Document Detail


Temporal profile of arginine vasopressin release from the neurohypophysis in response to hypertonic saline and hypotension measured using a fluorescent fusion protein.
MedLine Citation:
PMID:  21855574     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Methods currently employed to study the release of hormones such as arginine vasopressin (AVP), while sensitive, suffer from a low temporal resolution such that the monitoring of AVP release on a moment-to-moment basis is not possible. Here, we describe a new approach to indirectly monitor the temporal profile of AVP release from the neurohypophysis of transgenic rats expressing an AVP-eGFP fusion gene. Using fibre-optic probes (termed 'optrodes') we were able to indirectly monitor AVP release via a reporter moiety in real-time. This method is a major advance over current methods used to monitor AVP release. Intravenous administration of hypertonic saline (3M NaCl) induced a rapid (latency of 2-3s) increase in fluorescence detected in the neurohypophysis that lasted on average for 60s - a response that was highly reproducible. Infusion of sodium nitroprusside induced a rapid fall in blood pressure accompanied by a rapid, stimulus-locked increase in fluorescent signal that returned to baseline with the recovery of blood pressure to pre-stimulus levels - again this response was highly reproducible. Withdrawal of blood (to simulate haemorrhage) also resulted in a stimulus-locked increase in fluorescence that return to baseline after the withdrawn blood was returned to the animal. In conclusion, we developed a highly sensitive approach that allows the indirect measurement of AVP release via the monitoring of a reporter gene in real-time. This technology can be adapted to permit the study of a whole array of neurohormones/chemicals in transgenic animals expressing a fluorescent reporter construct.
Authors:
Song T Yao; Vagner R Antunes; Peter M J Bradley; Sergey Kasparov; Yoichi Ueta; Julian F R Paton; David Murphy
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-8-10
Journal Detail:
Title:  Journal of neuroscience methods     Volume:  -     ISSN:  1872-678X     ISO Abbreviation:  -     Publication Date:  2011 Aug 
Date Detail:
Created Date:  2011-8-22     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7905558     Medline TA:  J Neurosci Methods     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2011. Published by Elsevier B.V.
Affiliation:
Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Bristol BS1 3NY, UK.
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