Thyroid hormone regulates abrupt skin morphogenesis during zebrafish postembryonic development.

Thyroid hormone is a key regulator of post-embryonic vertebrate development. Skin is a biomedically important thyroid hormone target organ, but the cellular and molecular mechanisms underlying skin pathologies associated with thyroid dysfunction remain obscure. The transparent skin of zebrafish is an accessible model system for studying vertebrate skin development. During post-embryonic development of the zebrafish, scales emerge in the skin from a hexagonally patterned array of dermal papillae, like other vertebrate skin appendages such as feathers and hair follicles. We show here that thyroid hormone regulates the rate of post-embryonic dermal development through interaction with nuclear hormone receptors. This couples skin development with body growth to generate a well ordered array of correctly proportioned scales. This work extends our knowledge of thyroid hormone actions on skin by providing in-vivo evidence that thyroid hormone regulates multiple aspects of dermal development.

Control of osteoblast regeneration by a train of Erk activity waves.

Regeneration is a complex chain of events that restores a tissue to its original size and shape. The tissue-wide coordination of cellular dynamics that is needed for proper morphogenesis is challenged by the large dimensions of regenerating body parts. Feedback mechanisms in biochemical pathways can provide effective communication across great distances1-5, but how they might regulate growth during tissue regeneration is unresolved6,7. Here we report that rhythmic travelling waves of Erk activity control the growth of bone in time and space in regenerating zebrafish scales, millimetre-sized discs of protective body armour. We find that waves of Erk activity travel across the osteoblast population as expanding concentric rings that are broadcast from a central source, inducing ring-like patterns of tissue growth. Using a combination of theoretical and experimental analyses, we show that Erk activity propagates as excitable trigger waves that are able to traverse the entire scale in approximately two days and that the frequency of wave generation controls the rate of scale regeneration. Furthermore, the periodic induction of synchronous, tissue-wide activation of Erk in place of travelling waves impairs tissue growth, which indicates that wave-distributed Erk activation is key to regeneration. Our findings reveal trigger waves as a regulatory strategy to coordinate cell behaviour and instruct tissue form during regeneration.

Multiple roles forlaccase2 in butterfly wing pigmentation, scale development, and cuticle tanning.

Lepidopteran wing scales play important roles in a number of functions including color patterning and thermoregulation. Despite the importance of wing scales, however, we still have a limited understanding of the genetic mechanisms that underlie scale patterning, development, and coloration. Here, we explore the function of the phenoloxidase-encoding gene laccase2 in wing and scale development in the nymphalid butterfly Vanessa cardui. Somatic deletion mosaics of laccase2 generated by CRISPR/Cas9 genome editing presented several distinct mutant phenotypes. Consistent with the work in other nonlepidopteran insect groups, we observed reductions in melanin pigmentation and defects in cuticle formation. We were also surprised, however, to see distinct effects on scale development including complete loss of wing scales. This study highlights laccase2 as a gene that plays multiple roles in wing and scale development and provides new insight into the evolution of lepidopteran wing coloration.

Ontogenetic scaling patterns of lizard skin surface structure as revealed by gel-based stereo-profilometry.

The skin surface structure of squamate reptiles varies greatly among species, likely because it plays a key role in a range of tasks, such as camouflage, locomotion, self-cleaning, mitigation of water loss and protection from physical damage. Although we have foundational knowledge about squamate skin morphology, we still know remarkably little about how intraspecific variation in skin surface structure translates to functional variation. This gap in our understanding can be in part traced back to: (i) our lack of knowledge on how body size determines skin surface structure; and (ii) the lack of means to perform high-throughput and detailed analysis of the three-dimensional (3D) anatomy of reptilian skin surfaces in a non-destructive manner. To fill this gap, we explored the possibilities of a new imaging technique, termed gel-based stereo-profilometry, to visualize and quantify the 3D topography of reptilian skin surface structure. Using this novel approach, we investigated intra-specific and intra-individual variation in the skin surface morphology of a focal lizard species, Anolis cristatellus. We assessed how various characteristics of surface topography (roughness, skew and kurtosis) and scale morphology (area, height, width and shape) scale with body size across different body regions. Based on an ontogenetic series of A. cristatellus males, we show that skin roughness increases with body size. Skin patches on the ventral body region of lizards were rougher than on the dorsum, but this was a consequence of ventral scales being larger than dorsal scales. Dorsal surface skew and kurtosis varied with body size, but surfaces on the ventral skin showed no such relationship. Scale size scaled isometrically with body size, and while ventral scales differed in shape from dorsal scales, scale shape did not change with ontogeny. Overall, this study demonstrates that gel-based stereo-profilometry is a promising method to rapidly assess the 3D surface structure of reptilian skin at the microscopic level. Additionally, our findings of the explanatory power of body size on skin surface diversity provide a foundation for future studies to disentangle the relationships among morphological, functional and ecological diversity in squamate reptile skin surfaces.

Oxytocin/vasopressin-like peptide inotocin regulates cuticular hydrocarbon synthesis and water balancing in ants.

Oxytocin/vasopressin-like peptides are important regulators of physiology and social behavior in vertebrates. However, the function of inotocin, the homologous peptide in arthropods, remains largely unknown. Here, we show that the level of expression of inotocin and inotocin receptor are correlated with task allocation in the ant Camponotus fellah Both genes are up-regulated when workers age and switch tasks from nursing to foraging. in situ hybridization revealed that inotocin receptor is specifically expressed in oenocytes, which are specialized cells synthesizing cuticular hydrocarbons which function as desiccation barriers in insects and for social recognition in ants. dsRNA injection targeting inotocin receptor, together with pharmacological treatments using three identified antagonists blocking inotocin signaling, revealed that inotocin signaling regulates the expression of cytochrome P450 4G1 (CYP4G1) and the synthesis of cuticular hydrocarbons, which play an important role in desiccation resistance once workers initiate foraging.

Experimental investigation of the effects of temperature and feeding regime on scale growth in Atlantic salmon Salmo salar post-smolts.

Salmo salar post-smolts were reared in seawater under controlled laboratory conditions for 12 weeks. The fish were exposed to three constant temperature treatments (15, 10.5 and 6°C) and four feeding treatments (constant feeding, food withheld for 7 days, food withheld for 14 days and food withheld intermittently for four periods of 7 days). Scale growth was proportional to fish growth across all treatments, justifying the use of scale measurements as a proxy for growth during the early marine phase. The rate of circuli deposition was dependant on temperature and feeding regime and was generally proportional to fish growth but with some decoupling of the relationship at 15°C. Deposition rates varied from 4.8 days per circulus at 15°C (constant feeding) to 15.1 days per circulus at 6°C (interrupted feeding). Cumulative degree day (° D) was a better predictor of circuli number than age, although the rate of circuli deposition ° D-1 was significantly lower at 6°C compared with 15 and 10.5°C. Inter-circuli distances were highly variable and did not reflect growth rate; tightly packed circuli occurred during periods without food when growth was depressed, but also during periods of rapid growth at 15°C. The results further current understanding of scale growth properties and can inform investigations of declining marine growth in S. salar based on interpretations of scale growth patterns.

Identification and characterization of microRNAs involved in scale biomineralization in the naked carp Gymnocypris przewalskii.

The mineralized scale derived from skin plays a protective role for the fish body and also possesses important application values in the biomaterial field. However, little is known about fish scale biomineralization and related molecular regulatory mechanisms. Here, we used a comparative microRNA sequencing approach to identify and characterize differentially expressed microRNAs (DEMs) involved in scale biomineralization in the naked carp Gymnocypris przewalskii. A total of 18, 43, and 66 DEMs were obtained from skin tissues covered with initial, developing, and mature scales (IS, DS, and MS) compared with scale-uncovered skin. The target genes of these DEMs were significantly enriched in a sole biomineralization-related sphingolipid signaling pathway. Seven DEMs (dre-miR-124-3p, dre-miR-133a-2-5p, dre-miR-184, dre-miR-206-3p, novel_33, novel_56 and novel_75) were common in IS, DS, and MS. Dre-miR-124-3p, dre-miR-206-3p, and novel_33 were predicted to be able to target biomineralization-related genes. Stem-loop real-time quantitative PCR further confirmed that the common DEMs had higher expression levels in scale-covered skin tissues than that in the gill, intestine, and brain, except for dre-miR-133a-2-5p. Our results suggest that these identified microRNAs may play a role in scale biomineralization in G. przewalskii, and the obtained microRNAs are expected to be candidates in understanding the molecular mechanism of scale biomineralization in fish species.

Using scale and feather traits for module construction provides a functional approach to chicken epidermal development.

Gene co-expression network analysis has been a research method widely used in systematically exploring gene function and interaction. Using the Weighted Gene Co-expression Network Analysis (WGCNA) approach to construct a gene co-expression network using data from a customized 44K microarray transcriptome of chicken epidermal embryogenesis, we have identified two distinct modules that are highly correlated with scale or feather development traits. Signaling pathways related to feather development were enriched in the traditional KEGG pathway analysis and functional terms relating specifically to embryonic epidermal development were also enriched in the Gene Ontology analysis. Significant enrichment annotations were discovered from customized enrichment tools such as Modular Single-Set Enrichment Test (MSET) and Medical Subject Headings (MeSH). Hub genes in both trait-correlated modules showed strong specific functional enrichment toward epidermal development. Also, regulatory elements, such as transcription factors and miRNAs, were targeted in the significant enrichment result. This work highlights the advantage of this methodology for functional prediction of genes not previously associated with scale- and feather trait-related modules.