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.

     

Hypothalamohypophysial vascular and neurosecretory link in the teleost Bagarius bagarius (Ham.).

A three-dimensional picture of the hypothalamohypophysial neurosecretory system of B. bargarius could be obtained by adopting in situ staining techniques. The paired NPO give rise to left and right neurosecretory tracts which, although they approximate as a common tract, maintain their individuality till entering the pars intermedia. The ventral hypothalamic and the hypophysial arteries take their origin from the internal carotid artery. The former contributes to the formation of the primary plexus of the median eminence and the latter enters the pituitary directly, giving rise to the neuroadreno-interface vasculature. The vasculature of the median eminence lies in close contact with the long common neurosecretory tract. Morphological evidence suggests that in B. bagarius there are three pathways of hypothalamic control of the hypophysis: (1) A direct neuroglandular pathway, where the neurosecretory axons come directly in contact with the adrenohypophysial cells. (2) An indirect neurovasculoglandular pathway, where the blood exposed to the NSM at the median eminence comes in contact with gland cells. (3) Another indirect neurovasculoglandular pathway, where the blood is exposed to NSM at the neurohypophysis prior coming in contact with the gland cells. Thus, B. bagarius has the 'median eminence equivalent' neuroadeno-interface vasculature typical of the teleosts and the median eminence comparable to the Elasmobranchs, Holocephali and primitive Actinopterygians. This shows that the pituitary portal system of teleosts is in general agreement with that of other fish groups, except that in some it is restricted to the neuroadreno-interface, whereas in others like B. bagarius it extends to the hypothalamus also. These may be termed anterior and posterior median eminence.     

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.     

Light- and electron microscopic investigations on the median eminence of the pigeon after TSH and PTU treatment

Summary 1. The effects of TSH and PTU treatment with thyroid stimulating hormone and propylthiouracil on the structure of median eminence of the pigeon have been studied by light- and electron microscopy.2. By light microscopy it has been found that the aldehydefuchsin positive material of the external layers of the anterior median eminence differs functionally from that of the neural lobe.3. The light- and electron microscopic investigations revealed that the anterior and posterior median eminence differ both structurally and functionally. Specifically after the experimental interventions the considerable reaction can be observed only in the anterior median eminence.4. Electron microscopically it appears possible that the aldehyde-fuchsin positive material of the anterior median eminence may be associated with the 1200–1500 Å—size granules.5. The quantitative changes observed in the granular composition after the treatments show a close correlation with thyroid function, namely after treatment with thyroid stimulating hormone the relative quantity of the 1200–1500 Å—size granules increases markedly, while administration of propylthiouracil has the opposite effect and increases the relative quantity of the 1000 Å—size dense-core vesicles. They presumably may play a role in the control of TSH-secretion.Abbreviations TSH thyroide stimulating hormone - PTU propylthiouracil - ME median eminence - AME anterior median eminence - PME posterior median eminence - AF+ aldehyde-fuchsin positive - AF– aldehyde-fuchsin negative     

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.     

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.     

Distribution of LHRH in the rat and mouse brain with special reference to the tanycytes

Summary The distribution of luteinizing hormone-releasing hormone (LHRH) was studied in the rat and mouse brain by means of light and electron microscopic immunohistochemistry using the peroxidase-antiperoxidase method. An immunoreactive product to LHRH antiserum was found near the blood vessels of the vascular organ of the lamina terminalis. In the arcuate nucleus-median eminence region, an immunoreactive material occurred bilaterally in the hypothalamic tissue around the tuberoinfundibular sulci. Electron microscopy revealed that immunoreactive fibers observed light microscopically contain numerous granules 100–130 nm in diameter. No immunoreactive product was located in the tanycytes of the median eminence, the perikarya of hypothalamic neurons, and the parenchyma of several circumventricular organs (subfornical organ, subcommissural organ, pineal organ, area postrema).Supported by grants from the Ministry of Education of Japan and the Ford Foundation     

Melanin-concentrating hormone stimulates the release of luteinizing hormone-releasing hormone and gonadotropins in the female rat acting at both median eminence and pituitary levels

The purpose of this study was to investigate whether melanin-concentrating hormone (MCH) acts directly on the median eminence and on the anterior pituitary of female rats regulating LHRH and gonadotropin release. In addition, immunohistochemistry was used to examine the density and distribution of MCH-immunoreactive fibers in the median eminence of proestrous rats. MCH-immunoreactive fibers were found in both the internal and external layers of the median eminence and in close association with hypophysial portal vessels. In the first series of in vitro experiments, median eminences and anterior pituitaries were incubated in Krebs-Ringer bicarbonate buffer containing two MCH concentrations (10(-10) and 10(-8) M). The lowest MCH concentration (10(-10) M) increased (P < 0.01) LHRH release only from proestrous median eminences. Anterior pituitaries incubated with both MCH concentrations also showed that 10(-10) M MCH increased gonadotropin release only from proestrous pituitaries. In the second series of experiments, median eminences and pituitaries from proestrous rats were incubated with graded concentrations of MCH. MCH (10(-10) and 10(-9) M) increased (P < 0.01) LHRH release from the median eminence, and only 10(-10) M MCH increased (P < 0.01) LH and FSH release from the anterior pituitary. The effect of MCH on the stimulation of both gonadotropins from proestrous pituitaries was similar to the effect produced by LHRH. Simultaneous incubation of pituitaries with MCH and LHRH did not modify LH but increased the FSH release induced by LHRH. The present results suggest that MCH could be involved in the regulation of preovulatory gonadotropin secretion.     

Simultaneous monoamine histofluorescence and neuropeptide immunocytochemistry: I. Localization of catecholamines and gonadotropin-releasing hormone in the rat median eminence

Adjacent tissue sections through the rat median eminence were examined for the distribution of gonadotropin-releasing hormone (GnRH) and catecholamines (CA). A simultaneous visualization technique was employed for this correlative neuroanatomical analysis. At rostral and mid-central levels of the median eminence the majority of GnRH terminals do not appear in coexistence with CA terminals; the latter were confined to the outer-most 10 μm of the median eminence while the densest concentration of GnRH terminals was located internal to this layer. However, individual GnRH fibers appeared to penetrate the outer CA zone wherein they were found in juxtaposition to portal capillaries. At caudal levels of the median eminence, there was an extensive overlap of CA and GnRH varicosities adjacent to the tubero-infundibular sulcus. In addition, numerous GnRH terminals were seen adjacent to portal vessels. The differences in the positions of CA and GnRH terminals between rostral and caudal median eminence may provide a morphological basis for the hypothesis of separate regulatory mechanisms for CA upon GnRH secretion at these two levels of the median eminence.     

Immunohistochemical localization of thyrotropin-releasing factor in the rat median eminence

Summary Antibodies raised in rabbits against a synthetic preparation of thyrotropin-releasing factor (TRF) were used in association with the immunoperoxidase histochemical technique to localize TRF in the rat median eminence. Using coronal sections cut through the mid-arcuate region, specific immunoreactive material was observed in the medial and lateral regions of the superficial layer of the external median eminence.This work was financed by a grant from the Medical Research Council of New Zealand     

The hypothalamo-hypophyseal system of Rana temporaria under osmotic stimulation with special reference to the hypothalamo-median eminence-distal lobe axis

The secretory activity of the hypothalamo-hypophyseal system in the frog Rana temporaria under conditions of dehydration has been studied histologically using two parameters, the amount of stained neurosecretory material and the amount of neurosecretory material labelled by a radioactive precursor. The results are indicative of an increased secretory activity of the hypothalamo-neural lobe system as a result of water deprivation. In addition the involvement of the preoptic nucleus-median eminence axis in osmoregulation is confirmed. The pattern of distribution of autoradiographic silver grains over the median eminence together with data from literature support the idea that mesotocine is involved in the regulation of the activity of the adenohypophysis.     

Ultrastructure of the rat median eminence after superfusion

Summary Isolated medio-basal hypothalami of adult rats were continuously superfused in a chamber with controllable inputs and outputs, for periods from 30 to 240 min. The median eminence was prepared for transmission electron microscopy under carefully controlled conditions by immersion fixation with osmium tetroxide. The ultrastructure of superfused median eminence was compared with that of directly fixed, non-superfused median eminence. Even after 4h of superfusion, the median eminence displays remarkably well preserved histological and cytological patterns; cytomembranes, cell organelles, intercellular relationships, and extracellular spaces were remarkably similar in superfused and non-superfused tissues. As a consequence of osmium tetroxide fixation, microtubules were not observable. The ultrastructural information obtained from unstimulated rat median eminence superfused in vitro provides a basis for future morphofunctional correlations in the study of neurosecretion.     

Differential regional distribution and release of two forms of gonadotropin-releasing hormone in the chicken brain

The functional significance of two molecular forms of gonadotropin-releasing hormone (GnRH) in the chicken brain was investigated. The differential distribution of [Gln8]GnRH (chicken GnRH-I, cGnRH-I) and [His5,Trp7,Tyr8]GnRH (chicken GnRH-II, cGnRH-II) was determined using high performance liquid chromatography and radioimmunoassay with region-specific antisera. Potassium-stimulated release of immunoreactive cGnRH-I and cGnRH-II from brain regions was assessed in tissue incubations. cGnRH-I and cGnRH-II varied independently in different brain areas. The concentration of cGnRH-I was highest in the median eminence of the hypothalamus, and a small quantity was also detected in the midbrain and cerebrum. cGnRH-II was more widely distributed throughout the brain, with highest concentrations in areas of the hypothalamus outside the median eminence and in the medulla. Potassium stimulated the release of cGnRH-I from the median eminence 4-fold, while cGnRH-II release was not detectable. Neither cGnRH-I nor cGnRH-II was released from the medulla. These data suggest: 1) cGnRH-I is the prime regulator of gonadotropin release from the pituitary, and 2) cGnRH-II may have a neurotransmitter or neuromodulator role in areas of the brain outside the median eminence.     

Different populations of granules and their distribution in the hypothalamo-neurohypophysial tract of the rat under various experimental conditions

In connection with previous studies (Krisch, 1974), a morphologic analysis of the median eminence of male, female, pregnant, and thirsting rats has been made, in order to evaluate: 1. whether the median eminence is an additional area where oxytocin and vasopressin may be released from neurosecretory axons in the zona interna; 2. whether it is possible to distinguish different populations of axons according to the mean diameters of their granules in the external zone of the median eminence. In the zona interna of the male and the female control animals, the oxytocin- and vasopressin-containing granules have already reached their final diameters, i.e. their size remains constant until they reach and are stored in the neurohypophysis. However, the relative proportions of these populations in the fibre layer of the median eminence indicate certain differences between male and female animals, concerning the turnover of oxytocin vasopressin. In pregnant, as well as in thirsting animals, there is an apparent decrease of the mean granule diameters in oxytocin- and vasopressin-containing axons. In particular in the latter group, ballooned axons in the fibre layer of the median eminence, containing disintegrating granules, tubular structures filled with electron dense material, and fine-flocculent material in the axoplasm, together with the morphometric results, support our concept that in case of highly increased peripheral demand the contents of the mature, large granules are released into the axoplasm of the fibre layer in the median eminence. In addition, axons containing granules smaller than 115 nm may be classified into four populations with mean diameters of 84 nm, 96 nm, 103 nm, and 100n. The same populations are found in the palisade zone of the median eminence as well as in the pericapillary zone, but in contrast to the zona interna there is a fifth population of axons containing very small granules with a mean diameter of about 70-75 nm. The relative proportions of these five populations differ between the zona palisadica and the pericapillary zone on the one hand and between the experimental groups on the other hand. Some morphological findings concerning the tanycytes are discussed with respect to previously published data.     

The tubero-infundibular neuron system: a component of the photoperiodic control mechanism of the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary To assess the roles of the hypothalamic neurosecretory and tubero-infundibular neuron systems in the mechanism of photoperiodic control of testicular growth in Zonotrichia leucophrys gambelii, midline electrolytic lesions were created in the median eminence, in its individual divisions, and in the region of the infundibular nucleus. Radiography was employed to facilitate the stereotaxic placement of lesions. Extensive damage to the neurosecretion-rich anterior division of the median eminence neither prevented the initiation of testicular growth in photosensitive, photostimulated birds nor induced gonadal regression in birds in which gonadal growth had previously been initiated by natural photoperiodic stimulation. Likewise, there was no impairment of the gonadotropin release mechanism when damage was restricted primarily to the neurosecretion-deficient posterior division of the median eminence. However, in birds in which the zone of damage included both divisions of the median eminence, the photoperiodic testicular response was abolished or markedly suppressed; if testicular growth had been initiated prior to electrocoagulation of the median eminence, testicular regression was induced. Gonadotropic insufficiency comparable to that induced by lesions in the median eminence was caused also by large lesions in the region of the infundibular nucleus or by smaller ones restricted primarily to its median, basal portion. Zones of damage that impair gonadotropic function thus correspond to (a) the chief nucleus of origin of the tubero-infundibular tract, (b) the principal route of entry of tubero-infundibular fibers into the anterior and posterior divisions of the median eminence, and (c) the terminal distribution of tubero-infundibular fibers in the eminential zones of neurovascular contact. These observations suggest that the tubero-infundibular neuron system is an essential component of the photoperiodic control mechanism of Z. leucophrys gambelii and are consistent with an hypothesis that assigns to this parvicellular neuron system the production of a neurohormone that regulates the release of a growth-stimulating gonadotropin from the pars distalis. The failure of anterior median eminence lesions to eliminate gonadotropin release is inconsistent with the hypothesis that the eminential component of the hypothalamic neurosecretory system is an essential element of the mechanism that controls photoperiodic testicular growth.This investigation was supported by a research grant (NB 01353) to Professor Donald S. Farner from the National Institutes of Health. A portion of the research was conducted while the author held the William T. Porter Fellowship of The American Physiological Society. I am grateful to Professor Farner for his suggestions and criticisms.This paper is based on a thesis submitted in partial fulfillment of the requirements for the Ph. D. in Zoophysiology at Washington State University. Portions of this study have been published previously in abstract form (F. E. Wilson and Farner, 1965).     

Anterior and posterior groups of portal vessels in the avian pituitary: incidence in forty nine species.

The hypophyseal portal vessels were studied in forty nine species of birds. The primary capillary plexus in the median eminence is single or divided into an anterior and a posterior plexus. Irrespective of whether the primary capillary plexus is single or divided, distinct, non-interconnected anterior and posterior groups of portal vessels are present in all the species investigated. The anterior group of portal vessels originates in the anterior region of the median eminence and breaks up into capillaries in the cephalic lobe of the pars distalis; the posterior group of portal vessels originates in the posterior region of the median eminence and breaks up into capillaries in the caudal lobe of the pars distalis. This type of regional distribution of portal vessels appears to be of general occurrence in the avian pituitary. The median eminence in the species investigated shows an AF-positive anterior region and an AF-negative posterior region. The pars distalis is differentiated into histologically distinct cephalic and caudal lobes. The arrangement of the portal vessels into anterior and posterior groups provides morphological basis for the view that the functions of the cephalic lobe may be controlled by the anterior median eminence, whereas those of the caudal lobe may be controlled by the posterior median eminence. However, experimental data available to date do not suggest a physiological significance to the widespread incidence of the regional distribution of portal vessels in the avian pituitary.     

Autoradiographische Untersuchungen über das Wachstum des Reissnerschen Fadens von Cyprinus carpio (L.)

Summary The incorporation of³⁵S-l-cysteine into the hypothalamic-hypophyseal neurosecretory system was studied in normal male albino rats and in similar rats which had received thiouracil or excess thyroxine during 18 days. The study was performed by autoradiography. Labelled cysteine was administered by intraperitoneal injections. The animals were sacrificed 30 min, 3 and 24 hours after injection. From the autoradiographs the grain counts were determined in the supraoptic nucleus, paraventricular nucleus, median eminence and infundibular process. The thyroxine-treated rats in the 30 minute group displayed a statistically almost significantly higher grain count in the supraoptic nucleus than the corresponding controls; the same was observed in the 3 hour group in the median eminence and infundibular process. In the 24 hour group, however, the grain count was clearly lower at all sites examined than in the controls. With the rats that had been under thiouracil treatment statistically significant differences were only established in the 24 hour group. Their grain count was higher at all sites than that of the corresponding controls. From the results the inference is drawn that thyroxine accelerates the protein synthesis in the ganglion cells of the supraoptic nucleus and accelerates the flow of NSM to the infundibular process and its release there into the blood flow. Thiouracil, again, retards the flow and the release.This work was supported by a grant from the Sigrid Jusélius Foundation, Helsinki.     

Immunohistochemical and electron microscopic study of the rat hypothalamic nuclei and cell clusters under various experimental conditions

Summary In untreated, pregnant and thirsting rats the neurosecretory hypothalamic areas were investigated by means of the immunoperoxidase technique in order to demonstrate vasopressin- and oxytocin containing elements at the light- and electron microscopic level. In addition, chromalum-hematoxylinphloxin (CHP) staining and conventional double staining of ultrathin sections were used. The areas investigated included the anterior and posterior supraoptic nuclei, the paraventricular nuclei, the numerous accessory cell clusters in the region between the tractus opticus and the third ventricle as well as the median eminence. In all nuclei and in the accessory cell clusters, the number of vasopressin-reactive neurons exceeds that of oxytocin-reactive neurons. Compared with the anterior supraoptic nucleus, the posterior supraoptic nucleus and the accessory cell clusters react more heavily to prolonged thirst. In the median eminence the neurosecretory axons display close contacts with the portal vessels not only in its lateral portion but in thirsting animals also around the mid-line. There the internal layer is broadened and vasopressin-positive tanycytic processes reach the external zone. Parasagittally, fine vasopressin-positive material can be traced from the internal layer to small deposits at the portal vessels. In long term thirsting animals the typical feature of swollen axons exhibits a characteristic distribution in the median eminence and renders a distinct positive reaction to anti-vasopressin. The release of peptide hormones from the perikarya and from the axons within the nuclei as well as the mode of release within the median eminence are discussed. The significance of the positive immunostaining of the ependymal tanycytes and of some perikarya of the suprachiasmatic nucleus must be reconsidered by further studies.Supported by the Deutsche Forschungsgemeinschaft (Grant Nr. Kr 569/1) and Stiftung VolkswagenwerkDedicated to Professor Berta Scharrer on the occasion of her 70th birthdayThe author wishes to express her special gratitude to Dr. L.A. Sternberger for supplying the peroxidaseantiper oxidase-complex and to Dr. H. Stein (Pathologisches Institut der Universität Kiel) for supplying Anti-IgG. The skilful technical assistance of Mrs. H. Prien and Mrs. H. Schöning is thankfully acknowledged     

Hypothalamic neurosecretion and photoinduced testicular growth in the tree sparrow,Spizella arborea

Summary To assess the role of the hypothalamic neurosecretory system in the mechanism of photoperiodic control of testicular growth in Spizella arborea, we have examined the effect of (1) photostimulation on the density of paraldehyde-fuchsin-stainable (PAF+) material in the neurovascular zone of the median eminence, and (2) interruption of the hypothalamo-hypophysial (neurosecretory) tract on the photoperiodic testicular response. Birds were captured during midwinter and retained on 8-hr daily photoperiods several months prior to the experiment. Interruption of the neurosecretory tract was effected by a small knife fashioned from stainless steel wire; radiography was employed to facilitate stereotaxic placement of the knife assembly.Extending the daily photoperiod from 8 to 20 hrs induced rapid testicular growth in photosensitive Spizella arborea, but had no consistent effect on the density of PAF+ material in the palisade layer of the anterior median eminence. Whereas some terminal controls showed moderate depletion after 21 days' exposure to 20-hr daily photoperiods, others retained as much or more stainable material than initial controls in which testicular growth was not induced. Moreover, testicular growth was induced at essentially identical rates whether the PAF+ innervation of the anterior median eminence was intact or eliminated to varying degrees. The finding that the rate of photoperiodically induced testicular growth is independent of the density or pattern of accumulation of PAF+material in the neurovascular zone of the median eminence, coupled with the inconstant effect of photostimulation on the density of stainable material therein, strongly suggests that the eminential component of the hypothalamic neurosecretory system is not a part of the mechanism that controls photoperiodic testicular growth in S. arborea.Contribution No. 980, Division of Biology, Kansas Agricultural Experiment Station, Manhattan 66502. This investigation was supported in part by the Undergraduate Research Participation Program (GY-771) of the National Science Foundation. We are grateful to French's Pet Bird Laboratory, Rochester, N.Y., for the parakeet foods, to Richard S. Donham, Michael D. Ruff, Thomas G. Shane, and Philip G. Watt for assisting with field operations, to Mr. Gary R. Dierking for preparing the drawing, and to Mr. Don Pihlaja for assisting with the photomicrography.     

Immunohistochemical localization of gonadotropin-releasing hormone (GnRH) in the fetal and early postnatal mouse brain.

The objectives were to (a) determine the age in development when GnRH is first detectable in the brain and (b) observe the distribution of GnRH throughout the fetal and early postnatal period. GnRH was localized immunohistochemically in fetal (15, 16, 17 and 19 days of gestation) and early postnatal (1- and 7-day-old) mice with the peroxidase-antiperoxidase (PAP) method of Sternberger. In the organum vasculosum of the lamina terminalis (OVLT) and in the median eminence of the fetus, GnRH was first detected at 17 days of gestation. In the OVLT, GnRH was found ventral to the preoptic recess of the third ventricle near the ventral surface of the brain. In addition, GnRH was located adjacent to the superficial portal capillaries near the surface of the median eminence. At 19 days of gestation, the distribution of GnRH was similar to that observed at 17 days and there was a marked increase in amount. In the newborn mouse, GnRH was undetectable in the OVLT and its content in the median eminence was decreased as compared to that observed in the fetus. By the seventh postnatal day, a considerable accumulation of GnRH had occurred in the OVLT and median eminence. In the OVLT, it was associated with capillaries ventral to the preoptic recess, and its distribution in the median eminence was similar to that in the adult mouse. In both the OVLT and median eminence of the fetal and early postnatal mouse GnRH appeared to be stored in axons and axon endings, but was not detectable in nerve cell bodies or ependymal cells. These observations suggest that the potential for neuroendocrine control of gonadotropin secretion exists in the fetal mouse early as 17 days of gestation.