title: Expression of Niemann-Pick type C transcript in rodent cerebellum in vivo and in vitro.
abstract: This study assesses the developmental expression of the Niemann-Pick type C mRNA in vivo and in vitro in rat cerebellum. NPC is an autosomal recessive neurovisceral lipid storage disease associated with an alteration in cholesterol trafficking. In the mouse model of NPC and in the early onset form of human NPC, Purkinje neurons are among the first neurological targets, suffering stunted growth during postnatal development and dying, leading to ataxia. Recently, the genes responsible for human (NPC1) and mouse (Npc1) NPC disease have been cloned. Based on a highly homologous domain, we designed primers to look for levels of Npc1 mRNA with a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) approach using cyclophilin as an internal standard. Total RNA was isolated from various postnatal developmental stages of the rat cerebellum as template for the analyses. Npc1 transcripts were observed at postnatal day 0 and at later stages of development, both in vivo and in vitro from primary cerebellar cultures. To identify the location of Npc1 inside the cerebellum, we performed immunostaining with an anti-Npc1 antibody in primary rat cerebellar cultures identifying reactive Purkinje neurons by double-labeling with the Purkinje specific marker calbindin and sub-populations of glial cells. In summary, Npc1 is expressed in rat cerebellum in vivo and in vitro and is expressed during early postnatal development as well as in the adult cerebellum. Since Npc1 is expressed at similar levels throughout development, the vulnerability of Purkinje neurons to this disease is likely to involve disruption of an interaction with other developmentally-regulated proteins.
authors.affiliation: Department of Physiology, University of Arizona College of Medicine, Tucson, AZ 85724-5051, USA. tfalk@u.arizona.edu
authors.forename: T
authors.initials: T
authors.lastname: Falk
authors.forename: W S
authors.initials: WS
authors.lastname: Garver
authors.forename: R P
authors.initials: RP
authors.lastname: Erickson
authors.forename: J M
authors.initials: JM
authors.lastname: Wilson
authors.forename: A J
authors.initials: AJ
authors.lastname: Yool
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D064026:Calbindins; D002478:Cells, Cultured; D002531:Cerebellum / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D008818:Mice, Neurologic Mutants; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics; D000077296:Niemann-Pick C1 Protein; D009542:Niemann-Pick Diseases / Q000235:genetics* / Q000473:pathology; D019696:Peptidylprolyl Isomerase / Q000235:genetics; D011689:Purkinje Cells / Q000378:metabolism / Q000473:pathology; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D064030:S100 Calcium Binding Protein G / Q000235:genetics; D017386:Sequence Homology, Amino Acid
chemical_list: D064026:Calbindins; D008562:Membrane Glycoproteins; C573152:NPC1 protein, rat; D009419:Nerve Tissue Proteins; D000077296:Niemann-Pick C1 Protein; D064030:S100 Calcium Binding Protein G; D019696:Peptidylprolyl Isomerase