Hypodermal Cells Research Articles

Endogenous chondroitin extends lifespan by inhibiting VHA-7-mediated tubular lysosome formation

Chondroitin extends lifespan and healthspan in C. elegans, but the relationship between extracellular chondroitin and intracellular anti-aging mechanisms is unknown. The basement membrane (BM) that contains chondroitin proteoglycans is anchored to cells via hemidesmosomes (HDs), and it accumulates damage with aging. In this study, we found that chondroitin regulates aging through the formation of HDs and inhibition of tubular lysosomes (TLs). Reduction of chondroitin due to a mutation in sqv-5/Chondroitin synthase (ChSy) causes the earlier and excessive formation of TLs and leakage of the lysosomal nuclease in a manner dependent on VHA-7, the a-subunit of V-type ATPase. VHA-7, whose mutation suppresses the short lifespan of the sqv-5 mutant, is initially localized to the basal side of the hypodermal cells and transported to lysosomes with aging. These results demonstrate that endogenous chondroitin suppresses aging by inhibiting the earlier excessive formation of TLs. This is a novel anti-aging mechanism that is controlled by the BM.

A New Pattern of Phytomelanin Deposition in Cypselae of Bahieae (Asteraceae, Heliantheae Alliance)

ABSTRACTPhytomelanin is the dark and highly inert substance found in the fruit walls of the Heliantheae alliance clade within Asteroideae (Asteraceae). Phytomelanin deposition pattern in the cypselar wall was studied in two taxa of the Bahieae—Bahia pedata A. Gray and Palafoxia arida B. L. Turner. The pericarp anatomy studies have revealed that the phytomelanin deposits in the schizogenous space between the hypodermis and fiber layer. Our findings reveal that phytomelanin deposition is associated with fiber and has also brought out a new pattern of phytomelanin deposition between inner (fibrous zone) and outer sclerified layers (hypodermal) in the schizogenous space. The study recorded a first‐time report of a second layer of phytomelanin outer to the sclerified hypodermal cells at maturity. Our observations point to phytomelanin precursors being synthesized as pliable fluid substances associated with fiber cells and solidifies later in the intercellular space. The study also recorded the first report for the occurrence of tracheoidal cells in the pericarp wall of the Bahieae within the Heliantheae alliance. Our study has provided insights to decipher evolutionary patterns and the significance of phytomelanin in the phytomelanic fruit clade (PFC), and it could help to resolve long‐standing debates on deposition patterns and the site of synthesis of phytomelanin.

The structural organization of the outer tissues in the gametophytic stem of the umbrella moss Hypnodendron menziesii optimizes load bearing.

The ultrastructural design and biochemical organization of the significantly thickened outer tissues of the gametophytic stem of Hypnodendron menziesii optimizes load bearing of the stem. Hypnodendron menziesii is a bryoid umbrella moss growing in high humid conditions on the forest floors of New Zealand. The erect gametophyte bears up to eight whorls of branches in succession, spreading across the stem that bears the heavy weight of branches with highly hydrated leaves. Our investigation using a combination of light microscopy, transmission electron microscopy (TEM), scanning electron microscopy (SEM), and TEM-immunolabeling techniques provided novel information on the structural design and biochemical organization of greatly thickened cell walls of epidermal, hypodermal, and outermost cortical tissues, comparing underlying thin-walled cortical tissues in the gametophytic stem. Probing into the ultrastructure of the cell wall architecture of these target tissues by TEM and SEM revealed the cell walls to display a multilamellar organization, in addition to demonstrating the presence of an electron-dense substance in the cell wall, presumably flavonoids. The pattern of distribution and concentration of rhamnogalacturonan, homogalacturonan, and heteromannan, as determined by immunogold labeling, suggests that it is the combination of structural and molecular design of the cell wall that may optimize the mechanical function of the epidermal, hypodermal, and outer cortical tissues. Statistical relationships between the overall thickness of epidermal, hypodermal, and outer cortical cell walls, the lumen area of cells and the percentage area of cell wall occupied in these tissues at different heights of the stem, and thickness of secondary cell wall layers (L1-L4/5) were explored. The results of these analyses unequivocally support the contribution of outer tissues to the mechanical strength of the resilient stem.

Hyperspectral and Chlorophyll Fluorescence Analyses of Comparative Leaf Surfaces Reveal Cellular Influences on Leaf Optical Properties in Tradescantia Plants.

The differential effects of cellular and ultrastructural characteristics on the optical properties of adaxial and abaxial leaf surfaces in the genus Tradescantia highlight the intricate relationships between cellular arrangement and pigment distribution in the plant cells. We examined hyperspectral and chlorophyll a fluorescence (ChlF) kinetics using spectroradiometers and optical and electron microscopy techniques. The leaves were analysed for their spectral properties and cellular makeup. The biochemical compounds were measured and correlated with the biophysical and ultrastructural features. The main findings showed that the top and bottom leaf surfaces had different amounts and patterns of pigments, especially anthocyanins, flavonoids, total phenolics, chlorophyll-carotenoids, and cell and organelle structures, as revealed by the hyperspectral vegetation index (HVI). These differences were further elucidated by the correlation coefficients, which influence the optical signatures of the leaves. Additionally, ChlF kinetics varied between leaf surfaces, correlating with VIS-NIR-SWIR bands through distinct cellular structures and pigment concentrations in the hypodermis cells. We confirmed that the unique optical properties of each leaf surface arise not only from pigmentation but also from complex cellular arrangements and structural adaptations. Some of the factors that affect how leaves reflect light are the arrangement of chloroplasts, thylakoid membranes, vacuoles, and the relative size of the cells themselves. These findings improve our knowledge of the biophysical and biochemical reasons for leaf optical diversity, and indicate possible implications for photosynthetic efficiency and stress adaptation under different environmental conditions in the mesophyll cells of Tradescantia plants.

Cutinsomes of Malus Mill. (Rosaceae) leaf and pericarp: genesis, localization, and transport

New data were obtained on specific bionanostructures, cutinsomes, which are involved in the formation of cuticles on the surface of leaf blades and pericarp of Malus domestica Borkh (Malus Mill., Rosaceae)introduced to the mountains at the altitudes of 1200 and 1700 m above sea level. Cutinsomes, which are electron-dense structures of spherical shape, have been identified by transmission electron microscopy. It was demonstrated that plastids can be involved in the synthesis of their constituent nanocomponents. The greatest number of nanoparticles was observed in the granal thylakoid lumen of the chloroplasts in palisade mesophyll cells and pericarp hypodermal cells. The transmembrane transport of cutinsomes into the cell wall cuticle proper by exocytosis has been visualized for the first time. The plasma membrane is directly involved in the excretion of nanostructures from the cell. Nanoparticles of cutinsomes in the form of necklace-like formations line up in a chain near cell walls, merge into larger conglomerates and are loaded into plasmalemma invaginations, and then, in membrane packing, they move into the cuticle, which covers both outer and inner cell walls of external tissues. The original materials obtained by us supplement the ideas about the non-enzymatic synthesis of cuticle components available in the literature and expand the cell compartment geography involved in this process.

Histological and single-nucleus transcriptome analyses reveal the specialized functions of ligular sclerenchyma cells and key regulators of leaf angle in maize

Leaf angle (LA) is a crucial factor that affects planting density and yield in maize. However, the regulatory mechanisms underlying LA formation remain largely unknown. In this study, we performed a comparative histological analysis of the ligular region across various maize inbred lines and revealed that LA is significantly influenced by a two-step regulatory process involving initial cell elongation followed by subsequent lignification in the ligular adaxial sclerenchyma cells (SCs). Subsequently, we performed both bulk and single-nucleus RNA sequencing, generated a comprehensive transcriptomic atlas of the ligular region, and identified numerous genes enriched in the hypodermal cells that may influence their specialization into SCs. Furthermore, we functionally characterized two genes encoding atypical basic-helix–loop–helix (bHLH) transcription factors, bHLH30 and its homolog bHLH155, which are highly expressed in the elongated adaxial cells. Genetic analyses revealed that bHLH30 and bHLH155 positively regulate LA expansion, and molecular experiments demonstrated their ability to activate the transcription of genes involved in cell elongation and lignification of SCs. These findings highlight the specialized functions of ligular adaxial SCs in LA regulation by restricting further extension of ligular cells and enhancing mechanical strength. The transcriptomic atlas of the ligular region at single-nucleus resolution not only deepens our understanding of LA regulation but also enables identification of numerous potential targets for optimizing plant architecture in modern maize breeding.

Cacao Procyanidins-Induced Lifespan Extension in Caenorhabditis elegans in a Nervous System and CaMKII-Dependent Manner.

Procyanidins are gaining attention due to their potential health benefits. We found that cacao liquor procyanidin (CLPr) from Theobroma cacao seeds increased the lifespan of Caenorhabditis elegans, a representative model organism for aging studies. The genetic dependence of the lifespan-extending effect of CLPr was consistent with that of blueberry procyanidin, which is dependent on unc-43, osr-1, sek-1, and mev-1, but not on daf-16, sir-2.1, or skn-1. The lifespan-extending effect of CLPr was inhibited by neuron-specific RNA interference (RNAi) targeting unc-43 and pmk-1, and in worms with loss-of-function mutations in the odr-3, odr-1, or tax-4 genes, which are essential in sensory neurons, including AWC neurons. It was also inhibited in worms in which AWC neurons or AIB interneurons had been eliminated, and in worms with loss-of-function mutations in eat-4 or glr-1, which are responsible for glutamatergic synaptic transmission. These results suggest that the lifespan-extending effect of CLPr is dependent on the nervous system. In addition, it also requires unc-43 and pmk-1 expression in nonneuronal cells, as demonstrated by the experiments with RNAi in wild-type worms, the neuronal cells of which are not affected by systemic RNAi. The osr-1 gene is expressed in hypodermal and intestinal cells and regulates the response to osmotic stress along with unc-43/calcium/calmodulin-dependent protein kinase II and the p38 mitogen-activated protein kinase pathway. Consistent with this, CLPr improved osmotic stress tolerance in an unc-43- and pmk-1-dependent manner, and it was also dependent on AWC neurons. The lifespan-extending and osmotic-tolerance-improving activities were attributed to procyanidins with a tetrameric or higher-order oligomeric structure.

An RNAi screen for conserved kinases that enhance microRNA activity after dauer in Caenorhabditis elegans.

Gene regulation in changing environments is critical for maintaining homeostasis. Some animals undergo a stress-resistant diapause stage to withstand harsh environmental conditions encountered during development. MicroRNAs are one mechanism for regulating gene expression during and after diapause. MicroRNAs downregulate target genes posttranscriptionally through the activity of the microRNA-induced silencing complex. Argonaute is the core microRNA-induced silencing complex protein that binds to both the microRNA and to other microRNA-induced silencing complex proteins. The 2 major microRNA Argonautes in the Caenorhabditis elegans soma are ALG-1 and ALG-2, which function partially redundantly. Loss of alg-1 [alg-1(0)] causes penetrant developmental phenotypes including vulval defects and the reiteration of larval cell programs in hypodermal cells. However, these phenotypes are essentially absent if alg-1(0) animals undergo a diapause stage called dauer. Levels of the relevant microRNAs are not higher during or after dauer, suggesting that activity of the microRNA-induced silencing complex may be enhanced in this context. To identify genes that are required for alg-1(0) mutants to develop without vulval defects after dauer, we performed an RNAi screen of genes encoding conserved kinases. We focused on kinases because of their known role in modulating microRNA-induced silencing complex activity. We found RNAi knockdown of 4 kinase-encoding genes, air-2, bub-1, chk-1, and nekl-3, caused vulval defects and reiterative phenotypes in alg-1(0) mutants after dauer, and that these defects were more penetrant in an alg-1(0) background than in wild type. Our results implicate these kinases as potential regulators of microRNA-induced silencing complex activity during postdauer development in C. elegans.

A taxonomic revision of Rhizophora L. (Rhizophoraceae) in Thailand.

A taxonomic revision of Rhizophora L. (Rhizophoraceae) in Thailand is presented. Two species, R. apiculata Blume and R. mucronata Poir., are enumerated with updated morphological descriptions, illustrations and a taxonomic identification key, together with notes on distributions, habitats and ecology, phenology, conservation assessments, etymology, vernacular names, uses, and specimens examined. Three names in Rhizophora, are lectotypified: R. apiculata and two associated synonyms of R. mucronata (i.e., R. latifolia Miq. and R. macrorrhiza Griff.). R. longissima Blanco, a synonym of R. mucronata, is neotypified. All two Rhizophora species have a conservation assessment of Least Concern (LC). Based on the morphological identification, these two species can be distinguished from one another by the shape and width of the leaf laminae and the length of a terminal stiff point of the leaf laminae; the type and position of the inflorescences and the number of flowers per inflorescence; the character and color of the bracteoles; the presence or absence of the flower pedicels; the shape of the mature flower buds; the shape, color, and texture of the sepals; the shape, character, and the presence or absence of hairs of the petals; the number of stamens per flower; the size of the fruits; the color and size of the hypocotyls; the color and diameter of the cotyledonous cylindrical tubes; and the color of the colleters and exudate. The thick cuticles, sunken stomata, large hypodermal cells, and cork warts are adaptive anatomical features of leaves in Rhizophora that live in the mangrove environments. The pollen grains of Thai Rhizophora species are tricolporate, prolate spheroidal or oblate spheroidal shapes, small-sized, and reticulate exine sculpturing.

Anatomical Characteristics of Rhizophora’s Leaves as Mangrove Plant Adaptation at Banyuurip Mangrove Center

Rhizophora is a genus of mangrove plants that dominates the Banyuurip Mangrove Center. This plant certainly has an anatomical structure as a form of adaptation to high salinity environments. The purpose of this study was to describe the anatomical characteristics of the leaves of the three Rhizophora species as an adaptation form of mangrove plants in the Banyuurip Mangrove Center and the anatomical variations of the leaves between species. This type of research was descriptive observational in the form of leaf anatomy observations using the whole mount method for longitudinal incisions and the paraffin method for transverse incisions. The results showed that there were anatomical variations among the three Rhizophora species in the Banyuurip Mangrove Center including epidermal cell size, number of epidermal cells, cork warts, number of hypodermis cell layers, hypodermis cell size, stomata size, number of stomata, stomata index, and stomata density. The anatomical characteristics of the leaves of the three Rhizophora species which act as a form of adaptation to mangrove plants are the presence of hypodermis tissue which functions to store water and salt to remove salt content in plants when they abort their leaves, low density of stomata and the presence of cuticles on the adaxial surface of leaves which play a role in reducing the rate of transpiration thus maintaining water to support plant development in saline conditions, cork warts which function as a medium for expelling salt on leaves.

Comparative foliar anatomical study of six different varieties of Piper betle L.

The present investigation on a few selected cultivars of Piper betle L. was undertaken mainly to understand the anatomy and tissue components of the stem, leaves and petiole. This was accomplished by using basic anatomy techniques such as free hand and microtome sectioning. The varieties selected were: HY1, HY2, JB, KB, LV and SG. Major aspects of the present inquiry were - macro morphology, anatomy and histochemistry. The main focus of the study was on the internal structure of leaves and petioles to understand the role of specialised cells like enlarged hypodermal cells and mucilage cavities in retaining moisture and thereby preserving the shelf life of harvested leaves.

Ultrastructural characteristics of the reproductive organs of adult male helminth Heterakis dispar (Nematoda: Heterakidae)

Nematodes belonging to the Heterakidae family are similar in appearance and their identification causes certain difficulties. For this reason, there is a need to define new taxonomic characteristics. On the other hand, despite the fact that the nematode Heterakis dispar Schrank, 1790, included in the same family, has a wide distribution area, there is little information about the ultrastructure of the parasite. Research on the ultrastructure of the reproductive system of this helminth hase not been conducted. Parasitic worms collected from the caeca of birds were identified, following which, based on the accepted methods of electron microscopy, blocks were prepared, cut, stained, pictures and electrograms were prepared and described. In the present study, the ultrastructural characteristics of the reproductive organs (testis, seminal vesicle, vas deferens and ejaculatory duct) of the adult nematode H. dispar, a specific parasite of domestic waterfowl, were studied with the help of light and electron microscopic methods. The obtained results were compared with the structure of other studied species (H. gallinarum Gmelin, 1790 and H. spumosa Schneider, 1866) of the Heterakidae family. Firstly, it was revealed that the reproductive organs of the male nematode H. dispar are located in the pseudocoelomic cavity, which is considered a taxonomic sign of this family. As in other species of the family, the testis, seminal vesicle, and wall of the vas deferens of the male nematode H. dispar consist of a basement membrane and an epithelial layer. In addition to the basement membrane, muscular and epithelial layers, the wall of the ejaculatory duct is also composed of glandular cells in the front part of the duct. It was revealed that, in the lumen of the tubular reproductive organs of the adult male nematode H. dispar, germ-cells turn into spermatogonia in the germinal zone and spermatocytes in the growth zone of the testis, spermatids in the seminal vesicle, incomplete formed spermatozoa in the vas deferens, and fully formed spermatozoa in the ejaculatory duct of the parasite. The spicules of the parasite nematode, which are the secondary sexual organs consist of a thick cuticle on the outside, and hypodermal origin cells and processes of nerve cells (dendrites and axons) on the inside. Two processes are found on the spicules, which is not observed in other nematodes of the same family.

NHR-23 activity is necessary for C. elegans developmental progression and apical extracellular matrix structure and function.

Nematode molting is a remarkable process where animals must repeatedly build a new apical extracellular matrix (aECM) beneath a previously built aECM that is subsequently shed. The nuclear hormone receptor NHR-23 (also known as NR1F1) is an important regulator of C. elegans molting. NHR-23 expression oscillates in the epidermal epithelium, and soma-specific NHR-23 depletion causes severe developmental delay and death. Tissue-specific RNAi suggests that nhr-23 acts primarily in seam and hypodermal cells. NHR-23 coordinates the expression of factors involved in molting, lipid transport/metabolism and remodeling of the aECM. NHR-23 depletion causes dampened expression of a nas-37 promoter reporter and a loss of reporter oscillation. The cuticle collagen ROL-6 and zona pellucida protein NOAH-1 display aberrant annular localization and severe disorganization over the seam cells after NHR-23 depletion, while the expression of the adult-specific cuticle collagen BLI-1 is diminished and frequently found in patches. Consistent with these localization defects, the cuticle barrier is severely compromised when NHR-23 is depleted. Together, this work provides insight into how NHR-23 acts in the seam and hypodermal cells to coordinate aECM regeneration during development.

Glandular trichome development, morphology, and maturation are influenced by plant age and genotype in high THC-containing cannabis (Cannabis sativa L.) inflorescences

BackgroundGlandular capitate trichomes which form on bract tissues of female inflorescences of high THC-containing Cannabis sativa L. plants are important sources of terpenes and cannabinoids. The influence of plant age and cannabis genotype on capitate trichome development, morphology, and maturation has not been extensively studied. Knowledge of the various developmental changes that occur in trichomes over time and the influence of genotype and plant age on distribution, numbers, and morphological features should lead to a better understanding of cannabis quality and consistency.MethodsBract tissues of two genotypes—“Moby Dick” and “Space Queen”—were examined from 3 weeks to 8 weeks of flower development using light and scanning electron microscopy. Numbers of capitate trichomes on upper and lower bract surfaces were recorded at different positions within the inflorescence. Observations on distribution, extent of stalk formation, glandular head diameter, production of resin, and extent of dehiscence and senescence were made at various time points. The effects of post-harvesting handling and drying on trichome morphology were examined in an additional five genotypes.ResultsTwo glandular trichome types—bulbous and capitate (sessile or stalked)—were observed. Capitate trichome numbers and stalk length were significantly (P = 0.05) greater in “Space Queen” compared to “Moby Dick” at 3 and 6 weeks of flower development. Significantly more stalked-capitate trichomes were present on lower compared to upper bract surfaces at 6 weeks in both genotypes, while sessile-capitate trichomes predominated at 3 weeks. Epidermal and hypodermal cells elongated to different extents during stalk formation, producing significant variation in length (from 20 to 1100 μm). Glandular heads ranged from 40 to 110 μm in diameter. Maturation of stalked-capitate glandular heads was accompanied by a brown color development, reduced UV autofluorescence, and head senescence and dehiscence. Secreted resinous material from glandular heads appeared as droplets on the cuticular surface that caused many heads to stick together or collapse. Trichome morphology was affected by the drying process.ConclusionCapitate trichome numbers, development, and degree of maturation were influenced by cannabis genotype and plant age. The observations of trichome development indicate that asynchronous formation leads to different stages of trichome maturity on bracts. Trichome stalk lengths also varied between the two genotypes selected for study as well as over time. The variability in developmental stage and maturation between genotypes can potentially lead to variation in total cannabinoid levels in final product. Post-harvest handling and drying were shown to affect trichome morphology.

Large vesicle extrusions from C. elegans neurons are consumed and stimulated by glial-like phagocytosis activity of the neighboring cell.

Caenorhabditis elegans neurons under stress can produce giant vesicles, several microns in diameter, called exophers. Current models suggest that exophers are neuroprotective, providing a mechanism for stressed neurons to eject toxic protein aggregates and organelles. However, little is known of the fate of the exopher once it leaves the neuron. We found that exophers produced by mechanosensory neurons in C. elegans are engulfed by surrounding hypodermal skin cells and are then broken up into numerous smaller vesicles that acquire hypodermal phagosome maturation markers, with vesicular contents gradually degraded by hypodermal lysosomes. Consistent with the hypodermis acting as an exopher phagocyte, we found that exopher removal requires hypodermal actin and Arp2/3, and the hypodermal plasma membrane adjacent to newly formed exophers accumulates dynamic F-actin during budding. Efficient fission of engulfed exopher-phagosomes to produce smaller vesicles and degrade their contents requires phagosome maturation factors SAND-1/Mon1, GTPase RAB-35, the CNT-1 ARF-GAP, and microtubule motor-associated GTPase ARL-8, suggesting a close coupling of phagosome fission and phagosome maturation. Lysosome activity was required to degrade exopher contents in the hypodermis but not for exopher-phagosome resolution into smaller vesicles. Importantly, we found that GTPase ARF-6 and effector SEC-10/exocyst activity in the hypodermis, along with the CED-1 phagocytic receptor, is required for efficient production of exophers by the neuron. Our results indicate that the neuron requires specific interaction with the phagocyte for an efficient exopher response, a mechanistic feature potentially conserved with mammalian exophergenesis, and similar to neuronal pruning by phagocytic glia that influences neurodegenerative disease.

Factors determining the mechanical properties of banana fruit skin during induced ripening

Postharvest peel splitting in banana impairs visual quality and exacerbates mass loss, fruit fly infestation and microbial infection. The aim was to determine the mechanistic basis of peel splitting. We therefore monitored the change in physical and mechanical properties of the peel as affected by different storage conditions. Peel splitting occurred axially, in the fruit corners. It began 4 d after ripening induction. Splitting was preceded by radial cracks in the inner peel, that propagated towards the outside. In fruit stored at 98 % relative humidity (RH), splitting occurred in 38 % of fruit; there was no splitting in fruit stored at 14 or 60 % RH. The percentages of fruit with radial cracks in the inner peel were 63 %, 100 %, and 100 % at 14 %, 60 % and 98 % RH, respectively. The fracture force (Fmax) of the peel decreased rapidly from 53.0 ± 1.6 N to 2.1 ± 0.2 N during ripening. Simulated radial cracks (cuts) in the inner peel decreased Fmax from 22.8 ± 2.2 N to 15.9 ± 0.9 N Paring off the outer peel decreased Fmax from 18.9 ± 1.7 to 4.4 ± 0.8 N, but paring off the inner peel had no effect. Osmotic water uptake by peel discs incubated in glucose solutions decreased with increasing concentration. Water was lost to hypertonic glucose solutions causing discs to shrink. Epidermal and hypodermal cell wall thickness both increased due to cell wall swelling. Our results show: (1) the outer peel layers represent the mechanical backbone of the peel, (2) radial cracks in the inner peel are the first stage of peel splitting and (3) peel splitting is caused by local stress concentrations due to radial cracking and is exacerbated by cell wall swelling. Key to reducing peel splitting at retail is prompt removal of fruit from the high RH environment of the plastic-lined cartons.

An nhr-23::mScarlet::3xMyc knock-in allele for studying spermatogenesis and molting.

C. elegans NHR-23 is a nuclear hormone receptor transcription factor involved in molting, apical extracellular matrix structure, and spermatogenesis. To determine NHR-23 expression dynamics, we previously tagged the endogenous nhr-23 locus with a GFP::AID*::3xFLAG tag. To allow co-localization of NHR-23 with green fluorescent protein-tagged factors of interest, we generated an equivalent strain carrying an mScarlet::3xMyc tag to produce a C-terminal fusion. Similar to the GFP::AID*::3xFLAG knock-in, NHR-23 ::mScarlet::3xMyc was expressed in seam and hypodermal cells, vulval precursor cells, and the spermatogenic germline. We also observed a diffuse NHR-23::mScarlet expression pattern in spermatids and residual bodies after NHR-23 ceased to localize on chromatin. Further examination of this novel localization may provide insight into NHR-23 regulation of spermatogenesis.

An atonal homolog, lin-32, regulates hypodermal morphogenesis in Caenorhabditis elegans.

The transcription factor atonal contributes to patterning and cell fate determination in specialized epithelial cells in various animals, but its function in hypodermis is unknown. Here, we analyzed the atonal homolog lin-32 in C. elegans to clarify whether atonal acts in hypodermal development. The lin-32 null mutants exhibited bulges and cavities in their head, which were prevented by LIN-32 expression. Fluorescent protein was expressed in hypodermis cells at the embryonic stage by the lin-32 promoter. These results certify that atonal plays an essential role in the development of a broader range of tissues as hypodermis than initially thought.

The long isoform of the C. elegans ELT-3 GATA factor can specify endoderm when overexpressed.

The C. elegans elt-3 gene encodes a GATA transcription factor that is expressed in the hypodermis and has roles in hypodermal specification and regulation of collagen and stress response genes. The gene encodes short and long isoforms, ELT-3A and ELT-3B respectively, that differ upstream of their DNA-binding domains. Previous work showed that ELT-3A can specify hypodermal cell fates when forcibly overexpressed throughout early embryos. We recently showed that the ELT-3B orthologue from the distantly related species C. angaria can specify endodermal fates when forcibly overexpressed in C. elegans. Here, we show that C. elegans ELT-3B can also specify endoderm.

The novel gene, mtre-1 , is expressed downstream of MAB-3 and DMD-3 in the male tail tip at the termination of male tail tip retraction.

The development of the adult C. elegans male tail involves an extensive remodeling during the last larval stage where the pointed tail of the L4 male is converted into the blunt-ended adult tail with its collection of mechano-sensitive rays. The first step in this remodeling is the retraction of the four hypodermal cells of the tail tip to generate the blunt-ended tail. Male tail tip retraction is an excellent model for characterizing how upstream regulatory networks interact with the downstream cell biological effectors that drive morphogenetic changes in all animals. Previously, we've shown that two DM-domain transcription factors, MAB-3 and DMD-3 , are central regulators of male tail tip retraction. Using a microarray-based approach we have identified ~400 genes that are more highly expressed in the L4 male tail tip relative to the hermaphrodite L4 tail tip. The uncharacterized gene T05H10.3 , which we've named mtre-1 , was highly over-represented in the male tail tip vs. the hermaphrodite tail tip and was under-represented in mab-3 ; dmd-3 mutant male tail tips vs. wild-type male tail tips. A transcriptional reporter for mtre-1 shows clear expression in the male tail tip cells for a short period (~3 hours) at the end of retraction. This expression is dependent on the activity of MAB-3 and DMD-3 , since expression is reduced in dmd-3 single mutant males and absent in mab-3 ; dmd-3 mutant males. Finally, males homozygous for a putative null allele of mtre-1 display a phenotypically wild-type adult male tail, indicating that mtre-1 is not essential for male tail morphogenesis.