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Fine structure and function of isolated gonadotropic cells as revealed from pituitaries of immature rainbow trout, Salmo gairdneri, by means of a new enzymatic dispersion technique

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Summary

A procedure has been developed for dissociating pituitary glands of juvenile rainbow trout, Salmo gairdneri, producing a preparation of single dispersed pituitary cells in which morphological and functional integrity is preserved. The pituitaries are dispersed by sequential treatment with 0.1% collagenase, 0.04% ethylene-diamine-tetra-acetic acid (EDTA) and 0.125% dispase. The cell yield is 0.3–0.35 × 106 cells per pituitary with a cell viability percentage of 95±1 % and single cell percentage of 87±4%.

The isolated cells are kept in a suspension system and the gonadotropic cells are identified by the double antibody immuno-enzyme-cytochemical technique using anti-carp β-gonadotropin as first antibody. Secretory activity is estimated by measuring the gonadotropin content in cells and culture media by radioimmunoassay. Isolated cells show an autonomy of gonadotropin secretion.

17α-Methyltestosterone both in vivo and in vitro stimulates the production of gonadotropin in the cells and seems to inhibit its release from the cells.

It is concluded that this in vitro system can be used as a model for studying the control of gonadotropic cells in juvenile rainbow trout.

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Authors and Affiliations

  1. Zoological Laboratory, Research Group for Comparative Endocrinology, State University of Utrecht, Utrecht, The Netherlands

    Goverdina E. Fåhræus-van Ree, Stephan E. F. Guldenaar & Jeff Th. Gielen

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  1. Goverdina E. Fåhræus-van Ree
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  2. Stephan E. F. Guldenaar
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  3. Jeff Th. Gielen
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Additional information

The authors are indebted to Prof. Dr. D.R. Idler (Marine Sciences Research Laboratory, St. John's Newfoundland, Canada) and to Dr. H.J.Th. Goos (Zoological Laboratory, Utrecht, The Netherlands) for supplying the Con AII salmon gonadotropin and antibody to salmon gonadotropin, respectively, and to Dr. E. Burzawa-Gérard (Laboratoire de Physiologie générale et comparée du Muséum national d'Histoire naturelle, Paris, France) for the antibody to carp β-gonadotropin used in this study. Appreciation is expressed to Mrs. M.G.A. de Mol Moncourt-de Bruyn for her support in carrying out the electron microscopical study. Thanks are also due to Prof. Dr. P.G.W.J. van Oordt and Prof. Dr. W.A. de Voogd van der Straaten for critically reviewing the manuscript, to Mr. J.A.F.W. Kleijne and Mr. C.J. Panis for excellent technical assistance, to Mr. M.J. van Oosterum and his group for care in keeping the animals, and to Mr. H. van Kooten and his colleagues for photographic help

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Fåhræus-van Ree, G.E., Guldenaar, S.E.F. & Gielen, J.T. Fine structure and function of isolated gonadotropic cells as revealed from pituitaries of immature rainbow trout, Salmo gairdneri, by means of a new enzymatic dispersion technique. Cell Tissue Res. 226, 641–653 (1982). https://doi.org/10.1007/BF00214791

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