Antisense oligonucleotides in neuroendocrinology: enthusiasm and frustration

Antisense oligodeoxynucleotides (ODN) offer the potential advantage to manipulate neuropeptide or neuropeptide receptor expression within the brain transiently and site-specifically, thus providing a tool for neuroendocrinological research into the physiological function of a particular neuropeptide system. In this study, various approaches are introduced which reveal that antisense ODN may exert acute, short-term effects on neuronal responsiveness to afferent stimuli, as well as long-term effects on neuropeptide/receptor protein availability in a given system depending on the duration of treatment. Short-term effects were seen in that oxytocin (OXT) and vasopressin (AVP) antisense ODN affected electrophysiological and secretory parameters of oxytocinergic and vasopressinergic neurons, respectively, as well as their ability to express the Fos protein in response to afferent stimulation a few hours after a single infusion into the hypothalamic supraoptic nucleus. In this study, two methodological approaches to study long-term effects of the antisense ODN are exemplified, in which antisense ODN directed against the mRNA coding for the neuropeptide itself or its receptor were used. The repeated infusion of corticotropin releasing hormone (CRH) antisense ODN into the paraventricular nucleus resulted in reduced immunoreactive CRH, but not AVP, in the external zone of the median eminence. Furthermore, in order to evaluate the receptor-mediated effects of CRH and AVP released locally within the paraventricular nucleus on adrenocorticotropin (ACTH) release from the pituitary, CRH receptor (and also AVP receptor) antisense ODN were repeatedly infused into the hypothalamic nuclei; this treatment resulted in an elevation of stimulated, but not basal, ACTH release into the blood. However, in addition to these obvious antisense effects, results are discussed which demonstrate sequence-unspecific effects of phosphorothioated ODN, suggesting that some of their mechanisms of action are not yet understood.