The PKD Rodent Model Drug-Testing Core will provide reagents and services to Kidney Institute investigators and to investigators at KUMC and other institutions in established PKD labs or labs new to the PKD field. A number of mouse PKD lines are available for breeding experiments and for tissue harvest. The Core will also provide expertise and experimental support in handling PKD mouse models and in processing and analyzing tissue specimens.
Many of the reagents required for mouse PKD studies are difficult to acquire and/or expensive to maintain. The PKD Rodent Model Drug-Testing Core will increase the efficiency of PKD research in the KI by providing mouse lines, experimental expertise, and tissues to new and established PKD investigators.
Dr. James Calvet, PhD - Director
Brenda Magenheimer, Senior Scientist - Core Co-Director
Use of the Core will be arranged on a case-by-case basis. Requests should be submitted by email to firstname.lastname@example.org and cc'd to email@example.com. Consideration will be based on the rationale and feasibility of the study, IACUC approval, time commitment required for the service, and costs. Arrangements can be made to transfer mice and/or provide tissues, and/or can involve collaboration.
There are two routes a user can take to obtain mouse lines or tissues. 1) Request transfer of breeders to an IACUC approved protocol, or 2) Submit an addendum to the Core Director's ACUP to engage in a collaborative study.
It is costly for individual labs to maintain mouse colonies only for intermittent use, and it is often difficult to obtain mouse lines from an outside source to pilot-test an idea. It is usually a time-consuming process to bring new mice to KUMC and it is always a long wait before they can be used in experiments. The PKD Mouse and Tissue Resource Core maintains several commonly used mutant mouse lines. These lines can be rapidly expanded, making it possible to start experiments with minimal delay.
Many experiments require expertise from experienced users of mouse models. The Core will provide advice on mouse husbandry, breeding, colony maintenance, record-keeping, genotyping, tissue harvest, and phenotyping, including help with cutting and processing frozen tissue sections. The Core will also provide embryonic mouse kidneys for metanephric organ culture and/or expert guidance in doing experiments with embryonic kidneys. The Core will also help users design and implement drug treatment studies using mouse models. Core users will have the option of learning procedures for their own use or having the Core do experiments on a collaborative basis.
PKD Mouse Lines in the Core
The Core currently has four Pkd1 mutant mouse lines (and has the potential to generate others).
This mouse was developed by David Beier by ENU mutagenesis (Herron et al. Nat Genet. 2002 30:185-189). The mutation causes a single amino acid nonconservative substitution in the first transmembrane domain of polycystin-1, acting as a null. This cryo-archived mouse is currently available from the MMRRC, but would take 3-4 months to ship and currently costs $2,022. A breeding colony is currently maintained in the Core on a C57 background. Its pathogenic mechanism is currently being investigated by the Core Director in a collaborative study.
Pkd1 deltaL and Pkd1 NEO
This mouse was developed at KUMC by the Core Director's lab (publication pending) by knock-in mutagenesis, causing a single leucine deletion in the C-terminal tail of polycystin-1 mimicking a PKD patient mutation. While protein production is normal, the mutation acts as a null. The pathogenic mechanism appears to be disruption of heterotrimeric G-protein signaling. A Pkd1 deltaL precursor mouse line, Pkd1 NEO, is also maintained by the Core. These mice retain a Neo selectable cassette in Pkd1 intron 45 that disrupts splicing, causing no protein production. Both mice will be freely available to Kidney Institute and KUMC investigators (pending publication of the initial findings) on a collaborative basis. Both Pkd1 deltaL and Pkd1 NEO mouse lines are being stabilized on a C57 background.
Pkd1 m1bei has been bred to other mutant mouse lines, which are being maintained as double heterozygotes.
Pkd1B (Pkd1 m1Bei)
Pkd1B:Pkd2 null (Pkd2tm2Som)
Pkd1B:Cftr (Cftrm1unc) (S489X)
Pkd1B:Nfatc2 (Nfatc2tm1Glm) (on a Balb/c background)
Pkd1 deltaL has been bred with a floxed Pkd1 conditional mouse line together with Hoxb7-Cre for postnatal expression of the PKD phenotype.
Pkd1 deltaL (deltaL)
Pkd1 NEO (deltaL containing the Neo selectable cassette in Pkd1 intron 45)
Pkd1 deltaL:Pkd1C (conditional):Hoxb7-Cre (Pkd1tm2Ggg)
The following mouse lines are also being maintained.
EIIa-Cre Tg (EIIa-Cre) C5379Lmgd
Hoxb7-Cre [stock Tg (Hoxb7-Cre) 13Amc/J]
Nfat-Luc reporter 9X Tg (Myh6/NFAT-Luc) 1Jmol
The Core will consider maintaining additional PKD mutant mouse cell lines with mutations in Pkd1 or Pkd2, or other recessive mouse models of PKD . If costs are deemed too high for the Core budget, users will be requested to consider working with the Core Director to fashion a pilot grant proposal. The Core Director will also help in the design and submission of an addendum for submission to IACUC, or any necessary MTA's for permission to use the mice.