title: Immunohistochemical detection of the neuronal connexin36 in the mouse central nervous system in comparison to connexin36-deficient tissues.
abstract: Investigating the spatial and temporal expression of connexin36 (Cx36) protein in neuronal tissue is of prime importance to understand the molecular mechanisms underlying extensive electrical coupling. Although Cx36 mRNA was shown to be expressed in neurons of the central nervous system in different studies, only the determination of Cx36 protein expression allows a correlation between localization and its functional role in gap junction-mediated neuronal coupling. After the initial use of antibodies recognizing the skate connexin35 protein, antibodies directed to the mammalian Cx36 sequence allowed the detailed investigation of Cx36 cellular localization. However, results on Cx36 protein distribution still remained controversial in some areas of the central nervous system. In the present study, we have investigated: (a) the distribution of Cx36 protein in various areas of the central nervous system and (b) determined the specificity in the immunohistochemical staining of two polyclonal antibodies comparing wildtype and Cx36-deficient mice. In some areas of the central nervous system, for example in the retina and the inferior nuclear olivary complex, Cx36 antibodies were highly specific, and in the cerebellar cortex, Cx36 protein expression was partly specific. In other regions, particularly in pyramidal cells of the hippocampal formation, non-specific staining was prevalent, indicating that Cx36 antibodies also recognize proteins other than Cx36 in these tissues. The present results argue for a re-evaluation of many documented immunohistochemical protein distribution patterns and require, not only in connexin research, their assessment using null-mutant animals.
authors.affiliation: Department of Neuroanatomy and Molecular Brain Research, Ruhr-University Bochum, Universitätsstrasse 150, MA6/159, Germany. Carola.Meier@ruhr-uni-bochum.de
authors.forename: Carola
authors.initials: C
authors.lastname: Meier
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Petrasch-Parwez
authors.forename: Hans-Werner
authors.initials: HW
authors.lastname: Habbes
authors.forename: Barbara
authors.initials: B
authors.lastname: Teubner
authors.forename: Martin
authors.initials: M
authors.lastname: Güldenagel
authors.forename: Joachim
authors.initials: J
authors.lastname: Degen
authors.forename: Goran
authors.initials: G
authors.lastname: Söhl
authors.forename: Klaus
authors.initials: K
authors.lastname: Willecke
authors.forename: Rolf
authors.initials: R
authors.lastname: Dermietzel
mesh_terms: D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D001923:Brain Chemistry; D002490:Central Nervous System / Q000737:chemistry / Q000166:cytology*; D017630:Connexins / Q000172:deficiency / Q000276:immunology / Q000378:metabolism*; D003429:Cross Reactions; D005136:Eye Proteins / Q000276:immunology; D007150:Immunohistochemistry; D051379:Mice; D009474:Neurons / Q000737:chemistry*; D012160:Retina / Q000737:chemistry; D014018:Tissue Distribution; D000097004:Gap Junction delta-2 Protein
chemical_list: D000906:Antibodies; D017630:Connexins; D005136:Eye Proteins; D000097004:Gap Junction delta-2 Protein; C100585:connexin 35 protein, vertebrate