Immunohistochemical identification of cells in the pars distalis of the pituitary of the lizard anolis carolinensis

Summary Immunocharacteristics of the pars distalis cells of the pituitary of the male lizard A. carolinensis are determined by employing the immunoperoxidase technique with antisera to mammalian pituitary hormones. On the basis of their immunoreactivity, 5 different cell types with characteristic anatomical distribution are recognized. ACTH cells are found in the rostral half of the pars distalis, and PRL cells in the rostral two thirds of the pars distalis. GH and TSH cells are located in the caudal half of the pars distalis. GTH cells are distributed throughout the gland. When consecutive sections are stained with antiserum to ovine FSH or its -subunit and to ovine LH, the same cells show immunoreactivity to all the three antisera. None of the GTH cells show positive immunoreactivity to ovine -LH antiserum. The results suggest the existence of one gonadotropic cell type in the pituitary of this lizard.Supported by U.S. Council for International Exchange of Scholars (to D.R.N.) and PHS Grant NS09914     

Ontogeny of gonadotropin releasing hormone and gonadotropin immunoreactivity in brain and pituitary of normal and estrogen-treated guppies,Poecilia reticulata Peters

Summary Gonadotropin releasing hormone (GnRH) and gonadotropic hormone (GTH) were identified by immunohistochemistry in the brains and pituitaries of neonate, juvenile and adult guppies. GTH was present in some cells of the pars intermedia (pi) and proximal pars distalis (ppd) of all animals. GnRH was found in the perikarya of the nucleus olfactoretinalis. In the pituitaries of juvenile 30-day-old guppies, GnRH-immunoreactive cells existed in a juvenile pattern, whereas in adult animals GnRH was recognized in only a few cells. GnRH-immunoreactive fibers were seen in the pituitaries of animals that were 30 days or older. In adult guppies, the ventral and lateral ppd (the gonadotropic region) contained a dense network of GnRH-immunoreactive fibers. Pituitary cells staining for either GnRH or GTH were located in different places. After immunohistochemical double staining of adult pituitaries, none of the GnRH-immunoreactive cells were LH-immunoreactive, although both cell types were often found in close proximity. After 20 days or more of ethinylestradiol treatment, less immunoreactive GnRH was detected in the pituitary cells of juvenile guppies, and fewer animals exhibited the juvenile pattern of GnRH-immunoreactive pituitary cells, when compared with untreated controls. The results indicate that GnRH-immunoreactive pituitary cells in the guppy are distinct from gonadotropes and that these cells are involved in regulatory processes along the juvenile brain-pituitary-gonad axis.     

Immunocytochemical localization of S-100 protein in stellate cells (folliculo-stellate cells) of the adenohypophysis in the monkeys Macaca irus and Cercopithecus aethiops

Summary With the use of an antibody against bovine S-100 protein, it was possible to reveal a characteristic cell type in the pars distalis and the pars tuberalis of the monkey Macaca irus. In the adenohypophysis of Cercopithecus aethiops, labeled cells were present in the pars distalis, pars tuberalis, and pars intermedia. These cells, so-called folliculo-stellate cells, were found in all pituitaries studied. Surprisingly, an antibody against human S-100 protein did not label the stellate cells of the adenohypophysis. However, in Macaca irus, this antibody gave a strong positive reaction with various other cell types (interstitial cells of the pineal gland, Müller cells of the retina, autonomic ganglionic cells, glial cells of the central nervous system, Schwann cells, Bergmann glia of the cerebellum, fat cells, reticular cells of lymphoid organs). By use of double immunoenzymatic labeling, it was evident that stellate cells are spatially related either to somatotropes, prolactin cells, corticotropes, or to glycoprotein-containing cells. Thus, a specific relationship to a particular endocrine-cell type could not be observed.Dr. M.P. Dubois died in Tours (France) on March 30, 1986, aged 65     

Demonstration of gonadotropic cells in the adenohypophysis (pars distalis and pars tuberalis) of the monkey Macacus irus. Immunofluorescence study using human anti-beta-FSH and ovine anti-beta-LH antibodies

Indirect immunofluorescence technique with anti-beta FSH and anti-beta oLH antisera has allowed us to detect "gonadotropic cells" in the pars distalis and in the pars tuberalis of the adenohypophysis of the monkey Macacus irus. In the pars distalis, 85-90 % of the "gonadotropic cells" react simultaneously with these two antisera ; 10-15 % of these cells react only either with anti-beta hFSH or anti-beta oLH antisera. The gonadotropic cells are dispersed in the whole pars distalis, amid the other cellular types ; indeed, in the female, there is a "gonadotropic zone" in the median zone of the lateral lobes of the gland. In the pars tuberalis, we have observed "gonadotropic cells" which react only with anti-beta oLH antiserum. These results are compared with observations of some authors.     

Immunohistochemical identification of cells in the pars distalis of the pituitary of the lizard Anolis carolinensis

Immunocharacteristics of the pars distalis cells of the pituitary of the male lizard A. carolinensis are determined by employing the immunoperoxidase technique with antisera to mammalian pituitary hormones. On the basis of their immunoreactivity, 5 different cell types with characteristic anatomical distribution are recognized. ACTH cells are found in the rostral half of the pars distalis, and PRL cells in the rostral two thirds of the pars distalis. GH and TSH cells are located in the caudal half of the pars distalis. GTH cells are distributed throughout the gland. When consecutive sections are stained with antiserum to ovine FSH or its beta-subunit and to ovine LH, the same cells show immunoreactivity to all the three antisera. None of the GTH cells show positive immunoreactivity to ovine beta-LH antiserum. The results suggest the existence of one gonadotropic cell type in the pituitary of this lizard.     

The presence of gonadotropic and thyrotropic cells in the pituitary pars tuberalis of the monkey (Macaca mulatta).

Immunocytochemistry was utilized to determine if pars tuberalis cells in the pituitary of the monkey (Macaca mulatta) have the potential to elaborate gonadotropic and thyrotropic hormones normally secreted by the pars distalis. A total of 7 males and females were studied. The hormones were localized by the peroxidase-antiperoxidase method of Sternberger, and utilized with antisera to the following human hormones: somatotropin, mammotropin, beta(1-24)-corticotropin, chorionic gonadotropin, and the beta-subunits of follicle stimulating hormone and thyrotropin. Many of the parenchymal cells in the pars tuberalis of the median eminence were composed of gonadotropic cells, probably containing luteinizing hormone and follicle stimulating hormone, and thyrotropic cells. Corticotropic and somatotropic cells were seen only rarely, and mammotropic cells were undetectable. The results indicate that the pars tuberalis is able to secrete luteinizing hormone, follicle stimulating hormone, and thyrotropin.     

Changes in the pituitary of the migrating European eel during its journey from rivers to the sea

Summary The present studies demonstrate that, as migrating European silver eels enter the sea, important changes occur in the pars distalis of the pituitary. A decrease of Type A neurosecretory material in all nerve tracts penetrating the pars distalis was observed, as was an increased activity of Type B neurosecretion in the tracts of the proximal pars distalis.In the proximal pars distalis, gonadotropic cells show striking changes, and in the rostral pars distalis thyrotropic and prolactin cells. It is discussed whether these changes are related to migration to breeding grounds or to changes in the environment.Dedicated to Dr. Berta Scharrer on her 60th birthday.     

Effects of Methallibure (ICI 33,828) on the pars distalis of pituitary,testis and thumb pad of the green frog,Rana esculenta L.

Summary The influence of ICI compound 33,828 (1--methylallylthiocarbamoyl-2-methylthiocarbamoylhydrazine = Methallibure) on the pars distalis of pituitary, testes and thumb pads was investigated in the intact adult male green frog, Rana esculenta. Methallibure affected the gonadotropic basophils (particularly the B2 cells) of the pars distalis, which showed varying degrees of degranulation and underwent a notable decrease in their percentage. Within the testis this compound caused the arrest of spermatogenesis. The most uniform effect of Methallibure was observed in the thumb pads, which invariably showed regression of the epidermis and glandular epithelium. No histological changes occurred in the thyroid and adrenal glands and the B1 and A1 cells of the pars distalis remained unchanged cytologically. It is concluded, in concordance with the available data, that Methallibure is a non-steroidal antigonadotropic compound. The important question about its mode of action has been brought into discussion.Work supported by the National Research Council of Italy, and the Population Council (Grant No. M70.082 C) of New York.     

CXCL14-like immunoreactivity in growth hormone-containing cells of urodele pituitaries

Immunohistochemical techniques were employed to investigate the distribution of a chemokine, namely, CXCL14-like immunoreactivity in the axolotl (Ambystoma mexicanum) and Japanese black salamander (Hynobius nigrescens) pituitaries. CXCL14-immunoreactive cells concentrated at an area of the pars distalis adjacent to the pars intermedia. We found that these cells correspond to the cells immunoreactive to an antibody against rat growth hormone (GH). Immunoelectron microscopy indicated that the CXCL14-like substance and GH coexisted on the secretory granules in the axolotl pituitary. Western blot analysis of axolotl pituitary extracts revealed the anti-human CXCL14 antibody labeled an approximately 16.6-kDa band that was not labeled by the anti-GH antibody. The CXCL14-like substance in the pars distalis may participate in GH functions in these species.     

Gonadotropin-releasing hormone-immunoreactive neuronal structures in the brain and pituitary of the African catfish,Clarias gariepinus (Burchell)

Summary The distribution of gonadotropin-releasing hormone (GnRH) immunoreactivity was studied in the African catfish, Clarias gariepinus, by means of immunofluorescence and immunoperoxidase techniques. Immunoreactive neurons were found throughout the preoptic nucleus (NPO). However, only a portion of the secretory perikarya in the NPO showed a positive reaction by use of an anti-LHRH serum. Numerous immunoreactive fibres were found to enter the pituitary and to terminate in its proximal pars distalis, the site of concentration of the gonadotropic cells. Since GnRH is present in the brain and pituitary of the African catfish, the lack of spontaneous ovulation in captivity is apparently due to an insufficient release of GnRH.     

Immunocytochemistry of a “private” luteinizing-hormone-releasing hormone system in the pituitary

Summary Immunocytochemistry of paraffin sections of Bouin-fixed rat pituitaries with antiserum to luteinizinghormone-releasing hormone (LHRH) revealed two types of cells. Type I cells exhibit granular staining throughout their cytoplasm. The immunoreactivity of type II cells is confined to a much smaller area of the cytoplasm. Type I cells are located in the ventral margin of the pars intermedia, the region between the pars intermedia and the pars distalis, and the pars distalis adjacent to this region. Type II cells have a broader distribution in the pars distalis, but tend to concentrate in the region of the pars distalis near the pars intermedia. Type I cells are distinct from gonadotropes. Type II cells appear to comprise a subgroup of gonadotropes. Staining in type I, but not type II, cells in pituitary explants, maintained in serum-free media for seven days, is as intense as that in normal pituitary tissue. The data suggest that the type I cells are producing an intrinsic LHRH-like material and may be responsible, in part, for the regulation of some gonadotropes.Supported by NIH grants HD12932, NS15843 and NS15809 (LAS), National Science Foundation grant BNS 82-05643 (LAS), and a grant from the Phillippe Foundation (JYL)     

Peptides with NH2-terminal tryptophan in adrenocorticotrophic hormone and melanocyte-stimulating hormone granules of adenohypophysis.

Fluorescence microscopy has demonstrated formaldehyde-ozone-induced fluorescence in the pars intermedia cells (melanocyte-stimulating hormone cells) and in certain cells of the pars distalis of the mammalian pituitary. From histochemical and chemical evidence the fluorescence is believed to reflect the presence of peptides with NH2-terminal tryptophan. In the pars distalis of hamster, cat and pig pituitary, the cells that exhibit formaldehyde-ozone-induced fluorescence have now been identified as adrenocorticotrophic hormone (ACTH) cells by immunohistochemistry. Granules from pig pituitaries were purified by passage through a succession of Millipore filters followed by centrifugation on a continuous sucrose gradient. Two granular fractions were identified by electron microscopy and found to contain high concentrations of peptides with NH2-terminal tryptophan as well as high ACTH bioactivity. These fractions, when pelleted and analyzed histochemically, displayed formaldehyde-ozone-induced fluorescence and ACTH-like immunoreactivity.     

Localization of 3H-dihydrotestosterone in the pituitary gland of the rhesus monkey

Summary The uptake and retention of radiolabelled dihydrotestosterone by the pituitary gland was examined in the rhesus monkey. Two animals were given an intravenous injection of 1.0g/kg ³H-dihydrotestosterone (DHT) alone while one monkey received both the labelled androgen and 100g/kg of unlabelled steroid. One and a half hours later, they were sacrificed. The pituitary glands were removed and processed for autoradiography and immunocytochemistry. Autoradiographic localization of DHT was discernible in the partes nervosa, intermedia and distalis, albeit the highest concentration of radiolabelled cells was noted in the pars distalis. Immunocytochemical staining with antibodies to rat PRL, human TSH and ovine LH revealed a population of steroid-concentrating cells that contained TSH and a second group that contained LH. None of the cells that reacted with the anti-PRL serum were radiolabelled.     

Immunohistochemical detection of adenohypophyseal cells containing hormones in Rana ridibunda

Summary The pars distalis of the pituitary of Rana ridibunda captured throughout the spring and summer was examined with immunofluorescence techniques using antisera to mammalian pituitary hormones. On the basis of their immunoreactivity, four different cell types are recognized: 1) cells immunoreactive to anti-bovine LTH, 2) cells immunoreactive to anti-bovine STH, 3) cells immunoreactive to anti-ovine LH, and 4) cells immunoreactive to anti-synthetic ACTH (1–24). Their distribution and morphology as well as their staining characteristics (classical histological techniques) are reported in this study.     

Diurnal rhythm of hypophyseal cells in mice. IV. Pars Distalis∗

Abstract

Using various staining techniques and methods, the following cells were differentiated from pars distalis of the pituitary gland in the mouse: corticotropic, somatotropic, lactotropic, thyrotropic and gonadotropic cells.

Diurnal activity of the differentiated cells in pars distalis of the pituitary gland was determined by the method of karyometry.

The karyometric analysis showed the existence of a distinct diurnal rhythm in nuclear volume in all five types of the differentiated cells in both sexes.     

Histology of the pituitary gland of the grey mullet, Mugil cephalus L.

The present communication deals with a histological study of the pituitary gland of the teleost fish Mugil cephalus , found in the estuarine waters of Cochin area. Six different cell types were identified in the pituitary gland on the basis of their grouping, distribution and staining properties. The prolactin and the TSH cells (thyroid stimulating hormone producing cells or thyrotrops) were identified in the rostral pars distalis and the ACTH cells (adrenocorticotropic hormone producing cells or corticotrops) in the interphase between the neurohypophysis and the rostral pars distalis. The STH cells (somatotropic hormone producing cells or somatotrops) and the gonadotropic cells were distinguished in the proximal pars distalis and the MSH cells (melanin stimulating hormone producing cells or melanotrops) in the pars intermedia.     

Cloning and Comparison of Factor X from Rhesus Monkey (Macaca mulatta)

The reliability of the rhesus monkey as an important experimental animal depends on its genetic concordance with human. During our assessment of the rhesus monkey as a preclinical model for coagulation-related research, we cloned the full-length cDNA of rhesus monkey factor X (FX) and compared its genetic characteristics and coagulation activity with those of human FX. The full-length cDNA of rhesus monkey FX was 1683 bp in length, corresponding to 487 coding amino acids and sharing 94.71% nucleotide identity and 93.65% amino acid identity with human FX. When FX sequences from different animals were compared with that of human FX, rhesus monkey and baboon FX showed similar degrees of homology to human FX, which were less than that between human and chimpanzee FX sequences but remarkably higher than those of another 2 monkey species, bovine, pig, and rodents. Comparison of functional sites between human and rhesus monkey FX revealed high similarities between their amino acids sequences and 3-dimensional structures. The average coagulation activity of FX from 24 rhesus monkeys was in the normal range of that of healthy humans. The rhesus monkey therefore may be a suitable animal model for research addressing coagulation factor X.Abbreviations: 3D, 3-dimensional; FX, factor X; SMART, simple modular architecture research tool; TFPI, tissue factor pathway inhibitorRhesus monkeys (Macaca mulatta) continue to be widely used research animals in many biologic fields.¹⁵ Because of their close genetic relationship to humans, there is increasing interest in the use of rhesus monkeys for gene therapy, stem cell, infectious disease researches, and reproductive biology.¹² The effective application of rhesus monkey as an important experimental animal is dependent on the genomic and proteomic concordance between rhesus and human.¹⁷ The coagulation system plays an essential role in many physiologic and pathologic processes, including hematologic, cardiovascular, and liver diseases and transplantation. Clearly, the establishment of genetic information and reference activity values of rhesus monkey coagulation factors is requisite to interpretation of the data from preclinical coagulation-related research using rhesus monkeys. Factor X (FX) is a vitamin-K–dependent protein that is 1 of the most critical factors in the coagulation scheme; its activation is the convergence of the extrinsic and intrinsic activation pathways and leads to the final stages of hemostasis.⁶ Human FX has been investigated extensively during the past century, but there have been few reports regarding the characterization of monkey FX.¹⁴The ineffective binding of porcine tissue factor pathway inhibitor (TFPI) to human FX was proposed to be an important contributor to coagulopathy in pig-to-human xenotransplantation.¹¹ Many in vitro studies¹¹ have investigated the interaction between porcine endothelial cells and human coagulation factors, whereas in vivo studies⁸ have addressed the interaction between porcine endothelial cells and monkey coagulation factors. The concordance of the results from these in vitro and in vivo studies therefore reflects high homology, if not identity, between human FX and monkey FX. During our assessment of whether rhesus monkey is a reliable model for studying coagulation disorders in xenotransplantation, we cloned the full-length cDNA of rhesus monkey FX and compared its nucleotide and amino-acid sequences and coagulation activities with those of human FX.     

Life span changes in the presence of alpha-melanocyte-stimulating-hormone-containing cells in the human pituitary.

Since recent circumstantial evidence has suggested possible functions of alpha-MSH in intrauterine growth and labour, the presence of this hormone in the human pituitary was determined by means of the indirect immunofluorescence procedure during development and adulthood. Cross reaction of the antibodies with other peptides was measured after which they were purified by solid phase absorption. Experiments on the rat pituitary showed that staining of alpha-MSH- and ACTH-containing cells could be obtained well until 48 h after death. In the pars distalis the ability of ACTH-containing cells to take up stain increased during the period of post-mortem storage. In the youngest human fetus studied (15 weeks) only alpha-MSH-containing cells were found in the pars intermedia and no ACTH-containing cells were observed. In the other fetal pituitaries a distinct pars intermedia containing more alpha-MSH cells than ACTH cells was found. In the pars distalis of the fetuses more ACTH- than alpha-MSH-containing cells were observed. From birth to 19 years, progressively fewer alpha-MSH containing cells could be detected in the 'zona intermedia' and pars distalis, while in adults only a few such cells were found in either area. Irrespective of age, sex, cause of death or therapy, alpha-MSH-containing cells were found in all pituitaries throughout life. The number of ACTH containing cells gradually increased in the zona intermedia and pars distalis and reached a high adult level in the latter structure. In the pituitaries of seven anencephalics, no alpha-MSH-containing cells were present. The presence of alpha-MSH in the fetal pars intermedia, the change in the ratio of the alpha-MSH/ACTH cells during the course of development, and the absence of alpha-MSH in anencephaly all support the possibility that human fetal pituitary alpha-MSH is involved in both intrauterine growth and fetal adrenal function and thus also in parturition.     

Immunoreactivity to gonadotropin-releasing hormone and gonadotropic hormone in the brain and pituitary of the rainbow trout Salmo gairdneri

Summary Immunoreactivity to gonadotropin-releasing hormone (GnRH) and gonadotropic hormone (GTH) was studied at the light-microscopical level in the brain and pituitary of rainbow trout at different stages of the first reproductive cycle using antisera against synthetic mammalian GnRH and salmon GTH. GnRH perikarya were localized exclusively in the preoptic nucleus, both in the pars parvicellularis and the pars magnocellularis. A few somata contacted the cerebrospinal fluid. Not all neurosecretory cells were GnRH-positive, indicating at least a bifunctionality of the preoptic nucleus. We recorded no differences between sexes or stages of gonadal development in the location of GnRH perikarya, whereas gradual changes were found in staining intensity during the reproductive cycle. GnRH fibres ran from the partes parvicellularis and magnocellularis through the hypothalamus and merged into a common tract at the transverse commissure before entering the pituitary. In the pituitary, GnRH was localized in the neural tissue of the neurointermediate lobe and, to a lesser extent, in the neural protrusions penetrating the proximal pars distalis. The bulk of GTH-positive cells was situated in the proximal pars distalis. Some cells were found more rostrally amidst prolactin cells or in the neurointermediate lobe. Only a limited number of GTH cells appeared to be in close contact with GnRH-positive material.     

An immunocytochemical study of the pituitary gland of the white seabream (Diplodus sargus)

The adenohypophysis of the white seabream (Diplodus sargus) was studied using histochemical and immunocytochemical techniques. The adenohypophysis was composed of rostral pars distalis, proximal pars distalis and pars intermedia. Prolactin (anti-chum salmon prolactin positive) and adrenocorticotropic (anti-human ACTH positive) cells were found in the rostral pars distalis. Prolactin cells were organized into follicles, while ACTH cells were arranged in cords around neurohypophyseal tissue branches that penetrated the rostral pars distalis. In the proximal pars distalis, somatotropic (anti-chum salmon and anti-gilthead seabream growth hormone positive), gonadotropic (anti-chum salmon -gonadotrophin II and anti-carp -gonadotrophin II positive, but anti-chum salmon -gonadotrophin I negative) and thyrotropic (anti-human -thyrotropin positive) cells were observed. Growth hormone cells were restricted to the dorsal and ventral part of the proximal pars distalis. They were clustered or surrounded the neurohypophyseal branches. Only one type of gonadotrophin cell was identified and they were clustered or isolated in the proximal pars distalis. Scattered groups of thyrotropin cells were located throughout the proximal pars distalis. In the pars intermedia somatolactin (anti-chum salmon and anti-gilthead seabream somatolactin positive) and melanotropic (anti--melanotropic hormone positive) cells were localized. In addition, gonadotrophin cells surrounded the pars intermedia or distributed evenly between somatolactin and melanotropic hormone cells. Somatolactin cells were periodic acid-Schiff negative and surrounded the neurohypophyseal branches intermingled with melanotropic cells. These cells were also immunoreactive to anti-human ACTH antiserum.