Corticotropin releasing factor (CRF) is a major mediator of central and peripheral responses to environmental stressors, and antagonism of its receptors (CRF-R1, -R2) is an active area of pharmacotherapeutic research for stress-related disorders. Stress responses include CRF activation of the hypothalamus-pituitary-adrenal axis and behavioural inhibition. Valid in vivo models for the study of these neuro-endocrine and -behavioural CRF pathways and their central-peripheral antagonism are important. The aims of this study in C57BL/6 mice were to describe the acute effects of intracerebroventricular (ICV) CRF using plasma ACTH-CORT titres and locomotor activity as readouts, and to study the impact on these readouts of central versus peripheral pretreatment with the CRF-R1/2 antagonist, astressin. The following experiments were performed: Effects of (i) serial blood sampling (SBS) per se, (ii) physical confinement + SBS, (iii) ICV saline infusion + SBS, on plasma titres of ACTH-CORT. (iii) ICV saline infusion led to a marked increase in plasma ACTH, possibly due to assay crossreactivity with "washed out" pituitary peptides, and a mild increase in plasma CORT. (iv) ICV CRF (0.001-1 µg) induced no further increase in plasma ACTH versus vehicle, and induced dose-dependent increased plasma CORT. 1 µg ICV CRF also reduced locomotor activity. (v) ICV CRF induced dose-dependent increased plasma CRF. (vi) ICV astressin failed to block ICV CRF-induced increased plasma CORT, whereas IP astressin did do so. (vii) ICV CRF-induced locomotor inactivity was blocked by ICV astressin, but not by IP astressin. Therefore, ICV CRF induced a dose-dependent increase in plasma CORT via a peripheral pathway and a reduction in locomotion via a central pathway, indicated by the double dissociation in the ability of astressin to antagonize these effects relative to its route of administration, IP or ICV, respectively. The preparation described here could be readily used to provide initial indications on the central and peripheral activity of CRF-R antagonists, including pharmacokinetics following peripheral administration.Dear Ms. Yeates, Dear Prof. Markou, Thank you very much to you and the reviewers for the very constructive and helpful criticism of the manuscript. All of the comments have been taken into account and the manuscript has been revised accordingly. The details of the revisions undertaken are stated point-by-point below. The changes made in the revised manuscript relative to the original are highlighted in bold font in the revision.My co-authors and I look forward to your reply on the suitability of the revised manuscript for publication in Neuropharmacology in due course.
Thank you and yours sincerely, Christopher PryceCover letter
Responses to ReviewersEditor's comments:Both reviewers found your work mostly well conducted and of potential interest to the field. There are, however, some issues that the reviewers raised that need to be addressed with specific changes in the manuscript. For example, the limitation of dr...