Fujio Yoshimura

Pituitary folliculo-stellate cells immunostained with S-100 protein antiserum in postnatal, castrated and thyroidectomized rats.

SummaryPituitary folliculo-stellate cells and associated cells were studied immunohistochemically throughout the life of male and female rats. The marginal layer cells of the pars distalis and intermedia as well as the folliculo-stellate cells were immunostained with anti-S 100 protein serum. The immunostained folliculo-stellate cells in one section were identified as themselves by their ultrastructural properties on the adjacent section. Corticotrophs, thyrotrophs and prolactin cells, characterized by the stellate shape, were not immunostained with anti-S 100 protein serum. Reactivity for S-100 protein appeared on postnatal Day 6 in the marginal layer cells of the pars intermedia, and appeared on Day 10 in those cells of the pars distalis. No immunostained folliculostellate cells appeared before Day 6, but a few of them appeared on Day 10. Thereafter they increased in number, having more intense immunostaining with advancing age. In castrated rats, the immunostained folliculo-stellate cells appeared quite numerously, with branched cytoplasmic processes surrounding the gonadotrophs. In the thyroidectomized rats, however, folliculo-stellate cells lacked ramified cytoplasmic processes, and their topographic affinity for thyrotrophs was negligible.

Immunohistochemical and electron microscopical studies of mitotic adenohypophysial cells in different ages of rats

SummaryMitotic rates of the six types of immunohistochemically identifiable adenohypophysial cells were histometrically calculated in colchicine-pretrated male rats 5, 17, 30 and 70 days old. Sections were stained with the antisera against rLH, rFSH, rTSH, oGH, rPRL and pACTH1–39. The mitotic growth rate of the anterior pituitary gland at 30 days of age was much higher than at other times. Mitotic growth rates of GH and PRL cells increased with advancing age, while those of ACTH-, TSH- and immunonegative cells decreased with advancing age. LH/FSH cells showed no variation in mitotic growth rate with age. Mitotic cells can be classified into six cell types based on their fine structural properties: (1) agranular cells associated with the folliculo-stellate cells; (2) ambiguous cells with scanty minute secretory granules (50–150 nm in diameter); (3) basophils with a number of small secretory granules (130–200 nm); (4) immature acidophils whose large secretory granules (130–300 nm) are sporadically scattered; (5) acidophils with numerous spherical larger secretory granules (200–300 nm); and (6) prolactin cells with large polymorphic granules. At day 5 there was a high mitotic rate of the agranular and ambiguous cells [types (1) and (2)]; at day 70 a high mitotic rate was found in immature and mature acidophils [types (4) and (5)]. The mitotic rate of basophils (type 3) was high only at day 17 and low at all other times. The mitotic rate of prolactin cells (type 6) showed a slight increment with advancing age. It is concluded that the mitotic rates of the six cell types are age-dependent.

Prolactin immunoreactivity of acidophils of the small granule type

SummaryThe fine structure of some oval anterior pituitary cells of the adult male rats immunostained with an antiserum to rat prolactin was investigated electron microscopically on the adjacent thin sections. Their fine structural appearance is identical with that of acidophils of the small granule type (Yoshimura et al. 1974) resembling the Kurosumi-Oota LH gonadotrophs. The secretory granules of the oval cells are spherical in shape, ranging from 130 to 200 nm in diameter. Large polymorphic granules, which are generally believed to be characteristic of prolactin cells, are absent from their cytoplasm. It is concluded that the acidophil of the small granule type with a similar fine structure to the Kurosumi-Oota LH gonadotroph is a prolactin secreting cell.

Fine structural criteria for identifying rat corticotrophs.

SummaryThe fine structural characteristics of normal rat corticotrophs stained with anti-porcine ACTH1–39 serum were studied. At the ultrastructure level immunoreactive corticotrophs appear to comprise four distinct cell types: (1) large stellate cells (Siperstein cells) containing granules (170–250 nm in diameter) arranged in a peripheral row and usually embracing an acidophil; (2) elongate spindle-shaped cells (Moriarty cells) in which the secretory granules (170–250 nm in diameter) are distributed in a row or in small clusters in the peripheral cytoplasm; (3) oval or polygonal cells filled only with small secretory granules (130–170 nm in diameter), resembling the “acidophil of small granules type” (Yoshimura et al. 1974); and (4) polygonal or stellate cells filled with secretory granules of varying diameters (180–300 nm in diameter) and occasionally embracing an acidophil. The first type is the most common, but the others are infrequent. It is concluded that the criteria of Siperstein and Miller (1970) do not necessarily include all categories of rat corticotrophs.

Granulated folliculo-stellate cells and growth hormone cells immunostained with anti-S 100 protein serum in the pituitary glands of the goat

SummaryGoat pituitary glands were immunohistochemically studied with antisera for bovine S-100 protein, rat LHβ, FSH, TSHβ, prolactin, ovine GH, and porcine ACTH1–39 by use of the superimposition technique on adjacent sections. Folliculo-stellate (F-S) cells were divided into two categories on the basis of ultrastructural properties: One consisted of a mass of agranular cells in which the pseudolumina were equipped with microvilli and cilia. Elongate gap junctions were often observed among these cells. The other was a group of granulated cells with or without pseudolumina. In this group the gap junctions were shown to be disintegrated. The dense granules 150–250 nm in diameter began to accumulate in the cells. However, neither type of these F-S cells was immunostained for S-100 protein. On the other hand, numerous polygonal, elongate, irregular or stellate cells containing S-100 protein were distributed throughout the gland. Most of them were immunohistochemically identical with the GH cells laden with the secretory granules 250–450 nm in diameter, but some of them were identical to TSH and prolactin cells which immunostained faintly for S-100 protein. This appears to be the first demonstration of GH cells intensely immunostained for S-100 protein.

A whole range of fine structural criteria for immunohistochemically identified LH cells in rats

SummaryPituitaries from normal, young and adult male rats were fixed either in sublimate-formalin or in glutaraldehyde-osmium. In adjacent Paraplast sections, almost all the gonadotrophs were immunostained with both LH and FSH antisera. The rat LHβ and FSH antisera used were shown to be highly specific by the absorption test and by double antibody radioimmunoassay. Thin and thick adjacent Epon sections were prepared for EM and immunohistochemical examination. Cells stained with the rat LHβ antiserum were identified by LM, and then observed in detail by EM. On the basis of these observations we suggest that the LH cells are arranged in a sequence of basophils, i.e., Types II/III, III, III/IV and IV: Type II/III basophils are elongate with a cytoplasmic process and less vesiculated. They have morphological features of Type II (classical thyrotrophs) and also of Type III basophils. Type III basophils are oval in shape and moderately vesiculated. Both Types II/III and III basophils can be divided into two classes of cell characterized mainly by the existence of only small secretory granules (150–220 nm in diameter) (Type A) or by the coexistence of small and large (350–500 nm) (Type B). Type III/IV basophils are cells intermediate between types III and IV basophils, and moderately vesiculated with an abundance of secretory granules (150–300 nm in diameter). Type IV basophils are large, spherical or oval cells whose RER cisternae are conspicuously dilated; they contain less numerous secretory granules (150–300 nm in diameter). It is concluded that LH cells are not a single cell type, but include a wide range of subtypes.

Fine structural and immunohistochemical studies of goat adenohypophysial cells.

SummaryThe fine structure of each type of anterior pituitary cell in the male goat was studied through the application of a superimposition technique in which adjacent thick sections were used to identify individual cells beforehand by light-microscopic immunohistochemistry. A cone of the pars intermedia protrudes into the pars anterior, being surrounded by the narrow pituitary cleft; the immunohistochemical appearances of the cells forming the cone resemble those of the pars anterior. Several follicles appear in the pars anterior. Ultrastructurally GH cells resemble prolactin cells. The secretory granules of both types are spherical; the diameter of the former is about 340 nm, whereas that of the latter is about 440 nm. ACTH cells are polygonal in shape with secretory granules, about 180 nm in diameter, scattered throughout the cytoplasm. TSH cells, which are spherical in shape, contain the smallest secretory granules, 150 nm in diameter. The highly electron-dense LH cells contain numerous secretory granules about 210 nm in diameter. Their nuclei are irregular with incisures. Thus, the anterior pituitary cells of the goat are ultrastructurally characteristic and species-specific.

Immunohistochemical study of the postnatal development of pituitary thyrotrophs in the rat, with special reference to cluster formation

SummaryThe postnatal development of rat pituitary thyrotrophs was investigated immunohistochemically on days 1, 3, 5, 10, 15 and 25. Fetal thyrotrophs are strongly immunoreactive. In the postnatal period, however, weakly immunoreactive thyrotrophs increase in number to constitute clusters on days 3–5. The numbers and dimensions of the clusters reach a maximum on day 10. Thereafter the clusters break down to give rise to single, scattered neogenic thyrotrophs. Thyrotrophs in clusters on day 10 were investigated by electron microscopy in adjacent sections. They can be characterized as an immature type of basophil, according to the classification of Yoshimura et al. (1977): 1) Type I basophils, which are irregularly shaped with elongate processes, and characterized by rows of secretory granules about 100 nm in diameter. 2) Type I/II basophils, i.e., forms intermediate between Types I and II, containing less numerous secretory granules about 100–150 nm in diameter. Type II basophils which correspond to the classical thyrotrophs are not fully developed on day 10. Thus, most thyrotrophs develop from the clusters in the neonatal period. Such neogenic thyrotrophs retain the immature characteristics of Type I and I/II cells and may develop into Type II cells during subsequent maturation.

Immunohistochemical and ultrastructural study of anterior pituitary cells in the female Afghan pika, Ochotona rufescens rufescens

SummaryAn immunohistochemical study of the anterior pituitary gland of the female Afghan pika was carried out to distinguish the ultrastructural features of GH, PRL, ACTH, TSH and LH cells. The histochemically identified GH cells resembled ultrastructurally oval or round GH cells of the rat laden with large, dense secretory granules. PRL cells were divided into three subtypes based on differences in the diameter of their spherical secretory granules. They lacked polymorphic or irregularly shaped secretory granules. ACTH cells resembled ultrastructurally, in some respects, Sipersteins “corticotrophs” of the rat with peripheral arrangement of secretory granules. However, they were not always stellate, but elongate or angular in shape. The dense secretory granules were concentrated in the peripheral area of cytoplasm. TSH cells were non-stellate, but usually oval in shape, containing the smallest spherical secretory granules (100–200 nm in diameter). Almost all LH cells reacted also with FSH antiserum. They were irregular in shape, sometimes in contact with or surrounded the GH cells. They contained an abundance of medium-sized secretory granules (140–260 nm in diameter) which were larger than those in the LH cells of the female rat throughout the estrous cycle. Large secretory granules in the LH cells of the female pika seemed to be related to the endocrine state of persistent estrus.

Relationship between characteristics of immunoreactive LH/FSH cells and the levels of gonadotropin in the female rat.

Morphological and functional changes of pituitary LH/FSH cells in the female rat were investigated using the parameters on the radioimmunoassay, immunohistochemistry and ultrastructure. Changes in immunostainability, populations of intensely immunostained LH and FSH cells and total volume of secretory granules were correlated with the changes in pituitary LH and FSH contents during the estrous cycle. The immunohistochemical feature of gonadotropin release is the transformation of intensely immunostained gonadotrophs into the weakly stained ones. Secretory granules of small diameter (less than 150 nm) were numerous just before LH and FSH surges then sharply declined along with LH and FSH surges. The number of secretory granules of large diameter (larger than 150 nm) also decreased when LH and FSH surges took place. Then the number increased progressively until 17.00 h on the day of diestrus, corresponding to the increase in pituitary LH and FSH contents. It is suggested that small secretory granules are a release pool while large ones are a reserve pool.