Pavement Cells Research Articles

Measuring Biotic Ligand Model (BLM) Parameters in Vitro: Copper and Silver Binding to Rainbow Trout Gill Cells as Cultured Epithelia or in Suspension.

Biotic ligand models (BLMs) for metals are useful for risk assessment. The modeling of metal complexation by the biotic ligand has received little attention relative to the modeling of organic and inorganic complexation of metals in solution. We used ion selective electrodes (ISEs) to directly characterize copper and silver binding to rainbow trout gill cells, either as cultured reconstructed epithelia, or dispersed in suspension. Preparations were composed of pavement cells (PVCs) alone or mixtures of PVCs (≈85%) and mitochondria-rich cells (MRCs, ≈15%). Mixed cells showed up to an order of magnitude greater binding for both metals, indicating that MRCs were much more important than PVCs. Also, cell orientation had a dramatic effect; cells cultured as epithelia exhibited much greater binding than cells in suspension. Silver and copper demonstrated generally similar binding behavior, with stronger (logK ≈ 10 or greater) and weaker binding sites (logK ≈ 8). Comparisons to existing BLM calibrations show good agreement, but reveal that selection of analytical window can impact which binding sites are titrated. We conclude that cultured gill epithelia in vitro provide a powerful approach to studying metal complexation directly at the biotic ligand.

Pavement cells and the topology puzzle.

D'Arcy Thompson emphasised the importance of surface tension as a potential driving force in establishing cell shape and topology within tissues. Leaf epidermal pavement cells grow into jigsaw-piece shapes, highly deviating from such classical forms. We investigate the topology of developing Arabidopsis leaves composed solely of pavement cells. Image analysis of around 50,000 cells reveals a clear and unique topological signature, deviating from previously studied epidermal tissues. This topological distribution is established early during leaf development, already before the typical pavement cell shapes emerge, with topological homeostasis maintained throughout growth and unaltered between division and maturation zones. Simulating graph models, we identify a heuristic cellular division rule that reproduces the observed topology. Our parsimonious model predicts how and when cells effectively place their division plane with respect to their neighbours. We verify the predicted dynamics through in vivo tracking of 800 mitotic events, and conclude that the distinct topology is not a direct consequence of the jigsaw piece-like shape of the cells, but rather owes itself to a strongly life history-driven process, with limited impact from cell-surface mechanics.

An examination of nucleotypic effects in diploid and polyploid cotton

Nucleotypic effects are phenotypic changes related to the total nuclear DNA amount per cell. These effects are commonly observed among and within genera for certain cell types, and the generality of the positive correlation between genome size and cell size has been well established. However, there are few studies of nucleotypic effects which incorporate into the analysis both ploidy level and genome size (given as Mbp determined by 2C values). To test the hypothesis that cell size scales with genome size and ploidy, we measured the guard cell length, epidermal pavement cell surface area, and pollen grain diameter using individuals of multiple species and accessions of the cotton genus (Gossypium), in which different species exhibit three-fold variation in genome size. We measured cell sizes using calibrated microscopic image analysis. Significant relationships were found between genome size and cell size, with stronger correlations between guard cell length and genome size than with epidermal pavement cell surface area. We also found a relationship between pollen grain diameter and genome size. These results indicate that nucleotypic effects occur within Gossypium, scale with ploidy level, and are stronger in less variable cell types.

Exogenous Cellulase Switches Cell Interdigitation to Cell Elongation in an RIC1-dependent Manner in Arabidopsis thaliana Cotyledon Pavement Cells.

Pavement cells in cotyledons and true leaves exhibit a jigsaw puzzle-like morphology in most dicotyledonous plants. Among the molecular mechanisms mediating cell morphogenesis, two antagonistic Rho-like GTPases regulate local cell outgrowth via cytoskeletal rearrangements. Analyses of several cell wall-related mutants suggest the importance of cell wall mechanics in the formation of interdigitated patterns. However, how these factors are integrated is unknown. In this study, we observed that the application of exogenous cellulase to hydroponically grown Arabidopsis thaliana cotyledons switched the interdigitation of pavement cells to the production of smoothly elongated cells. The cellulase-induced inhibition of cell interdigitation was not observed in a RIC1 knockout mutant. This gene encodes a Rho-like GTPase-interacting protein important for localized cell growth suppression via microtubule bundling on concave cell interfaces. Additionally, to characterize pavement cell morphologies, we developed a mathematical model that considers the balance between cell and cell wall growth, restricted global cell growth orientation, and regulation of local cell outgrowth mediated by a Rho-like GTPase-cytoskeleton system. Our computational simulations fully support our experimental observations, and suggest that interdigitated patterns form because of mechanical buckling in the absence of Rho-like GTPase-dependent regulation of local cell outgrowth. Our model clarifies the cell wall mechanics influencing pavement cell morphogenesis.

Arabidopsis myrosinases link the glucosinolate-myrosinase system and the cuticle.

Both physical barriers and reactive phytochemicals represent two important components of a plant’s defence system against environmental stress. However, these two defence systems have generally been studied independently. Here, we have taken an exclusive opportunity to investigate the connection between a chemical-based plant defence system, represented by the glucosinolate-myrosinase system, and a physical barrier, represented by the cuticle, using Arabidopsis myrosinase (thioglucosidase; TGG) mutants. The tgg1, single and tgg1 tgg2 double mutants showed morphological changes compared to wild-type plants visible as changes in pavement cells, stomatal cells and the ultrastructure of the cuticle. Extensive metabolite analyses of leaves from tgg mutants and wild-type Arabidopsis plants showed altered levels of cuticular fatty acids, fatty acid phytyl esters, glucosinolates, and indole compounds in tgg single and double mutants as compared to wild-type plants. These results point to a close and novel association between chemical defence systems and physical defence barriers.

Growth increase of Arabidopsis by forced expression of rice 45S rRNA gene.

Forced expression of rice 45S rRNA gene conferred ca. 2-fold increase of above-ground growth in transgenic Arabidopsis . This growth increase was probably brought by cell proliferation, not by cell enlargement. Recent increase in carbon dioxide emissions is causing global climate change. The use of plant biomass as alternative energy source is one way to reduce these emissions. Therefore, reinforcement of plant biomass production is an urgent key issue to overcome both depletion of fossil energies and emission of carbon dioxide. Here, we created transgenic Arabidopsis with a 2-fold increase in above-ground growth by forced expression of the rice 45S rRNA gene using the maize ubiquitin promoter. Although the size of guard cells and ploidy of leaf-cells were similar between transgenic and control plants, numbers of stomata and pavement cells were much increased in the transgenic leaf. This data suggested that cell number, not cell expansion, was responsible for the growth increase, which might be brought by the forced expression of exogenous and full-length 45S rRNA gene. The expression level of rice 45S rRNA transcripts was very low, possibly triggering unknown machinery to enhance cell proliferation. Although microarray analysis showed enhanced expression of ethylene-responsive transcription factors, these factors might respond to ethylene induced by abiotic/biotic stresses or genomic incompatibility, which might be involved in the expression of species-specific internal transcribed spacer (ITS) sequences within rice 45S rRNA transcripts. Further analysis of the mechanism underlying the growth increase will contribute to understanding the regulation of the cell proliferation and the mechanism of hybrid vigor.

Plant Responses to Brief Touching: A Mechanism for Early Neighbour Detection?

In natural habitats plants can be exposed to brief and light contact with neighbouring plants. This mechanical stimulus may represent a cue that induces responses to nearby plants. However, little is known about the effect of touching on plant growth and interaction with insect herbivores. To simulate contact between plants, a soft brush was used to apply light and brief mechanical stimuli to terminal leaves of potato Solanum tuberosum L. The number of non-glandular trichomes on the leaf surface was counted on images made by light microscope while glandular trichomes and pavement cells were counted on images made under scanning electronic microscope. Volatile compounds were identified and quantified using coupled gas chromatography–mass spectrometry (GC-MS). Treated plants changed their pattern of biomass distribution; they had lower stem mass fraction and higher branch and leaf mass fraction than untouched plants. Size, weight and number of tubers were not significantly affected. Touching did not cause trichome damage nor change their total number on touched terminal leaves. However, on primary leaves the number of glandular trichomes and pavement cells was significantly increased. Touching altered the volatile emission of treated plants; they released higher quantities of the sesquiterpenes (E)-β-caryophyllene, germacrene D-4-ol and (E)-nerolidol, and lower quantities of the terpenes (E)-ocimene and linalool, indicating a systemic effect of the treatment. The odour of touched plants was significantly less preferred by the aphids Macrosiphum euphorbiae and Myzus persicae compared to odour of untouched plants. The results suggest that light contact may have a potential role in the detection of neighbouring plants and may affect plant-insect interactions.

Cytokinin activity increases stomatal density and transpiration rate in tomato.

Previous studies on cytokinin (CK) and drought have suggested that the hormone has positive and negative effects on plant adaptation to restrictive conditions. This study examined the effect of CK on transpiration, stomatal activity, and response to drought in tomato (Solanum lycopersicum) plants. Transgenic tomato plants overexpressing the Arabidopsis thaliana CK-degrading enzyme CK oxidase/dehydrogenase 3 (CKX3) maintained higher leaf water status under drought conditions due to reduced whole-plant transpiration. The reduced transpiration could be attributed to smaller leaf area and reduced stomatal density. CKX3-overexpressing plants contained fewer and larger pavement cells and fewer stomata per leaf area than wild-type plants. In addition, wild-type leaves treated with CK exhibited enhanced transpiration and had more pavement cells and increased numbers of stomata per leaf area than untreated leaves. Manipulation of CK levels did not affect stomatal movement or abscisic acid-induced stomatal closure. Moreover, we found no correlation between stomatal aperture and the activity of the CK-induced promoter Two-Component Signaling Sensor (TCS) in guard cells. Previous studies have shown that drought reduces CK levels, and we propose this to be a mechanism of adaptation to water deficiency: the reduced CK levels suppress growth and reduce stomatal density, both of which reduce transpiration, thereby increasing tolerance to prolonged drought conditions.

Changes in cuticular wax coverage and composition on developing Arabidopsis leaves are influenced by wax biosynthesis gene expression levels and trichome density.

Wax coverage on developing Arabidopsis leaf epidermis cells is constant and thus synchronized with cell expansion. Wax composition shifts from fatty acid to alkane dominance, mediated by CER6 expression. Epidermal cells bear a wax-sealed cuticle to hinder transpirational water loss. The amount and composition of the cuticular wax mixture may change as organs develop, to optimize the cuticle for specific functions during growth. Here, morphometrics, wax chemical profiling, and gene expression measurements were integrated to study developing Arabidopsis thaliana leaves and, thus, further our understanding of cuticular wax ontogeny. Before 5days of age, cells at the leaf tip ceased dividing and began to expand, while cells at the leaf base switched from cycling to expansion at day 13, generating a cell age gradient along the leaf. We used this spatial age distribution together with leaves of different ages to determine that, as leaves developed, their wax compositions shifted from C24/C26 to C30/C32 and from fatty acid to alkane constituents. These compositional changes paralleled an increase in the expression of the elongase enzyme CER6 but not of alkane pathway enzymes, suggesting that CER6 transcriptional regulation is responsible for both chemical shifts. Leaves bore constant numbers of trichomes between 5 and 21days of age and, thus, trichome density was higher on young leaves. During this time span, leaves of the trichome-less gl1 mutant had constant wax coverage, while wild-type leaf coverage was initially high and then decreased, suggesting that high trichome density leads to greater apparent coverage on young leaves. Conversely, wax coverage on pavement cells remained constant over time, indicating that wax accumulation is synchronized with cell expansion throughout leaf development.

The composition of surface wax on trichomes of Arabidopsis thaliana differs from wax on other epidermal cells.

To protect plants against biotic and abiotic stress, the waxy cuticle must coat all epidermis cells. Here, two independent approaches addressed whether cell-type-specific differences exist between wax compositions on trichomes and other epidermal cells of Arabidopsis thaliana, possibly with different protection roles. First, the total waxes from a mutant lacking trichomes (gl1) were compared to waxes from wild type and a trichome-rich mutant (cpc tcl1 etc1 etc3). In the stem wax, compounds with aliphatic chains longer than 31 carbons (derived from C32 precursors) increased in relative abundance in cpc tcl1 etc1 etc3 over gl1. Similarly, the leaf wax from the trichome-rich mutant contained higher amounts of C32+ compounds as compared to gl1. Second, leaf trichomes were isolated, and their waxes were analyzed. The wax mixtures of the trichome-rich mutant and the wild type were similar, comprising alkanes and alkenes as well as branched and unbranched primary alcohols. The direct analyses of trichome waxes confirmed that they contained relatively high concentrations of C32+ compounds, compared with the pavement cell wax inferred from analysis of gl1 leaves. Finally, the cell-type-specific wax compositions were put into perspective with expression patterns of wax biosynthesis genes in trichomes and pavement cells. Analyses of published transcriptome data (Marks etal., ) revealed that core enzymes involved in elongation of wax precursors to various carbon chain lengths are expressed differentially between epidermis cell types. By combining the chemical and gene expression data, we identified promising gene candidates involved in the formation of C32+ aliphatic chains.

The FOUR LIPS (FLP) and MYB88 genes conditionally suppress the production of nonstomatal epidermal cells in Arabidopsis cotyledons.

The Arabidopsis MYB transcriptional factors FOUR LIPS (FLP) and MYB88 are predicted to play a broader role than just limiting the number of guard cells per stoma in the epidermis based on their expression patterns. Analysis of the numbers of all epidermal cells in cotyledons in flp mutants and the flp-1 myb88 double mutant may substantiate the functions of FLP and MYB88 in nonstomatal epidermal cells. The number of all types of abaxial epidermal cells in mature cotyledons were determined under different growth conditions in flp-1, flp-3, flp-8, and tmm-1 single mutants, flp-1 tmm-1 and flp-1 myb88 double mutants, and accessions Columbia-0 (Col, control for flp-1, flp-3, and flp-1 myb88) and Landsberg erecta (Ler, control for flp-8). In soil-grown plants, the number of pavement cells and meristemoids per cotyledon were not statistically different between the flp mutants and their respective controls and between flp-1 and flp-1 myb88, except sometimes for the number of meristemoids or pavement cells between flp-1 and Col. In contrast, the same comparisons yielded statistically significant differences in medium-grown plants, i.e., more cells in the flp mutants and flp-1 myb88, except for meristemoids between flp-8 and Ler and pavement cells between flp-1 and flp-1 myb88. No significant difference was detected for nonstomatal epidermal cells between tmm-1 and flp-1/tmm-1 under the two conditions. FLP and MYB88 inhibit the production of nonstomatal epidermal cells largely in a growth-condition-dependent manner. The tmm-1 mutation is epistatic to the flp-1 mutation in the production of nonstomatal epidermal cells in the cotyledon.

The Dehydratase ADT3 Affects ROS Homeostasis and Cotyledon Development.

During the transition from seed to seedling, emerging embryos strategically balance available resources between building up defenses against environmental threats and initiating the developmental program that promotes the switch to autotrophy. We present evidence of a critical role for the phenylalanine (Phe) biosynthetic activity of AROGENATE DEHYDRATASE3 (ADT3) in coordinating reactive oxygen species (ROS) homeostasis and cotyledon development in etiolated Arabidopsis (Arabidopsis thaliana) seedlings. We show that ADT3 is expressed in the cotyledon and shoot apical meristem, mainly in the cytosol, and that the epidermis of adt3 cotyledons contains higher levels of ROS Genome-wide proteomics of the adt3 mutant revealed a general down-regulation of plastidic proteins and ROS-scavenging enzymes, corroborating the hypothesis that the ADT3 supply of Phe is required to control ROS concentration and distribution to protect cellular components. In addition, loss of ADT3 disrupts cotyledon epidermal patterning by affecting the number and expansion of pavement cells and stomata cell fate specification; we also observed severe alterations in mesophyll cells, which lack oil bodies and normal plastids. Interestingly, up-regulation of the pathway leading to cuticle production is accompanied by an abnormal cuticle structure and/or deposition in the adt3 mutant. Such impairment results in an increase in cell permeability and provides a link to understand the cell defects in the adt3 cotyledon epidermis. We suggest an additional role of Phe in supplying nutrients to the young seedling.

Plant-Herbivore Interaction: Dissection of the Cellular Pattern of Tetranychus urticae Feeding on the Host Plant.

The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), is one of the most polyphagous herbivores feeding on cell contents of over 1100 plant species including more than 150 crops. It is being established as a model for chelicerate herbivores with tools that enable tracking of reciprocal responses in plant-spider mite interactions. However, despite their important pest status and a growing understanding of the molecular basis of interactions with plant hosts, knowledge of the way mites interface with the plant while feeding and the plant damage directly inflicted by mites is lacking. Here, utilizing histology and microscopy methods, we uncovered several key features of T. urticae feeding. By following the stylet path within the plant tissue, we determined that the stylet penetrates the leaf either in between epidermal pavement cells or through a stomatal opening, without damaging the epidermal cellular layer. Our recordings of mite feeding established that duration of the feeding event ranges from several minutes to more than half an hour, during which time mites consume a single mesophyll cell in a pattern that is common to both bean and Arabidopsis plant hosts. In addition, this study determined that leaf chlorotic spots, a common symptom of mite herbivory, do not form as an immediate consequence of mite feeding. Our results establish a cellular context for the plant-spider mite interaction that will support our understanding of the molecular mechanisms and cell signaling associated with spider mite feeding.

Functional characterization of the ribosome biogenesis factors PES, BOP1, and WDR12 (PeBoW), and mechanisms of defective cell growth and proliferation caused by PeBoW deficiency in Arabidopsis

The nucleolar protein pescadillo (PES) controls biogenesis of the 60S ribosomal subunit through functional interactions with Block of Proliferation 1 (BOP1) and WD Repeat Domain 12 (WDR12) in plants. In this study, we determined protein characteristics and in planta functions of BOP1 and WDR12, and characterized defects in plant cell growth and proliferation caused by a deficiency of PeBoW (PES-BOP1-WDR12) proteins. Dexamethasone-inducible RNAi of BOP1 and WDR12 caused developmental arrest and premature senescence in Arabidopsis, similar to the phenotype of PES RNAi. Both the N-terminal domain and WD40 repeats of BOP1 and WDR12 were critical for specific associations with 60S/80S ribosomes. In response to nucleolar stress or DNA damage, PeBoW proteins moved from the nucleolus to the nucleoplasm. Kinematic analyses of leaf growth revealed that depletion of PeBoW proteins led to dramatically suppressed cell proliferation, cell expansion, and epidermal pavement cell differentiation. A deficiency in PeBoW proteins resulted in reduced cyclin-dependent kinase Type A activity, causing reduced phosphorylation of histone H1 and retinoblastoma-related (RBR) protein. PeBoW silencing caused rapid transcriptional modulation of cell-cycle genes, including reduction of E2Fa and Cyclin D family genes, and induction of several KRP genes, accompanied by down-regulation of auxin-related genes and up-regulation of jasmonic acid-related genes. Taken together, these results suggest that the PeBoW proteins involved in ribosome biogenesis play a critical role in plant cell growth and survival, and their depletion leads to inhibition of cell-cycle progression, possibly modulated by phytohormone signaling.

LobeFinder: A Convex Hull-Based Method for Quantitative Boundary Analyses of Lobed Plant Cells.

Dicot leaves are composed of a heterogeneous mosaic of jigsaw puzzle piece-shaped pavement cells that vary greatly in size and the complexity of their shape. Given the importance of the epidermis and this particular cell type for leaf expansion, there is a strong need to understand how pavement cells morph from a simple polyhedral shape into highly lobed and interdigitated cells. At present, it is still unclear how and when the patterns of lobing are initiated in pavement cells, and one major technological bottleneck to addressing the problem is the lack of a robust and objective methodology to identify and track lobing events during the transition from simple cell geometry to lobed cells. We developed a convex hull-based algorithm termed LobeFinder to identify lobes, quantify geometric properties, and create a useful graphical output of cell coordinates for further analysis. The algorithm was validated against manually curated images of pavement cells of widely varying sizes and shapes. The ability to objectively count and detect new lobe initiation events provides an improved quantitative framework to analyze mutant phenotypes, detect symmetry-breaking events in time-lapse image data, and quantify the time-dependent correlation between cell shape change and intracellular factors that may play a role in the morphogenesis process.

Homologs of SCAR/WAVE complex components are required for epidermal cell morphogenesis in rice.

Filamentous actins (F-actins) play a vital role in epidermal cell morphogenesis. However, a limited number of studies have examined actin-dependent leaf epidermal cell morphogenesis events in rice. In this study, two recessive mutants were isolated: less pronounced lobe epidermal cell2-1 (lpl2-1) and lpl3-1, whose leaf and stem epidermis developed a smooth surface, with fewer serrated pavement cell (PC) lobes, and decreased papillae. The lpl2-1 also exhibited irregular stomata patterns, reduced plant height, and short panicles and roots. Molecular genetic studies demonstrated that LPL2 and LPL3 encode the PIROGI/Specifically Rac1-associated protein 1 (PIR/SRA1)-like and NCK-associated protein 1 (NAP1)-like proteins, respectively, two components of the suppressor of cAMP receptor/Wiskott-Aldrich syndrome protein-family verprolin-homologous protein (SCAR/WAVE) regulatory complex involved in actin nucleation and function. Epidermal cells exhibited abnormal arrangement of F-actins in both lpl2 and lpl3 expanding leaves. Moreover, the distorted trichomes of Arabidopsis pir could be partially restored by an overexpression of LPL2 A yeast two-hybrid assay revealed that LPL2 can directly interact with LPL3 in vitro Collectively, the results indicate that LPL2 and LPL3 are two functionally conserved homologs of the SCAR/WAVE complex components, and that they play an important role in controlling epidermal cell morphogenesis in rice by organising F-actin.

Osteoblastoma in the retromolar region: a case report

ABSTRACT Osteoblastoma is a rare benign tumor that accounts for less than 1% of all bone tumors. About 10% of osteoblastomas are found in the skull bones and almost half of these cases involve the mandible, especially the posterior segments. This report describes the case of a 27-year-old female patient with a unilocular radiopaque expansive lesion in the left mandible, which had well-delimited margins and caused mild bone expansion. Microscopic analysis revealed the presence of mineralized material in the form of irregularly arranged vital trabeculae at different stages of mineralization. Osteocytes were trapped inside these trabeculae, which contained osteoclast-like multinucleated cells and voluminous pavement cells with hyperchromatic nuclei, sometimes interpreted as osteoblasts. Many bone-producing lesions have clinical, radiologic and histopathologic features that resemble osteoblastoma. The understanding and correlation of these findings are extremely important since they contribute to the correct diagnosis and appropriate treatment of this rare entity, improving its prognosis.

Cell wall accumulation of fluorescent proteins derived from a trans-Golgi cisternal membrane marker and paramural bodies in interdigitated Arabidopsis leaf epidermal cells

In most dicotyledonous plants, leaf epidermal pavement cells develop jigsaw puzzle-like shapes during cell expansion. The rapid growth and complicated cell shape of pavement cells is suggested to be achieved by targeted exocytosis that is coordinated with cytoskeletal rearrangement to provide plasma membrane and/or cell wall materials for lobe development during their morphogenesis. Therefore, visualization of membrane trafficking in leaf pavement cells should contribute an understanding of the mechanism of plant cell morphogenesis. To reveal membrane trafficking in pavement cells, we observed monomeric red fluorescent protein-tagged rat sialyl transferases, which are markers of trans-Golgi cisternal membranes, in the leaf epidermis of Arabidopsis thaliana. Quantitative fluorescence imaging techniques and immunoelectron microscopic observations revealed that accumulation of the red fluorescent protein occurred mostly in the curved regions of pavement cell borders and guard cell ends during leaf expansion. Transmission electron microscopy observations revealed that apoplastic vesicular membrane structures called paramural bodies were more frequent beneath the curved cell wall regions of interdigitated pavement cells and guard cell ends in young leaf epidermis. In addition, pharmacological studies showed that perturbations in membrane trafficking resulted in simple cell shapes. These results suggested possible heterogeneity of the curved regions of plasma membranes, implying a relationship with pavement cell morphogenesis.

Cinnamon clownfish Amphiprion melnaopus의 이소토신 유전자 구조와 삼투압 조절이 미치는 영향

Isotocin (IT), a nonapeptide homolog of oxytocin in mammals, has been suggested to be involved in physiological processes including social behaviors, stress responses, and osmoregulation in teleost fish. To study its structure and function, the gene encoding the IT precursor was cloned from the genomic DNA and brain cDNA of the cinnamon clownfish, Amphiprion melanopus. The IT precursor gene consists of three exons separated by two introns, and encodes an open reading frame of 156 amino acid (aa) residues, comprising a putative signal peptide of 19 aa, a mature IT protein of 9 aa, a proteolytic processing site of 3 aa, and 125 aa of neurophysin. Tissue-specific analysis of the IT precursor transcript indicated its expression in the brain and gonads of A. melanopus. To examine its osmoregulatory effects, the salinity of the seawater (34 ppt) used for rearing A. melanopus was lowered to 15 ppt. Histological analysis of the gills indicated the apparent disappearance of an apical crypt on the surface of the gill lamella of A. melanopus, as pavement cells covered the surface upon acclimation to the lower salinity. The level of Na + /K + -ATPase activity in the gills was increased during the initial stage of acclimation, followed by a decrease to its normal level, suggesting its involvement in osmoregulation and homeostasis. The only slight increase in the level of IT precursor transcript in the A. melanopus brain upon low-salinity acclimation suggested that IT played a minor role, if any, in the process of osmoregulation.

On growth and formins

ABSTRACTDevelopment of the plant aerial organs epidermis involves a complex interplay of cytoskeletal rearrangements, membrane trafficking-dependent cell surface expansion, and intra- and intercellular signaling, resulting in a pattern of perfectly interlocking pavement cells. While recent detailed in vivo observations convincingly identify microtubules rather than actin as key players at the early stages of development of pavement cell lobes in Arabidopsis, mutations affecting the actin-nucleating ARP2/3 complex are long known to reduce pavement cell lobing, suggesting a central role for actin. We have now shown that functional impairment of the Arabidopsis formin FH1 enhances both microtubule dynamics and pavement cell lobing. While formins are best known for their ability to nucleate actin, many members of this old gene family now emerge as direct or indirect regulators of the microtubule cytoskeleton, and our findings suggest that they might co-ordinate action of the two cytoskeletal systems during pavement cell morphogenesis.