Abstract
Corticotropin‐releasing factor (CRH) neurons in the paraventricular nucleus (PVN) are the essential component of the hypothalamus‐pituitary‐adrenal axis play a critical role in the regulation of autonomic, endocrine and behavioral responses to stress. Assessment of the CRH neurons provides valuable information on cellular mechanisms during physiological and disease process. However, CRH neurons in the rats cannot be reliably identified in live tissues since the identification of CRH neurons requires neuronal labeling and post‐mortem immunohistochemistry. To efficiently identifying CRH neurons in live tissue, we constructed a CRH promoter‐GFP plasmid and integrated this sequence into an adeno‐associated virus (AAV) vector. This AAV‐CRH promoter‐GFP vector was delivered into the rat PVN through microinjection (100 nl). The hypothalamic slices containing GFP‐tagged neurons was immunostained with a specific antibody against CRH. We found that 76.4% CRH immunoreactivity positive PVN neurons colocalized with GFP and 92.8% GFP‐tagged PVN neurons expressed CRH immunoreactivity. Whole‐cell recordings were performed on these GFP‐tagged CRH neurons in hypothalamic slice to assess their electrophysiological properties. Therefore, this AAV‐CRH‐GFP reliably targets CRH neurons in rat brain for in vitro assessment.Grant Funding Source: NIH: MH096086
Published Version
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call