial tissues were negative. Although HMGA1 expression was frequently detected in clinically non‐functioning adenomas – 90% of silent adrenocorticotropic hormone (ACTH), 76.2% of follicle‐stimulating hormone/luteinizing hormone and 100% of null cell adenomas – it was also detected in 48.1% of growth hormone (GH), 60% of mixed GH/prolactin (PRL), 62.5% of PRL, 66.6% of thyroid‐stimulating hormone and 37.5% of ACTH adenomas. HMGA1 expression was significantly higher in invasive adenomas or macroadenomas than in non‐invasive adenomas or microadenomas (invasive versus non‐invasive, P < 0.05; macroadenoma versus microadenoma, P < 0.05). In addition, HMGA1 showed the highest level in grade IV, more aggressive pituitary adenomas, than in grades I, II and III (IV versus I, P = 0.01; IV versus II, P = 0.01; IV versus III, P = 0.07). Furthermore, a significant correlation between HMGA1 expression and MIB‐1 labelling index was observed (R = 0.368, P < 0.0002). Conclusions: These findings suggest that HMGA1 up‐regulation has an important oncogenic role in pituitary tumorigenesis, as well as being a novel molecular marker of tumour proliferation and invasiveness.     

视交叉侧动脉显微解剖特点与垂体瘤患者视功能障碍的关系

目的 通过对视交叉侧动脉特点的显微解剖学研究,探讨其在垂体瘤致视功能障碍中的意义.方法 10例福尔马林固定的国人成人头颅标本,经双侧颈内动脉和椎动脉灌注红色乳胶后取脑,手术显微镜下剥离鞍区蛛网膜,暴露颈内动脉颅内段、Willis环及其穿动脉,以及垂体柄、视交叉、大脑脚、乳头体等鞍区重要结构.观察视交叉侧动脉的起始、走行、分支和分布,测量其起始部的直径并显微摄影. 结果 视交叉侧动脉自颈内动脉C<,2>段内侧壁发出,在蛛网膜下腔迂曲走行,起始部位靠上,几乎与视交叉处于同一水平,直径(0.18±0.06)mm,在到达视交叉之前发出2-3条亚支,呈"分水岭"样分布于视交叉侧缘、视交叉侧部的上面和下面(以下面为主)靠外侧的部分以及视神经近视交叉处外侧半.视交叉侧动脉具有较长的脑外游离段,各穿动脉主干之间不吻合,而在软膜上与邻近穿动脉之间存在广泛的终末吻合. 结论 垂体瘤不易累及视交叉侧动脉可能是垂体瘤患者鼻上象限视野往往可以长时间保留的原因.     

Effects of atrial natriuretic factor on anterior pituitary hormone secretion in normal man

Summary The effects of intravenous human atrial natriuretic factor ANF(99-126) administration on anterior pituitary hormone secretion have not been extensively investigated in humans. We repeatedly studied 10 healthy volunteers (5 female, 5 male, aged 28±2 years) on 2 occasions, 3 days apart. In randomized, single blind order, subjects received pretreatment with either placebo or intravenous ANF(99–126) (bolus 100 g/kg, 30-min infusion of 0.1 g/kg-min). Subsequently, on both occasions subjects received a combined intravenous bolus injection of pituitary releasing hormones (200 pg thyrotropin releasing hormone, 100 g gonadotropin releasing hormone, 50 g growth hormone releasing hormone and 100 g human adrenocorticotropin releasing hormone; Bissendorf, Hannover, FRG). Plasma concentrations of adrenocorticotropic hormone (ACTH), cortisol, luteinizing hormone (LH), follicle-stimulating hormone (FSH), growth hormone (GH), thyrotropin (TSH), prolactin, ANF and cyclic guanosine monophosphate (GMP) were determined by radioimmunoassay. ANF(99–126) treatment induced a significant reduction in basal ACTH plasma concentrations and tended to decrease basal plasma cortisol. The TSH response to combined releasing hormone administration was significantly diminished after ANF(99-126) pretreatment. In women, the releasing hormone induced prolactin increase was reduced after ANF(99–126) pretreatment. With the present study design, ANF(99–126) did not alter the basal or releasing hormone stimulated plasma concentrations of cortisol, LH, FSH and GH. Releasing hormone administration did not affect ANF and cyclic GMP plasma levels. In humans, effects of natriuretic peptides on anterior pituitary hormone secretion may have to be considered with investigational or therapeutic administration of ANF analogues or agents interfering with the ANF metabolism.Abbreviations ANF(99–126) human atrial natriuretic factor - ACTH adrenocorticotropic hormone - LH luteinizing hormone - FSH follicle-stimulating hormone - GH growth hormone - TSH thyrotropin - PRL prolactin - cyclic GMP cyclic guanosine monophosphate - TRH thyrotropin releasing hormone - GnRH gonadotropin releasing hormone - GHRH growth hormone releasing hormone - CRH adrenocorticotropin releasing hormone     

Pars intermedia of the human pituitary revisited: Morphologic aspects and frequency of hyperplasia of POMC-peptide immunoreactive cells

The human pituitary has no distinct pars intermedia (PI). Instead, the proopiomelanocortin (POMC) producing, adrenocorticotropin (ACTH)- and β-endorphin-immunoreactive PI cells are incorporated within the pars anterior, thereby participating in the formation of the pars distalis. Two hundred fifty autopsy pituitaries (156 males, 94 females) have been studied by histology and immunohistochemistry to determine the frequency of clinically non-functioning ACTH- and β-endorphin immunoreactive (POMC) cell hyperplasia/adenoma probably attributable to PI-derived cells. Such hyperplasia occurred in 29% of men and 14% of women; 80% of the male and 77% of the female subjects were over 50 yr of age. In two of the women, POMC cell adenoma was present as well. Except for hypothyroidism, none of the cases with POMC cell hyperplasia had endocrine disorder. No obvious correlation was evident between POMC cell hyperplasia/adenoma and clinical presentation. Hyperplasia of PI-derived POMC cells may be suspected by virtue of differential intraglandular localization of these cells. Except for cases of glucocorticoid treatment, leading to Crooke’s hyalinization of ACTH cells, but not affecting PI cells, no conclusive separation of various POMC-producing subsets is possible at present. The PI-derived cells probably give rise to silent “corticotroph” adenoma subtype 1 and subtype 2.     

Immunocytochemical analysis of the synaptic proteins SNAP-25 and Rab3A in human pituitary adenomas. Overexpression of SNAP-25 in the mammosomatotroph lineages

SNAP-25 and Rab3A were originally identified as synaptic proteins involved in neuronal membrane traffic. Recently, both proteins have been detected in several mammalian endocrine cell types and have been proposed as essential components of the exocytotic pathway in neuroendocrine cells. In this study, the expression of SNAP-25 and Rab3A was analysed in biopsied human anterior pituitary tumours (21 cases) by immunocytochemical methods. No differences in Rab3A immunoreactivity were observed between tumour and normal pituitary cells. Strong SNAP-25 immunoreactivity was detected in tumour cells of prolactinomas (n=3). Several growth hormone (GH)/prolactin (PRL) tumours also displayed intense SNAP-25 immunolabelling (n=3), whereas the remaining GH-secreting adenomas (n=4) exhibited moderate to weak SNAP-25 immunoreactivity. In contrast, SNAP-25 near-background immunostaining was observed in tumour cells of adrenocorticotrophic hormone (ACTH)-secreting tumours (n=4) and non-secreting tumours (n=7), as well as in normal pituitary cells. Since SNAP-25 and Rab3A have been shown to be involved in exocytotic events in rodent endocrine cells, overexpression of SNAP-25 protein in PRL and GH/PRL tumour cells might be implicated in the mechanism of exocytosis of the neoplastic human mammosomatotroph lineages. © 1997 John Wiley & Sons, Ltd.     

Sparsely granulated prolactin cell adenomas of the pituitary gland

Summary The possible relationship between the preoperative plasma prolactin levels of patients having a sparsely granulated prolactin cell adenoma of the pituitary gland and the morphology of the tumors was studied by means of quantitative electron microscopy. To this end, a number of ultrastructural variables were chosen which are generally regarded to be indicative of cellular activity and which could be determined in a quantitative or semiquantitative way. These variables were determined in 19 adenomas from 17 patients and plotted against the corresponding prolactin levels. It appeared that marked endocrine activity was associated with a small number of granules per cell, a high frequency of exocytosis, and a marked development of the rough endoplasmic reticulum. Granule size and development of Golgi apparatus and lysosomes were not at all, or only poorly correlated with the plasma hormone levels. Finally, the number of mitochondria per cell showed a totally unexpected inverse correlation with endocrine activity. Due to the close mutual correlation existing between several of the variables investigated, combining them in a multivariate analysis did not significantly improve the correlation with the hormone level.     

The origin of ciliated cell cysts of the anterior pituitary

Summary Rats were decapitated and the heads were stored at 4° C for 24 h. The anterior pituitaries were then removed and incubated for 5 days. On the 1st, 3rd and 5th days of incubation, explants were studied by electron microscopy and immunohistochemistry using antiserum against S-100 protein. In the explant many granulated cells underwent necrosis; folliculo-stellate (FS) cells formed many cyst-like structures (CLSs) and became squamous epithelioid cells (CLS-forming cells). After incubation for 5 days the explants were isotransplanted under the renal capsules of male rats in order to observe morphological changes in the CLSs. Immediately after transplantation, the CLSs were encircled by a basement membrane, but from the 8th to 14th day, ciliation occured in CLS-forming cells and ciliated cell cysts were formed. The ciliated cells were immunostained with antiserum against S-100 protein. The present study suggests that FS cells are related to CLS formation and have the potential to trans-differentiate to ciliated cells.