On the relations between the hypothalamus and the anterior pituitary in Rana temporaria

Summary Dawson (1952) showed that, in Rana, many nerve terminals end round the capillaries of the venous portal system of the median eminence. Our observations not only confirm these facts but show as well that, besides the posterior lobe, the outer zone of the median eminence is also a centre of accumulation of neurosecretory material derived from the pre-optic nucleus. The amount of this neurosecretory material is moreover subject to a certain variation in connection with the seasons and with experimental conditions. The idea is put forward that the outer zone of the median eminence of Rana has to be considered as an analogical organ of the posterior lobe. Indeed, in the median eminence, the same morphological facts can be found, which are advanced for the conception that the posterior lobe is a centre of accumulation and a place of release of neurosecretory material originating from the pre-optic nucleus. On the basis of our observations we presume that the hypothalamo-hypophyseal neurosecretory system of Rana can be divided in two sub-units. System I ends in the posterior lobe; system II in the outer zone of the median eminence. The data of the literature indicate that the two systems also probably exist in the higher vertebrates. From these facts it is obvious that the neurosecretory material is transported from the pre-optic nucleus, via the tract, towards the outer zone of the median eminence.     

Retrograde axoplasmic transport of neurosecretory material

The axonal transport of neurosecretory material was studied in neurosecretory axons of the supraoptico-posthypophyseal system after in-situ transection of the median eminence. Two hours, 8 h, and 18 h after the lesion, both vasopressin and oxytocin antibodies revealed progressive accumulations of immunoreactive material not only in the proximal but also in the distal stumps of the transected axons. The electron-microscopic examination of these axonal portions revealed that such intense immunopositive labelings could be correlated, in both stumps, to a conspicuous accumulation of neurosecretory granules. It is concluded that, under normal physiological conditions, a significant amount of axoplasmic neurosecretory material is transported in retrograde direction and that such a retrograde transport mainly involves neurosecretory granules.     

The distribution of monoamine oxidase and acetylcholinesterase in the hypothalamus and its relation to the hypothalamo-hypophysial neurosecretory system in the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary The distribution of monoamine-oxidase and acetylcholinesterase activities in the hypothalamus of the White-crowned Sparrow has been studied in relation to the hypothalamohypophysial neurosecretory system. The enzyme activities, as revealed by the methods employed, are unaffected during photoperiodically induced testicular growth. Monoamine oxidase has a distribution distinctly different from that of the aldehyde-fuchsin positive neurosecretory material in that there is high activity in the peripheral palisade layers of both the anterior and posterior divisions of the median eminence. Intimate contact is made between these areas with the primary vessels of the hypophysial portal system. A second concentration of activity lies in a layer between the ependymal cells and the neurosecretory material of the fiber tract. In general, monoamine oxidase appears to be associated with glial elements and non-neurosecretory axons. The pars nervosa has little or no monoamine-oxidase activity. The distribution of acetylcholinesterase activity in the anterior division of the median eminence is very similar to that of the aldehyde-fuchsin positive neurosecretory neurons; however, acetylcholinesterase also occurs in the posterior division without associated neurosecretory fibers. These distribution of enzyme activities are considered in relation to possible adrenergic and cholinergic mechanisms in the median eminence.Dedicated to Professor Berta Scharrer in honor of her 60th birthday.Supported by grant NB-01353 from the National Institutes of Health to Professor Farner. This investigation was conducted while Doctor Kobayashi was a National Science Foundation Senior Foreign Scientist at Washington State University. We are indebted to Doctor Christian Da Lage, Laboratoire de Histologie, Falculté de Médecine de Paris, for the preliminary development of some of the techniques used in these investigations.     

Fine structure of the median eminence and the pars nervosa of the bullfrog,Rana catesbeiana

Summary The hypothalamic neurosecretory system of the bullfrog, Rana catesbeiana, was studied with light- and electron microscopy. The median eminence is roughly divided into two portions. The upper portion mostly consists of ependymal cells, glial cells and preoptico-hypophysial nerve tract, whereas in the lower portion, neurosecretory axons, glial cells, processes of glial and ependymal cells, and fine blood vessels of the hypothalamic portal vein are located. A part of the neurosecretory axons of the preoptico-hypophysial tract proceeds to the lower portion of the median eminence. These axons are arranged perpendicularly to the capillaries of the hypothalamic portal vein. The glial cells are densely located in the area of the median eminence where neurosecretory material is abundant. The neurosecretory material in the neurosecretory cells, their axons, the median eminence and the pars nervosa of the bullfrog shows a positive reaction to PAS treatment.The neurohemal area of the median eminence is occupied by many neurosecretory and non-neurosecretory axons, containing neurosecretory granules and/or synaptic vesicles. The axonal portions with the synaptic vesicles which are considered to be the nerve endings abut on the capillaries of the portal system. The size of synaptic vesicles in the axon terminals containing few neurosecretory granules is larger than those in the endings with many neurosecretory granules. Infrequently glial and ependymal processes are interposed between the nerve endings and the capillary wall.In the hilar region of the infundibulum, synapses are frequently observed between the thin fibers with or without neurosecretory granules and dendrites of non-neurosecretory neurons. The probable functions of these synapses are briefly discussed on the basis of our findings. Both in the hilar region of the infundibulum and in the pars nervosa, electron-dense neurosecretory granules of two different sizes were observed. The median eminence contains only one type of granules.The fine structure of the pars nervosa shows similar structures to those of the median eminence. Both in the median eminence and the pars nervosa, the fenestrated endothelium of the capillaries was frequently observed. The thick perivascular connective tissue space containing fibroblasts and collagen fibrils was observed both in the median eminence and the pars nervosa. Vesicles in the cytoplasm of the endothelial cells which appear to take a part in the transendothelial transport were observed.This investigation was supported in part by United States Public Health Service Research Grant, No. A-3678, to Hideshi Kobayashi from the National Institute of Arthritis and Metabolic Diseases and partly by a grant for Fundamental Scientific Research from the Ministry of Education of Japan. The authors wish to express their thanks to Prof. K. Takewaki for his kind encouragement.     

Premigratory Changes in the Hypothalamo-Hypophysial Neurosecretory System in the Red-winged Blackbird (Agelaius phoeniceus)*

Abstract

• 1 The hypothalamo-hypophysial neurosecretory system in the Red-winged Blackbird (Agelaius phoeniceus) has been studied with special reference to changes during the period prior to fall migration.
• 2 In September the supraoptic and paraventricular cells were found to be considerably larger with substantial amounts of aldehyde fuchsin positive neurosecretory material (NSM) accumulating in the perikaryon. Large amounts of the NSM were found also in the anterior median eminence and pars nervosa.
• 3 Towards migration time (November), the cells in the hypothalamic nuclei contained very little NSM. A marked decline in the amount of NSM was also seen in the anterior median eminence and probably in the pars nervosa too.
• 4 The release of NSM toward migration time has been discussed as an important event in the sequence of events that control and regulate migratory activity.

     

The hypothalamo-hypophysial system in the ground squirrels, Citellus erythrogenys Brandt and Citellus undulatus Pallas. I. Microanatomy and cytomorphology of the gomori-positive neurosecretory system with special reference to its state during hibernation.

Microanatomy of the peptidergic hypothalamo-hypophysial neurosecretory system in the ground squirrels, C. erythrogenys and C. undulatus, is similar to that of other rodents, but differs in details. Bundles of neurosecretory fibres with single neurosecretory cells along their course connect the hypothalamic neurosecretory centers into the one system. The dorso-caudal portions of the NPV reach the level of the columna fornix. The NPO is a special indepedent formation and appears to have some specific functions in the HHNS. The infundibular recess penetrates deeply ihto the hypophysial stem, but does not reach the posterior pituitary. The glandular epithelium of the tuberal portions of the adenohypophysis surrounds the median eminence and a considerable part of the lateral surface of the tuber cinereum. Different functional types of neurosecretory cells reflect certain phases of their secretory cycle. Correlations between their size, structure and the content of NSM are found. A low activity of the HHNS is observed in both C. erythrogenys and C. undulatus during December-January. A higher activity of the HHNS in torpid C. undulatus than in C. erythrogenys is established. This correlates with a difference in deepness of torpor in C. erythrogenys and C. undulatus. An important role of the HHNS in hibernation, i.e. in realization of adaption of species in ontogenesis is suggested.     

Hypothalamo-neurohypophysial complex in leukemia L 1210-bearing mice.

Mice of the DBA strain were inoculated with 4.5 x 10(6) L1210 ascites cells and the time response of the hypothalamo-neurohypophysial complex was studied. The amount of neurosecretory material was determined in the neuroendocrine cells of supraoptic and paraventricular nuclei, throughout the hypothalamo-neurohypophysial tract and in the neural lobe of the hypophysis. Three methods of response evaluation were employed: the amoount of neurosecretory material was estimated according to an arbitrary scale, by cytophotometry while for the hypothalamic cell-nuclei the karyometric method was used. The existence of a functional interrelation between the presence of leukemic cells and the activity of the hormonogenic sites in the hypothalamus is suggested. A clear response to the tumor inoculation was established both in the amount of stainable substance and in the nuclear volume of the secretory neurones. It is suggested that an accelerated synthesis and release of neurosecretory material in tumor recipient mice reduces renal excretion in order to compensate a dehydration caused by the production of ascitic fluid.     

Neuroendocrine control of oocyte development in Poecilocera picta (Insecta: Orthoptera)

The neuroendocrine control of oocyte development in Poecilocera picta Fab. has been described. The secretory activity of the A type of neurosecretory cells has been correlated with ovarian development. In P. picta during the first four days after emergence the neurosecretory material is seen passing down the axons but the cells are largely devoid of neurosecretory material. When the oocytes are developed the A type cells are with stored neurosecretory material.
In P. picta the mature males do not appear to accelerate the process of maturation in females. The females which are reared without males or with castrated males also mature at the same time as the females which are reared with males. The corpus allatum also enlarges and decreases at the same period. The number of resorptive bodies is much more in the females which are reared with castrated males or without males. There appears to be some correlation in the secretion of the neurosecretory material, copulation, and the appearance of resorptive bodies. In P. picta the role of the mature male is only in copulation which very likely allows the cell to synthesize and secrete and release a large amount of neurosecretory material whose discharge in the haemolymph enables a successful development of the oocyte. Corpus allatum appears to be controlled by a precursor from the brain.
Cautery of the cerebral neurosecretory cells, allatectomy and sectioning of the nerves have been done to find out the role of neurosecretory material.     

Mitotic Figures in the Median Eminence of the Hypothalamus

The median eminence of the hypothalamus is part of the avenue by which neurosecreted hormones from the hypothalamic nuclei reach the pars nervosa (neural lobe) of the pituitary and eventually the bloodstream. Lithium treatment and osmotic stress increases the transport of neurosecretory hormones to the pituitary in the adult rat. Specialized astrocytes termed pituicytes in the pars nervosa of the pituitary participate in the secretory process and also develop considerable mitotic activity. The present work reveals similar mitotic figures in cells within the median eminence following 3 days of lithium treatment. The location and appearance of these mitoses add to the evidence that pituicytes are present in the median eminence. Moreover, mitoses occur within the ependymal (tanycyte) layer of the median eminence. Thus, the present results suggest that the tanycyte layer may contain pituicytes, indicating that the hypothalamus possesses specialized cells for modulating neurosecretion in response to osmotic challenges.     

Changes in the hypothalamo-hyophyseal neurosceretory system of mammals following pituitary stalk transection and hypophysectomy]

Hypophysectomy and pituitary stalk section result in dramatic morpho-functional changes in all parts of mammalian hypothalamo-hypophyseal neurosecretory system. Reorganization of the hypophyseal stalk consists of several interconnected but differing in time processes. Simultaneously with the developing traumatic changes (degeneration of the sectioned neurosecretory fibers, secretory disorders) proliferation of pituicytes with characteristic phagocytic activity is observed. A little bit later, intensive mitotic division of endothelial cells and capillary formation piercing the stalk periphery begins. At the same time, a new way for blood outflow from the capillaries of the primary portal plexus into the synuses of the brain pias is restored. Degenerated neurosecretory fibers are gradually substituted by regenerating fibers forming a dense network in heavily vascularizated stalk parts. As differentiation of endothelial cells and regeneration of neurosecretory fibers procede, axovasal contacts are gradually forming. At that time the hypophyseal stalk begins functioning as a neurohumoral organ but morpho-functionally less perfect than the posterior hypophyseal lobule. In the median eminence of the operated animals, unlike the intact ones, neurosecrete is accumulating around the capillaries of the portal plexus. Mechanical damage of neurosecretory fibers during the operation results in degeneration of a greater number of neurosecretory cells in the supraoptical and paraventricular nuclei. Preserved cells have an increased functional activity because of neurohormonal deficiency in the organism. As a result of the structural changes mentioned, diabetes mellitus develops, subsiding gradually with time course.     

Licht- und elektronenmikroskopische Untersuchung von neurosekretorischen Zellen im Gehirn der Eintagsfliege Ephemera danica Müll. (Ephemeroptera: Ephemeridae)

In the brains of the males the amount of stained secretory material was nearly constant during the last four instars. In the females a decrease in this neurosecretory product in the last nymphal and subimaginal stage could be observed, followed by an increase in the imagines. In the final nymphal stage four types of neurosecretory cells (nsc) were found in the medial protocerebral cell group, showing differences in shape, size, and the contents of the cytoplasm, especially of the secretory granules. In addition to the medial nsc, some cells in the frontal part of the brain and in the deutocerebrum are described. They contain electron-opaque granules and probably have a neurosecretory function. The secretory product of the medial nsc is transported along the axons of the nervus corporis cardiaci 1 (ncc1), an unpaired nerve tract on the ventral side of the brain. Leaving the brain the ncc1 immediately enters the corpus cardiacum. Connections between secretory granules and neurotubuli point to an important role for the neurotubuli in the transport of secretory material.     

Regeneration of the supraoptico-hypophyseal and paraventriculo-hypophyseal tracts in the hypophysectomized rat

Summary The effect of hypophysectomy on the nerve fiber pattern in the median eminence and infundibular stem of the rat has been investigated by a slightly modified Bodian technique. Postoperative changes in the distribution of neurosecretory material and connective tissue and changes in vascularity have also been studied.Extensive regeneration of the fibers of the supraoptico-hypophyseal and paraventriculo-hypophyseal tract could be demonstrated. It is most pronounced at the distal extremity of the infundibular stem but occurs also in the rostral part of the infundibular stem and in the median eminence. Regeneration starts in the second postoperative week and is completed about four weeks later. The nervous regeneration observed in the pituitary area after hypophysectomy is more extensive than is usually encountered after lesions elsewhere in the central nervous system.It could be demonstrated moreover that neurosecretory material accumulates at the same sites in which the terminals of the regenerated nerve fibers can be found. Hypophysectomy also causes an increase in capillary density and connective tissue content of the infundibular stem. Accumulations of neurosecretory material are always found in areas showing a high capillary density and a considerable amount of connective tissue.Factors which might be responsible for the extensive nervous regeneration in the pituitary area are discussed as are the factors determining the pattern of outgrowth of the regenerating nerve fibers. Morphological aspects of storage of posterior pituitary hormones are considered in the light of data in the literature and the results of the present work.Partly supported by a U.S. Public Health grant to Dr. E. Scharrer and a travel grant to the author from the Netherlands Organization for pure scientific Research (ZWO).     

Neurosecretory cells in Dysdercus similis (Hemiptera)

The neurosecretory cells of Dysdercus similis have been described. "A", "B", "C" and "D" types of neurosecretory cells are present. The "A" type of cells of the pars-intercerebralis show cyclical secretion. When these cells show secretory activity during one to three days of emergence, they have scattered granules. The cells are seen packed with clumps of neurosecretory material when they are not secreting, and this is interpreted as a storage stage. The axons of these cells supply the corpora cardiaca and some neurosecretory material also reaches the corpus allatum. The release of this neurosecretory material can be correlated both with moulting in the young stages and later with reproduction in the adults.     

Identification of adenohypophysiotropic neurohormone producing neurosecretory cells in Rana temporaria

Summary By means of electron microscopy, in the median eminence of Rana temporaria, the terminal arborizations of axons of six different types of neurosecretory cells, located in the pars ventralis of the tuber cinereum, were identified. In addition, phenomena connected with the release of neurosecretory material from the axon terminals of these neurosecretory cells into the blood capillaries of the median eminence are described.Preliminary results suggested the existence, in the median eminence, of additional different neurosecretory axon types which could also belong to corresponding neurosecretory cell types probably located in the apical part of the pars ventralis of the tuber cinereum. Moreover, in the external region of the normal median eminence, separate monoaminergic nerve fibres were tentatively identified. Arguments are adduced which plead (1) against the assumption that the ependyma or the pituicytes of the median eminence could produce adenohypophysiotropic hormones; (2) against the inference that the ependymal cells of the median eminence might be involved in the transport of adenohypophysiotropic hormones from the cerebrospinal fluid into the blood capillaries of the median eminence.     

鲫鱼尾部神经分泌系统显微和亚显微结构的季节性变化

鲫鱼尾部神经分泌系统的神经分泌细胞和它的轴突中可观察到各种不同电子密度的颗粒。在性腺各个不同的发育阶段,该系统的分泌物具有累积、充满、释放和恢复这样一种周期性变化,由此说明鲫鱼的尾部神经分泌系统和它的生殖有关。     

Influence of neurosecretion on the activity of median eminence and pars intermedia in hereditary nephrogenic diabetes insipidus mice with bilateral supraoptic lesions

Summary The extremely hypertrophied hypothalamo-hypophysial neurosecretory system of the hereditary nephrogenic diabetes insipidus (DI Os/+) mice, shows distinctly hypertrophied median eminence (ME) indicated by the hypertrophied ependymal cells, increased number of Herring bodies and a great number of neurosecretory fibers loaded with secretion converged towards blood capillaries and towards pars tuberalis. The other distinct feature of the DI Os/+ mice is the marked hypertrophy and hyperplasia of the pars intermedia (PI) infiltrated with many beaded neurosecretory fibers. The majority of the PI glandular cells are of the large light type rather than the small type, the latter being comparatively numerous in normal mice. Very often these large light glandular cells and the marginal cuboidal epithelial cells of the hypophysial cleft contain or are surrounded by AF⁺ granules and/or colloid.When the SON is destroyed bilaterally by means of electrolytic lesions, loss of NSM from the ME, infundibulum and the pars nervosa, is accompanied by the disappearance of AF⁺ granules in the PI, and the shrinkage of all the above mentioned parts. The changes also occur in the three glandular cell types. The large light glandular cells, decrease in number as well as in size, while the granularity of the dark cells increases. The marginal cuboidal cells of the hypophysial cleft and the large light glandular cells loose most of the AF⁺ material from the cells as well as from the surroundings. Wherever the AF⁺ material does not disappear in the PI, the PI cells remain much big, as in the controls. The colloidal secretion on the margin of the PI and from the cleft may be the secretion of PI glandular cells, as a result of the stimulation of the neurosecretory fibers in the PI of DI Os/+ mice. This colloid disappears when the neurosecretory fibers are destroyed from the PI as a result of lesioning. Therefore, it suggests that this secretory material may belong to SON, or possibly to some other secretory cells residing in the anterior hypothalamic region. Or more probable explanation could be that with the disappearance of supraoptic neurons and/or anterior hypothalamic neurons due to electryolytic lesions, the secretory activity of the PI cells and its AF⁺ material, might have been hampered due to lack of stimulation of these fibers in PI.Therefore, from the previous work and from the present one, it is suggested that the NSM may contain some substance which stimulates the glandular cells of the PI and this substance may be either vasopressin and/or a type of CRF. However, it is concluded that the activated neurosecretory system plays an important role in the hypertrophy of the ME and PI and certain changes in PI cell types. The impairment of catecholamine fibers and their lack of influence on synthesis and/or release in PI parenchyma cells is discussed.This study was supported by MRC of Canada Grant No. MA-3759. A preliminary report on a part of this work appeared (Naik, 1970b) in Proc. Can. Fed. Biol. 13, 147.     

Cytophysiological study of neurosecretory and pheromonal influences on sexual development in Ophryotrocha puerilis (Polychaeta,Dorvilleidae)

Summary

Prominent secretory nerve endings are found at the posterior margin of the supraesophageal ganglion in the protandric polychaete, Ophryotrocha puerilis. Solitary juveniles developing as primary males, and then as females, accumulate neurosecretory material in the nerve endings which thereby swell and become filled with granules. Females maintained in mass culture have similar terminals, whereas in secondary males (males which had been females before), these axon terminals are very small and contain no material. When such males are isolated, they accumulate neurosecretory material within the nerve endings and become females. When formerly isolated females are put together, their stores of neurosecretory material are rapidly discharged. Subsequently they lay egg masses and switch to the male state. These effects are mediated by a pheromone released during social contact of formerly isolated females. The complexity of the relationship between neurosecretory activity and sexual state is indicated by the situation in animals maintained in pairs, when both male and female partners have swollen nerve endings packed with secretory material.     

Cholinesterases in the hypothalamo-hypophysial neurosecretory system of the White-crowned Sparrow,Zonotrichia leucophrys gambelii

Summary The distribution of cholinesterases in hypothalamo-hypophysial neurosecretory system of the White-crowned Sparrow has been examined histochemically. The perikarya of the neurosecretory cells of the paraventricular and supraoptic nuclei have a high acetylcholinesterase activity. Acetylcholinesterase activity also occurs in the cells of the infundibular nucleus. The proximal parts of the axons of the cells of the neurosecretory and infundibular nuclei have strong acetylcholinesterase activity and weak non-specific cholinesterase activity. In the median eminence, the activity of acetylcholinesterase is strongest in the palisade layer. In the pars nervosa, there is definite, although weak, acetylcholinesterase activity.This investigation was supported by grants from the National Institutes of Health to Professor Farner (B-1353) and to Dr. Kobayashi (A-3678).     

Immuno-cytochemical demonstration,in the external region of the amphibian median eminence,of separate vasotocinergic and mesotocinergic nerve fibres

Summary By means of single and double immuno-enzyme cytochemical staining techniques, it was shown that the external region of the amphibian median eminence contains separate vasotocinergic and mesotocinergic nerve fibres. Moreover, it was demonstrated that the vasotocinergic fibres also contain neurophysin. In animals in which the hypothalamic magnocellular neurosecretory preoptic nuclei had been completely removed, the immuno-reactive vasotocinergic and mesotocinergic fibres of the median eminence had disappeared. From this result, it is concluded that, at least the great majority of the vasotocinergic and mesotocinergic fibres of the external region of the amphibian median eminence are processes of neurosecretory perikarya located in the hypothalamic magnocellular preoptic nuclei. On the other hand, our results do not exclude the possibility that a minority of these neurosecretory fibres originate from small immuno-reactive perikarya which were found in the tuber cinereum. The observation that both kinds of processes accumulate around blood capillaries of the hypophysial portal system strongly suggests that they play a role in the control of the activity of the pars distalis of the hypophysis.This investigation was supported by a grant from the Belgian Nationaal Fonds voor Geneeskundig Wetenschappelijk Onderzoek     

Influence of light and darkness on the hypothalamo-hypophysial system of Rana temporaria L.

Summary The influence of a long daily photoperiod and continuous darkness was studied on the hypothalamo-hypophysial system of Rana temporaria. After six weeks of treatment (December-January 1968/1969) the secretory activity of the hypothalamo-hypophysial system of the light-treated animals was enhanced.The amount of aldehyde-fuchsin positive material in the preoptic nucleus increased as a result of the light treatment, and decreased in the median eminence, whereas in the posterior lobe no difference could be observed between the light- and dark-treated animals.The karyometric results showed an increase of the nuclear volume of the cells of the preoptic nucleus after light treatment; small differences were noticed between distinct areas of the preoptic nucleus.Autoradiographic data especially showed that light influenced the hypothalamo-hypophysial system to a high degree. Half an hour after the injection of ³⁵S-cysteine, the incorporation of the isotope in the preoptic nucleus of the light-treated animals was five times as much as in the dark-treated animals. In the median eminence and in the posterior lobe, the autoradiographic results confirmed the results found with regard to the amount of aldehyde-fuchsin positive material. Labelled material is released only from the median eminence and not from the posterior lobe.The author is indebted to Prof. Dr. J. C. van de Kamer and Dr. F. C. G. van de Veerdonk for their interest and helpful encouragement. Thanks are also due to Ir. J. J. Bezem for his assistance in solving the statistical problems and to Mr. H. van Kooten and his staff for making the illustrations. The co-operation of the student Mr. E. Schenk is gratefully acknowledged. Dr. L. Boomgaart was so kind to check and amend the English writing.