Distribution of corticotropin-releasing factor neurons in the mouse brain: a study using corticotropin-releasing factor-modified yellow fluorescent protein knock-in mouse
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We examined the morphological features of corticotropin-releasing factor (CRF) neurons in a mouse line in which modified yellow fluorescent protein (Venus) was expressed under the CRF promoter. We previously generated the CRF-Venus knock-in mouse, in which Venus is inserted into the CRF gene locus by homologous recombination. In the present study, the neomycin phosphotransferase gene (Neo), driven by the pgk-1 promoter, was deleted from the CRF-Venus mouse genome, and a CRF-Venus∆Neo mouse was generated. Venus expression is much more prominent in the CRF-Venus∆Neo mouse when compared to the CRF-Venus mouse. In addition, most Venus-expressing neurons co-express CRF mRNA. Venus-expressing neurons constitute a discrete population of neuroendocrine neurons in the paraventricular nucleus of the hypothalamus (PVH) that project to the median eminence. Venus-expressing neurons were also found in brain regions outside the neuroendocrine PVH, including the olfactory bulb, the piriform cortex (Pir), the extended amygdala, the hippocampus, the neocortices, Barrington’s nucleus, the midbrain/pontine dorsal tegmentum, the periaqueductal gray, and the inferior olivary nucleus (IO). Venus-expressing perikarya co-expressing CRF mRNA could be observed clearly even in regions where CRF-immunoreactive perikarya could hardly be identified. We demonstrated that the CRF neurons contain glutamate in the Pir and IO, while they contain gamma-aminobutyric acid in the neocortex, the bed nucleus of the stria terminalis, the hippocampus, and the amygdala. A population of CRF neurons was demonstrated to be cholinergic in the midbrain tegmentum. The CRF-Venus∆Neo mouse may be useful for studying the structural and functional properties of CRF neurons in the mouse brain.
KeywordsHomologous recombination Immunofluorescence In situ hybridization Glucocorticoids Stress
This work was supported in part by the Research Grants from JSPS (K.I., K.S., M.W.), JST (K.I., K.S., M.W.), and Comprehensive Brain Research Network (K.I., K.S., M.W.). Anti-CRF antibodies were generously donated by Drs. Wylie Vale and Paul Sawchenko, Salk Institute, Ca, USA, and Dr. Tamotsu Shibasaki, Nippon Medical School, Tokyo, Japan. Anti-thyrotropin-releasing hormone antibody was a gift from Dr. Fekete, Hungarian Academy of Sciences. We thank Dr. Sadayoshi Ito, Tohoku University, and Shinji Ohara, Matsumoto Medical Center, for encouragements. Address any requests to K.I. for mice generated in his laboratory and presented in the present manuscript. The CRF-Venus mouse is also available from Experimental Animal Division, RIKEN BRC (Access Number: RBRC06519).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.
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