Generation of two modified mouse alleles of the Hic1 tumor suppressor gene

HIC1 (hypermethylated in cancer 1) is a tumor suppressor gene located on chromosome 17p13.3, a region frequently hypermethylated or deleted in human neoplasias. In mouse, Hic1 is essential for embryonic development and exerts an antitumor role in adult animals. Since Hic1-deficient mice die perinatally, we generated a conditional Hic1 null allele by flanking the Hic1-coding region by loxP sites. When crossed to animals expressing Cre recombinase in a cell-specific manner, the Hic1 conditional mice will provide new insights into the function of Hic1 in developing and mature tissues. Additionally, we used gene targeting to replace sequence-encoding amino acids 186-893 of Hic1 by citrine fluorescent protein cDNA. We demonstrate that the distribution of Hic1-citrine fusion polypeptide corresponds to the expression pattern of wild-type Hic1. Consequently, Hic1-citrine "reporter" mice can be used to monitor the activity of the Hic1 locus using citrine fluorescence.     

A mart-1::Cre transgenic line induces recombination in melanocytes and retinal pigment epithelium

The number of transgenic mouse lines expressing Cre in either type of pigment cells (melanocytes and retinal pigment epithelium, RPE) is limited, and the available lines do not always offer sufficient specificity. In this study, we addressed this issue and we report on the generation of a MART‐1::Cre BAC transgenic mouse line, in which the expression of Cre recombinase is controlled by regulatory elements of the pigment cell‐specific gene MART‐1 (mlana). When MART‐1::Cre BAC transgenic mice were bred with the ROSA26‐R reporter line, ß‐galactosidase expression was observed in RPE from E12.5 onwards, and in melanocyte precursors from E17.5, indicating that the MART‐1::Cre line provides Cre recombinase activity in pigment‐producing cells rather than in a particular lineage. In addition, breeding of this mouse line to mice carrying a conditional allele of RBP‐Jκ corroborated the reported phenotypes in both pigment cell lineages, inducing hair greying and microphthalmia. Our results thus suggest, that the MART‐1::Cre line may serve as a novel and useful tool for functional studies in melanocytes and the RPE.genesis 49:403–409, 2011. © 2011 Wiley‐Liss, Inc.     

Suppression of herbivory by macroalgal density: a critical feedback on coral reefs?

Coral reefs globally are in decline, with some reefs undergoing phase shifts from coral-dominance to degraded states dominated by large fleshy macroalgae. These shifts have been underpinned by the overharvesting of herbivorous fishes and represent a fundamental change in the physical structure of these reefs. Although the physical structure provided by corals is regarded as a key feature that facilitates herbivore activity, the influence of the physical structure of macroalgal stands is largely unknown. Using transplanted Sargassum, the largest coral reef macroalga, we created habitat patches of predetermined macroalgal density (0.25-6.23 kg m(-2)). Remote video cameras revealed both grazing and browsing fishes avoided high density patches, preferring relatively open areas with low macroalgal cover. This behaviour may provide a positive feedback leading to the growth and persistence of macroalgal stands; increasing the stability of phase shifts to macroalgae.     

Genetic parsimony: a factor in the evolution of complexity, order and emergence

Two conjectures, drawn from Gregory Chaitin's Algorithmic Information Theory, are examined with respect to the relationship between an algorithm and its product; in particular his finding that, where an algorithm is minimal, its length provides a measure of the complexity of the product. Algorithmic complexity is considered from the perspective of the relationship between genotype and phenotype, which Chaitin suggests is analogous to other algorithm-product systems. The first conjecture is that the genome is a minimal set of algorithms for the phenotype. Evidence is presented for a factor, here termed 'genetic parsimony', which is thought to have helped minimize the growth of genome size during evolution. Species that depend on rapid replication, such as prokaryotes which are generally r -selected are more likely to have small genomes, while the K -strategists accumulate introns and have large genomes. The second conjecture is that genome size could provide a measure of organism complexity. A surrogate index for coding DNA is in agreement with an established phenotypic index (number of cell types), in exhibiting an evolutionary trend of increasing organism complexity over time. Evidence for genetic parsimony indicates that simplicity in coding has been selected, and is responsible for phenotypic order. It is proposed that order evolved because order in the phenotype can be encoded more economically than disorder. Thus order arises due to selection for genetic parsimony, as does the evolution of other 'emergent' properties.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 88 , 295–308.     

The C-value enigma in plants and animals: a review of parallels and an appeal for partnership

• Aims Plants and animals represent the first two kingdomsrecognized, and remain the two best-studied groups in termsof nuclear DNA content variation. Unfortunately, the traditionalchasm between botanists and zoologists has done much to preventan integrated approach to resolving the C-value enigma, thelong-standing puzzle surrounding the evolution of genome size.This grand division is both unnecessary and counterproductive,and the present review aims to illustrate the numerous linksbetween the patterns and processes found in plants and animalsso that a stronger unity can be developed in the future. • Scope This review discusses the numerous parallels thatexist in genome size evolution between plants and animals, including(i) the construction of large databases, (ii) the patterns ofDNA content variation among taxa, (iii) the cytological, morphological,physiological and evolutionary impacts of genome size, (iv)the mechanisms by which genomes change in size, and (v) thedevelopment of new methodologies for estimating DNA contents. • Conclusions The fundamental questions of the C-valueenigma clearly transcend taxonomic boundaries, and increasedcommunication is therefore urged among those who study genomesize evolution, whether in plants, animals or other organisms.     

Production of CETD transgenic mouse line allowing ablation of any type of specific cell population

Diphtheria toxin A-chain (DT-A) is a potent inhibitor of protein synthesis. As little as a single molecule of DT-A can result in cell death. DT-A gene driven by a tissue-specific promoter is used to achieve genetic ablation of a particular cell lineage. However, this transgenic approach often results in aberrant depletion of unrelated cells. To avoid this, we established a method for specific depletion of a cell population by controlled expression of the DT-A gene via the Cre-loxP system. We produced five transgenic mice carrying CETD construct containing loxP-flanked enhanced green fluorescent protein (EGFP) cDNA and the DT-A gene. Transfection of primary cultured cells derived from CETD transgenic fetus with Cre expression plasmid resulted in extensive cell loss, as expected. Bigenic (double transgenic) offspring obtained by crossbreeding between CETD and MNCE transgenic mice in which Cre expression is controlled by the myelin basic protein (MBP) promoter exhibited embryonic lethality, suggesting expression of Cre at embryonic stages. Intravenous injection of Cre expression vector to CETD mice led to generation of glomerular lesions, probably due to predominant depletion of glomerular epithelial cells. This Cre-loxP-based cell ablation technology is powerful and convenient method of generating mice lacking any chosen cell population.     

Alternative reproductive tactics and status-dependent selection

The status-dependent selection model on alternative reproductivetactics predicts a single switch-point in status: usually allplayers above some status (e.g., competitive ability) shouldpractice the tactic with the higher average payoff, while thosebelow that point should make the "best of a bad job" by practicingthe alternative, lower payoff tactic. Many empirical studiesindeed show a relationship between status and tactic choice,but they do not conform to this single switch-point prediction.I modify the status-dependent selection model by consideringstatus-dependent fitness that is mediated, at least in part,by resource acquisition (e.g., status-based competition forterritories or nuptial gifts). With variation in resource quality,predicted tactic-choice distributions change: a high-statusmale may be territorial on a high-quality territory, a lowerstatus male may practice an alternative tactic, and an evenlower status male may be territorial on a low-quality territory.Tactic choice thus alternates as in many empirical studies andcan appear to be but is not actually stochastic. As the numberof theoretically predicted switch-points increases, however,mixed or mixed-conditional strategies should become more prevalent.While alternative tactics will likely usually differ in meanpayoff, viewing alternative reproductive tactics as inherently"better" or "worse" (e.g., viewing cuckoldry as "worse"—thebest of a bad job) is misleading if not tempered with awarenessthat payoff can vary greatly within tactics and overlap betweentactics.     

Conditional knockout mice to study alternative splicing in vivo

Analysis of genomes has revealed that the total number of human genes is comparable to those of simpler organisms, and thus, the number of genes does not correlate with the complexity and functional diversity of different organisms. Multiple mechanisms, including alternative splicing, are believed to contribute to the molecular complexity in higher eukaryotes. Given the fact that more than half of human genes undergo alternative splicing, however, little is known about the biological relevance of most alternative splicing events and their regulatory mechanisms. Recent work has highlighted the power of reverse genetic approaches in addressing regulated splicing in animal models. Here, we focus on the conditional knockout approach adapted for splicing research with the intention to provide a general guide to the generation of mouse models to study regulated splicing in development and disease.     

Temperature,Herbivory and Epibiont Acquisition as Factors Controlling the Distribution and Ecological Role of an Invasive Seaweed

The invasive canopy alga, Codium fragile ssp. tomentosoides, first observed at the Isles of Shoals in 1983, has become the dominant canopy species to 8 m throughout the islands. Codium populations are replacing themselves at most sites in what appears to be a new, climax, canopy species. However, Codium densities have declined in protected Gosport Harbor areas where it first became established. Codium has only slowly expanded its presence in adjacent nearshore subtidal habitats. Recent studies suggest a combination of factors that may be influencing the relative success of populations between habitats. The herbivorous sea slug, Placida dendritica, may be reducing populations in protected areas in spite of predators such as the green crab, Carcinus maenas, while surge may inhibit herbivore buildup in exposed habitats. Temperature instability due to localized, wind-driven upwelling may be slowing the buildup of subtidal Codium populations in nearshore sites. The combination of Codium dominance and the acquisition of increasing epibiont diversity are producing a new, potentially more complex community state than the previous kelp-dominated climax typical of the Gulf of Maine.