Prosaposin: threshold rescue and analysis of the "neuritogenic" region in transgenic mice.
Prosaposin is the precursor of four glycoprotein activators (saposins) for lysosomal hydrolases. Intact prosaposin also has lipid transfer properties in vitro as well as neuritogenic effects ex vivo and in vivo. Such "neuritogenic" effects of saposin C were evaluated in vivo using transgenic mice with prosaposin cDNAs having normal (PS-N) or mutated neuritogenic region. The mutant prosaposin cDNA (PS-CBC) encoded a chimeric saposin C that contained the non-neuritogenic sequence of saposin B, but retained acid beta-glucosidase (GCase) activation effects. When driven by the PGK (3-phosphoglycerate kinase) promoter, transgene expression was highest in the cerebrum for any of the transgenes (range from 15% to 42% of wild-type). Low levels were in visceral tissues. Prosaposin knock-out (PS-/-) mice expressing N or CBC transgenes, even at low levels, had delayed onset of neurologic signs and neuropathology, and significant lengthening of life span (from 1.7- to 7-fold) with age dependent partial correction of GlcCer and LacCer accumulation in the brain. Neuropathologic progression and neuronal glycosphingolipid storage were related directly to the transgene expression levels in the brain. Purkinje cell loss was age dependent. Gross brain and neuronal organizations were indistinguishable in PS-/- mice with or without the various transgenes, albeit the phenotype appeared later in the mice with transgenes. These studies show the degree of neuropathologic manifestations in each transgenic line depended on expression level rather than on the nature of the transgene. These studies also show in vivo localization of the GCase activation region to the carboxy terminal half of saposin C and the lack of a significant gross trophic effect of saposin C on CNS organization in vivo.
Sun
Ying
Y
The Division and Program in Human Genetics, Children's Hospital Research Foundation, Cincinnati, OH 45229-3039, USA.
Qi
Xiaoyang
X
Witte
David P
DP
Ponce
Elvira
E
Kondoh
Keiji
K
Quinn
Brian
B
Grabowski
Gregory A
GA
D000818:Animals; D001921:Brain / Q000473:pathology; D018076:DNA, Complementary; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D006028:Glycosphingolipids / Q000378:metabolism; D017403:In Situ Hybridization; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D004676:Myelin Basic Protein / Q000235:genetics*; D010525:Peripheral Nerves / Q000473:pathology; D011498:Protein Precursors / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D049231:Saposins; D013116:Spinal Cord / Q000473:pathology
D018076:DNA, Complementary; D006023:Glycoproteins; D006028:Glycosphingolipids; D004676:Myelin Basic Protein; D011498:Protein Precursors; C483344:Psap protein, mouse; D049231:Saposins