Scott Lozanoff

Initiation and patterning of the snake dentition are dependent on Sonic Hedgehog signaling

Here we take the first look at cellular dynamics and molecular signaling in the developing snake dentition. We found that tooth formation differs from rodents in several respects. The majority of snake teeth bud off of a deep, ribbon-like dental lamina rather than as separate tooth germs. Prior to and after dental lamina ingrowth, we observe asymmetries in cell proliferation and extracellular matrix distribution suggesting that localized signaling by a secreted protein is involved. We cloned Sonic hedgehog from the African rock python Python sebae and traced its expression in the species as well as in two other snakes, the closely-related Python regius and the more derived corn snake Elaphe guttata (Colubridae). We found that expression of Shh is first confined to the odontogenic band and defines the position of the future dental lamina. Shh transcripts in pythons are progressively restricted to the oral epithelium on one side of the dental lamina and remain in this position throughout the prehatching period. Shh is expressed in the inner enamel epithelium and the stellate reticulum of the tooth anlagen, but is absent from the outer enamel epithelium and its derivative, the successional lamina. This suggests that signals other than Shh are responsible for replacement tooth formation. Functional studies using cyclopamine to block Hh signaling during odontogenesis prevented initiation and extension of the dental lamina into the mesenchyme, and also affected the directionality of this process. Further, blocking Hh signaling led to disruptions of the inner enamel epithelium. To explore the role of Shh in lamina extension, we looked at its expression in the premaxillary teeth, which form closer to the oral surface than elsewhere in the mouth. Oral ectodermal Shh expression in premaxillary teeth is lost soon after the teeth form reinforcing the idea that Shh is controlling the depth of the dental lamina. In summary, we have found diverse roles for Shh in patterning the snake dentition but, have excluded the participation of this signal in replacement tooth formation.

Morphometry of the Cranial Base in Subjects with Class III Malocclusion

The significance of the cranial base in the development of Class III malocclusion remains uncertain. The purpose of this study was to determine whether the form of the cranial base differs between prepubertal Class I and Class III subjects. Lateral cephalographs of 73 children of European-American descent aged between 5 and 11 years with Class III malocclusion were compared with those of their counterparts with a normal, Class I molar occlusion. The cephalographs were traced, checked, and subdivided into seven age- and sex-matched groups. Average geometries, scaled to an equivalent size, were generated based on 13 craniofacial landmarks by means of Procrustes analysis, and these configurations were statistically tested for equivalence. Bivariate and multivariate analyses utilizing 5 linear and angular measurements were undertaken to corroborate the Procrustes analysis. Graphical analysis, utilizing thin-plate spline and finite element methods, was performed for localization of differences in cranial base morphology. Results indicated that cranial base morphology differed statistically for all age-wise comparisons. Graphical analysis revealed that the greatest differences in morphology occurred in the posterior cranial base region, which generally consisted of horizontal compression, vertical expansion, and size contraction. The sphenoidal region displayed expansion, while the anterior regions showed shearing and local increases in size. It is concluded that the shape of the cranial base differs in subjects with Class III malocclusion compared with the normal Class I configuration, due in part to deficient orthocephalization, or failure of the cranial base to flatten during development.

Canalization and developmental stability in the Brachyrrhine mouse

The semi‐dominant Br mutation affects presphenoid growth, producing the facial retrognathism and globular neurocranial vault that characterize heterozygotes. We analysed the impact of this mutation on skull shape, comparing heterozygotes to wildtype mice, to determine if the effects are skull‐wide or confined to the sphenoid region targeted by the mutation. In addition, we examined patterns of variability of shape for the skull as a whole and for three regions (basicranium, face and neurocranium). We found that the Br mice differed significantly from wildtype mice in skull shape in all three regions as well as in the shape of the skull as a whole. However, the significant increases in variance and fluctuating asymmetry were found only in the basicranium of mutant mice. These results suggest that the mutation has a significant effect on the underlying developmental architecture of the skull, which produces an increase in phenotypic variability that is localized to the anatomical region in which the mean phenotype is most dramatically affected. These results suggest that the same developmental mechanisms that produce the change in phenotypic mean also produce the change in variance.

Location and dimensions of the mental foramen: a radiographic analysis by using cone-beam computed tomography

INTRODUCTION The mental foramen (MF) is an important landmark in dentistry. Knowledge of its position is central to perform block anesthesia of the mental nerve or to avoid nerve damage during surgical procedures in the premolar area of the mandible. The present radiographic study aimed at evaluating the location and dimension of the MF and measuring distances to neighboring structures by using limited cone-beam computed tomography (CBCT). METHODS Sagittal, axial, and coronal CBCT images of 142 patients (26 bilateral and 116 unilateral cases) were retrospectively screened to determine the location of the MF with respect to adjacent teeth and to take linear measurements of the size of the MF and its distances to the upper and lower borders of the mandible. In addition, the course and angulation of the mental canal exiting the MF were assessed. RESULTS The majority of MF (56%) were located apically between the 2 premolars, and another 35.7% of MF were positioned below the second premolar. On average, the MF was localized 5.0 mm from the closest root of the adjacent tooth (range, 0.3-9.8 mm). The mean size of the MF showed a height of 3.0 mm and a length of 3.2 mm; however, individual cases showed large differences in height (1.8-5.1 mm) and in length (1.8-5.5 mm). All mental canals exiting the MF demonstrated an upward course in the coronal plane, with 70.1% of the mental canal presenting an anterior loop (AL) in the axial view. The mean extension of AL in cases with an AL was 2.3 mm. CONCLUSIONS This study is consistent with previous radiographic studies regarding size and location of MF and distances between MF and adjacent anatomic structures. The assessed bilateral cases showed a high intraindividual concordance for certain features when comparing right and left sides.

Misexpression of Six2 is associated with heritable frontonasal dysplasia and renal hypoplasia in 3H1 Br mice

A radiation‐induced mouse mutant, Brachyrrhine (Br), exhibits frontonasal dysplasia and renal hypoplasia, two malformations associated with deficiencies in mesenchymal condensation. The purpose of this study was to resolve the Br locus, evaluate possible candidate genes, and identify developmental defects in the mutant chondrocranium. Linkage analysis mapped the Br mutation to a critical region distal to D17Mit76, which contains only one gene, the transcription factor Six2. Sequence analysis of the Six2 gene, including 1.5 kb of the promoter, failed to reveal the Br mutation. However, homozygous Br/Br embryos showed almost complete absence of Six2 mRNA and protein in craniofacial and renal tissues while heterozygous Br/+ embryos displayed intermediate Six2 levels. Mutant embryos displayed malformations of neural crest‐derived structures of the anterior cranium where Six2 is normally expressed. These data suggest a mutation in a novel cis‐acting regulatory region inhibits Six2 expression and is associated with frontonasal dysplasia and renal hypoplasia. Developmental Dynamics 237:1767–1779, 2008.

SPLINE ANALYSIS OF THE MANDIBLE IN HUMAN SUBJECTS WITH CLASS III MALOCCLUSION

This study determines deformations that contribute to a Class III mandibular morphology, employing thin-plate spline (TPS) analysis. A total of 133 lateral cephalographs of prepubertal children of European-American descent with either a Class I molar occlusion or a Class III malocclusion were compared. The cephalographs were traced and checked, and eight homologous landmarks on the mandible were identified and digitized. The datasets were scaled to an equivalent size and subjected to statistical analyses. These tests indicated significant differences between average Class I and Class III mandibular morphologies. When the sample was subdivided into seven age and sex-matched groups statistical differences were maintained for each group. TPS analysis indicated that both affine (uniform) and non-affine transformations contribute towards the total spline, and towards the average mandibular morphology at each age group. For non-affine transformations, partial warp 5 had the highest magnitude, indicating large-scale deformations of the mandibular configuration between articulare and pogonion. In contrast, partial warp 1 indicated localized shape changes in the mandibular symphyseal region. It is concluded that large spatial-scale deformations affect the body of the mandible, in combination with localized distortions further anteriorly. These deformations may represent a developmental elongation of the mandibular corpus antero-posteriorly that, allied with symphyseal changes, leads to the appearance of a Class III prognathic mandibular profile.

Early morphogenesis of heterodont dentition in minipigs.

The minipig provides an excellent experimental model for tooth morphogenesis because its diphyodont and heterodont dentition resemble that of humans. However, little information is available on the processes of tooth development in the pig. The purpose of this study was to classify the early stages of odontogenesis in minipigs from the initiation of deciduous dentition to the late bell stage when the successional dental lamina begins to develop. To analyze the initiation of teeth anlagens and the structural changes of dental lamina, a three-dimensional (3D) analysis was performed. At the earliest stage, 3D reconstruction revealed a continuous dental lamina along the length of the jaw. Later, the dental lamina exhibited remarkable differences in depth, and the interdental lamina was shorter. The dental lamina grew into the mesenchyme in the lingual direction, and its inclined growth was underlined by asymmetrical cell proliferation. After the primary tooth germ reached the late bell stage, the dental lamina began to disintegrate and fragmentize. Some cells disappeared during the process of lamina degradation, while others remained in small islands known as epithelial pearls. The minipig can therefore, inter alia, be used as a model organism to study the fate of epithelial pearls from their initiation to their contribution to pathological structures, primarily because of the clinical significance of these epithelial rests.

Deficiency in Six2 during prenatal development is associated with reduced nephron number, chronic renal failure, and hypertension in Br/+ adult mice

The Br/+ mutant mouse displays decreased embryological expression of the homeobox transcription factor Six2, resulting in hertitable renal hypoplasia. The purpose of this study was to characterize the renal physiological consequences of embryonic haploinsuffiency of Six2 by analyzing renal morphology and function in the adult Br heterozygous mutant. Adult Br/+ kidneys weighed 50% less than those from wild-type mice and displayed glomerulopathy. Stereological analysis of renal glomeruli showed that Br/+ kidneys had an average of 88% fewer glomeruli than +/+ kidneys, whereas individual glomeruli in Br/+ mice maintained an average volume increase of 180% compared with normal nephrons. Immunostaining revealed increased levels of endothelin-1 (ET-1), endothelin receptors A (ET(A)) and B (ET(B)), and Na-K-ATPase were present in the dilated renal tubules of mutant mice. Physiological features of chronic renal failure (CRF) including elevated mean arterial pressure, increased plasma creatinine, and dilute urine excretion were measured in Br/+ mutant mice. Electron microscopy of the Br/+ glomeruli revealed pathological alterations such as hypercellularity, extracellular matrix accumulation, and a thick irregular glomerular basement membrane. These results indicate that adult Br/+ mice suffer from CRF associated with reduced nephron number and renal hypoplasia, as well as glomerulopathy. Defects are associated with embryological deficiencies of Six2, suggesting that proper levels of this protein during nephrogenesis are critical for normal glomerular development and adult renal function.

Procrustes, Euclidean and cephalometric analyses of the morphology of the mandible in human Class III malocclusions

The role of mandibular phenotype in the development of Class III malocclusion remains unclear. The purpose of this study was to determine whether the form of the mandible differed between prepubertal individuals with Class I and Class III malocclusions. Lateral cephalographs of 73 children of European-American descent aged between 5-11 years with Class III malocclusion were compared to those of 60 counterparts with a normal, Class I molar occlusion. The cephalographs were traced and checked, and eight homologous mandibular landmarks were digitized. Average mandibular geometries, scaled to an equivalent size, were generated using Procrustes superimposition. Euclidean distance matrix analysis (EDMA) was undertaken to corroborate the Procrustes analysis, and bivariate analysis utilizing eight linear and five angular measurements was also performed. Residuals and F-values from Procrustes analysis indicated that mandibular configurations differed statistically for Class I and Class III types. EDMA confirmed that the Class I and Class III geometries were significantly different, revealing that the greatest differences in morphology arose in the anterior-most mandibular regions. As well, most variables showed statistically significant differences when the Class I and Class III mandibular types were compared. When the sample was subdivided into seven age- and sex-matched groups, nearly all age-based comparisons were significantly different. It is concluded that the morphology of the mandible differs in individuals with Class III malocclusions when compared to the normal Class I configuration, and that these alterations may indicate dichotomous postnatal mandibular ontogeny.

Molecular signaling along the anterior–posterior axis of early palate development

Cleft palate is a common congenital birth defect in humans. In mammals, the palatal tissue can be distinguished into anterior bony hard palate and posterior muscular soft palate that have specialized functions in occlusion, speech or swallowing. Regulation of palate development appears to be the result of distinct signaling and genetic networks in the anterior and posterior regions of the palate. Development and maintenance of expression of these region-specific genes is crucial for normal palate development. Numerous transcription factors and signaling pathways are now recognized as either anterior- (e.g., Msx1, Bmp4, Bmp2, Shh, Spry2, Fgf10, Fgf7, and Shox2) or posterior-specific (e.g., Meox2, Tbx22, and Barx1). Localized expression and function clearly highlight the importance of regional patterning and differentiation within the palate at the molecular level. Here, we review how these molecular pathways and networks regulate the anterior–posterior patterning and development of secondary palate. We hypothesize that the anterior palate acts as a signaling center in setting up development of the secondary palate.