Parkinson's disease model and methods
This application provides a novel mouse model (PLA2g6 KO Ex2 ) in which genetic deletion of the N terminus of PLA2g6 results in a loss of dopaminergic (DA) neurons in substantia nigra (SN), and development of PD-like motor deficits that can be significantly improved by L-DOPA. Based in part on experimental results demonstrated with this model, this disclosure provides genetically modified animals and genetically modified animal cells that comprise a mutant allele of PLA2g6 and in which store-operated Ca 2+ entry (SOCE) is impaired and ER Ca 2+ stores are depleted. This disclosure also provides methods of screening a compound for an effect on the SOCE pathway and/or ER Ca 2+ by administering the compound to such a genetically modified animal or genetically modified animal cell. This disclosure also provides methods of treating or preventing PD-related deficit(s) in an animal by characterizing a compound as a SOCE activator using the screening methods and then administering an effective amount of the compound to an animal. This disclosure also provides methods of restoring normal store-operated Ca 2+ entry (SOCE) pathway and ER Ca 2+ in a cell, comprising introducing a caspase-3 cleavage-resistant PLA2g6 protein into the cell. This disclosure also provides methods of treating or preventing a PD-related deficit(s) in an animal, comprising administering a caspase-3 cleavage-resistant PLA2g6 protein to the animal.
1. A genetically modified mammalian cell comprising: a first recombinant allele of PLA2g6 that comprises a deletion of exon 2 and encodes a catalytically active PLA2g6 protein deleted for the N-terminal 178 amino acids of the protein; and a second recombinant allele of PLA2g6 that comprises a deletion of exon 2 and encodes a catalytically active PLA2g6 protein deleted for the N-terminal 178 amino acids of the protein;
wherein store-operated Ca 2+ entry (SOCE) and activation of SOCE by depletion of endoplasmic reticulum (ER) Ca 2+ stores are impaired in the genetically modified mammalian cell.
2. The genetically modified mammalian cell of claim 1 , wherein the first and second recombinant alleles of PLA2g6 are the same.
3. The genetically modified mammalian cell of claim 1 , wherein the mammalian cell is a human cell.
4. The genetically modified mammalian cell of claim 1 , wherein the mammalian cell is a mouse cell.
5. The genetically modified mammalian cell of claim 2 , wherein the mammalian cell is a mouse cell.
6. A method of screening a compound for an effect on the SOCE pathway, comprising:
providing the compound to a genetically modified mammalian cell according to claim 1 ; and
determining the effect of the compound on SOCE in the animal cell.
7. The method of claim 6 , wherein the compound activates SOCE in the genetically modified mammalian cell and is thereby identified as a SOCE activator.
8. A genetically modified mammalian cell comprising: a first recombinant allele of PLA2g6 that encodes a catalytically active PLA2g6 protein that comprises an N-terminal deletion of from 151 to 178 amino acids of the protein; and a second recombinant allele of PLA2g6 that encodes a catalytically active PLA2g6 protein that comprises an N-terminal deletion of from 151 to 178 amino acids of the protein;
wherein store-operated Ca 2+ entry (SOCE) and activation of SOCE by depletion of endoplasmic reticulum (ER) Ca 2+ stores are impaired in the genetically modified mammalian cell.
9. The genetically modified mammalian cell of claim 8 , wherein the first and second recombinant alleles of PLA2g6 are the same.