Characterization of Newly Formed and Aged Granules in the Neurohypophysis

Neurosecretory granules from the rat and bovine neurohypophysis were isolated and some of their biochemical and biophysical properties studied. Neurosecretory granules (NSG) from rat neurohypophysis were labeled, in vivo, with [35S]cysteine and isolated on isoosmotic gradients. Whereas 1 day after labeling most of the radioactivity was found in the lower part of the gradient, 35 days later the isotope was also located in the lighter NSG-containing fraction. Different analytical procedures showed that the lighter fraction, both in bovine and rat NSG, contain more subpopulations of neurophysin-like material than the heavier fraction. The first material to be released during stimulation of secretion, in vivo or in vitro, is mobilized from the heavy NSG. Isolation of rat NSG, at different times during and after dehydration of the animals, reveals that the newly synthesized material is found in the heavy NSG-containing fraction. Furthermore, the results indicate that the newly synthesized NSG are more resistant to lysis than the lighter granules. The results are discussed in relation to the maturation and degradation processes of the granule content and to the functional state of the NSG.     

Distribution of FMRFamide-like immunoreactivity in the brain and neurohypophysis of the lamprey,Lampetra japonica

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.     

Correlation between microtubular number and transport activity of hypothalamo-neurohypophyseal secretory neurons

Summary The lower pituitary stalk has been studied in rats given hypertonic saline for four days and in rats with congenital diabetes insipidus (GDI), that is, in two groups of animals in which there is evidence of an increase in the transport of polypeptides down the axons of the tractus hypophyseus.A quantitative ultrastructural analysis of this tract, the first yet made, has demonstrated that in normal animals it is composed of unusually small axons, over half of which are less than 300 nm in diameter.In this tract there is a significant increase in axonal diameter and microtubular content in animals given hypertonic saline for four days. In adult rats with CDI, these changes are even more marked, a feature which is correlated with the fact that in these animals the transport of polypeptides has probably been abnormal since birth.An alteration in the microtubular content of neurons has not previously been observed either in physiologically stressed animals or in diseased animals; such alterations suggest the participation of microtubules in polypeptide transport. The narrow diameter of axons in the tractus hypophyseus of the normal animal raises the possibility of the extravesicular transport of such polypeptides.Abbreviations used ADH antidiuretic hormone - AF aldehyde fuchsin - CDI congenital diabetes insipidus - HNS hypothalamo-neurohypophysiel system - NSM neurosecretory material - NSV neurosecretory vesicle - PVN paraventricular nucleus - SON supraoptic nucleus This paper is dedicated to Professor W. Bargmann.We are grateful to Miss Pauline Chapman for technical assistance.This project is supported by the Medical Research Council.     

The fine structure of the neural lobe of the mouse after transplantation under the kidney capsule

Summary The neurohypophysis of donor mice was implanted under the renal capsule of the recipients. The pituicytes survived while the neurosecretory axons disappeared. The ultrastructure of the glial cells was observed seven and nine weeks after transplantation. There were no signs of phagocytotic activity although remnants of axons were still present at seven weeks. The numerous processes of the pituicytes form a network with intercellular spaces wide in younger and narrower in older implants. The cells are connected by desmosomes and gap junctions. Pituicytes as well as blood vessels preserve their organotypic appearance. The transplant thus represents an experimental model for investigations on pituicytes in vivo in the absence of neurosecretory axons.Fellow of the Alexander von Humboldt-Stiftung. On leave from Department of Histology and Embryology, Institute of Biostructure, Academy of Medicine, Pozna, Poland     

Organization and development of the perivascular space system in the neurohypophysis of the laboratory mouse

Summary The organization of the system of perivascular space around the capillaries in the neurohypophysis was studied in the adult and developing laboratory mouse by the use of histological silver impregnation and electron microscopical techniques.In the median eminence short and long extensions, arising mainly from the shallow space around capillary loops of the primary plexus of the portal system, formed radiations into the adjacent neural tissue of the external zone. The tissue of the neural lobe was separable into non-vascular regions dominated by undilated portions of neurosecretory nerve fibres and pituicytes, and neurovascular regions with perivascular space extensions forming an extensive system of connections between neighbouring capillaries.In the median eminence, the system of extensions of the perivascular space was estimated to increase the neurovascular contact surface area by at least 50%, implying an increased efficiency of the organ without a notable increase of its volume. The possibility that the ramifications of the perivascular space imply an enhanced uptake rate into the bloodstream and a subsequent increased concentration of the neurohormones in the portal blood, was discussed.During development of the median eminence, differentiation of perivascular space extensions of the adult type started in the juvenile of about three weeks of age, when shallow capillary loops had been formed. In the neural lobe, perivascular space ramifications were already present when the internal capillaries were formed and were fairly frequent in ten-day young. At the age of three to four weeks the organization of the system was similar to that of the adult animal.     

The function of Ca channel subtypes in exocytotic secretion: new perspectives from synaptic and non-synaptic release

By mediating the Ca²⁺ influx that triggers exocytotic fusion, Ca²⁺ channels play a central role in a wide range of secretory processes. Ca²⁺ channels consist of a complex of protein subunits, including an ₁ subunit that constitutes the voltage-dependent Ca²⁺-selective membrane pore, and a group of auxiliary subunits, including β, γ, and ₂–δ subunits, which modulate channel properties such as inactivation and channel targeting. Subtypes of Ca²⁺ channels are constituted by different combinations of ₁ subunits (of which 10 have been identified) and auxiliary subunits, particularly β (of which 4 have been identified). Activity-secretion coupling is determined not only by the biophysical properties of the channels involved, but also by the relationship between channels and the exocytotic apparatus, which may differ between fast and slow types of secretion. Colocalization of Ca²⁺ channels at sites of fast release may depend on biochemical interactions between channels and exocytotic proteins. The aim of this article is to review recent work on Ca²⁺ channel structure and function in exocytotic secretion. We discuss Ca²⁺ channel involvement in selected types of secretion, including central neurotransmission, endocrine and neuroendocrine secretion, and transmission at graded potential synapses. Several different Ca²⁺ channel subtypes are involved in these types of secretion, and their function is likely to involve a variety of relationships with the exocytotic apparatus. Elucidating the relationship between Ca²⁺ channel structure and function is central to our understanding of the fundamental process of exocytotic secretion.     

Microtubules in the neurosecretory neurones of the posterior pituitary of the rat

Summary The microtubules in the neurosecretory neurones of the posterior pituitary were studied using different electron microscopical techniques. Tannic acid staining indicated that the microtubules had a 13 protofilament substructure similar to that described for microtubules from other tissues and organisms; the dimensions of the microtubules were also similar to that previously reported. Albumen pretreatment clearly showed the microtubules running across axonal swellings, but not continuing across the nerve endings. The only organelles showing possible association with the microtubules were small vesicles and smooth endoplasmic reticulum, no association between hormone granules and microtubules could be seen. Acknowledgement: The author is grateful to P. Wilks for skilled technical assistance.     

Glial cells positive for glial fibrillary acidic protein in the neurohypophysis of the Djungarian hamster (Phodopus sungorus)

Summary The presence and distribution of the glial fibrillary acidic protein (GFAP; an astrocytic marker protein associated with glial filaments) in the neurohypophysis of the Djungarian hamster (Phodopus sungorus) were investigated immunohistochemically. Our study revealed characteristic GFAP-staining patterns within the median eminence, infundibular stem and neural lobe. In the whole neurohypophysis, few glial cells showed immunoreactivity. In the neural lobe, immunopositive pituicytes appeared preferentially in the periphery. At the ultrastructural level, we found some pituicytes containing filaments, most notably in their processes. We thus demonstrated that, in contrast to the GFAP-immunoreactivity of cultured pituicytes, pituicytic GFAP-expression in vivo coincides with the presence of electron-microscopically detectable filaments.     

On the fine structure of the perivascular cells in the neural lobe of rats

Summary The purpose of this study is to investigate the ultrastructural features of perivascular cells as found in the neural lobe of the hypophysis. In particular, an inquiry was made into the nature of the relationship of such cells to neurosecretory fibers and endings. The latter, in fact, are often invaginated within the perivascular cells and enveloped by their processes; furthermore, they often reveal a certain number of empty granules as well as characteristics of degenerative nature. In the course of this study the localization of the perivascular cells has been investigated as well as that of their processes within the extensive interlobular network typical of the hypophysial neural lobe of rodents. Based on the data gathered, the hypothesis is put forward that the perivascular cells play an important role in the turnover of neurosecretory endings, both under physiological and experimental conditions, contributing thereby to the release of post-hypophysial hormones.With the technical assistance of Vincenzo Panetta.