Secretion of CRF-41, AVP and SRIF by Fetal Hypothalamic Neurons Cultured in Defined Medium

  • G. Gillies
  • M. Clarke
Conference paper
Part of the Neuroendocrine Perspectives book series (NEUROENDOCRINE, volume 6)

Abstract

The precise mechanisms regulating the secretion of the 41 amino acid corticotrophin-releasing factor (CRF-41) and arginine vasopressin (AVP) into the hypothalamo-hypophysial portal circulation remain undetermined. This is attributable in part to the inherent complications of in vivo studies on parameters particularly sensitive to external manipulations and to the indirect methods of CRF measurements both in vivo and in vitro. These rely on biological activities, e.g. changes in levels of adrenocorticotrophic hormone (ACTH), corticosteroids or adrenal ascorbic acid, which tell us nothing of the individual components of the hypothalamic CRF complex (1,2). In order to study its control, therefore, we established a system of long-term primary culture of rat hypothalamic neurons coupled with the direct measurement of released CRF-41 and AVP by immunoassay. As an internal control, we simultaneously measured somatostatin (SRIF). Initially, the neurons were grown in a serum-containing medium (3). Detection of released and extracted CRF-41 and AVP depended first, on the concentration of very small amounts of peptides on silica resins; second, on the maintenance of the cultures beyond two weeks; and third, on highly sensitive assays for CRF-41 (4) and AVP. A large number of reports have concentrated on the release of SRIF from similar serum-grown cultures between days 10 and 14 in vitro, when it is present in abundance (5–7). However, serum is likely to represent a nonphysiological component of the central neuron environment. Therefore, we have investigated the responses of neurons grown in a defined medium based on those used by Bottenstein and Sato (8) and by Faivre-Bauman and colleagues (9).

Keywords

Corticosteroid Selenium Serotonin Penicillin Arginine 

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Copyright information

© Springer-Verlag New York Inc. 1989

Authors and Affiliations

  • G. Gillies
    • 1
  • M. Clarke
    • 1
  1. 1.Protein Hormone UnitSt. Bartholomew’s HospitalLondonUK

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