Expanded carrier screening for inherited genetic disease using exome and genome sequencing.

The goal of this study was to assess the feasibility of using exome (ES) and genome sequencing (GS) in guiding preconception genetic screening (PCGS) for couples who are planning to conceive by creating a workflow for identifying risk alleles for autosomal recessive (AR) and X-linked (XL) disorders without the constraints of a predetermined, targeted gene panel. There were several limitations and challenges related to reporting and the technical aspects of ES and GS, which are listed in the discussion. We selected 150 couples from a cohort of families (trios) enrolled in a research protocol where the goal was to define the genetic etiology of disease in an affected child. Pre-existing, de-identified parental sequencing data were analyzed to define variants that would place the couple at risk of having a child affected by an AR or XL disorder. We identified 17 families who would be selected for counseling about risk alleles. We noted that only 3 of these at-risk couples would be identified if we limited ourselves to the current ACMG-recommended expanded carrier screening gene panel. ES and GS successfully identified couples who are at risk of having a child with a rare AR or XL disorder that would have been missed by the current recommended guidelines. Current limitations of this approach include ethical concerns, difficulties in reporting results including variant calling due to the rare nature of some of the variants, determining which disorders to report, as well as technical difficulties in detecting certain variants such as repeat expansions.

Bristles formation in adhesive pads and sensilli of the gecko Tarentola mauritanica derive from a massive accumulation of corneous material in Oberhautchen cells of the epidermis.

Among lizards, geckos possess special digital scales modified as hairy-like lamellae that allow attachment to vertical substrates for the movement using adhesive nanoscale filaments called setae. The present study shows new ultrastructural details on setae formation in the gecko Tarentula mauritanica. Setae derive from the special differentiation of an epidermal layer termed Oberhauchen and can reach 30-60 µm in length. Oberhautchen cells in the adhesive pad lamellae becomes hypertrophic and rest upon 2 layers of non-corneous and pale cells instead of beta-cells like in the other scales. Only 1-2 beta-layers are formed underneath the pale layer. Setae derive from the accumulation of numerous roundish and heterogenous beta-packets with variable electron-density in Oberhautchen cells, possibly indicating a mixed protein composition. Immunofluorescence and immunogold labeling for CBPs show that beta-packets merge at the base of the growing setae forming long corneous bundles. Pale cells formed underneath the Oberhautchen layer contain small vesicles or tubules with a likely lipid content, sparse keratin filaments and ribosomes. In mature lamellae these cells merge with Oberhautchen and beta-cells forming a thin electron-paler layer located between the Oberhautchen and the thin beta-layer, a variation of the typical sequence of epidermal layers present in other scales. The formation of a softer pale layer and of a thin beta-layer likely determines a flexible corneous support for the adhesive setae. The specific molecular mechanism that stimulates the cellular changes observed during Oberhautchen hypertrophy and the alteration of the typical epidermal stratification in the pad epidermis remains unknown.

Micro-ornamentation patterns in different areas of the epidermis in the gecko Tarentola mauritanica reflect variations in the accumulation of corneous material in Oberhautchen cells.

Micro-ornamentations characterize the surface of scales in lepidosaurians and are summarized in four main patterns, i.e., spinulated, lamellated, lamellate-dentate, and honeycomb, although variations of these patterns are present in different species. Although geckos are known to possess a spinulated pattern derived from the Oberhautchen layer, also other pattern variations of the spinulated micro-ornamentation are present such as those indicated as dendritic ramification, corneous belts, and small bare patches. The present study mainly describes the variation of micro-ornamentations present in scales of different skin regions in the Mediterranean gecko Tarentula mauritanica using scannig and transmission electron microscopy. The study reports that the accumulation of corneous material in Oberhautchen cells is not homogenous in different areas of body scales and, when mature, this process gives rise to different sculpturing on the epidermal surface generating not only spinulae but also transitional zones leading to the other main patterns. It is hypothesized that spinulae formation derives from the vertical and lateral symmetric growth of tubercolate, non-overlapped scales of geckos. Sparse areas also result smooth or with serpentine-ridges likely revealing the beta-layer located underneath and merged with the Oberhautchen. The eco-functional role of this variable micro-ornamentation in the skin of lizards however remains largely speculative.