Tetsutaro Yamaguchi

A Common Variation in EDAR Is a Genetic Determinant of Shovel-Shaped Incisors

Shovel shape of upper incisors is a common characteristic in Asian and Native American populations but is rare or absent in African and European populations. Like other common dental traits, genetic polymorphisms involved in the tooth shoveling have not yet been clarified. In ectodysplasin A receptor (EDAR), where dysfunctional mutations cause hypohidrotic ectodermal dysplasia, there is a nonsynonymous-derived variant, 1540C (rs3827760), that has a geographic distribution similar to that of the tooth shoveling. This allele has been recently reported to be associated with Asian-specific hair thickness. We aimed to clarify whether EDAR 1540C is also associated with dental morphology. For this purpose, we measured crown diameters and tooth-shoveling grades and analyzed the correlations between the dental traits and EDAR genotypes in two Japanese populations, inhabitants around Tokyo and in Sakishima Islands. The number of EDAR 1540C alleles in an individual was strongly correlated with the tooth-shoveling grade (p = 7.7 x 10(-10)). The effect of the allele was additive and explained 18.9% of the total variance in the shoveling grade, which corresponds to about one-fourth of the heritability of the trait reported previously. For data reduction of individual-level metric data, we applied a principal-component analysis, which yielded PC1-4, corresponding to four patterns of tooth size; this result implies that multiple factors are involved in dental morphology. The 1540C allele also significantly affected PC1 (p = 4.9 x 10(-3)), which denotes overall tooth size, and PC2 (p = 2.6 x 10(-3)), which denotes the ratio of mesiodistal diameter to buccolingual diameter.

Effects of an Asian-specific nonsynonymous EDAR variant on multiple dental traits.

Dental morphology is highly diverse among individuals and between human populations. Although it is thought that genetic factors mainly determine common dental variations, only a few such genetic factors have been identified. One study demonstrated that a nonsynonymous single-nucleotide polymorphism (370V/A, rs3827760) in the ectodysplasin A receptor gene (EDAR) is associated with shoveling and double-shoveling grades of upper first incisors and tooth crown size. Here, we examined the association of EDAR 370V/A with several dental characters in Korean and Japanese subjects. A meta-analysis that combined analyses of Korean and Japanese subjects revealed that the Asian-specific 370A allele is associated with an increase in the grades of shoveling and double shoveling, as previously found. We also showed a highly significant association between EDAR 370V/A genotype and crown size, especially mesiodistal diameters of anterior teeth. Moreover, we found that the 370A allele was associated with the presence of hypoconulids of lower second molars. These results indicated that the EDAR polymorphism is responsible, in part, for the Sinodonty and Sundadonty dichotomy in Asian populations, and clearly demonstrated that the EDAR polymorphism has pleiotropic effects on tooth morphology. As the 370A allele is known to be a most likely target of positive selection in Asian populations, some phenotypes associated with the variant may be ‘hitchhiking phenotypes’, while others may be actual targets of selection.

Exome resequencing combined with linkage analysis identifies novel PTH1R variants in primary failure of tooth eruption in Japanese

Massively parallel sequencing of target regions, exomes, and complete genomes has begun to increase the opportunities for identifying genetic variants underlying rare and common diseases dramatically. Here we applied exome resequencing to primary failure of tooth eruption (PFE) to identify the genetic causality of the disease. Two Japanese families having PFE were recruited and examined by genome‐wide linkage study and subsequently exome analyses. Linkage analyses of these two families comprising eight affected individuals and two unaffected individuals revealed linkage signals at 10 loci with a maximum LOD score of 1.5. Four affected individuals in one family were pooled and further processed for exome analysis, followed by massive parallel sequencing. After three‐step filtering including annotation and functional expectation, three variants were found to be candidates for PFE. Among the three variants, only a novel variant of parathyroid hormone 1 receptor gene (PTH1R), R383Q, was cosegregated in the first PFE family. Accordingly, we screened the gene for variants at all coding exons and the respective intron‐exon boundaries in the second family and two sporadic individuals with PFE. We also identified a novel missense variant, P119L, cosegregating in the second family and missense variants P132L and R147C in the sporadic cases. These variants all were in the highly conserved region across zebrafish to chimpanzee and not observed in 192 unrelated controls, supporting the pathogenicity of the variants. The combination of linkage and exome analyses employed in this study provides a powerful strategy for identifying genes responsible for Mendelian disorders.

Effects of mastication on mandibular growth evaluated by microcomputed tomography.

It is well known that mastication has a significant influence on mandibular growth and development, but the mechanism behind this effect has not yet been clarified. Furthermore, no studies have examined the effects of changes in mastication on the three-dimensional (3D) morphometry of the mandible. The aim of the present study was to investigate the influences of changes in mastication on mandibular growth and morphology. Twenty-five 3-week-old (at the time of weaning) imprinting control region mice were randomly divided into three groups: mice fed a hard diet (HD), mice fed a soft diet (SD), and mice alternately fed hard and soft diets (HSDs) every week for 4 weeks. The morphometry of the mandible was analysed using 3D microcomputed tomography (muCT). Statistical analysis was undertaken using a t-test. muCT analysis showed that the condylar width was significantly greater in the HD group than in the SD group after 1 week. After 4 weeks, mandibular length was significantly longer and ramus height was greater in the HSD group than in the other two groups. Bone volume was significantly less in the SD group than in the other two groups after 4 weeks. These findings suggest that changes in mastication markedly affect mandibular condylar cartilage growth and mandibular morphology. It is considered that dietary education at an early age is important in order to prevent disruption of the development of the mandible.

Further evidence for an association between mandibular height and the growth hormone receptor gene in a Japanese population

INTRODUCTION Craniofacial morphology has a significant genetic component. It has recently been reported that single nucleotide polymorphisms (SNPs) in the growth hormone receptor gene (GHR) are associated with mandibular height. To confirm these findings, we genotyped SNPs in healthy Japanese subjects, about 1.7 times the number analyzed in previous reports. METHODS By using 5 SNPs in exon 10 of the GHR, we studied the relationships between genotypes and craniofacial linear measurements. The allelic frequencies in a multi-ethnic population (Han Chinese, African American, European American, and Hispanic) were also observed. RESULTS There was a significant association between SNPs and mandibular ramus height (P = 0.029), confirming our previous report of an association between polymorphism P561T and mandibular ramus height. Moreover, the SNP, C422F, was in linkage disequilibrium with P561T. Subjects with genotype CC of polymorphism P561T and genotype GG of polymorphism C422F had significantly greater mandibular ramus height than those with genotypes CA and GT. Four of the 5 SNPs were found almost exclusively in Asians, with the frequencies in other populations extremely small. CONCLUSIONS Our results indicate that the GHR polymorphisms P561T and C422F are associated with mandibular ramus height in Japanese population and suggest that the SNPs of the GHR associated with the Japanese are likely to be different in other ethnic groups. This might partly explain the differing craniofacial morphology among different ethnicities.

Association of the growth hormone receptor gene polymorphisms with mandibular height in a Korean population

Growth hormone receptor gene (GHR) is one of the likely candidates for determining morphological traits, because GH is a key regulator of bone growth. The genetic association of GHR in exon 10 with mandibular ramus height has been found in different populations, Japanese and Chinese. On the other hand, two common isoforms of GHR, one full-length (fl-GHR) and the other lacking the extracellular domain encoded by exon 3 (d3-GHR), are associated with differences in responsiveness to GH. The purpose of this study involving 159 Korean subjects was to study the associations between a GHR polymorphism (d3/fl-GHR) that results in genomic deletion of exon 3 and craniofacial morphology, and to study the associations between GHR genotypes in exon 10 and craniofacial morphology. Moreover, the allelic frequencies in a multi-ethnic population (24 Han Chinese, 24 African-Americans, 24 European-Americans, and 24 Hispanics) in a GHR polymorphism (d3/fl-GHR) were compared in this study. The five craniofacial linear measurements (cranial base length, maxillary length, overall mandibular length, mandibular corpus length, and mandibular ramus height) obtained from lateral cephalograms were examined as craniofacial morphology. We found that the d3/fl-GHR polymorphism had no association for any measurements, and a statistically significant association (P=0.024) between the GHR polymorphisms P561T and C422F in exon 10 and mandibular ramus height. Neither SNPs besides P561T and C422F polymorphisms in exon 10 nor the measurements besides mandibular ramus height have statistically significances. Both derived alleles at P561T and C422F SNPs were highly associated with only one haplotype, haplotype-4 in Korean population. As quantitative haplotype association, the results showed a significant difference in mandibular ramus height between individuals having one haplotype-4 and others without haplotype-4 (P=0.028). Moreover, we found that the d3/fl-GHR polymorphism showed diverse frequency in different population. Regarding GHR genotypes in exon 10, the present study mostly reflected the results obtained for a Japanese population, although our current study does not replicate the correlation between the I526L polymorphism of GHR and mandibular ramus height as was reported in a previous study of Han Chinese. The results of the present study suggest that the GHR exon 10 SNPs, not d3/fl-GHR, contribute to changes in the mandibular ramus height of Koreans.

Genome-wide SNP analysis reveals population structure and demographic history of the Ryukyu Islanders in the southern part of the Japanese Archipelago

The Ryukyu Islands are located to the southwest of the Japanese archipelago. Archaeological evidence has revealed the existence of prehistoric cultural differentiation between the northern Ryukyu islands of Amami and Okinawa, and the southern Ryukyu islands of Miyako and Yaeyama. To examine a genetic subdivision in the Ryukyu Islands, we conducted genome-wide single nucleotide polymorphism typing of inhabitants from the Okinawa Islands, the Miyako Islands, and the Yaeyama Islands. Principal component and cluster analyses revealed genetic differentiation among the island groups, especially between Okinawa and Miyako. No genetic affinity was observed between aboriginal Taiwanese and any of the Ryukyu populations. The genetic differentiation observed between the inhabitants of the Okinawa Islands and the Miyako Islands is likely to have arisen due to genetic drift rather than admixture with people from neighboring regions. Based on the observed genetic differences, the divergence time between the inhabitants of Okinawa and Miyako islands was dated to the Holocene. These findings suggest that the Pleistocene inhabitants, whose bones have been found on the southern Ryukyu Islands, did not make a major genetic contribution, if any, to the present-day inhabitants of the southern Ryukyu Islands.

Gene Expression Profiling of Mouse Condylar Cartilage during Mastication by Means of Laser Microdissection and cDNA Array

Little is known about the mechanisms of mandibular condylar growth. In this study, gene expression in the mandibular condylar cartilage of young post-natal mice was monitored by means of a cDNA microarray, real-time PCR, and laser microdissection before and after the initiation of mastication (newborn, 7 days, 21 days, initiation of mastication, and 35 days). Insulin-like growth factor-1 (IGF-I), transforming-growth-factor-beta-2 (TGFbeta2), and aggrecan mRNAs were clearly expressed at 21 days, while the expression of osteopontin mRNAs was most clear at 35 days. Parathyroid-hormone-related protein (PTHrP), Indian-hedgehog (Ihh), and insulin-like growth factor-2 (IGF-2) mRNAs were clearly expressed during lactation (newborn and 7 days). Heat-shock-protein 84 (HSP-84) and heat-shock-protein 86 (HSP-86) were clearly expressed at 35 days. These results revealed that gene expression changed during mandibular condylar cartilage growth, and that, interestingly, these changes coincided with the initiation of mastication.

Effectiveness of piezoelectric surgery in reducing surgical complications after bilateral sagittal split osteotomy.

Our aim was to investigate the effectiveness of piezoelectric surgery, where the osteotomy is made using ultrasonic vibration, in reducing surgical complications after bilateral sagittal split osteotomy (BSSO). Fifty-nine patients with skeletal mandibular prognathism who had mandibular setback with BSSO between January 2009 and April 2011 were included in the study. Piezosurgery was used in 29 cases, and the bone was split using a separator. In the remaining 30 cases, a Lindeman bur was used for the osteotomy and a chisel was used to split the bone. The amount of intraoperative bleeding and the Semmes Weinstein test scores were used as objective variables to evaluate the degree of neurosensory disturbance, and sex, age, use of piezosurgery, degree of setback, operating time, and method of fixation were used as explanatory variables. We used analysis of covariance (ANCOVA) to assess the significance of differences. Intraoperative bleeding was significantly less with age (p=0.003), and longer when operating time was prolonged (p=0.017), and was not influenced by the use of piezosurgery. The Semmes Weinstein test score significantly increased with age (p=0.01), and was significantly greater when piezoelectric surgery was used (p=0.008), and at 3 months, there were signs of more neurosensory disturbance in older patients and those who had had piezoelectric surgery. In this retrospective non-random study piezoelectric surgery reduced neither blood loss nor the incidence of neurosensory disturbance in BSSO.

Association of common PAX9 variants with permanent tooth size variation in non-syndromic East Asian populations

Studies on the heredity of dental characteristics in humans have indicated that the variance in many dental traits results from genetic variation. However, the genetic factors that influence commonly occurring dental variants are poorly understood. Paired domain box 9 (PAX9) codes a transcription factor that is important in tooth development. We investigated whether PAX9 polymorphisms are associated with normal variations in tooth agenesis and morphology. The study subjects were 273 Japanese and 223 Korean adults. Single-nucleotide polymorphisms (SNPs) in PAX9 (rs2295222, rs4904155, rs2073244, rs12881240 and rs4904210) were genotyped, and third molar agenesis and mesiodistal and buccolingual diameters were measured. We found that four of the five SNPs were significantly associated with the crown size. However, no SNP was associated with third molar agenesis. In additional analyses on non-metric dental traits, we found significant associations of PAX9 SNPs with shoveling of upper first incisors. In summary, common variants in PAX9 contributed to morphological variation in permanent teeth in humans.