Secretion of melanophore-stimulating hormone (MSH) in long-term cultures of pituitary neurointermediate lobes

Summary Neurointermediate lobes from pituitaries of the frog, Rana berlandieri forreri (Rana pipiens, sensu lato), were maintained in organ culture in media with and without serum for up to six months. The cultured tissues were examined periodically by light microscopy and transmission electron microscopy and by bioassay of the melanophore-stimulating hormone (MSH) secreted and present in the culture media. Light-microscopic observations revealed a high degree of preservation of the pars intermedia at four weeks with isolated areas of some glands maintaining histological integrity for the entire six months. Similarly, at the ultrastructural level the cells appeared morphologically intact and to be actively synthesizing and secreting hormone. Bioassays showed the glands to be continuously secreting MSH; however, larger yields of hormone were obtained in media lacking serum. No significant ultrastructural differences between cells grown in the presence or absence of serum were detected. The difference in concentration of MSH between the two groups therefore apparently results from enzymatic degradation of the hormone by the serum. Organ culture of the vertebrate neurointermediate lobe may provide a unique method for the production of large quantities of MSH and for the study of other melanotropic and opiate peptides as they may be synthesized and secreted by the pars intermedia.     

Immunofluorescence of somatostatin-producing sites in the hypothalamus of the tadpole,Alytes obstetricans Laur.

Summary Three sites of somatostatin-synthesizing perikarya, or a related antigen, were determined by immunofluorescence in the hypothalamus of the tadpole, Alytes obstetricans (Amphibia, Anura). Two sites of neurosecretory perikarya were localized in the preoptic nuclei of the anterior hypothalamus; the axons extended either to the anterior diencephalon or to the median eminence and the pituitary. The third site was found in the posterior hypothalamus. These neurosecretory cells showed a strong immunofluorescent reaction; their axons all terminated at the level of the median eminence. Somatostatin cells were only found in intact or hypophysectomized tadpoles given somatotropin (STH). The strong reaction observed in hypophysectomized tadpoles was possibly due to the loss of the terminal portion of the neurosecretory pathway (median eminence and pituitary) by which the agent is transported to the site of discharge.     

Studies on bone marrow histogenesis: Morphometric and autoradiographic studies of regenerating marrow stroma in extramedullary autoimplants and after evacuation of marrow cavity

Summary The marrow cavity of the rat tibia was mechanically evacuated and autoimplanted to the subcutaneous tissue. The regenerative process which restored the integrity of marrow stroma and hemopoiesis, was morphometrically evaluated in whole mount of tibia. Following evacuation, the clot filled the cavity. The granulation tissue then appeared and expanded, penetrating and replacing the clot. The fibroblasts of the granulation tissue differentiated into osteoblasts forming osteoid bone. Within its interstices, the primordial marrow consisting of loose connective tissue and vascular sinuses appeared and hemopoiesis resumed. Expansion of hemopoiesis resulted in the resorption of bone and within three weeks the tibial cavity was restored to the pre-evacuation state.Autoradiography indicated that the labeling index was initially high in fibroblasts and osteoblasts but was subsequently reduced while it increased in osteocytes, cells of Haversian canals, stromal and hemopoietic cells of marrow. The finding is in disagreement with the view that the regenerative process originates from the Haversian canal. When the label was introduced on day 4 post-operatively, it subsequently appeared in osteocytes, cells of Haversian canal, stromal elements of the marrow, but not in the hemopoietic cells. This indicates complete dissociation of marrow stroma and hemopoietic stem cell.Supported by NASA Contract NSG 9061. Mehdi Tavassoli is the recipient of a CRD Award AM-70551     

The eyes of mesopelagic crustaceans

Summary In Streetsia challengeri left and right eyes have fused and become a single cylindrical photoreceptor, which occupies the basal half of a forward directed head projection. This unusual compound eye consists of approximately 2500 ommatidia, which are arranged in such a way that the animal has almost circumferential vision, but cannot look ahead or behind. It is thought that the eye operates on light-guide principles, and that the crystalline cones are the major dioptric component. Ommatidia in anterior-posterior rows show a greater overlap of visual fields than dorso-ventrally arranged ommatidia. Cone layer and retinula are separated by a 4 m thick screen-membrane, which contains tiny pigment granules of 0.15 m diameter. Cells of unknown function and origin, containing unusual multitubular organelles, are regularly found near the proximal ends of the crystalline cone threads. The twisted rhabdoms measure 18–20 m in diameter, and consist of microvilli 0.05 m in width, which belong to five retinula cells and which show no trace of disintegration. The position of interommatidial screening pigment, the density of retinula cell vesicles and inclusions, and the narrowness of the perirhabdomal space all suggest that the eyes have been light-adapted at the time of fixation for electron microscopy. The retinula cell nuclei lie on the proximal side of the heavily pigmented basement membrane. A tapetum or basal retinula cells are not developed. It is concluded that the eye optimally combines acuity with sensitivity, and that for distance estimation parallax may be important.Address until January 25th 1978: Scott Base, Ross Dependency, Antarctica (C/-Chief Post Office, Christchurch, New Zealand)     

The rhombencephalic recess in the rat

Summary The rhombencephalic recess, an ependymal organ, has been studied for the first time by light- and electron microscopy. It is situated mediosagittally on the floor of the rhomboid fossa at the level of the colliculus facialis. The recess and the superimposed tissue are built up by tanycytes, their apices being connected by tight junctions. HRP, injected into the c.s.f., does not penetrate into the intercellular clefts of the recess area. The recess area reveals a certain autonomy regarding its supply with arteries and capillaries. A bloodbrain barrier exists, but shows slight leakage in circumscribed areas as a result of intense transendothelial vesicular transport. The organization of the recess area is compared with that of other ependymal organs, especially circumventricular organs.The skilful technical assistance of Miss K. Bielenberg, Mrs. H. Prien, Mrs. E. Schöngarth and Mrs. H. Schöning is thankfully acknowledgedSupported by the Deutsche Forschungsgemeinschaft (Kr 569/1 and SFB 34/D4) and Stiftung Volkswagenwerk

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Histogenesis of the pituitary in rainbow trout (Salmo gairdneri) in different ambient salinities with particular reference to the rostral pars distalis

Summary The pituitary gland is first evident in rainbow trout two weeks before hatching. Differentiation of prolactin and ACTH cells is not marked until 3–4 days after hatching when the follicular arrangement of the prolactin cells become apparent. There is no difference in the time of development of either prolactin or ACTH cells in larvae reared in different ambient salinities despite marked changes in tissue ion and water content. This suggests that prolactin and ACTH do not play the osmo- and iono-regulatory roles in larval rainbow trout that they are considered to play in adult salmonids.The work was supported by a grant-in-aid of research from the National Research Council of Canada     

Calcium and sulphur in neurosecretory granules and calcium in mitochondria as determined by electron microscope X-ray microanalysis

Summary Sections of neurosecretory cells fixed in glutaraldehyde and osmium tetroxide were studied by means of an EMMA-4 analytical microscope. Secretory granules in neurosecretory cells of the corpus cardiacum and of the brain, both in the desert locust Schistocerca and in the blowfly Calliphora, as well as neurosecretory granules in posterior pituitaries of the frog Rana and of the albino rat all contain a high concentration of calcium. A distinct sulphur peak was also a constant feature.In neurosecretory cells of the corpus cardiacum of Schistocerca the chromatin contained a high concentration of calcium. The mitochondria also contained much calcium, but part of this disappeared during preparation except when fixative and wash contained calcium chloride. By block staining with uranyl acetate most calcium is displaced from the mitochondria, whereas most of the calcium remains in the neurosecretory granules. Since the calcium peaks in spectra from neurosecretory granules appear of similar size, regardless of variations in the preparative procedure, this calcium must be firmly bound. The possible role of the calcium bound to the neurosecretory substance is discussed.The presence of sulphur in insect neurosecretory granules indicates the presence of a protein besides the hormone, i.e., an insect neurophysin.We wish to thank Dennis Greer for the operation of the analytical microscope. This work was supported by a Wellcome-Carlsberg Travelling Research Fellowship awarded to T.N. The EMMA-4 facility is supported by a grant from the British Science Research Council     

Kinetics of functional and morphological changes during decoupling and recoupling induced by glycerol in isolated muscle fibres of the crayfish

Summary The development of ultrastructural changes in the T-system of isolated muscle fibres of the crayfish by the glycerol procedure is described in correlation with the dissociation of excitation-contraction (E-C) coupling as well as with recoupling of the E-C link. The sequence of events in the process of disconnection of the tubules is as follows: dilation of the T-system tubules, disconnection of the constricted tubular segments from the surface membrane and from the T-system vesicle, disappearance of the lumen and its disintegration. The decoupled state is characterised by the presence of round vesicles uniformly distributed in the entire volume of the fibre. The volume of vesicles accounts well for the residual postglycerol volume increase (15%) of the muscle fibres. Functional and structural recovery can be induced by reapplication of glycerol to fibres decoupled and vesiculated with concentrations of glycerol300mmol · l^-1 in crayfish saline. The restitution starts with the organisation of the material of the disintegrated connecting segment of the T-system tubule into small vesicles which coalesce to form the tubule from the vesicular site. At the same time the surface membrane is invaginated toward the vesicle, thus forming the tubule from the surface membrane site. Recovery starts already in the first minute after application of glycerol and is completed within approximately 15min.     

Fine structure and composition of the submucous nerve plexus of the guinea-pig trachea

Summary The ultrastructure of the nerves forming the submucous plexus of cervical and thoracic parts of the trachea was studied in the guinea-pig. Specimens were obtained from 6 animals perfused with warm fixative and from 6 animals in which pieces of trachea were incubated in buffer containing 5-hydroxydopamine before being immersed in cold fixative. Of the two types of axonal terminal identified in the nerves, one contained mainly large dense-cored vesicles, and the second contained numerous small vesicles. In specimens incubated in 5-hydroxydopamine, the small vesicles of the latter terminals exhibited the electron-dense cores which are characteristic of adrenergic axonal terminals. Counts made on perfused specimens showed that, in both the thoracic and cervical parts of the trachea, the density of adrenergic terminals was higher than that of terminals containing mainly large dense-cored vesicles. Overall terminal density was, however, higher in the thoracic than in the cervical part of the trachea, and estimates of nerve size showed that this was associated with the presence in the thoracic plexus of a substantially greater proportion of nerves with less than 6 axons. The possible function of the nerves in the control of the calibre of the submucous blood vessels was discussed.     

The relative effectiveness of three techniques to induce the trophotropic response

The purpose of this study was to examine the relative effectiveness of electromyographic biofeedback training(EMG BFT), remeditation, and progressive muscle relaxation(PMR) in eliciting a relaxation or trophotropic response as measured by frontalis muscle tension, heart rate, electrodermal response, respiration rate, and skin temperature. Fifty-four college students were randomly assigned to one of five groups:(1) control,(2) placebo control,(3) EMG BFT,(4) meditation,(5) PMR. After baseline measures were obtained subjects were trained in 10 30-minute training sessions and posttested. Comparisons by ANOVAs indicated there was a significant decrease in muscle tension in the EMG BFT and meditation groups and significant decreases in respiration rate in the meditation and PMR groups. No other changes were attributed to treatment.