Structural changes in the pars intermedia of the cichlid teleost Sarotherodon mossambicus as a result of background adaptation and illumination

Summary The pars intermedia of Sarotherodon mossambicus (Tilapia mossambica) contains two cell types which can be differentiated at both the light and electron microscopic level. The predominant cell type is lead haematoxyline positive, and has been shown to be the MSH producing cell type by means of immunocytochemical staining at the ultrastructural level. The changes in cellular and nuclear volume, as well as the results of stereological measurements on the cytoplasmic organelles, show that the activity of MSH cells is high on a black background and low on a white background or in total darkness. In blinded fish under a normal day-night regime the activity of the MSH cell is as high as that in black adapted fish, whereas the activity is low when the blinded fish are kept in total darkness. From the observed differences in activity of the MSH cells between the experimental groups, it is concluded that the MSH cells are not activated by the absence of reflected light, but by a high ratio between direct and reflected light. A second light-sensitive organ, supposedly the pineal gland, is also involved in the background response of the MSH producing cells.     

Structural changes in the pars intermedia of the cichlid teleost Sarotherodon mossambicus as a result of background adaptation and illumination

Summary The MSH producing cells in the pars intermedia of Sarotherodon mossambicus have been shown to be involved in background adaptation processes. Reflected light received by the eyes affects the activity of these cells. In the present study the hypothesis has been tested that also the pineal organ, as a second photoreceptor, is involved in regulation of the metabolic activity of the MSH cells. The pineal organ appears to contain photoreceptor cells and is considered to be capable of transferring information about light conditions to the animal. Removal of the pineal organ of fish kept on a black background has no effect on activity of MSH cells, whereas the activity of these cells in fish kept in darkness is increased. Thus it seems that the pineal organ exercises its influence on MSH cells only in darkness and that this influence results in a reduced activity of these cells. It is therefore concluded that the metabolic activity of MSH cells is inhibited not only by reflected light received by the eyes, but also by the action of the pineal organ as a result of the absence of illumination.No structural signs of secretory activity can be observed in the pineal, which might indicate synthesis or release of substances like melatonin. However, administration of melatonin reduces the activity of MSH cells. Neither pinealectomy nor treatment with melatonin has any influence on the second cell type of the pars intermedia, the PAS positive cells.     

Responses of the PAS-positive pars intermedia cells in the cichlid fish Sarotherodon mossambicus to ambient calcium and background adaptation

Summary The function of the PAS-positive pars intermedia cells in fish has been connected with control of background adaptation and of plasma calcium levels. Since background reflectivity and illumination influence calcium metabolism in S. mossambicus, we have tested the hypothesis that the effects of background reflectivity on the PAS-positive cells are mediated by changes in plasma calcium. However, total plasma calcium does not correlate with the activity of the PAS-positive cells as estimated by morphometrical criteria. Transfer of S. mossambicus to a white background leads to a drop in plasma calcium, and a marked reduction of the activity of the PAS-positive cells. Adaptation to low-calcium freshwater, on a neutral background, has the same effect on plasma calcium but has no effect on the PAS-positive cells. The characteristic structural features of highly active PAS-positive cells in fish from a black background are not due to the slight hypocalcemia that occurs in these fish, since addition of calcium to the water in concentrations that fully prevent the drop in plasma calcium does not suppress the PAS-positive cells. These findings make it very unlikely that these cells produce a hypercalcemie factor in S. mossambicus.     

The effect of water acidification on prolactin cells and pars intermedia PAS-positive cells in the teleost fish Oreochromis (formerly Sarotherodon) mossambicus and Carassius auratus

Summary Although exposure to acid water (pH 3.5) induces severe and prolonged reduction in plasma osmolarity and total plasma calcium concentration in tilapia (Oreochromis mossambicus) and goldfish (Carassius auratus), the responses of the hypophyseal cells are clearly different. In tilapia, the size of the rostral pars distalis of the pituitary gland is enlarged as a result of the increase in size and number of prolactin cells. The pars intermedia PAS-positive (PIPAS) cells are not noticeably changed. Conversely, in goldfish, prolactin cells are unaffected, whereas the number of enlarged PIPAS cells increases markedly. Stimulation of prolactin secretion may be responsible for the partial restoration of plasma osmolarity and calcium levels observed in tilapia after two weeks exposure to acid water. Prolactin cells apparently play a role in the adaptation to acid stress by counteracting osmoregulatory disturbances. Goldfish show no restoration of plasma osmolarity during the course of the experiment. Plasma calcium levels tend to increase. Although prolactin may have an osmoregulatory function in goldfish under steady state conditions, goldfish prolactin cells do not seem to participate in the physiological adaptation to environmental changes that disturb water and ion homeostasis. The function of PIPAS cells in tilapia remains unclear and is apparently unconnected with ion regulation. The observations on these cells in goldfish are consistent with the hypercalcemic activity suggested for them.     

Response of PAS-positive cells of the pituitary pars intermedia in the teleost Carassius auratus to acid water

Summary The PAS-positive or PIPAS cells in the pars intermedia of goldfish are activated after reduction of the pH of the ambient freshwater from 7.5 to 3.5. The cells increase in number and exhibit a five-fold increase in cell volume. Granular endoplasmic reticulum occupies most of the cytoplasm. Goldfish PIPAS cells (also termed calciumsensitive cells) are thought to have a hypercalcemic function. Therefore, their activation in acid water may be caused by the severe drop in plasma calcium concentration following exposure of the fish to low water pH. However, activation of the PIPAS cells in response to acidification of the water is not prevented when the calcium concentration of the water is increased to levels that result in hypercalcemia instead of hypocalcemia. Activation of the PIPAS cells occurs also in fish exposed to acidified freshwater enriched with NaCl to an osmolarity similar to that of the blood. This prevents the reduction in plasma osmolarity and Na⁺ and Cl^- concentrations that follow exposure of goldfish to acidified normal freshwater. Our observations do not support the hypothesis that the PIPAS cells in goldfish produce a hypercalcemic hormone, or indeed any hormone involved in calcium metabolism or osmoregulation. The cells may be implicated in acid-base regulation (a characteristic of many types of fish when exposed to acidified water) but the evidence is indirect.     

A morphometric study on the pars intermedia of the hypophysis during impairment of the renin-angiotensin-aldosterone system in sodium depleted mice

Fine structural alterations were investigated in cells of the pars intermedia of the pituitary of mice treated for four weeks with (a) a sodium deficient diet, (b) a sodium deficient diet mixed with propranolol (renin-inhibitor), (c) a sodium deficient diet combined with propranolol and amino-glutethimide (corticosterone 18-hydroxylase inhibitor), and (d) a sodium deficient diet combined with propranolol, aminoglutethimide and dexamethasone. The number of secretory granules decreased from 5.0/mum2 in the normal control of 2.4/mum2 in all four experimental groups suggesting that the cells in treated groups had reached an equilibrium in the production and release of secretory granules during the chronic treatments. The number of immature Golgi granules per unit Golgi area was 0.91 in the control, while this value rose to 3.29 (3.62 fold of the control), 4.37 (4.8 fold), 4.94 (5.43 fold) and 5.16 (5.67 fold) respectively in the four experimental groups. In these groups a good correlation was observed between the number of immature granules and the percent volume of rough endoplasmic reticulum (r=0.985, p less than 0.01). The present study suggests that the pars intermedia contains an unidentified pituitary factor (or factors) essential for aldosterone biosynthesis.     

Cellular localization of somatolactin in the pars intermedia of some teleost fishes

Summary We report here on the cellular localization in the fish pituitary of somatolactin (SL), a putative new pituitary hormone related to growth hormone and prolactin, which has been recently identified in the piscine pituitary gland. Immunocytochemical staining, using anti-cod SL serum, revealed that in the cod pituitary gland, SL is produced by cells in the intermediate lobe, bordering the neural tissue. These cells, staining weakly with periodic-acid-Schiff (PAS), are distinct from the melanocyte stimulating hormone (MSH) cells which, as in all teleosts, are PAS-negative. SL-immunoreactivity was observed in the same location in all other teleost species examined: flounder, rainbow trout, killifish, molly, catfish and eel. In most fish the SL-immunoreactive cells are either strongly or weakly PAS-positive but in rainbow trout are chromophobic, indicating that the SL protein can probably exist in glycosylated and non-glycosylated forms. Thus, in demonstrating the cellular localization of SL, this study provides the first identification of the enigmatic, second cell-type of the fish pars intermedia.     

Structural relationships between parenchymal and stromal elements in the pars intermedia of the rat adenohypophysis as demonstrated by extracellular space markers

Summary The pars intermedia (PI) of the rat adenohypophysis was studied by light and transmission electron microscopy after conventional staining as well as ruthenium red staining, and after systemic injection of horseradish peroxidase. The studies disclose a complex and constant system of two channel types (Type I and Type II channels) formed by PI cells with specific relationships to a very rich nerve supply, to each other, and to a stellate cell type proposed here to represent an element of neuroglia. The channel system could perform a function in the movement of fluids and solutes within the PI which is virtually avascular in the rat as well as in other mammals.Supported by the Medical Research Council of Canada Grant # MA6445We thank Mr. J. Jones and Mr. A.R. Morris for their expert technical assistance     

Phagocytic activity of the stellate cells in the anuran pars intermedia

Summary In an attempt to study further the stellate cell and its functions, the ultrastructure of this cell type in the neurointermediate lobe of the bullfrog, Rana catesbeiana, was examined in both organ and dissociated-cell culture. The cytoplasmic activity of stellate cells from neurointermediate lobes incubated 3 1/2 or 5 1/2 h was greater than that of those in vivo. Mitochondria and bundles of cytoplasmic filaments were numerous, in addition to prominent, well-developed Golgi complexes with associated vesicles. The most striking ultrastructural feature was the presence of phagocytic vacuoles that contain cellular debris. The stellate cells were seen to form cytoplasmic processes that phagocytosed this extracellular debris identifiable as belonging to the secretory cells of the pars intermedia. The stellate cells from the dissociated-cell preparations were also seen to contain debris within phagocytic vacuoles. In those neurointermediate lobes transplanted for 3 1/2 to 4 days into the anterior chamber of the eye, the stellate cells demonstrated similar phagocytic ability, but the phagocytic vacuoles contained material that seemed to be at a later stage of degradation. In all three of these conditions, the stellate cells were not seen to release this cellular debris nor were they seen to undergo cell division. These glial-like stellate cells of the pars intermedia acted as macrophages in all three of these experiments. There is now, therefore, a need to determine under what conditions, if any, these stellate cells function in vivo as macrophages.Supported by NSF Program for Small College Faculty Engaged in Research at Larger Institutions and Department of Energy — Associated Western Universities Faculty Participation Program. The authors thank Dr. W. Ferris and Dr. J. Berliner for the use of the electron microscopy facilities at the University of Arizona and Nuclear Medicine Laboratory, UCLA, respectively. Warm thanks are due to Ms. Ruth Cole for technical assistance     

PAS-positive cells of the pars intermedia are calcium-sensitive in the goldfish maintained in a hyposmotic milieu

Summary Cytological changes in the pars intermedia of the goldfish were investigated after adding calcium to deionized water (DW). In fish maintained in DW, the PAS-positive cells are highly stimulated in comparison to cells of fish kept in fresh water (FW). In DW supplemented with calcium at the same concentration as in FW (2 mM/l), the hyperactivity of the PAS-positive cells is prevented. When calcium ions are added 60 h before the animals are sacrificed, the PAS positive cells start to show signs of regression and their granules are stored: the release of the granular material appears to be suppressed by calcium. In the goldfish, the PAS-positive cells, homologous to a similar cell type in the eel, react only very weakly with the PAS technique. The name calcium-sensitive cells appears to be more appropriate in the goldfish for this particular cell type, secreting an unknown factor. This factor, different from the prolactin produced in the rostral pars distalis of the hypophysis, might be an equivalent of a hypercalcin.     

The effect of change of background colour on the ultrastructure of the pars intermedia of the pituitary of the eel (Anguilla anguilla)

Summary Colour change in the eel resulted in marked ultrastructural changes in the pre-dominating (Type II) secretory cells of the pars intermedia of the pituitary. The effects on these cells of transferring eels from white to black backgrounds for periods of up to 56 days were: a) hypertrophy of the rough endoplasmic reticulum, which increased from 12 to 35% of the cytoplasmic volume; b) loss of secretory granules which decreased from 38 to 5% of the cytoplasmic volume; c) development of a system of fine (25–35 nm) tubules located especially at the secretory poles of the cells but also found in the region of the Golgi apparatus. The tubules were seen to connect with the plasma membrane, with the limiting membrane of the secretory granules, and in one instance to connect a granule with the plasma membrane. After glutaraldehyde fixation at pH 5, electron dense material similar to that found in the secretory granules was observed in the lumen of many of the tubules.The changes that occurred in black background eels are taken to indicate that the Type II cells of the pars intermedia are responsible for MSH secretion, particularly since these changes were reversed by returning eels to white backgrounds. The cytoplasmic tubules found in Type II cells may indicate a process for MSH release which does not involve granule extrusion, but rather direct transport of material from the Golgi apparatus to the cell membrane.The electron microscope facilities used in this investigation were funded by the Medical Research Council.     

Immunohistochemical localization of adrenocorticotropin and melanocyte stimulating hormone in pars intermedia of rat hypophysis

Summary The sites of production of adrenocorticotropin (ACTH) and melanocyte stimulating hormone (MSH) are studied by the immunoglobulin-peroxidase bridge technique, using antisera prepared against synthetic porcine 1–24 and 17–39 ACTH, and bovine MSH on the rat adenohypophysis. Presence of ACTH all over the pars intermedia (PI) is indicated by staining with antisera _p 1–24 and _p 17-3-9 ACTH. There are darkly stained ACTH cells in the PI and pars tuberalis (PT), similar to those in the pars distalis (PD). With higher dilutions of the ACTH antiserum, staining intensity disappears or reduces markedly in majority of the PI cells, whereas, the ACTH cells in the PI, PD and PT do not vary much in their staining intensity. Therefore, it is concluded that majority of the PI glandular cells (light glandular and dark cells) contain less corticotropin than the ACTH cells. From these observations, it seems to me that the major amount of corticotropin is supplied by the ACTH cells of the PD, PI and PT, and less by the light glandular and dark cells of the PI. The antiserum is ineffective after absorption, so the staining reaction appears to be specific for _p 1–24 and _b 17–39 ACTH.Presence of MSH all over the PI is indicated by staining with antisera to bovine MSH. Majority of the PI cells are highly stained even with higher dilution of the antiserum. The unstained cells in the PI seem to be ACTH cells and/or marginal cuboidal cells. The antiserum was ineffective after absorption, so the staining reaction appears to be specific for _b MSH.Control over the PD corticotropin through the median eminence portal circulation and the PI and PT control through nervous system is also discussed.This study was supported by MRC of Canada Grant nos. MA-3759, and MA-5160.The author gratefully wishes to thank Drs. P. Desaulles and W. Rittel (CIBA, Basle, Switzerland) for the synthetic _p 1–24 ACTH and _b MSH, Dr. R. F. Phifer for _p 17–39 ACTH, and Dr. S. S. Spicer for providing samples of rabbit anti-porcine 17–39 ACTH and anti-human ACTH sera, Drs. George Sétáló and Paul Nakane for their valuable advice. He also acknowledges the help of Mr. Shankar Nayak to prepare the antisera and the skilful technical assistance of Miss. Elise Poiré.     

Light and electron microscopic study of the pars intermedia of the lizard,Klauberina riversiana

Summary Two cell types can be distinguished in the pars intermedia of Klauberina: (1) Glandular cells, which form a single-layered columnar epithelium on the vascular septum which divides the pars nervosa from the pars intermedia. (2) Marginal cells which form a flattened epithelium over the glandular cells and line the hypophysial cleft. Occasional projections from the marginal cells extend between the glandular cells to contact the basement membrane of the vascular septum, and occasional projections of the vascular septum extend across the glandular epithelium to reach the marginal epithelium. Both cell types are AF negative. The granules of the glandular cells are strongly PAS positive, and acidophilic in response to Mallory's trichrome stain. In electron micrographs, the glandular cells contain large quantities of secretory granules. In one class of cells, they range from 2,000 to 2,500 Å in diameter, in the other, from 4,000 to 5,000 Å. Electron-dense granules 1,000 to 1,500 Å in diameter occur in the cytoplasm of the marginal cells in the region of contact with the vascular septum. Hence more than one active principle may be produced by the pars intermedia.No nerve endings of any kind are present in the pars intermedia. Therefore, synaptic contact of neurons with the secretory cells seems not to be necessary for the regulation of their secretory activity as appears to be the case in other vertebrate groups. It is suggested that regulatory factors are secreted in the pars nervosa and transported to the pars intermedia via the vascular septum.Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina.This investigation was supported in part by a Public Health Service fellowship 1 FZ HD 32, 949-01 REP from the National Institute of Child Health and Human Development. The authors wish to thank Professor H. Heller for his encouragement and kind cooperation during this study and Professor H. D. Dellmann for providing the facilities of his department. They are indebted to the officers and men of the Naval Ordinance Test Station, Pasadena and San Clemente Island, California, for their help in obtaining animals for this investigation.     

Corticotrophic cells in the rostral zone of the pars intermedia and in the adjacent neurohypophysis of the rat and mouse

Summary In the mouse, the rostral zone of the pars intermedia is almost exclusively composed of typical corticotrophic cells. They are located around and even within the neural stalk, at the level of transition between stalk and neural lobe. In the rat, the corticotrophic cells of the rostral zone are found in scattered islets among the MSH producing cells, and also in the neural lobe. In both the rat and mouse, these cells are in direct contact with various types of nerve terminals. Synaptoid contacts with aminergic and neurosecretory nerve fibers are observed. Furthermore they are also closely related to the hypophysial portal vessels. Following adrenalectomy, the cells located in the neurohypophysis always react more intensely than tose in the rostral zone. The functional significance of these corticotrophic cells which are subject to both humoral and neural regulation remains as yet hypothetical. Their participation in neurogenic stress response seems probable.     

Immuno-enzyme cytochemical demonstration of mesotocinergic nerve fibres in the pars intermedia of the amphibian hypophysis

Summary Immuno-enzyme cytochemical investigations showed that the whole amphibian pars intermedia of the hypophysis is innervated by an intercellular network of peptidergic varicose nerve fibres which contain mesotocin or (and) parts of the mesotocin molecule. The pars intermedia does not contain vasotocinergic fibres. The mesotocinergic fibres are branches of axons leaving the pituitary stalk and the neural lobe. In animals of which the hypothalamic magnocellular neurosecretory preoptic nuclei had been completely removed, the immuno-reactive mesotocinergic fibres of the pars intermedia had totally disappeared. From this result, it is concluded that the mesotocinergic fibres of the pars intermedia of the amphibian hypophysis are axons of neurosecretory perikarya located in the hypothalamic magnocellular neurosecretory preoptic nuclei.Dedicated to Professor Berta Scharrer on the accasion of her 70th birthdayThis investigation was supported by a grant from the Belgian Nationaal Fonds voor Geneeskundig Wetenschappelijk Onderzoek     

Light and electron microscopic studies on the pars intermedia of the chameleon

Summary The neurointermediate lobe of the hypophysis in the Chameleon (Chamaeleo dilepis) was examined with light and electron microscopic methods, with special reference to the cytology of the pars intermedia (PI). The PI is the largest lobe of the hypophysis consisting of (1) dark cells with secretory granules ranging from 200–600 nm; (2) light cells, far fewer in number, containing granules 150–300 nm in diameter; (3) stellate, non-secretory cells. The secretory cells abut onto the perivascular basal lamina of the capillary sinusoids while their apical part borders an intercellular space. This surface of the cells often bears a cilium. The granules arise from the Golgi cisternae while small detached vesicles are found between circumscribed sites of the cell membrane and the Golgi apparatus. No nervous elements were found in the pars intermedia and it is assumed that the regulation of this lobe is purely humoral. This is supported by the presence of three types of nerve terminals in the pars nervosa: (a) terminals with large secretory granules and small vesicles; (b) terminals with dense-core vesicles and small vesicles; (c) terminals with small vesicles only. All of these are secretory as indicated by the presence of the synaptic semidesmosomes formed with the perivascular basal lamina.I would like to thank Mr. W.N. Newton for his skill and aid in all aspects of this work, Mr. A. Ansary for expert photographic assistance and the Central Pathology Laboratory, University of Dar es Salaam, for the electron microscopic facilities provided. Research sponsored by the University of Zambia Grants J02-18-00 and Medic 74/6     

Mitotic activity of the pars intermedia in the female mouse: age-associated variations in proliferation rate and circadian periodicity

We previously reported daily variations in the mitotic activity of the endocrine cells in the pars intermedia of 21- and 28-day-old male mice. Since cellular proliferation might be affected by factors such as sex and age, we undertook the present experiments to study the mitotic activity of the pars intermedia from 14-, 28-, and 150-day-old female mice. Inbred C3H/S mice, grouped according to age, were housed under standard conditions (12h each of light and dark [LD 12:12]) for periodicity analysis and were killed in lots of 5-11 animals every 4h over a single 24h cycle, with each mouse receiving 2 μg/g of colchicine 4h before decapitation. Pituitaries were excised, extracted, fixed in buffered formaldehyde, embedded in celloidin-paraffin, sectioned at 5 μm, and stained with hematoxylin and eosin. We counted the total number of nuclei to estimate the total number of cells monitored and then calculated the mitotic index (metaphases/1000 nuclei). Differences were analyzed for statistical significance by the Student t test. While the 14-day-old animals manifested no significant changes in mitotic activity during the 24h cycle, the 28- and 150-day-old mice showed higher mitotic indices during the period of darkness. The average mitotic activity over the entire cycle, however, was higher in the two groups of younger animals than in the 150-day-old mice. Moreover, the averages for the 28-day-old females were higher than the corresponding values previously reported by us for male mice of the same age. (Chronobiology International, 17(6), 751-756, 2000)     

Specific effect of calcium ions on the calcium-sensitive cells of the pars intermedia in the goldfish

Summary Cytological changes in the calcium-sensitive (Ca-s) cells (formerly termed PAS-positive cells) of the pars intermedia were investigated in the goldfish after adaptation to deionized water (DW), with or without addition of sodium, potassium and magnesium. These ions were added as chloride salts at concentrations similar to those present in fresh water (FW). The marked stimulation of the Ca-s cells is not inhibited in DW supplemented with Na⁺ (0.35 mM/1), K⁺ (0.05 mM/1), and Mg²⁺ (0.2mM/1) for a period of 24 days. The inhibition of the response to DW with calcium chloride (2 mM/1) is reproduced with calcium formiate (2 mM/1). These data show that chloride ions are not responsible for the regression of the Ca-s cells observed in goldfish kept in DW supplemented with calcium chloride. The effect of calcium ions on the Ca-s cells appears to be specific. These results support the hypothesis that the Ca-s cells synthesize a factor (hypercalcin?) involved in calcium regulation, and that its release is influenced by the calcium content of the environment. The role of the pars intermedia in calcium metabolism is strengthened by the present results. Biochemical data suggest the presence of a hypercalcemic factor in the pituitary of fish (Parsons et al. 1978) and are in agreement with the present cytological findings.     

Electron microscopic-immunocytochemical localization of adrenocorticotropin and melanocyte stimulating hormone in the pars intermedia cells of rats and mice

Summary Electron microscopic localization of adrenocorticotropin (ACTH) and melanocyte stimulating hormone (MSH) in light, dark and ACTH cells in the pars intermedia (PI) of rats and mice is attempted by using antisera to _p 1–24, _p 17–39 ACTH and _b MSH with the immunoglobulin-peroxidase bridge technique. All of the PI parenchymatous cells (light, dark and ACTH cells), except the marginal cuboidal and the ependymal like cells, in rats and mice show very good localization of ACTH and MSH staining. In the light and dark cells, stain of varying intensity is seen on the secretory granules, vesicles and also in many places on the surface of the rough endoplasmic reticulum. There is no staining on the mitochondria, in the nuclei or in the granules inside and around the cisternae of the Golgi complex. Dark stained dense core granules become larger and larger as they appear farther and farther away from the Golgi complex. On the other hand, in the ACTH cells of the PI, ACTH antisera show stronger stained granules in the Golgi complex including the cisternae, similar to the pars distalis (PD) ACTH cells. From these observations it is concluded that the corticotropin in light and dark cells, is not packaged or condensed in the Golgi complex like that in the ACTH cells. MSH synthesis in light and dark cells also seems to be similar to that of ACTH synthesis. It is likely that the granules accumulate ACTH and MSH secretions after they are liberated from the Golgi cisternae, and thus become bigger and bigger in size. In case of ACTH cells of PI and PD, corticotropin may be packaged in Golgi cisternae and the size of the granule does not change much. This shows that there are distinct immunocytochemical differences between the light, dark and ACTH cells of the PI. At the moment, it is difficult to say whether ACTH and MSH are present in the same granule or not.The present and previous studies show that the ACTH and MSH secretion in the PI of rats and mice depends on the hypothalamic neural control.This study was supported by MRC of Canada Grant nos. MA-3759, and MA-5160.The author gratefully wishes to thank Drs. P. Desaulles and W. Rittel (CIBA, Basle, Switzerland) for the synthetic _p 1–24 ACTH and _b MSH, Dr. R. F. Phifer for _p 17–39 ACTH, and Dr. S. S. Spicer for providing samples of rabbit anti-porcine 17–39 ACTH and anti-human ACTH sera, Drs. George Sétáló and Paul Nakane for their valuable advice. He also acknowledge the help of Mr. Shankar Nayak to prepare the antisera and the skilful technical assistance of Miss. Elise Poiré. He wishes to acknowledge Mr. Gatson Lambert for his photography.