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Release in vitro of neurohypophysial proteins from neural lobe tissue slices and from isolated neurosecretory granules of the rat

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Summary

The release of neurophysin from neural lobe tissue slices and isolated neurosecretory granules of the rat was studied at various time intervals after injection of (35S) cysteine into the supraoptic nucleus. For hours after isotope injection the release of radioactive neurophysin from neural lobe tissue was increased by depolarizing concentration of potassium in the presence of calcium ions. Fourteen and 30 days after isotope injection the release of radioactive neurophysin was relatively decreased in a medium of high potassium concentration which might be explained by the heterogeneity of the pool of neurophysin within the neural lobe. Four h after isotope injection the “spontaneous” release of neurophysin from neural lobe tissue was higher in dehydrated rats than in controls, and the neural lobes of these animals did not respond with an increased release of radioactive neurophysin when exposed to high potassium concentration.

Eighteen h after isotope injection the predominating proportion of neurophysin-bound radioactivity was found in the neurosecretory granule fraction, whereas 14 days after injection a fairly equal amount of radioactivity was found in this fraction and in the “soluble” protein fraction. This indicates that with time an increasing amount of radioactive neurophysin passes from an intragranular to an extragranular pool.

The spontaneous release of radioactive neurophysin from isolated neurosecretory granules was higher, and the increase of release upon exposure to an ATP-regenerating system was higher 14 days after isotope injection than 18 h after injection. This may imply that the neurosecretory material undergoes an intragranular “maturation” process to become more easily releasable. The release of radioactive neurophysin was inhibited in the presence of AMP and EDTA, which demonstrates the dependence of the release process of an ATPase and, probably, of calcium ions.

Neurosecretory granules from dehydrated rats possessed a higher spontaneous release than those of control rats 18 h after injection, which may indicate an enhanced intragranular “maturation” process of the neurosecretory material due to osmotic stimulation.

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Abbreviations

NSG:

neurosecretory granule

NSM:

neurosecretory material

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  1. Institute of Neurobiology, University of Göteborg, Göteborg, Sweden

    Anders Norström

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  1. Anders Norström
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The present work was supported by grants from Carl-Bertel Nathhorsts vetenskapliga stiftelse, from “Svenska Livförsäkrings bolags nämnd för medicinsk forskning”, from Överläkare Albert Wallins fond” and from the Medical Faculty of the University of Göteborg. I am most indebted to Miss Gull Grönstedt for careful secreterial work.

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Norström, A. Release in vitro of neurohypophysial proteins from neural lobe tissue slices and from isolated neurosecretory granules of the rat. Z. Zellforsch 129, 114–139 (1972). https://doi.org/10.1007/BF00307114

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