The causative mutation in the ENU mutagenesis-derived recessive mutant mouse line HST011, a mouse line showing increased plasma urea levels as an indicator for kidney disease, was identified in the Pou3f3 gene. POU transcription factors regulate a variety of developmental processes. POU3F3 plays a role in neuronal development and is expressed in the developing neocortex, both in the late precursor cells and in the migrating neurons. A standardized, systemic phenotypic analysis of Pou3f3L423P homozygous mutant mice was carried out in the German Mouse Clinic.
Differences between wildtype and mutant mice were found in many screens and parameters. Some of these are named here.
- Compared to wild-type controls, Pou3f3L423P heterozygous mutant mice showed no differences in the guiding primary symptoms of plasma urea value and relative kidney weight.
- In the open field test mutant mice generally showed an increased locomotor activity in this environment with increased distance moved forward and increased average speed of movement.
- Body weights of mutant mice were lower than that of the controls.
- Significantly altered parameters of the SHIRPA analysis (at 8 weeks) were presence of lacrimation, presence of pelvic elevation during walking, and especially mutant mice showed trunk curl upon tail suspension and an impaired contact righting reflex when the mice were turned on their back.
- Evaluation of motor coordination and balance in three consecutive trials on the accelerating rotarod at the age of 9 weeks revealed a tendency to increased latencies (sec) on the rod in the light-weight mutants and significantly increased passive rotations. Since the mutants showed movement deficits hinting towards coordination or balance deficits, additional tests especially sensitive to balance impairments were carried out with the same group of mice at the age of 19 weeks. Balance beam, beam ladder, swim ability as well as gait analysis were performed with alterations detected in all tests.
- In mutants morphological alterations in the ear were identified. Acoustic startle reactivity and prepulse inhibition were significantly decreased in mutant mice at all sound pressure levels.
- In the hot plate assay female mutants showed a significant hypoalgesia.
- Besides increased plasma urea levels, mutants showed significantly increased values for creatinine, urea, chloride, potassium as well as alkaline phosphatase, aspartate aminotransferase and lactate dehydrogenase activity, and decreased values for glucose, albumin, total protein, cholesterol and triglycerides.
- In the hematology analysis mutants showed erythropenic anemia with a trend towards microcytosis. The metabolic cage analysis revealed that mutant mice harbored a lower body weight and showed an increased urine volume in the males as well as decreased urinary uric acid levels indicating kidney dysfunction, also together with the other parameters showing altered urine levels by tendency in both sexes when compared to the controls.
- In addition, an intraperitoneal glucose tolerance test (IpGTT) showed alterations glucose metabolism.
- Analysis of the energy metabolism revealed lower metabolic rate.
- Genome-wide transcriptome profiling of brain, kidney and spleen identified differentially expressed genes. The regulated genes in the brain are functionally associated with movement disorders, Huntington’s disease, necrosis and RNA transcription, the regulated genes in kidney with concentration of metal ions, lipids, proteins and steroids as well as renal impairment and failure and the regulated genes in spleen are functionally associated with inflammatory response including proliferation/migration/movement of leukocytes, phagocytes, granulocytes and lymphocytes.
Sudhir Kumar, Birgit Rathkolb, Elisabeth Kemter, Sibylle Sabrautzki, Dian Michel, Thure Adler, Lore Becker, Johannes Beckers, Dirk H. Busch, Lillian Garrett, Wolfgang Hans, Sabine M. Hölter, Marion Horsch, Martin Klingenspor, Thomas Klopstock, Ildikó Rácz, Jan Rozman, Ingrid Liliana Vargas Panesso, Alexandra Vernaleken, Andreas Zimmer, Helmut Fuchs, Valérie Gailus-Durner, Martin Hrabě de Angelis, Eckhard Wolf, Bernhard Aigner: Generation and Standardized, Systemic
Phenotypic Analysis of Pou3f3L423P Mutant Mice. 2016; PLOS ONE 11(3) DOI:10.1371/journal.pone.0150472