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Bodies are often made of repeated units, or serial homologs, that develop using the same core gene regulatory network. Local inputs and modifications to this network allow serial homologs to evolve different morphologies, but currently we do not understand which modifications allow these repeated traits to evolve different levels of phenotypic plasticity. Here we describe variation in phenotypic plasticity across serial homologous eyespots of the butterfly Bicyclus anynana, hypothesized to be under selection for similar or different functions in the wet and dry seasonal forms. Specifically, we document the presence of eyespot size and scale brightness plasticity in hindwing eyespots hypothesized to vary in function across seasons, and reduced size plasticity and absence of brightness plasticity in forewing eyespots hypothesized to have the same function across seasons. By exploring the molecular and physiological causes of this variation in plasticity across fore and hindwing serial homologs we discover that: 1) temperature experienced during the wandering stages of larval development alters titers of an ecdysteroid hormone, 20-hydroxyecdysone (20E), in the hemolymph of wet and dry seasonal forms at that stage; 2) the 20E receptor (EcR) is differentially expressed in the forewing and hindwing eyespot centers of both seasonal forms during this critical developmental stage; and 3) manipulations of EcR signaling disproportionately affected hindwing eyespots relative to forewing eyespots. We propose that differential EcR expression across forewing and hindwing eyespots at a critical stage of development explains the variation in levels of phenotypic plasticity across these serial homologues. This finding provides a novel signaling pathway, 20E, and a novel molecular candidate, EcR, for the regulation of levels of phenotypic plasticity across body parts or serial homologs., Author Summary One of the most exquisite types of organismal adaptations in nature occurs when organisms are able to change the way they develop in anticipation of the different selective environments they will experience as adults. This leads to variation in adult morphologies that are adaptive. Environmental cues experienced during development often lead to variation in hormonal titers that can have profound effect on the way genes are regulated and on the adult morphology. Here we examine the hormonal and molecular mechanisms that allow specific traits that are repeated in an organism (butterfly eyespots) to either be sensitive to environmental cues–and develop different morphologies—or insensitive to these cues and develop the same morphology. We discover that a specific gene, a hormone receptor, that is expressed in the sensitive eyespots but absent in the insensitive eyespots, is responsible for regulating the level of sensitivity of each of the eyespots to an environmental cue. We identify a molecule that is regulating levels of environmental sensitivity, or phenotypic plasticity, across repeated traits in an organism.

South Asia possesses a significant amount of genetic diversity due to considerable intergroup differences in culture and language. There have been numerous reports on the genetic structure of Asian Indians, although these have mostly relied on genotyping microarrays or targeted sequencing of the mitochondria and Y chromosomes. Asian Indians in Singapore are primarily descendants of immigrants from Dravidian-language–speaking states in south India, and 38 individuals from the general population underwent deep whole-genome sequencing with a target coverage of 30X as part of the Singapore Sequencing Indian Project (SSIP). The genetic structure and diversity of these samples were compared against samples from the Singapore Sequencing Malay Project and populations in Phase 1 of the 1,000 Genomes Project (1 KGP). SSIP samples exhibited greater intra-population genetic diversity and possessed higher heterozygous-to-homozygous genotype ratio than other Asian populations. When compared against a panel of well-defined Asian Indians, the genetic makeup of the SSIP samples was closely related to South Indians. However, even though the SSIP samples clustered distinctly from the Europeans in the global population structure analysis with autosomal SNPs, eight samples were assigned to mitochondrial haplogroups that were predominantly present in Europeans and possessed higher European admixture than the remaining samples. An analysis of the relative relatedness between SSIP with two archaic hominins (Denisovan, Neanderthal) identified higher ancient admixture in East Asian populations than in SSIP. The data resource for these samples is publicly available and is expected to serve as a valuable complement to the South Asian samples in Phase 3 of 1 KGP., Author Summary Indians of South Asia has long been a population of interest to a wide audience, due to its unique diversity. We have deep-sequenced 38 individuals of Indian descent residing in Singapore (SSIP) in an effort to illustrate their diversity from a whole-genome standpoint. Indeed, among Asians in our population panel, SSIP was most diverse, followed by the Malays in Singapore (SSMP). Their diversity is further observed in the population's chromosome Y haplogroup and mitochondria haplogroup profiles; individuals with European-dominant haplogroups had greater proportion of European admixture. Among variants (single nucleotide polymorphism and small insertions/deletions) discovered in SSIP, 21.69% were novel with respect to previous sequencing projects. In addition, some 14 loss-of-function variants (LOFs) were associated to cancer, Type II diabetes, and cholesterol levels. Finally, D statistic test with ancient hominids concurred that there was gene flow to East Asians compared to South Asians.

Obesity is characterized by accumulation of excess body fat, while lipodystrophy is characterized by loss or absence of body fat. Despite their opposite phenotypes, these two conditions both cause ectopic lipid storage in non-adipose tissues, leading to lipotoxicity, which has health-threatening consequences. The exact mechanisms underlying ectopic lipid storage remain elusive. Here we report the analysis of a Drosophila model of the most severe form of human lipodystrophy, Berardinelli-Seip Congenital Lipodystrophy 2, which is caused by mutations in the BSCL2/Seipin gene. In addition to reduced lipid storage in the fat body, dSeipin mutant flies accumulate ectopic lipid droplets in the salivary gland, a non-adipose tissue. This phenotype was suppressed by expressing dSeipin specifically within the salivary gland. dSeipin mutants display synergistic genetic interactions with lipogenic genes in the formation of ectopic lipid droplets. Our data suggest that dSeipin may participate in phosphatidic acid metabolism and subsequently down-regulate lipogenesis to prevent ectopic lipid droplet formation. In summary, we have demonstrated a tissue-autonomous role of dSeipin in ectopic lipid storage in lipodystrophy., Author Summary Obesity and lipodystrophy are medical conditions characterized by excess body fat or too little body fat, respectively. Interestingly, a common feature of both conditions is ectopic accumulation of lipids (fat) in cells where fat is not normally stored. This can cause tissue damage with health-threatening consequences. We are trying to understand how these two very different diseases lead to lipid storage in non-fat tissues. In this study, we used fruit flies (Drosophila melanogaster) with a mutation in the dSeipin gene as a lipodystrophy model to explore the mechanism of ectopic lipid storage. In dSeipin mutant flies, we found numerous lipid droplets in the salivary gland, a non-fat storage tissue, and reduced lipid storage in the fat body, an adipose tissue. Furthermore, we proved that dSeipin functions within salivary gland cells to prevent the formation of ectopic lipid droplets. We also found that dSeipin genetically interacts with other fat synthesis and metabolism genes in the formation of ectopic lipid droplets. The fruit fly dSeipin mutant provides an excellent model system for dissecting the mechanisms that regulate the storage of excess lipids.

Phosphoinositides represent important lipid signals in the plant development and stress response. However, multiple isoforms of the phosphoinositide biosynthetic genes hamper our understanding of the pivotal enzymes in each step of the pathway as well as their roles in plant growth and development. Here, we report that phosphoinositide-specific phospholipase C2 (AtPLC2) is the primary phospholipase in phosphoinositide metabolism and is involved in seedling growth and the endoplasmic reticulum (ER) stress responses in Arabidopsis thaliana. Lipidomic profiling of multiple plc mutants showed that the plc2-1 mutant increased levels of its substrates phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate, suggesting that the major phosphoinositide metabolic pathway is impaired. AtPLC2 displayed a distinct tissue expression pattern and localized at the plasma membrane in different cell types, where phosphoinositide signaling occurs. The seedlings of plc2-1 mutant showed growth defect that was complemented by heterologous expression of AtPLC2, suggesting that phosphoinositide-specific phospholipase C activity borne by AtPLC2 is required for seedling growth. Moreover, the plc2-1 mutant showed hypersensitive response to ER stress as evidenced by changes in relevant phenotypes and gene expression profiles. Our results revealed the primary enzyme in phosphoinositide metabolism, its involvement in seedling growth and an emerging link between phosphoinositide and the ER stress response., Author Summary Plant growth requires continuous signal transduction in response to ever-changing environment. Phosphoinositides represent primary lipid-derived signals that are involved in various plant responses to surrounding environment. However, enzymes that determine the complex phosphoinositide molecular profiles remained elusive due to the existence of a number of candidate genes for each step. Here, in Arabidopsis thaliana, we found AtPLC2 as the enzyme that is decisive in phosphoinositide metabolism by analytical lipidomics of the gene knockout study. Functional characterization of AtPLC2 knockout plant enabled us to find two novel roles of phosphoinositides: requirement in seedling growth and ER stress tolerance. Because economically important environmental stresses such as salinity and temperature upshift cause ER stress, our findings may open up a new avenue in addressing ER stress responses via phosphoinositide signaling.

Lipid droplets (LDs) are important cellular organelles that govern the storage and turnover of lipids. Little is known about how the size of LDs is controlled, although LDs of diverse sizes have been observed in different tissues and under different (patho)physiological conditions. Recent studies have indicated that the size of LDs may influence adipogenesis, the rate of lipolysis and the oxidation of fatty acids. Here, a genome-wide screen identifies ten yeast mutants producing “supersized” LDs that are up to 50 times the volume of those in wild-type cells. The mutated genes include: FLD1, which encodes a homologue of mammalian seipin; five genes (CDS1, INO2, INO4, CHO2, and OPI3) that are known to regulate phospholipid metabolism; two genes (CKB1 and CKB2) encoding subunits of the casein kinase 2; and two genes (MRPS35 and RTC2) of unknown function. Biochemical and genetic analyses reveal that a common feature of these mutants is an increase in the level of cellular phosphatidic acid (PA). Results from in vivo and in vitro analyses indicate that PA may facilitate the coalescence of contacting LDs, resulting in the formation of “supersized” LDs. In summary, our results provide important insights into how the size of LDs is determined and identify novel gene products that regulate phospholipid metabolism., Author Summary Lipid droplets (LD) are primary lipid storage structures that also function in membrane and lipid trafficking, protein turnover, and the reproduction of deadly viruses. Increased LD accumulation in liver, skeletal muscle, and adipose tissue is a hallmark of the metabolic syndrome. Enlarged LDs are often found in these tissues under disease conditions. However, little is known about how the size of LDs is controlled in eukaryotic cells. In this study, we use genetic and biochemical methods to identify important gene products that regulate the size of the LDs. Notably, a common feature among these mutants with “supersized” LDs is an increased level of phosphatidic acid (PA). We also show that a small amount of PA can increase the size of artificial LDs in vitro. Overall, our study identifies important lipids and proteins in determining LD size. These results provide valuable insights into how human cells/tissues handle abnormal influx of lipids in today's obesogenic environment.