Transgenic animal model of mood disorders
A non-human transgenic animal having a polynucleotide encoding a PTN polypeptide, which polynucleotide is operably linked to a promoter, wherein said transgenic animal has greater than wild-type expression of the PTN polypeptide in at least one brain region, as well as related vectors, methods of producing transgenic animals, in vitro and in vivo screening methods for potential therapeutic agents, and methods for treating and diagnosing neuropsychiatric illnesses, particularly anxiety and depression, are disclosed.
1. A transgenic mouse whose genome comprises a polynucleotide encoding a Pleiotrophin (PTN) polypeptide, which polynucleotide is operably linked to a neuron-specific promoter, wherein said transgenic mouse has greater than wild-type expression of the PTN polypeptide in at least one brain region, and wherein said neuron-specific promoter is selected from the group consisting of: a Thy1 gene promoter, Neuron specific enolase (NSE) gene promoter; Rhombotin I gene promoter; PGK gene promoter; Neurofilament Low (NF-L) gene promoter; dopamine beta-hydroxylase (DBH) gene promoter; and Synapsin-1 gene promoter, and wherein said transgenic mouse exhibits at least one behavior selected from the group consisting of reduced time spent in open arms of an elevated plus maze; reduced time in central area in an open-field test; reduced time in light in a light-dark box test; increased latency to feed in a Novelty Suppressed Feeding Test; increased immobility time in tail suspension test; and reduced sucrose intake in sucrose intake task.
2. The transgenic mouse according to claim 1 , wherein said polynucleotide encodes:
(i) a PTN polypeptide having an amino acid sequence having at least 80% amino acid sequence identity to the sequence of SEQ ID NO: 2;
(ii) a PIN polypeptide having the amino acid sequence of SEQ ID NO: 2
(iii) a PIN polypeptide having an amino acid sequence having at least 80% amino acid sequence identity to the sequence of SEQ ID NO: 4; or
(iv) a PIN polypeptide having the amino acid sequence of SEQ ID NO: 4 and wherein said PIN polypeptide of any one of (i)-(iv) is capable of binding to specific receptors RPTPβ/ζ, ALK or Syndecan 3 polypeptide.
3. The transgenic mouse according to claim 1 , wherein the neuron-specific promoter is a Thy1 gene promoter.
4. The transgenic mouse according to claim 3 , wherein the Thy1 promoter comprises a polynucleotide having at least 80% nucleic acid sequence identity to the sequence of SEQ ID NO: 5.
5. The transgenic mouse according to claim 1 , wherein said at least one brain region is selected from: cortex and hippocampus.
6. The transgenic mouse according to claim 1 , having at least 100% greater expression of the PTN polypeptide in said at least one brain region, as measured by Western blot, immunofluorescence and/or qPCR of an PTN mRNA.
7. The transgenic mouse according to claim 1 , wherein the mouse is a hybrid B6/SJL-F1J mouse.
8. An in vivo method for identifying an agent comprising:
administering a test agent to the transgenic mouse of claim 1 and subsequently assessing the presence and/or severity of one or more behaviours in the transgenic mouse relative to the same one or more behaviours in a control transgenic mouse, which has not been exposed to the test agent,
wherein said behaviours are selected from the group consisting of reduced motor activity in an open-field test; reduced time spent in open arms of an elevated plus maze; reduced time spend in the lit portion of a light-dark box; increased latency to feed in a Novelty Suppressed Feeding test; increased immobility time in tail suspension test; and decreased sucrose intake, and wherein the test agent is found to reduce the presence and/or severity of said one or more behaviours.
9. The method according to claim 8 , which further comprises isolating the test agent and, optionally, formulating the test agent into a pharmaceutical composition with at least one pharmaceutically acceptable salt, carrier or excipient.