Generation and Standardized, Systemic Phenotypic Analysis of <i>Pou3f3^L423P</i> Mutant Mice

<div><p>Increased levels of blood plasma urea were used as phenotypic parameter for establishing novel mouse models for kidney diseases on the genetic background of C3H inbred mice in the phenotype-driven Munich ENU mouse mutagenesis project. The phenotypically recessive mutant line HST011 was established and further analyzed. The causative mutation was detected in the POU domain, class 3 transcription factor 3 (<i>Pou3f3</i>) gene, which leads to the amino acid exchange <i>Pou3f3</i>^<i>L423P</i> thereby affecting the conserved homeobox domain of the protein. <i>Pou3f3</i> homozygous knockout mice are published and show perinatal death. Line <i>Pou3f3</i>^<i>L423P</i> is a viable mouse model harboring a homozygous <i>Pou3f3</i> mutation. Standardized, systemic phenotypic analysis of homozygous mutants was carried out in the German Mouse Clinic. Main phenotypic changes were low body weight and a state of low energy stores, kidney dysfunction and secondary effects thereof including low bone mineralization, multiple behavioral and neurological defects including locomotor, vestibular, auditory and nociceptive impairments, as well as multiple subtle changes in immunological parameters. Genome-wide transcriptome profiling analysis of kidney and brain of <i>Pou3f3</i>^<i>L423P</i> homozygous mutants identified significantly regulated genes as compared to wild-type controls.</p></div

Neurology analysis by modified SHIRPA of line <i>Pou3f3</i>^<i>L423P</i>.

<p>Neurology analysis by modified SHIRPA of line <i>Pou3f3</i>^<i>L423P</i>.</p

Morphological analysis of the inner ears of line <i>Pou3f3</i>^<i>L423P</i>.

<p>Morphological analysis of the inner ears using the whole mount clearing method showed abnormal superior semi-circular canal formation in the homozygous mutants of line <i>Pou3f3</i>^<i>L423P</i>. Left picture shows a control inner ear with normal semi-circular canals. Right picture shows the smaller inner ear of a <i>Pou3f3</i>^<i>L423P</i> homozygous mutant animal with abnormal superior semi-circular canal formation. Red arrow shows the constriction of the superior semi-circular canal. SSCC: superior semi-circular canal, PSCC: posterior semi-circular canal, ASCC: anterior semi-circular canal.</p

Clinical chemical analysis of line <i>Pou3f3</i>^<i>L423P</i>.

<p>Clinical chemical analysis of line <i>Pou3f3</i>^<i>L423P</i>.</p

Immunohistochemical localization of POU3F3 in kidneys.

<p>POU3F3 was detected in nuclei of numerous but not all tubule segments with weak up to strong staining intensity in a wild-type kidney. The immunostaining pattern and staining intensity of POU3F3 in the kidney of a <i>Pou3f3</i>^<i>L423P</i> homozygous mutant mouse was similar to the findings observed in the wild-type control kidney. DAB (brown color); nuclear staining: haemalum (blue color). Age of mice analyzed: 22 months. Bars represent 100 μm.</p

Urine analysis of line <i>Pou3f3</i>^<i>L423P</i> (in an additional group of mice not transferred to the German Mouse Clinic).

<p>Urine analysis of line <i>Pou3f3</i>^<i>L423P</i> (in an additional group of mice not transferred to the German Mouse Clinic).</p

Dual energy X-ray absorption (DXA) analysis of bone- and weight-related parameters in line <i>Pou3f3</i>^<i>L423P</i>.

<p>Dual energy X-ray absorption (DXA) analysis of bone- and weight-related parameters in line <i>Pou3f3</i>^<i>L423P</i>.</p

Analysis of energy metabolism in line <i>Pou3f3</i>^<i>L423P</i>.

<p>Analysis of energy metabolism in line <i>Pou3f3</i>^<i>L423P</i>.</p

Hematological analysis of line <i>Pou3f3</i>^<i>L423P</i>.

<p>Hematological analysis of line <i>Pou3f3</i>^<i>L423P</i>.</p

Open field behavioral analysis of line <i>Pou3f3^L423P</i>.

<p>Open field behavioral analysis of line <i>Pou3f3^L423P</i>.</p