Localization of transforming growth factors, TGFbeta1 and TGFbeta3, in hypothalamic magnocellular neurones and the neurohypophysis.

The distribution of transforming growth factor beta (TGFbeta) in the rat and human hypothalamus and neurohypophysis was investigated by immunocytochemical techniques using rabbit polyclonal antisera against TGFbeta(1) and TGFbeta(3). Colocalization of TGFbeta(1) or TGFbeta(3) and arginine vasopressin (AVP) in the rat hypothalamus was studied by double immunolabelling in light microscopy, while their subcellular localization in the rat neurohypophysis was investigated by immunoelectron microscopy. TGFbeta(1) and TGFbeta(3) immunoreactivity was demonstrated in the cell bodies and processes of neurones in the supraoptic nucleus (SON) and paraventricular nucleus (PVN). The TGFbeta-immunoreactive cells were more numerous in the SON compared to the PVN. TGFbeta/AVP double-labelled cells were seen in both nuclei, but some neurones in the SON were labelled for TGFbeta(1) or TGFbeta(3), although not for AVP. In the rat and human neurohypophysis, TGFbeta(3) immunolabelling was more diffuse and stronger than TGFbeta(1) immunolabelling. TGFbeta(1) expression was seen in axonal vesicles and in neurosecretory granules of the axonal endings, while TGFbeta(3) was observed in axonal fibres. Colocalization of TGFbeta(3) or TGFbeta(1) and AVP was observed in some neurosecretory granules, but many were either single-labelled for TGFbeta or AVP or unlabelled. Our results demonstrate, for the first time, the colocalization of TGFbeta and neurohypophysial hormones in magnocellular neurones. We suggest that TGFbeta secreted by the neurohypophysis regulates the proliferation and secretion of certain anterior pituitary cells.

[Prolactinoma in man: clinical and histological characteristics]. / Le prolactinome de l'homme: caractères cliniques et histologiques.

Prolactinoma usually occurs a small intrasellar tumor in women or as a large tumor in men. To determine whether the predominance of macroprolactinomas in men is due to a delay in diagnosis as has been suggested, or whether there is a sex-related difference in growth rate we conducted a retrospective study in 45 men and 51 women with prolactinomas. Preoperative prolactin level (PRL) was 2,789 573 ng/ml and mean tumor size was 26 2 mm. Prolactin levels and tumor size were significantly higher in women (292 74 ng/ml and 10 1 mm; p<0.01). There was no correlation with age at diagnosis or duration of symptoms. Giant tumors were only observed in men (n=8). Frequency of resistance to bromocriptine (30% vs 5%, p<0.01) and invasive tumors (52% vs 27%, p<0.001) were significantly higher in men than in women. Likewise, proliferation rate was higher for the prolactinomas in men (Ki-67: 2.6 1.1% positive nuclei vs 0.4 0.2%; p=0.08; PCNA: 5.0 2.3% vs 3.7 1.1%). In conclusion, prolactinomas in men are more aggressive than in women. They grow rapidly, often invade the cavernous sinus and are resistant to bromocriptine; proliferation rates can be increased.

Angulate lysosomes in skin biopsies of patients with degenerative neurological disorders: high frequency in neuronal ceroid lipofuscinosis.

Angulate lysosomes with intralysosomal trilamellar structures were first described in patients with metabolic peroxisomal disorders. In this ultrastructural study of skin biopsies of 139 patients with degenerative neurological disorders and 45 patients with static encephalopathies, we observed angulate lysosomes with similar ultrastructure exclusively in degenerative neurological disorders. They were found in only a few cases (8%), but especially in patients with degenerative metabolic disorders (72%). Because they were never observed in patients with static encephalopathies, angulate lysosomes in the skin would seem to be a sign of progressive encephalopathy. The great majority (75%) of angulate lysosomes were associated with neuronal ceroid-lipofuscinosis (NCL). Their presence in skin biopsy could suggest the diagnosis of NCL and eliminate a peroxisomal disorder. In the latter pathology, angulate lysosomes, numerous in the liver and in the brain, were never observed in the skin. As described in pigmentary retinopathy, a conspicuous feature of NCL, we suggest that in this lysosomal storage disorder, the angulate lysosomes in skin biopsies could result from the phagocytosis of melanin.

Mono- and plurihormonal thyrotropic pituitary adenomas: pathological, hormonal and clinical studies in 12 patients.

In a series of 12 patients (eight women and four men, aged between 20 and 62 years), operated on for a pituitary adenoma shown to be thyrotropic by immunocytochemistry, we performed a retrospective and comparative analysis of clinical and biological data, tumor studies including immunocytochemistry with double labeling, and proliferation marker (proliferative cell nuclear antigen (PCNA) and Ki-67) detection, electron microscopy and culture. Our study leads us to confirm that thyrotropic tumors are rare (12 of 1174 pituitary adenomas: 1%). The main points arising were that: (1) high or normal plasma TSH associated with an increase in plasma alpha-subunit and high thyroid hormone levels is the best criterion for diagnosis; (2) the failure of TSH to respond to TRH or Werner's test is not a reliable criterion for diagnosis; (3) thyrotropic adenomas may be 'silent', without clinical signs of hyperthyroidism and with only slight increase in TSH, tri-iodothyronine and thyroxine concentrations; (4) mitoses and nuclear atypies are frequently detected in large tumors, which are invasive in more than 50% of cases - the first analysis of two proliferation markers (PCNA and Ki-67) bears out the relative aggressiveness of thyrotropic adenomas; (5) thyrotropic adenomas are frequently plurihormonal. Immunocytochemical double labeling, complemented by in vitro study, showed that thyrotropic tumor cells sometimes can or sometimes cannot cosecrete TSH, GH or prolactin. The pathological identification of monohormonal and plurihormonal adenomas seems to be supported by clinical and biological differences.

Relationships between pathological diagnosis and clinical parameters in acromegaly.

From our series of 185 somatotropic adenomas with acromegaly, we found that sparsely granulated adenomas were more frequent (56%) than densely granulated ones. Immunocytochemistry detected growth hormone (GH) plurihormonal adenomas in 68% of patients. GH-alpha-subunit (alpha SU) and GH-alpha SU-prolactin (PRL) were more frequent (38%) than GH monohormonal adenomas (32%). The colocalization of GH and alpha SU in the same cell was obvious in many tumors. In contrast, colocalization of GH and PRL was demonstrated in only 25% of GH-PRL adenomas. The relationships between age, sex, tumor size, GH and PRL plasma levels, granularity, and percentage of GH-, alpha SU-, and PRL-immunoreactive cells were established in 105 acromegalic patients by three statistical methods, mainly by a principal component analysis. Correlations were found between the percentage of alpha SU- and GH-immunoreactive cells, and between densely granulated character and the percentage of GH-immunoreactive cells. Tumor size was not correlated with alpha SU, but was positively correlated with PRL plasma levels. Patients' age and percentage of GH-immunoreactive cells were inversely related to tumor size. Plurisecretion and sparsely granulated aspect are not related to age and tumor size.

Quantitative evaluation of the distribution of cell density. Application to human pituitary corticotroph cells.

OBJECTIVE: An image analysis program was developed to analyze cell density in a whole organ. The method was applied to the evaluation of adrenocorticotrophic hormone (ACTH-storing cells in normal human anterior pituitary in order to help the pathologist to evaluate hyperplasia on small surgical biopsies. STUDY DESIGN: ACTH cells were labeled using the immunoperoxidase technique. Data were systematically acquired on a composite image that made possible two-dimensional cell pattern representation in sequential sections of the organ and measurements of cell density in selected histologic parts. RESULTS: ACTH-storing cells were located more in the anterior than posterior part and more in the lower than in the upper pituitary part. The probability of observing high-density value even in normal cases was precisely evaluated zone by zone. Moreover, a model was proposed to evaluate mean cell density from small surgical pieces. CONCLUSION: The same methodology could be applied to other endocrine cell types in the thyroid, pancreas or intestinal tract when hyperplasia is considered the cause of certain clinical syndromes.

Mapping of corticotropic cells in the normal human pituitary.

We accomplished the first mapping of corticotropic cells in the whole human adult pituitary. Corticotropic cells were identified by immunocytochemistry (ICC) and quantified by image analysis on 12 pituitaries obtained from people who had died suddenly. An overall view of each pituitary was given by 15-21 sections (mean 18 sections) at 300-micron intervals on six slides. Each section was systematically treated by indirect immunoperoxidase using an anti-ACTH[17-39] polyclonal antiserum. All the measures were done with a x 6.3 objective lens, each field (0. 5 mm2) being considered as the unit area. The mean pituitary density (surface of labeled cells/total surface) of corticotropic cells (9.5 +/- 3.0% per 0. 5 mm2) is significantly higher in men (11.5 +/- 5.1%) than in women (7.0 +/- 1.3%). This difference is due to an inverse relationship between the corticotropic cell density and the weight of the pituitary, which is higher in women than in men. The mean diameter of corticotropic cells is 14.9 micron and their total number per pituitary is approximately 10(7) cells. We confirmed that the spatial distribution of corticotropic cells is nonuniform: they are mainly distributed in the anteromedian part of the anterior lobe. In addition, our results demonstrated that the inferior part of the pituitary contained three times more corticotropic cells than the superior part (mean density 18.0% vs 6.0%) and the anterior part twice as many as the posterior part (mean density 12.3% vs 6.8%). On the horizontal plane, the pituitary was divided into eight zones, in which the mean of area was 2.5-21.0%. The maximal cell density may reach 40-60%. The use of this map should help the pathologist to recognize if there is corticotropic hyperplasia in a small pituitary fragment surgically removed from a patient with Cushing's disease. On the basis of this study, we put forward some criteria for diagnosing corticotropic hyperplasia.