Morphometric Study Of The Skull In SLC28A3 Knockout Mice Based On EDMA

ObjectiveThe SLC28A3 gene encodes a nucleoside transporter protein that is primarily involved in transmembrane transport of purine and pyrimidine nucleosides in a variety of cellular processes in vivo and is essential for cell and tissue growth.Whole-exome sequencing studies have identified mutations at the rs10868138（NM₀01199638）locus of SLC28A3 that are closely associated with four facial phenotypes and one cranial phenotype in humans.In this study,we quantified the cranial morphology based on SLC28A3 knockout and wild-type C57BL/6 mouse models by Euclidean distance matrix analysis to investigate the effect of SLC28A3 knockout on the cranial morphology of mice.MethodsSLC28A3+/-mice purchased from Saiye Biotechnology Co.,Ltd.were paired and bred,and their offspring were genotyped by PCR and gene sequencing after expansion.Eight 8-week-old male SLC28A3^-/-mice,SLC28A3+/-mice and WT mice were selected and their skulls were scanned by Micro-CT,and the 3D reconstruction of the skulls was performed using VG studio 3.2 MAX software to collect the 3D coordinate data of biologically relevant markers on the skulls.Win EDMA 1.1 software was used to quantify and compare the cranial morphological differences of these marker points based on EDMA.ResultsThe results of the overall Euclidean distance matrix analysis of the mouse skull in each group were as follows: SLC28A3^-/-/WT group（T=1.49,P=0.6）,SLC28A3+/-/WT group（T=1.35,P=0.35）,SLC28A3^-/-/SLC28A3+/-group（T=1.67,P=0.37）;cranial left-right asymmetry in each group Analysis results: SLC28A3^-/-group（T=1.28,P=0.354）,SLC28A3+/-group（T=1.14,P=0.40）in WT group mice（T=1.06,P=0.62）;local Euclidean distance matrix analysis: nasal bone region SLC28A3^-/-/WT group（T=1.33,P=0.04<0.05）Difference line segments <0.95 were lmax-lmaxna,rmax-rmaxna,nsl-lmaxna,rmax-lmaxna,nsl-rmaxna,lmax-rmaxna,ids-lmaxna,nas-lpln,ids-rmaxna,nas-lmax,nas-rmax rmax-rpln,nsl-nas,lmax-rpln,with difference line segments >1.05 for lmaxna-lpln,nsl-rmax,ids-nsl.SLC28A3+/-/WT group（T=1.25,P=0.10）,SLC28A3^-/-/SLC28A3+/-group（T=1.23,P=0.26）;frontotemporal bone region SLC28A3^-/-/WT group（T=1.21,P=0.14）,SLC28A3+/-/WT group（T=1.22,P=0.23）,SLC28A3^-/-/SLC28A3+/-group（T=1.15,P=0.35）;parietal bone region SLC28A3^-/-/WT group（T=1.10,P=0.13）,SLC28A3+/-/WT group（T=1.07,P=0.58）,SLC28A3^-/-/SLC28A3+/-group（T=1.10,P=0.12）;SLC28A3^-/-/WT group in the parietal bone region（T=1.08,P=0.10）,SLC28A3+/-/WT group（T=1.06,P=0.09）,SLC28A3^-/-/SLC28A3+/-group（T=1.09,P=0.033<0.05）Difference line >1.05 for pari-rpto.Conclusions1、Euclidean distance matrix analysis,SLC28A3 knockout mice showed no statistically significant difference in overall cranial morphology compared with WT mice;there was no significant asymmetry between the left and right sides of the mice’s cranium.2、SLC28A3 knockout had some effect on the local morphology of mouse skull.Compared with WT group mice,the local Euclidean distance matrix analysis showed that the morphology of part of the nasal bone in SLC28A3^-/- group mice was changed,the length of nasal bone in anterior-posterior direction was slightly shorter,and the distance from the anterior nasal spine to the intersection of nasal bone in the midline was slightly longer;compared with SLC28A3+/-group mice,the intersection of parietal bone and midpoint of parietal bone to the intersection of right parietal bone,temporal bone and occipital bone in SLC28A3^-/-group mice was longer.