Journal: Molecular Neurobiology
Article Title: Functional and Anatomical Characterization of Corticotropin-Releasing Factor Receptor Subtypes of the Rat Spinal Cord Involved in Somatic Pain Relief
doi: 10.1007/s12035-021-02481-z
Figure Lengend Snippet: Immunofluorescence staining of CRF-R2 ( a – c ) and CRF-R1 ( d – f ) in the superficial laminae of the rat spinal dorsal horn as well as the brain. ( a – c) Immunohistochemical staining of the dorsal horn of the spinal cord ( a , b ) and of the hypothalamus of the brain ( c ) with the polyclonal rabbit anti- CRF-R2 antibody ( Texas red fluorescence ). Note specific CRF-R2 immunoreactivity in Rexed laminae I and II of the spinal dorsal horn ( a ) and in the immediate vicinity of the central canal ( b ), an area receiving descending projections from supraspinal nuclei. CRF-R2 immunoreactivity was also identified in neurons of the hypothalamus ( c ). ( d – f ) Immunohistochemical staining of the dorsal horn of the spinal cord ( d , e ) and of the hypothalamus of the brain ( f ) with the polyclonal rabbit anti-CRF-R1 antibody ( Texas red fluorescence ). In contrast to CRF-R2, CRF-R1 immunoreactivity was not identified in the spinal dorsal horn ( d ) and only scarce around the spinal central canal ( e ), whereas CRF-R1 was abundantly shown in neurons of the hypothalamus ( f ). Bar = 40 µm (A, D), Bar = 20 µm (B, C, E, F)
Article Snippet: Briefly, coronal or parasagittal spinal cord and brain sections were incubated with rabbit polyclonal antibody against CRF-R1 or CRF-R2 alone or in combination with guinea pig polyclonal antibody against CGRP (1:1000, Peninsula Laboratories, Belmont, CA, USA), mouse monoclonal antibody against ENK (1:1000) or guinea pig polyclonal antibody against MOR (1:1000, Chemicon International, MA, USA) as well as with guinea pig polyclonal antibody against CGRP in combination with rabbit anti-rat MOR (dilution of 1:1000) overnight at 4 °C.
Techniques: Immunofluorescence, Staining, Immunohistochemical staining, Fluorescence