title: Presynaptic localization of a truncated isoform of the type 2 corticotropin releasing factor receptor in the cerebellum.
abstract: It is now well established that corticotropin releasing factor is present in two major excitatory afferent systems to the cerebellum, namely climbing fibers and mossy fibers. Two major classes of corticotropin releasing factor receptors, each with unique binding characteristics, have been identified as type 1 and type 2. In this study we used an antibody made to the n-terminus of the type 2 corticotropin releasing factor receptor. Characterization of this antibody showed that it strongly labeled a protein with a molecular weight of 16-32 kDa and only faintly labels a 62-83 kDa protein. The lower molecular weight protein corresponds to the weight of a recently described truncated isoform of this receptor that is designated corticotropin releasing factor-type 2alpha-truncated isoform. We carried out transfection paradigms using corticotropin releasing factor-type 2alpha-truncated isoform constructs and confirmed that the antibody recognized the truncated isoform of the type 2 corticotropin releasing factor receptor. Further, light and electron microscopic studies were carried out in mice and rats to define the distribution of the truncated receptor. Immunoreactivity is evident in the basal region of many, but not all Purkinje cell bodies and their initial axonal segments, as well as the initial axonal segments of isolated Golgi cells, and cerebellar nuclear neurons. In addition, punctate elements in the molecular layer were immunolabeled. The localization of the receptor to the initial segment of Purkinje cells was confirmed with electron microscopy. Further, the punctate labeling in the molecular layer was localized to parallel fibers and their terminals. In conclusion, evidence has been presented to show that distinct isoforms of the corticotropin releasing factor receptor are present in the cerebellum. The complex interactions between corticotropin releasing factor and other members of the corticotropin releasing factor family of peptides with both pre- and postsynaptic receptors support a growing concept that corticotropin releasing factor plays an important role in modulating activity in cerebellar circuits and ultimately in controlling motor behavior.
authors.affiliation: Department of Neuroscience, The Ohio State University, 333 West 10th Avenue, Columbus, OH 43210, USA.
authors.forename: J B
authors.initials: JB
authors.lastname: Tian
authors.forename: X
authors.initials: X
authors.lastname: Shan
authors.forename: G A
authors.initials: GA
authors.lastname: Bishop
authors.forename: J S
authors.initials: JS
authors.lastname: King
mesh_terms: D000324:Adrenocorticotropic Hormone / Q000502:physiology; D000818:Animals; D002460:Cell Line; D002531:Cerebellum / Q000166:cytology / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007668:Kidney; D009830:Olfactory Bulb / Q000502:physiology; D017729:Presynaptic Terminals / Q000502:physiology* / Q000648:ultrastructure; D016548:Prosencephalon / Q000502:physiology; D020033:Protein Isoforms; D051381:Rats; D017207:Rats, Sprague-Dawley; D018019:Receptors, Corticotropin-Releasing Hormone / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection
chemical_list: C101531:CRF receptor type 2; D020033:Protein Isoforms; D018019:Receptors, Corticotropin-Releasing Hormone; D011994:Recombinant Proteins; D000324:Adrenocorticotropic Hormone