Basal Accumulation Research Articles

The promoted lateral root 1 (plr1) mutation is involved in reduced basal shoot starch accumulation and increased root sugars for enhanced lateral root growth in rice

Lateral roots (LRs) are indispensable for plant growth, adaptability and productivity. We previously reported a rice mutant, exhibiting a high density of thick and long LRs (L-type LRs) with long parental roots and herein referred to as promoted lateral root1 (plr1). In this study, we describe that the mutant exhibited decreased basal shoot starch accumulation, suggesting that carbohydrates might regulate the mutant root phenotype. Further analysis revealed that plr1 mutation gene regulated reduced starch accumulation resulting in increased root sugars for the regulation of promoted LR development. This was supported by the exogenous glucose application that promoted L-type LRs. Moreover, nitrogen (N) application was found to reduce basal shoot starch accumulation in both plr1 mutant and wild-type seedlings, which was due to the repressed expression of starch biosynthesis genes. However, unlike the wild-type that responded to N treatment only at seedling stage, the plr1 mutant regulated LR development under low to increasing N levels, both at seedling and higher growth stages. These results suggest that plr1 mutation gene is involved in reduced basal shoot starch accumulation and increased root sugar level for the promotion of L-type LR development, and thus would be very useful in improving rice root architecture.

Integrins regulate epithelial cell shape by controlling the architecture and mechanical properties of basal actomyosin networks

Forces generated by the actomyosin cytoskeleton are key contributors to many morphogenetic processes. The actomyosin cytoskeleton organises in different types of networks depending on intracellular signals and on cell-cell and cell-extracellular matrix (ECM) interactions. However, actomyosin networks are not static and transitions between them have been proposed to drive morphogenesis. Still, little is known about the mechanisms that regulate the dynamics of actomyosin networks during morphogenesis. This work uses the Drosophila follicular epithelium, real-time imaging, laser ablation and quantitative analysis to study the role of integrins on the regulation of basal actomyosin networks organisation and dynamics and the potential contribution of this role to cell shape. We find that elimination of integrins from follicle cells impairs F-actin recruitment to basal medial actomyosin stress fibers. The available F-actin redistributes to the so-called whip-like structures, present at tricellular junctions, and into a new type of actin-rich protrusions that emanate from the basal cortex and project towards the medial region. These F-actin protrusions are dynamic and changes in total protrusion area correlate with periodic cycles of basal myosin accumulation and constriction pulses of the cell membrane. Finally, we find that follicle cells lacking integrin function show increased membrane tension and reduced basal surface. Furthermore, the actin-rich protrusions are responsible for these phenotypes as their elimination in integrin mutant follicle cells rescues both tension and basal surface defects. We thus propose that the role of integrins as regulators of stress fibers plays a key role on controlling epithelial cell shape, as integrin disruption promotes reorganisation into other types of actomyosin networks, in a manner that interferes with proper expansion of epithelial basal surfaces.

Totten Glacier subglacial hydrology determined from geophysics and modeling

Aurora Subglacial Basin (ASB), which feeds Totten Glacier, is a marine basin lying below sea level and contains up to 3.5m of global sea level equivalent. Rates of future sea level rise from this area are primarily dependent on the stability of Totten Ice Shelf and the controls on ice flow dynamics upstream of the grounding line, both of which may be influenced by subglacial hydrology. We apply the GlaDS subglacial hydrology model to ASB to examine whether the spatial patterns of distributed and efficient drainage systems impact the dynamics of Totten Glacier. We determine the most appropriate model configuration from our series of sensitivity tests by comparing the modeled basal water pressure and water depth results with specularity content data. Those data are derived from ICECAP radar surveys over the same region and represent regions of basal water accumulation. The best match between simulated basal hydrology properties and specularity content shows a strong correspondence in regions of distributed water in the ASB troughs for both water depth and water pressure, but weak correspondence between water depth and specularity content near the grounding line. This may be due to the presence of several large channels draining over the grounding line into the head of Totten Ice Shelf, which are likely not as well represented in the specularity content data as distributed systems. These channels may have a significant impact on melt, and therefore the stability, of Totten Ice Shelf. Within ASB, regions of high water pressure and greater water accumulation correspond well with regions of faster ice flow, suggesting some control of basal hydrology on ice dynamics in this region.

The Lac au Renard Tephra Cluster: a record of Lochkovian (Lower Devonian) volcanism in the Indian Point Formation, Gaspe Peninsula, Quebec, Canada

Approximately twenty tephra beds, comprising the Lac au Renard Tephra Cluster (new), occur in the Rosebush Cove and Petit Portage members of the Indian Point Formation (Chaleurs Group) on the Gaspe Peninsula of Quebec, Canada. The tephra beds range in thickness from <1.0 cm to 82 cm and occur in a mudrock-dominated sequence with coarser tempestite interbeds. Mineralogically and texturally graded accumulations of phenocrysts mark the bases of the thickest tephra beds. Early diagenetic concretions in one composite tephra preserve pre-compaction fabrics and original hypocrystalline textures with microphenocrysts and devitrified glass shards. The presence of plagioclase, quartz, K-feldspar, biotite, apatite, and zircon suggest a rhyolitic source. The coarseness of the microphenocrysts in the basal accumulations, along with the abundance and thickness of the tephras, suggest that deposition in the Gaspe area was in a proximal position relative to the volcanic source. The zonal graptolites Monograptus praehercynicus and Monograptus aequabilis ssp. from the Petit Portage Member indicate a middle Lochkovian age for the Lac au Renard Tephra Cluster of the Indian Point Formation. Correlation with the tephra cluster that includes the Judds Falls Bentonite in the New Scotland Formation and other possible tephras in the Kalkberg Formation (Helderberg Group) of New York and the Corriganville and Mandata formations of Pennsylvania is likely. The graptolite fauna of the Indian Point and probable correlations to New York may provide additional biostratigraphic constraints on a U–Pb zircon radiometric age determination of 417.6 Ma from New York that has been used to estimate the age of the Silurian–Devonian boundary.

Loblolly pine outperforms slash pine in the southeastern United States – A long-term experimental comparison study

Loblolly pine (Pinus taeda L.) and slash pine (Pinus elliottii Engelm.) are the two most important commercial timber species in the southeastern U.S. A long-term experimental comparison study, in which loblolly and slash pine plots were paired for combinations of planting density (741, 2224, 3706 trees ha−1) and cultural intensity (operational versus intensive), was established in the lower Coastal Plain to investigate species differences in growth dynamics. Using age 2–21-year inventory plot data and destructive biomass/nutrient sampling data, species comparisons were conducted comprehensively, and the reasons of the species differences and management implications were discussed. When compared to slash pine, loblolly pine had higher stand basal area carrying capacity and maximum stand density index, lower fusiform rust infection rate and less wind damage, leading to its lower mortality. With no species difference in average DBH, loblolly pine consistently had greater average height and live crown length and higher level of tree size inequality. Loblolly pine had lower foliar biomass and lower foliar density, and higher crown length/width ratios due primarily to its longer crowns, suggesting perhaps a slightly greater shade tolerance or less dense canopy. As a result, loblolly pine outperformed slash pine in terms of stand basal area, total volume, aboveground biomass and carbon accumulation. Loblolly pine consistently accumulated more potassium, and this species difference increased with increasing stand age; while slash pine had slightly higher nitrogen and phosphorus contents before age 12. Species differences in mortality patterns and in nutrient accumulations in aboveground biomass emphasized the need to develop species-specific management strategies including different thinning and fertilization regimes.

Ultrastructural Characterization of Stem Cell-Derived Replacement Vestibular Hair Cells Within Ototoxin-Damaged Rat Utricle Explants.

ABSTRACTThe auditory apparatus of the inner ear does not show turnover of sensory hair cells (HCs) in adult mammals; in contrast, there are many observations supporting low‐level turnover of vestibular HCs within the balance organs of mammalian inner ears. This low‐level renewal of vestibular HCs exists during normal conditions and it is further enhanced after trauma‐induced loss of these HCs. The main process for renewal of HCs within mammalian vestibular epithelia is a conversion/transdifferentiation of existing supporting cells (SCs) into replacement HCs.In earlier studies using long‐term organ cultures of postnatal rat macula utriculi, HC loss induced by gentamicin resulted in an initial substantial decline in HC density followed by a significant increase in the proportion of HCs to SCs indicating the production of replacement HCs. In the present study, using the same model of ototoxic damage to study renewal of vestibular HCs, we focus on the ultrastructural characteristics of SCs undergoing transdifferentiation into new HCs. Our objective was to search for morphological signs of SC plasticity during this process. In the utricular epithelia, we observed immature HCs, which appear to be SCs transdifferentiating into HCs. These bridge SCs have unique morphological features characterized by formation of foot processes, basal accumulation of mitochondria, and an increased amount of connections with nearby SCs. No gap junctions were observed on these transitional cells. The tight junction seals were morphologically intact in both control and gentamicin‐exposed explants. Anat Rec, 303:506–515, 2020. © 2019 The Authors. The Anatomical Record published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists.

Species-Specific (Hyalella azteca and Lymnea stagnalis) Dietary Accumulation of Gold Nano-particles Associated with Periphyton.

Ecological effects of gold nano-particles (AuNP) are examined due to growing use in consumer and industrial materials. This study investigated uptake and movement of AuNPs through an aquatic food chain. Simple (single-species) and diverse (multi-species) periphyton communities were exposed to AuNP (0, 100, 500µg L-1 treatments). AuNP quickly aggregated and precipitated from the water column, suggesting it is an insignificant route of AuNP exposure even at elevated concentrations. Gold was measured in 100 and 500µg L-1 periphyton treatments. Gold accumulation was similar between periphyton treatments, suggesting physical processes were important for AuNP basal accumulation. Hyalella azteca and Lymnea stagnalis whole body tissue analysis indicated gold accumulation may be attributed to different feeding mechanisms, general versus selective grazing, respectively. Results suggest trophic transfer of AuNP is organism specific and aggregation properties of AuNP are important when considering fate of nano-particles in the environment and movement through aquatic food webs.

IL-1 induces p62/SQSTM1 and autophagy in ERα+ /PR+ BCa cell lines concomitant with ERα and PR repression, conferring an ERα- /PR- BCa-like phenotype.

Estrogen receptor α (ERα)low/- tumors are associated with breast cancer (BCa) endocrine resistance, where ERα low tumors show a poor prognosis and a molecular profile similar to triple negative BCa tumors. Interleukin-1 (IL-1) downregulates ERα accumulation in BCa cell lines, yet the cells can remain viable. In kind, IL-1 and ERα show inverse accumulation in BCa patient tumors and IL-1 is implicated in BCa progression. IL-1 represses the androgen receptor hormone receptor in prostate cancer cells concomitant with the upregulation of the prosurvival, autophagy-related protein, Sequestome-1 (p62/SQSTM1; hereinafter, p62); and given their similar etiology, we hypothesized that IL-1 also upregulates p62 in BCa cells concomitant with hormone receptor repression. To test our hypothesis, BCa cell lines were exposed to conditioned medium from IL-1-secreting bone marrow stromal cells (BMSCs), IL-1, or IL-1 receptor antagonist. Cells were analyzed for the accumulation of ERα, progesterone receptor (PR), p62, or the autophagosome membrane protein, microtubule-associated protein 1 light chain 3 (LC3), and for p62-LC3 interaction. We found that IL-1 is sufficient to mediate BMSC-induced ERα and PR repression, p62 and autophagy upregulation, and p62-LC3 interaction in ERα+ /PR+ BCa cell lines. However, IL-1 does not significantly elevate the high basal p62 accumulation or high basal autophagy in the ERα- /PR- BCa cell lines. Thus, our observations imply that IL-1 confers a prosurvival ERα- /PR- molecular phenotype in ERα+ /PR+ BCa cells that may be dependent on p62 function and autophagy and may underlie endocrine resistance.

Astaxanthin exerts anti-inflammatory and antioxidant effects in macrophages in NRF2-dependent and independent manners

Although anti-inflammatory effects of astaxanthin (ASTX) have been suggested, the underlying mechanisms have not been fully understood. Particularly, the modulatory action of ASTX in the interplay between nuclear factor E2-related factor 2 (NRF2) and nuclear factor κB (NFκB) to exert its anti-inflammatory effect in macrophages is unknown. The effect of ASTX on mRNA and protein expression of pro-inflammatory and antioxidant genes and/or cellular reactive oxygen species (ROS) accumulation were determined in RAW 264.7 macrophages, bone marrow-derived macrophages (BMDM) from wild-type (WT) and Nrf2-deficient mice, and/or splenocytes and peritoneal macrophages of obese mice fed ASTX. The effect of ASTX on M1 and M2 macrophage polarization was evaluated in BMDM. ASTX significantly decreased LPS-induced mRNA expression of interleukin 6 (Il-6) and Il-1β by inhibiting nuclear translocation of NFκB p65; and attenuated LPS-induced ROS with an increase in NRF2 nuclear translocation, concomitantly decreasing NADPH oxidase 2 expression in RAW 264.7 macrophages. In BMDM of WT and Nrf2-deficient mice, ASTX decreased basal and LPS-induced ROS accumulation. The induction of Il-6 mRNA by LPS was repressed by ASTX in both types of BMDM while Il-1β mRNA was decreased only in WT BMDM. Furthermore, ASTX consumption lowered LPS sensitivity of splenocytes in obese mice. ASTX decreased M1 polarization of BMDM while increasing M2 polarization. ASTX exerts its anti-inflammatory effect by inhibiting nuclear translocation of NFκB p65 and by preventing ROS accumulation in NRF2-dependent and -independent mechanisms. Thus, ASTX is an agent with anti-inflammatory and antioxidant properties that may be used for the prevention of inflammatory conditions.

14-3-3εa directs the pulsatile transport of basal factors toward the apical domain for lumen growth in tubulogenesis

The Ciona notochord has emerged as a simple and tractable in vivo model for tubulogenesis. Here, using a chemical genetics approach, we identified UTKO1 as a selective small molecule inhibitor of notochord tubulogenesis. We identified 14-3-3εa protein as a direct binding partner of UTKO1 and showed that 14-3-3εa knockdown leads to failure of notochord tubulogenesis. We found that UTKO1 prevents 14-3-3εa from interacting with ezrin/radixin/moesin (ERM), which is required for notochord tubulogenesis, suggesting that interactions between 14-3-3εa and ERM play a key role in regulating the early steps of tubulogenesis. Using live imaging, we found that, as lumens begin to open between neighboring cells, 14-3-3εa and ERM are highly colocalized at the basal cortex where they undergo cycles of accumulation and disappearance. Interestingly, the disappearance of 14-3-3εa and ERM during each cycle is tightly correlated with a transient flow of 14-3-3εa, ERM, myosin II, and other cytoplasmic elements from the basal surface toward the lumen-facing apical domain, which is often accompanied by visible changes in lumen architecture. Both pulsatile flow and lumen formation are abolished in larvae treated with UTKO1, in larvae depleted of either 14-3-3εa or ERM, or in larvae expressing a truncated form of 14-3-3εa that lacks the ability to interact with ERM. These results suggest that 14-3-3εa and ERM interact at the basal cortex to direct pulsatile basal accumulation and basal-apical transport of factors that are essential for lumen formation. We propose that similar mechanisms may underlie or may contribute to lumen formation in tubulogenesis in other systems.

Abstract 311: In vitro Regulation of G-protein Coupled Receptor Kinase 5 (GRK5) Induced Cardiac Hypertrophy by a Novel Interaction With the Phosphatase PHLPP2

PHLPP is a Ser/Thr phosphatase that has been shown to directly dephosphorylate several members of the AGC family of kinases, which are important regulators of cell growth, proliferation, and survival. We have previously demonstrated that removal of PHLPP1 increases Akt activity in neonatal rat ventricular myocytes (NRVMs) (Miyamoto et al 2010) as well as the heart (Moc et al 2015) leading to protection from pathological hypertrophy and damage, however the effects of PHLPP2 removal on cardiomyocyte hypertrophic growth are unknown. In NRVMs, we found that removal of PHLPP2 induced hypertrophic growth basally without altering Akt activity and immunoprecipitation experiments revealed a novel interaction with an AGC kinase, G-protein coupled receptor kinase 5 (GRK5). GRK5 acts as an HDAC kinase downstream of phenylephrine (PE) to regulate hypertrophy in NRVMs. Thus, our objective was to determine whether PHLPP2 regulates PE-induced cardiomyocyte hypertrophy through its interaction with GRK5. We found that knockdown of PHLPP2, not PHLPP1, potentiated PE-induced hypertrophy (siPH2+PE (109 ± 3.5μm) vs. siCon+PE (88 ± 1.9μm)), and increased mRNA expression of fetal genes (i.e. ANF and BNP). Conversely, overexpression of PHLPP2 attenuated PE-induced hypertrophic growth (PH2+PE (66.6 ±7.7μm) vs. GFP+PE (95.5 ± 2.7μm)) as well as fetal gene re-expression. This inhibition of hypertrophic growth was dependent on phosphatase activity since a PHLPP2 mutant lacking the phosphatase domain (αPP2C) reversed the effect of PHLPP2 overexpression. To determine whether removal of PHLPP2 alters GRK5 translocation, cells were fractionated, and we found that PHLPP2 removal significantly increased basal and PE-induced nuclear accumulation of GRK5 that was blocked by overexpression of PHLPP2. Finally, knockdown studies with siGRK5 revealed that GRK5 was indeed required for the hypertrophic response elicited by PHLPP2 removal. Overall, our data revealed for the first time that PHLPP2 plays a role in regulating PE-induced cardiac hypertrophy via a GRK5 dependent pathway. Understanding the biological function and signaling pathways altered by PHLPP2 in cardiomyocytes may help delineate therapeutic targets for cardiac hypertrophy.

Characterizing Forest Floor Fuels Surrounding Large Sugar Pine (Pinus lambertiana) in the Klamath Mountains, California

Fire exclusion has contributed to substantial basal accumulations of forest floor fuels in many historically fire-prone forests in western North America. These accumulations can contribute to undesired impacts (e.g., elevated tree mortality, more smoke production). Thus, managers interested in limiting some of these impacts require a better understanding of the spatial factors contributing to forest floor fuel variation in long-unburned forests. To address this need, we measured the forest floor mound size and depth of each layer at four distances from the tree (base, half, edge, and beyond) in each cardinal direction from the tree, and a subset of density measures for each layer and position around 25 large sugar pine (Pinus lambertiana) in the Klamath Mountains of California. Based on our study we found: 1) duff depths were greater than litter depths and both decreased with distance from tree; 2) duff densities were typically greater than litter but were highly variable with distance to tree; 3) litter and duff depths were positively correlated with loading; 4) use of a previously developed depth-to-loading equation performed well, but tended to underestimate litter loading and overestimate duff loading; and 5) tree size was the most strongly related to basal accumulations of forest floor fuels. Our results indicate there are consistent tree characteristics that can help predict the accumulation of forest floor fuels. This research can help provide managers with site-specific equations to inform fuel and fire treatments in areas with substantial basal accumulation in forests with large sugar pine.

Impact of moderate prenatal alcohol exposure on histaminergic neurons, histidine decarboxylase levels and histamine H2 receptors in adult rat offspring.

We have reported that moderate prenatal alcohol exposure (PAE) elevates histamine H3 receptor-mediated inhibition of glutamatergic neurotransmission in dentate gyrus (DG), and that the H3 receptor antagonist ABT-239 ameliorates PAE-induced deficits in DG long-term potentiation. Here, we investigated whether PAE alters other markers of histaminergic neurotransmission. Long-Evans rat dams voluntarily consumed either a 0% or a 5% ethanol solution 4h each day throughout gestation. Young adult female offspring from each prenatal treatment group were used in histidine decarboxylase (HDC) immunohistochemical studies of histamine neuron number in ventral hypothalamus, quantitative Western blotting studies of HDC expression in multiple brain regions, radiohistochemical studies of H2 receptor density in multiple brain regions, and in biochemical studies of H2 receptor-effector coupling in dentate gyrus. Rat dams consumed a mean of 1.90g of ethanol/kg/day during pregnancy. This level of consumption did not affect maternal weight gain, offspring birth weight, or litter size. PAE did not affect the number of HDC-positive neurons in ventral hypothalamus. However, HDC expression was reduced in frontal cortex, dentate gyrus, and cerebellum of PAE rats compared to controls. Specific [125I]-iodoaminopotentidine binding to H2 receptors was not altered in any of the brain regions measured, nor was basal or H2 receptor agonist-stimulated cAMP accumulation in DG altered in PAE rats compared to controls. These results suggest that not all markers of histaminergic neurotransmission are altered by PAE. However, the observation that HDC levels were reduced in the same brain regions where elevated H3 receptor-effector coupling was observed previously raises the question of whether a cause-effect relationship exists between HDC expression and H3 receptor function in affected brain regions of PAE rats. This relationship, along with the question of why these effects occur in some, but not all brain regions, requires more-detailed investigation.

Effects of six-week consumption of lard or palm oil on blood pressure and blood vessel H2S in middle-aged male rats

Background: Cardiovascular disease is the leading cause of death. The etiology of this disease is multifactorial, with unhealthy nutrition being one of the main risk factors. Diets high in animal fats and saturated fatty acid have been associated with an increased risk for cardiovascular diseases. However, the results of investigations on the effects of lard (LO)- and palm oil (PO) on cardiovascular risk are still controversial due to the dosages used and the age of the animals investigated. Objective: We investigated whether LO or PO consumption led to different effects on blood pressure, vascular functions, and lipid profiles within middle-aged rats. Methods: The study was performed in middle-aged male rats, n = 6 for each group. LO, PO, or distilled water (control) 1 or 3 ml/kg were orally gavaged once a day for 6 weeks. Basal blood pressure and heart rate were measured in anesthetized rats. Fasting serum lipids were measured by enzymatic methods. The vascular functions of isolated thoracic aorta were studied using pharmacological techniques in the absence or presence of N-nitro-L-arginine, a nitric oxide synthase (eNOS) inhibitor and or DL-propargylglycine (PAG), a cystothionine-γ-lyase (CSE) inhibitor. Additionally, the aortic wall eNOS and CSE protein expression were measured by Western blotting.Results: In comparison to the control group (distilled water, DW), no differences were observed in any of the parameters studied after the rats took 1 ml/kg of LO or PO. However, PO caused an increase in neutrophil/lymphocyte ratio and body fat. At 3 ml/kg dosage, LO caused increased basal blood pressure (LO, 153.4 ± 3.2; DW, 131.4 ± 3.2 mm Hg for systolic blood pressure and LO, 130.9 ± 2.5; DW, 107.9 ± 5.8 mmHg for diastolic blood pressure) body and liver cell lipid accumulation, while PO led to increased body fat and fasting serum triglyceride (PO, 131.5 ± 13.2; DW, 91.8 ± 4.8 mg %). Neither LO nor PO treatment had any effect on vascular contraction to phenylephrine, except in the presence of PAG which led to an increased contractile response to phenylephrine. PO but not LO treatment caused increased vascular wall CSE protein expression.Conclusion: The results document how both LO and PO at a dose of 3 ml/kg (corresponding to three servings of Thai fast food) cause increased cardiovascular risk factors. However, the blood vessel H2S production increased while the lower dose had a minimal effect.Keywords: Lard oil; Palm oil; blood vessel; liver lipid; NO; H2S

Autophagic Flux Is Regulated by Interaction Between the C-terminal Domain of PATCHED1 and ATG101

The Hedgehog (Hh) receptor Patched1 (PTCH1) is a well-known tumor suppressor that in its active form represses Smoothened (SMO) activity, inhibits proliferation, and induces apoptosis. The cytoplasmic C-terminal domain (CTD) regulates PTCH1 turnover and nucleates a proapoptotic complex. In this study, it was mechanistically determined that Autophagy-related 101 (ATG101), essential for mammalian autophagy, physically interacts with the CTD of PTCH1 and connects it to the ULK complex, which stimulates the autophagy machinery in response to changes in nutrient availability. This interaction results in a blockade of basal autophagic flux and accumulation of autophagosomes with undegraded cargo. Remarkably, this function of PTCH1 is independent of its repressive activity on SMO, as shown in SMO-deficient cells or in the presence of a SMO inhibitor, but is opposed by Sonic Hedgehog (SHH). These findings reveal a novel noncanonical function of PTCH1 that limits autophagy, mediated by ATG101, which could have therapeutic implications in Hh-dependent cancers.Implications: Loss-of-function of the tumor suppressor Patched1 might promote cancer cell fitness by increasing autophagic flux in response to metabolic or environmental stresses. Mol Cancer Res; 16(5); 909-19. ©2018 AACR.

The alpha-7 nicotinic acetylcholine receptor is involved in a direct inhibitory effect of nicotine on GnRH release: In vitro studies

The activation of nicotinic cholinergic receptors (nAChR) inhibits the reproductive axis; however, it is not clear whether nicotine may directly modulate the release of hypothalamic gonadotropin-releasing hormone (GnRH). Experiments carried out in GT1-1 immortalized GnRH neurons reveal the presence of a single class of high affinity α4β2 and α7 nAchR subtypes. The exposure of GT1-1 cells to nicotine does not modify the basal accumulation of GnRH. However, nicotine was found to modify GnRH pulsatility in perifusion experiments and inhibits, the release of GnRH induced by prostaglandin E1 or by K+-induced cell depolarization; these effects were reversed by D-tubocurarine and α-bungarotoxin. In conclusion, the results reported here indicate that: functional nAChRs are present on GT1-1 cells, the activation of the α-bungarotoxin-sensitive subclass (α7) produces an inhibitory effect on the release of GnRH and that the direct action of nicotine on GnRH neurons may be involved in reducing fertility of smokers.

Insights on Forest Structure and Composition from Long-Term Research in the Luquillo Mountains

The science of ecology fundamentally aims to understand species and their relation to the environment. At sites where hurricane disturbance is part of the environmental context, permanent forest plots are critical to understand ecological vegetation dynamics through time. An overview of forest structure and species composition from two of the longest continuously measured tropical forest plots is presented. Long-term measurements, 72 years at the leeward site, and 25 years at windward site, of stem density are similar to initial and pre-hurricane values at both sites. For 10 years post-hurricane Hugo (1989), stem density increased at both sites. Following that increase period, stem density has remained at 1400 to 1600 stems/ha in the leeward site, and at 1200 stems/ha in the windward site. The forests had similar basal area values before hurricane Hugo in 1989, but these sites are following different patterns of basal area accumulation. The leeward forest site continues to accumulate and increase basal area with each successive measurement, currently above 50 m2/ha. The windward forest site maintains its basal area values close to an asymptote of 35 m2/ha. Currently, the most abundant species at both sites is the sierra palm. Ordinations to explore variation in tree species composition through time present the leeward site with a trajectory of directional change, while at the windward site, the composition of species seems to be converging to pre-hurricane conditions. The observed differences in forest structure and composition from sites differently affected by hurricane disturbance provide insight into how particular forest characteristics respond at shorter or longer time scales in relation to previous site conditions and intensity of disturbance effects.

Deletion of inositol-requiring enzyme-1α in podocytes disrupts glomerular capillary integrity and autophagy.

Inositol-requiring enzyme-1α (IRE1α) is an endoplasmic reticulum (ER)-transmembrane endoribonuclease kinase that plays an essential function in extraembryonic tissues during normal development and is activated during ER stress. To address the functional role of IRE1α in glomerular podocytes, we produced podocyte-specific IRE1α-deletion mice. In male mice, deletion of IRE1α in podocytes resulted in albuminuria beginning at 5 mo of age and worsening with time. Electron microscopy revealed focal podocyte foot-process effacement in 9-mo-old male IRE1α-deletion mice, as well as microvillous transformation of podocyte plasma membranes. Compared with control, glomerular cross-sectional and capillary lumenal areas were greater in deletion mice, and there was relative podocyte depletion. Levels of microtubule-associated protein 1A/1B-light chain 3 (LC3)-II expression and c-Jun N-terminal kinase-1 phosphorylation were decreased in IRE1α-deletion glomeruli, in keeping with reduced autophagy. Deletion of IRE1α exacerbated glomerular injury in anti-glomerular basement membrane nephritis. In cell culture, IRE1α dominant-negative mutants reduced the physiological (basal) accumulation of LC3B-II and the size of autophagic vacuoles but did not affect ER-associated degradation. Thus IRE1α is essential for maintaining podocyte and glomerular integrity as mice age and in glomerulonephritis. The mechanism is related, at least in part, to the maintenance of autophagy in podocytes.

In vitro effect of leptin on anterior pituitary cells LH secretory activity during early pregnancy in pig.

Leptin modulates reproductive activity but its potential influence on LH secretion from anterior pituitary (AP) cells during implantation period in pigs (days 14-16 of pregnancy) remained unexplored. This study focused on determination whether leptin affects basal and GnRH-induced LH secretion and intracellular accumulation and whether leptin receptor (OB-Rb) mRNA is expressed in the AP gland during implantation in pigs. Four individual AP glands were developed into separate primary cultures. 2×105 cells/ml were preincubated (72 h) and next, for 3.5 h, experimentally treated with GnRH (100 ng/ml), leptin (10-11, 10-9, 10-7, 10-6 M) alone, or given in respective combinations with GnRH. In the AP gland, OB-Rb mRNA expression was determined by real-time PCR method. Leptin activated LH secretion and its concentration-dependent effect was observed as stimulation shown in a full range tested (culture 1) and exhibited only at 10-6 M (culture 2). A pooled data analysis revealed that basal LH secretion increased at 10-9, 10-7 and 10-6 M, but GnRH-induced LH release decreased at 10-6 M. Leptin down-regulated GnRH-induced LH secretion in all cultures, but only culture 3 exhibited sensitivity for all concentrations tested. Basal LH accumulation was activated in culture 1 (at 10-11 M) and inhibited in culture 4 (at 10-9 M). In the presence of GnRH leptin up-regulated LH accumulation with individual culture leptin-sensitivity (culture 1-3), while down-regulated LH accumulation in culture 4. Obtained data indicate that OB-Rb mRNA is expressed in the AP gland and leptin alone and in combination with GnRH specifically modulates LH activity during early pregnancy in pigs.

ROS accumulation and antiviral defence control by microRNA528 in rice.

MicroRNAs (miRNAs) are key regulators of plant-pathogen interactions. Modulating miRNA function has emerged as a new strategy to produce virus resistance traits1-5. However, the miRNAs involved in antiviral defence and the underlying mechanisms remain largely elusive. We previously demonstrated that sequestration by Argonaute (AGO) proteins plays an important role in regulating miRNA function in antiviral defence pathways6. Here we reveal that cleavage-defective AGO18 complexes sequester microRNA528 (miR528) upon viral infection. We show that miR528 negatively regulates viral resistance in rice by cleaving L-ascorbate oxidase (AO) messenger RNA, thereby reducing AO-mediated accumulation of reactive oxygen species. Upon viral infection, miR528 becomes preferentially associated with AGO18, leading to elevated AO activity, higher basal reactive oxygen species accumulation and enhanced antiviral defence. Our findings reveal a mechanism in which antiviral defence is boosted through suppression of an miRNA that negatively regulates viral resistance. This mechanism could be manipulated to engineer virus-resistant crop plants.