Alternative splicing of a protein domain indispensable for function of transient receptor potential melastatin 3 (TRPM3) ion channels.

TRPM3 channels form ionotropic steroid receptors in the plasma membrane of pancreatic ß and dorsal root ganglion cells and link steroid hormone signaling to insulin release and pain perception, respectively. We identified and compared the function of a number of TRPM3 splice variants present in mouse, rat and human tissues. We found that variants lacking a region of 18 amino acid residues display neither Ca(2+) entry nor ionic currents when expressed alone. Hence, splicing removes a region that is indispensable for channel function, which is called the ICF region. TRPM3 variants devoid of this region (TRPM3ΔICF), are ubiquitously present in different tissues and cell types where their transcripts constitute up to 15% of the TRPM3 isoforms. The ICF region is conserved throughout the TRPM family, and its presence in TRPM8 proteins is also necessary for function. Within the ICF region, 10 amino acid residues form a domain essential for the formation of operative TRPM3 channels. TRPM3ΔICF variants showed reduced interaction with other TRPM3 isoforms, and their occurrence at the cell membrane was diminished. Correspondingly, coexpression of ΔICF proteins with functional TRPM3 subunits not only reduced the number of channels but also impaired TRPM3-mediated Ca(2+) entry. We conclude that TRPM3ΔICF variants are regulatory channel subunits fine-tuning TRPM3 channel activity.

Accuracy of craniofacial measurements: computed tomography and three-dimensional computed tomography compared with stereolithographic models.

In a retrospective study, distance measurements of nine children with craniofacial malformation were analyzed. The accuracy of measurements was compared when measured on a workstation using a 16-slice multidetector spiral computed tomography and on a stereolithographic model. Three different methods of defining distances were investigated: 1) on the stereolithographic plastic models, 14 distances connecting landmarks were identified with a digitizer (Polaris Tracker); 2) the same distances were defined at axial, coronal, and sagittal reformats of the computed tomography data set and measured using a Philips MX View workstation; and 3) the same 14 distances were defined at three-dimensional virtual reality models of the skulls at the same workstation. All measurements were performed with all three methods by three different readers. The following conclusions could be drawn: stereolithographic models provide a highly exact reproduction of the skull in children with craniofacial malformations. They are a reliable basis for all analytic and probatory endeavors preparing complicated surgical corrections. Three-dimensional virtual reality display modes serve significantly better for exact distance measurements on the complex surface of the human skull than planar reformats of the same computed tomography data sets.

Comparison of left and right ventricular volume measurement using the Simpson's method and the area length method.

PURPOSE: To compare ventricular volume measurement using a volumetric approach in the three standard cardiac planes and ventricular volume estimation by a geometrical model, the Area-Length method (ALM). MATERIALS AND METHODS: Fifty-six healthy volunteers were examined (27 males, 29 females) on a 1.5T MR-unit with ECG-triggered steady state free precision (SSFP) Cine-MR sequences and parallel image acquisition. Multiple slices in standardized planes including the short-axis view (sa), 4-chamber view (4ch), left and right 2-chamber views (2ch) were used to cover the whole heart. End-systolic and end-diastolic ventricular volumes (EDV, ESV), stroke volume (SV), and ejection fraction (EF) were calculated with Simpson's rule in all planes and with ALM in the 2ch and 4ch planes. Global function parameters measured in the sa plane were compared with those obtained in the other imaging planes. RESULTS: A very good correlation is observed when comparing functional parameters calculated with Simpson's rule in all imaging planes: for instance, the mean EDV/ESV of the left and right ventricle of the female population group measured in sa, 4ch, and 2ch: left ventricle EDV/ESV 114.3/44.4, 120.9/46.5, and 117.7/45.3 ml; right ventricle EDV/ESV 106.6/46.0, 101.2/41.1, and 103.5/43.0 ml. Functional parameters of the left ventricle calculated with ALM in 2ch and 4ch correlate to parameters obtained in sa with Simpson's rule in the range of 5-10%: for instance, the EDV/ESV of the left ventricle of the male population group measured in the sa, 4ch, and 2ch: 160.3/63.5, 163.1/59.0, and 167.0/65.7 ml. Functional parameters of the right ventricle measured with ALM in 4ch are 40-50% lower and calculated in 2ch almost double as high as compared with the parameters obtained in sa with Simpson's rule: for instance, male right ventricular EDV/ESV measured in sa, 4ch, and 2ch: 153.4/68.1, 97.5/34.5, and 280.2/123.2 ml. The EF correlates for all imaging planes measured with the Simpson's rule in both ventricles and using ALM in the left ventricle except for males with an overestimation of less than 6%. The EF of the right ventricle is calculated higher using ALM in 4ch and 2ch compared to the EF calculated in sa: female/male EF of the right ventricle measured in the sa, 4ch, and 2ch: 56.8/55.7, 66.0/65.0, and 60.0/57.0%. CONCLUSION: In the setting of healthy volunteers the ALM method should not be used in 2ch and 4ch planes of the right ventricle because of lacking correlation of global functional parameters compared to those obtained in the sa plane. Using Simpson's rule functional parameters correlate well to each other in the different imaging planes.