ALBERT

Notes: Summary Peroxidase injected into the subarachnoid space in mice is absorbed by ependymal cells of the median eminence. The ependymal cells of the median eminence of the rat and Japanese quail absorb peroxidase injected into the third ventricle. The processes of these ependymal cells terminate at the capillaries of the primary plexus or those surrounding the ventromedial nucleus of the hypothalamus. In all three species, peroxidase is absorbed by the ependymal cells of the paraventricular organ and by those in close proximity to it. Some ependymal cells send processes to the capillaries in the lateral nucleus of the hypothalamus. These phenomena are discussed in relation to adenohypophysial function.

Notes: Summary Following the bilateral implantation of puromycin into the paraventricular nuclei of rats, the neurosecretory cells became atrophic and the amount of aldehyde-fuchsin (AF) positive material in the neural lobe decreased. In these rats, urine excretion and water intake increased remarkably. The supraoptic nuclei of the rats were not affected by this treatment. After the unilateral implantation of puromycin in the paraventricular nucleus, the neurosecretory cells of the implanted side became atrophic, while those of the unimplanted side hypertrophied. The neural lobe contained similar amounts of AF-positive material to those of the control rats with unilateral cholesterol implants. In the rats implanted bilaterally with puromycin immediately above the supraoptic nucleus, the neurosecretory cells of this nucleus contained little or no AF-positive material, and urine excretion and water intake increased greatly. The cells of the paraventricular nucleus remained unchanged in these rats.

Notes: Summary Antiserum generated against synthetic urotensin II of the goby, Gillichthys mirabilis, was used to localize urotensin II in the caudal neurosecretory system in six species of freshwater teleosts; Cyprinus carpio, Carassius auratus, Oreochromis mossambicus, Oreochromis niloticus, Salmo gairdneri and Plecoglossus altivelis, and six species of seawater teleosts: Acanthogobius flavimanus, Pagrus major, Paapristipoma trilineatum, Trachurus japonicus, Seriola dumerili and Seriola quinqueradiata. In the carp, urotensin II-immunoreactive perikarya were classified into three groups according to their size and shape. Small cells were located in the spinal cord dorsal to the urophysis, medium-sized cells immediately anterior to the urophysis, and large cells anterior to the medium-sized cells. In each group, a small number of nonreactive cells was found. Urotensin II-immunoreactive nerve fibers extended toward the urophysis and terminated around the blood vessels. Other species of teleosts showed a similar immunoreaction to that observed in the carp. The immunoreaction of the urophysis was stronger in seawater fish than freshwater fish. Urotensin II-immunoreactive elements could not be detected in the brains of the carp, goldfish and goby.

Notes: Abstract. The immunohistochemical localization of nine different neuropeptides was studied in the central nervous system of the amphioxus, Branchiostoma belcheri. In the brain, perikarya immunoreactive for urotensin I and FMRFamide were localized in the vicinity of the central canal. One of the processes of each of these perikarya was found to cross the dorsoventral slit-like lumen of the central canal. Oxytocin-immunoreactive short fibers, but not perikarya, were detected in the ventral part of the brain. Perikarya immunoreactive for arginine vasopressin/vasotocin, oxytocin and FMRFamide were widely distributed in the spinal cord. Arginine vasopressin/vasotocin-immunoreactive fibers often made contacts with Rohde cell axons. Angiotensin II-immunoreactive perikarya were observed in the posterior half of the spinal cord, and urotensin I-immunoreactive perikarya were found in the caudal region of the spinal cord. Cholecystokinin/gastrin-immunoreactive fibers, but not perikarya, were detected in the spinal cord; some extended as far as the ependymal layer of the cerebral ventricle. No colocalization of the peptides examined was observed. No immunoreactivity for atrial and brain natriuretic peptides nor for urotensin II was detected. The present study indicates that there are at least six separate neuronal systems that contain different peptides, respectively, in the central nervous system of the amphioxus. Their functions remain to be determined.

Notes: Abstract The immunohistochemical localization of nine different neuropeptides was studied in the central nervous system of the amphioxus, Branchiostoma belcheri. In the brain, perikarya immunoreactive for urotensin I and FMRFamide were localized in the vicinity of the central canal. One of the processes of each of these perikarya was found to cross the dorso ventral slit-like lumen of the central canal. Oxytocin-immunoreactive short fibers, but not perikarya, were detected in the ventral part of the brain. Perikarya immunoreactive for arginine vasopressin/vasotocin, oxytocin and FMRFamide were widely distributed in the spinal cord. Arginine vasopressin/vasotocin-immunoreactive fibers often made contacts with Rohde cell axons. Angiotensin II-immunoreactive perikarya were observed in the posterior half of the spinal cord, and urotensin I-immunoreactive perikarya were found in the caudal region of the spinal cord. Cholecystokinin/gastrin-immunoreactive fibers, but not perikarya, were detected in the spinal cord; some extended as far as the ependymal layer of the cerebral ventricle. No colocalization of the peptides examined was observed. No immunoreactivity for atrial and brain natriuretic peptides nor for urotensin II was detected. The present study indicates that there are at least six separate neuronal systems that contain different peptides, respectively, in the central nervous system of the amphioxus. Their functions remain to be determined.

Notes: Summary The ependymal linings of the median eminence were destroyed by electric cautery or intraventricular injection of picric acid in the rat and Japanese quail. In these animals the ventricular lumen near the median eminence disappeared due to adhesion of lesioned walls on both sides of the third ventricle. Electric lesions of the ependymal layer containing tanycytes did not induce appreciable disturbance in the estrous cycles. Rats in which tanycytes were lesioned by picric acid displayed 4-day estrous cycles after prolonged diestrus (10–22 days). After destruction of tanycytes in the quail, a photostimulated gonadal growth was observed. It is concluded that the tanycyte transport of the ventricular fluid to capillaries of the portal vessels appears unnecessary for maintenance of adenohypophysial gonadotrophic activities.

Notes: Summary The pituitary of the larval guppy, Lebistes reticulatus (Peters), was investigated cytologically in relation to the osmotic properties of the environment. In larvae of Stage I, removed from the mother about one week before birth, the sizes of the nuclei of the prolactin, STH, ACTH, and pars intermedia cells were small. The nuclei of these cell types became larger when Stage I larvae were transferred into fresh water (F. W.); however, these animals did not survice. In Stage-II larvae, removed from the mother just before birth, the nuclei of the abovementioned adenohypophysial cells were larger than those of the Stage-I larvae. Stage-II larvae were able to survive in F. W. Thus, it seem that osmoregulatory mechanisms in the larval guppy develop between Stages I and II. The nuclei of prolactin, STH, ACTH and pars intermedia cells decreased significantly in size after birth (0-hr). When larvae of the 0-hr group were immersed in 1/3 sea water (S. W.), only the nuclei of the ACTH cells increased in size. The neurohypophysis of Stage-I larvae contained a very small amount of aldehyde-fuchsin (AF)-positive material, while the neurohypophysis of Stage-II and 0-hr fish was rich in this material. When Stage-I larvae were immersed in F. W., the neurohypophysis showed as great an accumulation of AF-positive material as Stage-II and 0-hr fish.

Notes: Summary Distribution of molluscan cardio-excitatory tetrapeptide Phe—Met—Arg—Phe—NH2 (FMRFamide) was determined by means of immunohistochemistry in the brain and neurohypophysis of the lamprey, Lampetra japonica. Many FMRFamide-like immunoreactive neurons were found in the periventricular nuclear region and in a region near the mammillary recess. Neurons situated in the former region were larger. The immunoreactive cell groups were shown to be located at sites differing from those of the AF-positive cell groups. The fibers of immunoreactive neurons extended in all directions within the brain and towards the spinal cord, some reaching the third ventricle and capillaries. Thus, FMRFamide-like immunoreactive peptides appear to function as neurotransmitters or neuromodulators and possibly also as neurohormones. FMRFamide-like immunoreactive material was rarely observed in the posterior neurohypophysis (neural lobe), but was noted to be present to a limited extent in the caudal part of the anterior neurohypophysis (median eminence). It would thus follow that FMRFamide-like immunoreactive neurons may not necessarily be related to the hypothalamo-neural lobe system, but may possibly be associated with the hypothalamoadenohypophysial system. The pineal body showed no FMRFamide-like immunoreactivity.

Notes: Summary 1. The metabolic activity of the hypothalamo-hypophysial neurosecretory system of the White-crowned Sparrow, Zonotrichia leucophrys gambelii, has been investigated by means of quantitative estimates of acid- and alkaline-phosphatase activity. The activities of samples from the median eminence, samples from the supraoptic region, the adenohypophyses, and the neurohypophyses of photosensitive and refractory birds, both subjected to short and long daily photoperiods, have been compared. Histochemical demonstrations of the distribution of acid- and alkaline-phosphatase activity have been effected to provide a better basis for interpretation of the quantitative data. 2. The photosensitive birds which were subjected to 20-hour daily photoperiods showed the typical photoperiodically induced testicular growth and fat deposition. There was an increase in acid-phosphatase activity in the supraoptic region. This increase in acid-phosphatase activity may be correlated with increased activity of the neurosecretory cells as indicated by cytologic criteria, including an increased rate of formation of aldehyde-fuchsin stainable material. No such increase in acid phosphatase occurred in refractory birds on 20-hour photoperiods. In photosensitive birds, there was also an increase in acid-phosphatase activity in the median eminence, an increase which appears to be attributable primarily to changes in activity of the axons in the glandular layer although a contribution by the nucleus tuberis cannot be excluded. The change in acid-phosphatase activity in the median eminence coincides well with the observed increase in rate of disappearance of aldehyde-fuchsin-positive granules from the axons. Neither an increase in acid phosphatase nor a decrease in granules occurred in the median eminences of refractory birds subjected to 20-hour daily photoperiods. The acid-phosphatase activity of the adenohypophyses of photosensitive birds was greater than in refractory birds although neither was affected by long daily photoperiods. These results are consistent with the hypothesis that the photoperiodic response in Zonotrichia leucophrys gambelii involves, in the supraoptic region, an increase in acid-phosphatase activity associated with increased synthesis of aldehyde-fuchsin-positive material in the neurosecretory cells, and, in the axons of the glandular layer of the median eminence, an essentially simultaneous increase in acid-phosphatase activity associated with a humoral transfer to the capillaries of the hypophysial portal system. Our data suggest that the higher rate of production of gonadotropins in the adenohypophysis by photosensitive birds involves a higher acid-phosphatase activity but that the photoperiodically induced increase in release of these gonadotropins, mediated by a humoral agent in the hypophysial blood, does not involve a detectable further increase in acid-phosphatase activity. 3. The data on alkaline phosphatase in supraoptic-region, median-eminence region, and adenohypophysis give no indication that this enzyme is involved in the photoperiodically induced activity of the hypothalamo-hypophysial system. 4. In agreement with previous investigations in this laboratory (Oksche et al., 1959), the data obtained in this investigation on phosphatase activity indicate no participation by the neurohypophysis in the photoperiodic response. 5. The histochemical observations indicate that the neurosecretory cells of the paraventricular and supraoptic nuclei have acid- and alkaline-phosphatase activities similar to those of ordinary neurones. The axons of neurosecretory cells, except in the median eminence, are not identifiable by histochemical demonstration of phosphatase activity. In the fibers of the glandular layer of the median eminence there is a pronounced acid-phosphatase, but no alkaline-phosphatase reaction, in these fibers. Also the nuclei and nucleoli of cells of the mammillary and tuberal nuclei have a fairly strong acid-phosphatase activity. In the adenohypophysis there is a conspicuous acid-phosphatase and a strong alkalinephosphatase reaction in the nuclei and nucleoli, but only slight activities in the cytoplasm.

Notes: 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.