title: Non-specific immunostaining by a rabbit antibody against gustducin α subunit in mouse brain.
abstract: Gustducin is a guanosine nucleotide-binding protein functionally coupled with taste receptors and thus originally identified in taste cells of the tongue. Recently, bitter taste receptors and gustducin have been detected in the airways, digestive tracts and brain. The existing studies showing taste receptors and gustducin in the brain were carried out exclusively on frozen sections. In order to avoid the technical shortcomings associated with frozen sectioning, we performed immunofluorescence staining using vibratome-cut sections from mouse brains. Using a rabbit gustducin antibody, we could not detect neurons or astrocytes as reported previously. Rather, we found dense fibers in the nucleus accumbens and periventricular areas. We assumed these staining patterns to be specific after confirmation with conventional negative control staining. For the verification of this finding, we stained gustducin knockout mouse brain and tongue sections with the same rabbit gustducin antibody. Whereas negative staining was confirmed in the tongue, intensive fibers were constantly stained in the brain. Moreover, immunostaining with a goat gustducin antibody could not demonstrate the fibers in the brain tissue. The present study implies a cross immunoreaction that occurs with the rabbit gustducin antibody in mouse brain samples, suggesting that the conventional negative controls may not be sufficient when an immunostaining pattern is to be verified.
authors.affiliation: Division of Neurology, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania (GX, ASC)
authors.forename: Guoxiang
authors.initials: G
authors.lastname: Xiong
authors.affiliation: Monell Chemical Senses Center, Philadelphia, Pennsylvania (KR)
authors.forename: Kevin
authors.initials: K
authors.lastname: Redding
authors.affiliation: Departments of Otorhinolaryngology - Head and Neck Surgery, University of Pennsylvania, Philadelphia, Pennsylvania (BC, NAC)
authors.forename: Bei
authors.initials: B
authors.lastname: Chen
authors.affiliation: Division of Neurology, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania (GX, ASC)|Pediatrics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania (ASC)
authors.forename: Akiva S
authors.initials: AS
authors.lastname: Cohen
authors.affiliation: Departments of Otorhinolaryngology - Head and Neck Surgery, University of Pennsylvania, Philadelphia, Pennsylvania (BC, NAC)
authors.forename: Noam A
authors.initials: NA
authors.lastname: Cohen
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D007150:Immunohistochemistry / Q000379:methods*; D008297:Male; D051379:Mice; D009714:Nucleus Accumbens / Q000166:cytology / Q000378:metabolism; D011817:Rabbits; D014059:Tongue / Q000166:cytology / Q000378:metabolism; D015293:Transducin / Q000276:immunology*
chemical_list: D000906:Antibodies; C075192:gustducin; D015293:Transducin
keywords: axonal terminal; immunohistochemistry; peptide neutralization; polyclonal antibody; taste bud