Neural innervation as a potential trigger of morphological color change and sexual dimorphism in cichlid fish

Many species change their coloration during ontogeny or even as adults. Color change hereby often serves as sexual or status signal. The cellular and subcellular changes that drive color change and how they are orchestrated have been barely understood, but a deeper knowledge of the underlying processes is important to our understanding of how such plastic changes develop and evolve. Here we studied the color change of the Malawi golden cichlid (Melanchromis auratus). Females and subordinate males of this species are yellow and white with two prominent black stripes (yellow morph; female and non-breeding male coloration), while dominant males change their color and completely invert this pattern with the yellow and white regions becoming black, and the black stripes becoming white to iridescent blue (dark morph; male breeding coloration). A comparison of the two morphs reveals that substantial changes across multiple levels of biological organization underlie this polyphenism. These include changes in pigment cell (chromatophore) number, intracellular dispersal of pigments, and tilting of reflective platelets (iridosomes) within iridophores. At the transcriptional level, we find differences in pigmentation gene expression between these two color morphs but, surprisingly, 80% of the genes overexpressed in the dark morph relate to neuronal processes including synapse formation. Nerve fiber staining confirms that scales of the dark morph are indeed innervated by 1.3 to 2 times more axonal fibers. Our results might suggest an instructive role of nervous innervation orchestrating the complex cellular and ultrastructural changes that drive the morphological color change of this cichlid species.Peer reviewe

Developmental and Cellular Basis of Vertical Bar Color Patterns in the East African Cichlid Fish Haplochromis latifasciatus

The East African adaptive radiations of cichlid fishes are renowned for their diversity in coloration. Yet, the developmental basis of pigment pattern formation remains largely unknown. One of the most common melanic patterns in cichlid fishes are vertical bar patterns. Here we describe the ontogeny of this conspicuous pattern in the Lake Kyoga species Haplochromis latifasciatus. Beginning with the larval stages we tracked the formation of this stereotypic color pattern and discovered that its macroscopic appearance is largely explained by an increase in melanophore density and accumulation of melanin during the first 3 weeks post-fertilization. The embryonal analysis is complemented with cytological quantifications of pigment cells in adult scales and the dermis beneath the scales. In adults, melanic bars are characterized by a two to threefold higher density of melanophores than in the intervening yellow interbars. We found no strong support for differences in other pigment cell types such as xanthophores. Quantitative PCRs for twelve known pigmentation genes showed that expression of melanin synthesis genes tyr and tyrp1a is increased five to sixfold in melanic bars, while xanthophore and iridophore marker genes are not differentially expressed. In summary, we provide novel insights on how vertical bars, one of the most widespread vertebrate color patterns, are formed through dynamic control of melanophore density, melanin synthesis and melanosome dispersal.Peer reviewe

Functional conservation and divergence of color-pattern-related agouti family genes in teleost fishes

While color patterns are highly diverse across the animal kingdom, certain patterns such as countershading and stripe patterns have evolved repeatedly. Across vertebrates, agouti-signaling genes have been associated with the evolution of both patterns. Here we study the functional conservation and divergence by investigating the expression patterns of the two color-pattern-related agouti-signaling genes, agouti-signaling protein 1 (asip1) and agouti-signaling protein 2b (asip2b, also known as agrp2) in Teleostei. We show that the dorsoventral expression profile of asip1 and the role of the "stripe repressor" asip2b are shared across multiple teleost lineages and uncover a previously unknown association between stripe-interstripe patterning and both asip1 and asip2b expression. In some species, including the zebrafish (Danio rerio), these two genes show complementary and overlapping expression patterns in line with functional redundancy. Our results thus suggest how conserved and novel functions of agouti-signaling genes might have shaped the evolution of color patterns across teleost fishes.Peer reviewe

Stability Analysis and Reinforcement of the Existing Karst Cave Passing through Yujingshan Tunnel

High-speed Railway tunneling in karst terrain presents a huge challenge to the engineer including the identification, stability analysis and reinforcement of the karst cavities. The Cheng-Gui high-speed railway tunnel had to pass through the largest karst cave discovered in tunnel construction. To guaranteeing the tunnel construction safety, a series of corresponding prevention and control measures are put forward. To begin with, geological drilling, electromagnetic method and surface electrical resistivity tomography are adopted to detect and delineate the underground karst zone. Based on the detection results, this paper has put forward strategies to make the pre-support of karst cave and the main technical of those strategies include: the side-walls or in the crown was applied with shotcret (C40 steel fiber concrete); use expanding-shell pre-stressed hollow anchor rod and pre-stressed cable reinforcement; fix steel-mesh-bolting; the shotcrete sealing was applied. Moreover, if instabilities would develop in the side-walls, it should be sufficient to stabilize the cavities, to do dental cleaning of the broken rocks, and fill the voids with shotcrete or pumped lean concrete. At last, systematic grouting treatment around the excavated section, and was excavated with the layer-step method. The solutions presented here may provide guidance for the design and construction of high-speed railway tunnels to be implemented affected by karst processes. The technical validation of the proposed solutions was demonstrated by the successful completion of the Yujingshan tunnel. Please click Additional Files below to see the full abstract

Feature Proliferation -- the "Cancer" in StyleGAN and its Treatments

Despite the success of StyleGAN in image synthesis, the images it synthesizes are not always perfect and the well-known truncation trick has become a standard post-processing technique for StyleGAN to synthesize high-quality images. Although effective, it has long been noted that the truncation trick tends to reduce the diversity of synthesized images and unnecessarily sacrifices many distinct image features. To address this issue, in this paper, we first delve into the StyleGAN image synthesis mechanism and discover an important phenomenon, namely Feature Proliferation, which demonstrates how specific features reproduce with forward propagation. Then, we show how the occurrence of Feature Proliferation results in StyleGAN image artifacts. As an analogy, we refer to it as the" cancer" in StyleGAN from its proliferating and malignant nature. Finally, we propose a novel feature rescaling method that identifies and modulates risky features to mitigate feature proliferation. Thanks to our discovery of Feature Proliferation, the proposed feature rescaling method is less destructive and retains more useful image features than the truncation trick, as it is more fine-grained and works in a lower-level feature space rather than a high-level latent space. Experimental results justify the validity of our claims and the effectiveness of the proposed feature rescaling method. Our code is available at https://github. com/songc42/Feature-proliferation.Comment: Accepted at ICCV 202

Comparative ontogenetic and transcriptomic analyses shed light on color pattern divergence in cichlid fishes

Funding Information: This study was supported by the Baden-Württemberg Foundation (to Claudius F. Kratochwil), grants by the Deutsche Forschungsgemeinschaft (DFG) to Axel Meyer, Claudius F. Kratochwil (KR 4670/2-1 and KR 4670/4-1), and Paolo Franchini (FR 3399/1-1), and a stipend from the China Scholarship Council (CSC, to Yipeng Liang). We thank Max Grauvogl who conducted the qPCR experiments during his bachelor thesis. Open Access funding enabled and organized by Projekt DEAL. Funding Information: This study was supported by the Baden‐Württemberg Foundation (to Claudius F. Kratochwil), grants by the Deutsche Forschungsgemeinschaft (DFG) to Axel Meyer, Claudius F. Kratochwil (KR 4670/2‐1 and KR 4670/4‐1), and Paolo Franchini (FR 3399/1‐1), and a stipend from the China Scholarship Council (CSC, to Yipeng Liang). We thank Max Grauvogl who conducted the qPCR experiments during his bachelor thesis. Open Access funding enabled and organized by Projekt DEAL. Publisher Copyright: © 2022 The Authors. Evolution & Development published by Wiley Periodicals LLC.Stripe patterns are a striking example for a repeatedly evolved color pattern. In the African adaptive radiations of cichlid fishes, stripes evolved several times independently. Previously, it has been suggested that regulatory evolution of a single gene, agouti-related-peptide 2 (agrp2), explains the evolutionary lability of this trait. Here, using a comparative transcriptomic approach, we performed comparisons between (adult) striped and nonstriped cichlid fishes of representatives of Lake Victoria and the two major clades of Lake Malawi (mbuna and non-mbuna lineage). We identify agrp2 to be differentially expressed across all pairwise comparisons, reaffirming its association with stripe pattern divergence. We therefore also provide evidence that agrp2 is associated with the loss of the nonstereotypic oblique stripe of Mylochromis mola. Complementary ontogenetic data give insights into the development of stripe patterns as well as vertical bar patterns that both develop postembryonically. Lastly, using the Lake Victoria species pair Haplochromis sauvagei and Pundamilia nyererei, we investigated the differences between melanic and non-melanic regions to identify additional genes that contribute to the formation of stripes. Expression differences—that most importantly also do not include agrp2—are surprisingly small. This suggests, at least in this species pair, that the stripe phenotype might be caused by a combination of more subtle transcriptomic differences or cellular changes without transcriptional correlates. In summary, our comprehensive analysis highlights the ontogenetic and adult transcriptomic differences between cichlids with different color patterns and serves as a basis for further investigation of the mechanistic underpinnings of their diversification.Peer reviewe

Evolutionary Dynamics of Structural Variation at a Key Locus for Color Pattern Diversification in Cichlid Fishes

Color patterns in African cichlid fishes vary spectacularly. Although phylogenetic analysis showed already 30 years ago that many color patterns evolved repeatedly in these adaptive radiations, only recently have we begun to understand the genomic basis of color variation. Horizontal stripe patterns evolved and were lost several times independently across the adaptive radiations of LakeVictoria, Malawi, and Tanganyika and regulatory evolution of agouti-related peptide 2 (agrp2/asip2b) has been linked to this phenotypically labile trait. Here, we asked whether the agrp2 locus exhibits particular characteristics that facilitate divergence in color patterns. Based on comparative genomic analyses, we discovered several recent duplications, insertions, and deletions. Interestingly, one of these events resulted in a tandem duplication of the last exon of agrp2. The duplication likely precedes the EastAfrican radiations that started 8-12 Ma, is not fixed within any of the radiations, and is found to vary even within some species. Moreover, we also observed variation in copy number (two to five copies) and secondary loss of the duplication, illustrating a surprising dynamic at this locus that possibly promoted functional divergence of agrp2. Our work suggests that such instances of exon duplications are a neglected mechanism potentially involved in the repeated evolution and diversification that deserves more attention.Peer reviewe