Evolution de la radioactivité des cellules neurosécrétrices de la pars intercerebralis chez Locusta migratoria migratorioides (Insecte Orthoptère) après injection de cystéine S35

Résumé Après injection de cystéine S³⁵, les cellules A, B, C et les neurones banaux de la pars intercerebralis chez Locusta (femelles immatures et mûres) sont radioactifs. Le taux d' incorporation de la cystéine S³⁵ dans les cellules B est identique chez toutes les femelles et il est légèrement supérieur à celui des neurones banaux. Ces résultats confirment l'inactivité sécrétoire protéique des cellules B. Les cellules C incorporent 3 à 5 fois plus de cystéine S³⁵ que les neurones banaux. Elles synthétisent donc une ou plusieurs protéines contenant de la cystéine ce qui réaffirme leur activité neurosécrétrice chez Locusta. Les cellules A possèdent le taux d'incorporation de cystéine S³⁵ le plus élevé: 5 à 8 fois celui des neurones banaux.Chez toutes les femelles, les cellules A synthétisent plus de neurosécrétion et en éliminent proportionellement plus que les cellules C. La neurosécrétion A est élaborée sous sa forme figurée plus rapidement (30 min) que la neurosécrétion C (60 min). Le renouvellement de la neurosécrétion A est donc quantitativement plus important et plus rapide que celui de la neurosécrétion C. Chez les femelles immatures, les cellules A et C synthétisent plus de matériel et en éliminent proportionnellement plus que chez les femelles mûres. Le temps nécessaire à l'élaboration et à la vidange des grains de neurosécrétion A est identique chez toutes les femelles. Il en est de même pour le matériel C. Le renouvellement des neurosécrétions A et C est donc plus important chez les femelles immatures que chez les femelles mûres mais il n'est pas plus rapide. L'accumulation du matériel fuchsinophile dans les cellules A et C lors de la maturation ovarienne correspond à une réduction de leur fonction neurosécrétrice: elle résulte d'une diminution de l'activité d'élimination des cellules neurosécrétrices A et C supérieure à l'affaiblissement de leur activité de synthèse.

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Evolution of the radioactivity of the neurosecretory cells of pars intercerebralis in Locusta migratoria migratorioides (insect orthoptera) after injection of ³⁵S-cysteineAn autoradiographic study by optical and electronic microscopy

Summary After injection of ³⁵S-cysteine, the A, B, C cells and the ordinary neurones of pars intercerebralis in Locusta — immature and mature females — are radioactive. The rate of incorporation of ³⁵S-cysteine into the B cells is the same for all the females, and it is slightly higher than the rate of incorporation of ³⁵S-cysteine into the ordinary neurones. These results demonstrate the proteinic secretory inactivity of the B cells. The C cells incorporate 3 to 5 times more ³⁵S-cysteine than ordinary neurones. Thus, the C cells synthesize one protein or several proteins with cysteine; this observation confirms their neurosecretory activity in Locusta. The A cells have the highest rate of incorporation of ³⁵S-cysteine: 5 to 8 times the one of ordinary neurones.In all the females, the A cells produce and release proportionally more neurosecretion than the C cells. The production of granules is faster in A cells (30 min) than in C cells (60 min). The turnover of the A neurosecretion is consequently higher and quicker than the C neurosecretion. In immature females, the A and C cells synthesize and release proportionally more material than in mature females. The time necessary for production and release of the A neurosecretion is the same for all the females. It is so for the C material. The turnover of the A and C neurosecretions is thus more important in immature females than in mature females but it is not more rapid. The accumulation of stainable neurosecretory material in A and C cells at the time of ovarian maturation is associated with a reduction of their neurosecretory activity: it is due to a decrease of the rate of release of the A and C cells being more important than their rate of production.

     

Specific effect of calcium ions on the calcium-sensitive cells of the pars intermedia in the goldfish

Summary Cytological changes in the calcium-sensitive (Ca-s) cells (formerly termed PAS-positive cells) of the pars intermedia were investigated in the goldfish after adaptation to deionized water (DW), with or without addition of sodium, potassium and magnesium. These ions were added as chloride salts at concentrations similar to those present in fresh water (FW). The marked stimulation of the Ca-s cells is not inhibited in DW supplemented with Na⁺ (0.35 mM/1), K⁺ (0.05 mM/1), and Mg²⁺ (0.2mM/1) for a period of 24 days. The inhibition of the response to DW with calcium chloride (2 mM/1) is reproduced with calcium formiate (2 mM/1). These data show that chloride ions are not responsible for the regression of the Ca-s cells observed in goldfish kept in DW supplemented with calcium chloride. The effect of calcium ions on the Ca-s cells appears to be specific. These results support the hypothesis that the Ca-s cells synthesize a factor (hypercalcin?) involved in calcium regulation, and that its release is influenced by the calcium content of the environment. The role of the pars intermedia in calcium metabolism is strengthened by the present results. Biochemical data suggest the presence of a hypercalcemic factor in the pituitary of fish (Parsons et al. 1978) and are in agreement with the present cytological findings.     

Intercellular channels in the pars tuberalis of the rat hypophysis and their relationship to the subarachnoid space

Summary A system of intercellular channels is described in the pars tuberalis (PT) of the female rat. These spaces are lined by all types of cells found in the PT and are not sealed off by tight junctions. Ventrally and dorsally, the intercellular spaces open toward the basement membranes separating the PT from (i) the subarachnoid space, and (ii) the perivascular space of the portal capillaries, respectively. These intercellular channels differ from the follicles, which are also found in the PT, being lined by a particular type of cell.In a second group of female rats an epoxy mixture was injected into the third ventricle; 10 min thereafter horseradish peroxidase was infused into the cisterna magna. After processing the brain for the demonstration of exogenous peroxidase, it was found that the tracer had reached the subarachnoid space adjacent to the hypothalamus and entered into all ventricular cavities with the exception of the infundibular recess. Under these experimental conditions it was found that the tracer fills all intercellular channels of the PT, thus indicating that there is no barrier between the subarachnoid space and the PT. It is suggested that the subarachnoid space should be regarded as a probable route for the transport of trophic factor(s) and/or secretory product(s) of the PT.Supported by Grant S-80-13 from Directión de Investigaciones, Universidad Austral de Chile     

A morphometric study on the pars intermedia of the hypophysis during impairment of the renin-angiotensin-aldosterone system in sodium depleted mice

Fine structural alterations were investigated in cells of the pars intermedia of the pituitary of mice treated for four weeks with (a) a sodium deficient diet, (b) a sodium deficient diet mixed with propranolol (renin-inhibitor), (c) a sodium deficient diet combined with propranolol and amino-glutethimide (corticosterone 18-hydroxylase inhibitor), and (d) a sodium deficient diet combined with propranolol, aminoglutethimide and dexamethasone. The number of secretory granules decreased from 5.0/mum2 in the normal control of 2.4/mum2 in all four experimental groups suggesting that the cells in treated groups had reached an equilibrium in the production and release of secretory granules during the chronic treatments. The number of immature Golgi granules per unit Golgi area was 0.91 in the control, while this value rose to 3.29 (3.62 fold of the control), 4.37 (4.8 fold), 4.94 (5.43 fold) and 5.16 (5.67 fold) respectively in the four experimental groups. In these groups a good correlation was observed between the number of immature granules and the percent volume of rough endoplasmic reticulum (r=0.985, p less than 0.01). The present study suggests that the pars intermedia contains an unidentified pituitary factor (or factors) essential for aldosterone biosynthesis.     

Intracellular acetylation of desacetyl αMSH in the xenopus laevis neurointermediate lobe

High performance liquid chromatography (HPLC) followed by radioimmunoassay (RIA) of the chromatographic fractions were used to separate and quantify, respectively, the αMSH-like peptides stored in the neurointermediate lobe (NIL) of the Xenopus laevis (X. laevis) pituitary gland and released from the X. laevis NIL, in vitro. Immunoreactive (IR) material eluting with a similar HPLC retention time as desacetyl αMSH was the major IR peptide in the NIL. Material with a retention time similar to αMSH and immunological properties equivalent to αMSH was also present in the NIL. However, the retention times of the X. laevis and mammalian αMSH-like peptides were not identical, suggesting species difference in these peptides. Following incubation of NILs in the presence of [³H]-acetyl CoA, the X. laevis variant of αMSH was the major [³H]-labeled, immunoprecipitable material present. Following an incubation of NILs in the presence of [³H]-amino acids for 21 hours, immunoprecipitable [³H]-αMSH was detected in the NILs and the ratio of [³H]-desacetyl αMSH to [³H]-αMSH was similar to the ratio of IR-desacetyl αMSH to IR-αMSH. The X. laevis variant of αMSH was the major αMSH-like peptide released from the NILs into the incubation medium. Dopamine (50 μM) significantly inhibited the release of IR-αMSH but not IR-desacetyl αMSH. No net increase in total αMSH (sum of release and NIL content) was observed in the actively secreting (control) NIL group versus the dopaminetreated group. These results indicate that acetylation of desacetyl αMSH occurs intracellularly.     

Etude du lobe intermédiaire de l'hypophyse chez la souris normale et thyroïdectomisée: Volumétrie,histologie et incorporation de divers éléments radioactifs

Résumé Après radiothyroïdectomie chez la souris, l'augmentation volumétrique du lobe intermédiaire, son aspect cytologique, l'incorporation importante de ³H-Méthionine et de ³⁵SO₄Na₂ par ses cellules, plaident en faveur d'une hyperactivité sécrétrice et excrétrice parallèle à celle du métabolisme thyréotrope du lobe antérieur.

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Study of the pars intermedia of the hypophysis in normal and thyroidectomized mice: Volumetry, histology and incorporation of various radioactive elements

Summary After radiothyroidectomy in the mouse, the volumetric increase of the intermediate lobe, its cytological aspect, the considerable incorporation of ³H-Methionine and ³⁵SO₄Na₂ by its cells, speak in favour of a secretory and excretory hyperactivity paralleling the anterior lobe's thyreotrope metabolism.

Abréviations utilisées: LA Lobe antérieur de l'hypophyse. LI Lobe intermédiaire. LN Lobe nerveux. NSO Noyau supra-optique de l'hypothalamus. PAS Periodic acid Schiff. BA Bleu Alcian. AF Aldéhyde fuchsine.     

Etude au microscope électronique de la neurohypophyse de Rana esculenta L.

Résumé L'éminence médiane et la pars nervosa de Rana esculenta diffèrent du point de vue de leur structure.L'éminence médiane se compose de 2 zones différentes: la zone externe placée près du lobe distal et la zone interne située sous l'épendyme. Dans la zone externe, on distingue, d'après la taille et la forme des grains de sécrétion, 5 types de terminaisons.1. des terminaisons avec de fins granules sphériques denses de 800 à 1000 Å de diamètre; 2. des terminaisons avec des granules de 1000 à 1200 Å de diamètre; 3. des terminaisons avec des grains de forme irrégulière de diamètre supérieur aux précédents (1200 à 1600 Å); 4. des terminaisons avec de volumineux grains denses sphériques d'environ 1200 à 1800 Å de diamètre; 5. un petit nombre de terminaisons ne contenant que des vésicules. Les terminaisons des catégories 3 et 4 sont probablement du type neurosécrétoire.La zone interne contient de nombreuses fibres neurosécrétrices. Elles sont de 2 types, l'une avec de gros granules (1600–2400 Å), l'autre avec des granules moins volumineux (1300–2000 Å). Des fibres non neurosécrétrices ont également été observées.Dans la pars nervosa, on rencontre deux types principaux de fibres neurosécrétrices, l'une avec des grains denses de 1600 à 2400 Å de diamètre, l'autre avec des grains moins denses d'environ 1300 à 2000 Å de diamètre. Dans la zone externe bordant la pars intermedia des fibres aminergiques avec de fines granulations ont été observées.

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Electron microscopic study of the neurohypophysis of Rana esculenta L.

Summary The median eminence and the pars nervosa of Rana esculenta have a different structure.The median eminence has 2 different zones: the outer zone situated near the pars distalis and the inner zone under the ependyme. In the outer zone there are, according to the size and the shape of the granules, 5 types of nerve terminals.1. Endings containing spherical fine dense granules of 800 to 1000 Å in diameter; 2. Endings with spherical granules from 1000 to 1200 Å in diameter; 3. Endings with granules of irregular shape which are bigger than the former (1200 to 1600 Å); 4. Endings with spherical dense granules of about 1200 to 1800 Å in diameter; 5. A few endings containing only clear vesicles. Type 3 and type 4 endings are probably neurosecretory.The inner zone contains numerous neurosecretory fibres. They are of two types: one with big granules (1600–2400 Å), the second with smaller granules (1300–2000 Å). Non-neurosecretory fibres have also been observed.The pars nervosa contains two principal types of neurosecretory fibres: one with dense granules of 1600 to 2400 Å in diameter, the other with lighter granules of about 1300 to 2000 Å. In the external zone lining the pars intermedia, aminergic fibres with fine granules have been observed.

Je tiens à exprimer mes vifs remerciements, à Monsieur le Professeur E. Follenius pour l'intérêt constant qu'il prend à ce travail. Je remercie également Madame R. O. Clauss, collaboratrice technique et Madame Schwoerer, photographe, pour leur aide précieuse.     

Intracellular acetylation of desacetyl alpha MSH in the Xenopus laevis neurointermediate lobe

High performance liquid chromatography (HPLC) followed by radioimmunoassay (RIA) of the chromatographic fractions were used to separate and quantify, respectively, the alpha MSH-like peptides stored in the neurointermediate lobe (NIL) of the Xenopus laevis (X. laevis) pituitary gland and released from the X. laevis NIL, in vitro. Immunoreactive (IR) material eluting with a similar HPLC retention time as desacetyl alpha MSH was the major IR peptide in the NIL. Material with a retention time similar to alpha MSH and immunological properties equivalent to alpha MSH was also present in the NIL. However, the retention times of the X. laevis and mammalian alpha MSH-like peptides were not identical, suggesting species difference in these peptides. Following incubation of NILs in the presence of [3H]-acetyl CoA, the X. laevis variant of alpha MSH was the major [3H]-labeled, immunoprecipitable material present. Following an incubation of NILs in the presence of [3H]-amino acids for 21 hours, immunoprecipitable [3H]-alpha MSH was detected in the NILs and the ratio of [3H]-desacetyl alpha MSH to [3H]-alpha MSH was similar to the ratio of IR-desacetyl alpha MSH to IR-alpha MSH. The X. laevis variant of alpha MSH was the major alpha MSH-like peptide released from the NILs into the incubation medium. Dopamine (50 microM) significantly inhibited the release of IR-alpha MSH but not IR-desacetyl alpha MSH. No net increase in total alpha MSH (sum of release and NIL content) was observed in the actively secreting (control) NIL group versus the dopamine-treated group. These results indicate that acetylation of desacetyl alpha MSH occurs intracellularly.