Transcriptional Responses of Cultured Rat Sympathetic Neurons during BMP-7-Induced Dendritic Growth

Dendrites are the primary site of synapse formation in the vertebrate nervous system; however, relatively little is known about the molecular mechanisms that regulate the initial formation of primary dendrites. Embryonic rat sympathetic neurons cultured under defined conditions extend a single functional axon, but fail to form dendrites. Addition of bone morphogenetic proteins (BMPs) triggers these neurons to extend multiple dendrites without altering axonal growth or cell survival. We used this culture system to examine differential gene expression patterns in naïve vs. BMP-treated sympathetic neurons in order to identify candidate genes involved in regulation of primary dendritogenesis.To determine the critical transcriptional window during BMP-induced dendritic growth, morphometric analysis of microtubule-associated protein (MAP-2)-immunopositive processes was used to quantify dendritic growth in cultures exposed to the transcription inhibitor actinomycin-D added at varying times after addition of BMP-7. BMP-7-induced dendritic growth was blocked when transcription was inhibited within the first 24 hr after adding exogenous BMP-7. Thus, total RNA was isolated from sympathetic neurons exposed to three different experimental conditions: (1) no BMP-7 treatment; (2) treatment with BMP-7 for 6 hr; and (3) treatment with BMP-7 for 24 hr. Affymetrix oligonucleotide microarrays were used to identify differential gene expression under these three culture conditions. BMP-7 significantly regulated 56 unique genes at 6 hr and 185 unique genes at 24 hr. Bioinformatic analyses implicate both established and novel genes and signaling pathways in primary dendritogenesis.This study provides a unique dataset that will be useful in generating testable hypotheses regarding transcriptional control of the initial stages of dendritic growth. Since BMPs selectively promote dendritic growth in central neurons as well, these findings may be generally applicable to dendritic growth in other neuronal cell types

Retinoic Acids Potentiate BMP9-Induced Osteogenic Differentiation of Mesenchymal Progenitor Cells

As one of the least studied bone morphogenetic proteins (BMPs), BMP9 is one of the most osteogenic BMPs. Retinoic acid (RA) signaling is known to play an important role in development, differentiation and bone metabolism. In this study, we investigate the effect of RA signaling on BMP9-induced osteogenic differentiation of mesenchymal progenitor cells (MPCs).Both primary MPCs and MPC line are used for BMP9 and RA stimulation. Recombinant adenoviruses are used to deliver BMP9, RARalpha and RXRalpha into MPCs. The in vitro osteogenic differentiation is monitored by determining the early and late osteogenic markers and matrix mineralization. Mouse perinatal limb explants and in vivo MPC implantation experiments are carried out to assess bone formation. We find that both 9CRA and ATRA effectively induce early osteogenic marker, such as alkaline phosphatase (ALP), and late osteogenic markers, such as osteopontin (OPN) and osteocalcin (OC). BMP9-induced osteogenic differentiation and mineralization is synergistically enhanced by 9CRA and ATRA in vitro. 9CRA and ATRA are shown to induce BMP9 expression and activate BMPR Smad-mediated transcription activity. Using mouse perinatal limb explants, we find that BMP9 and RAs act together to promote the expansion of hypertrophic chondrocyte zone at growth plate. Progenitor cell implantation studies reveal that co-expression of BMP9 and RXRalpha or RARalpha significantly increases trabecular bone and osteoid matrix formation.Our results strongly suggest that retinoid signaling may synergize with BMP9 activity in promoting osteogenic differentiation of MPCs. This knowledge should expand our understanding about how BMP9 cross-talks with other signaling pathways. Furthermore, a combination of BMP9 and retinoic acid (or its agonists) may be explored as effective bone regeneration therapeutics to treat large segmental bony defects, non-union fracture, and/or osteoporotic fracture

Short communication. Biological fixation of nitrogen and N balance in soybean crops in the pampas region

Biological nitrogen fixation (BNF) is of key importance in the N balance of soybean (Glycine max) crops. A number of authors have suggested that a negative balance may occur under high yield conditions. Few studies have measured the contribution of BNF to soil N in the pampas region. The aims of the present study were to compare three BNF determination methods – two isotopic methods using sorghum or a non-nodulating soybean isoline as a reference crop, and one involving the calculation of the difference in N content between the nodulating and non-nodulating soybean isolines – and to estimate the N balance in soybean crops raised under conventional tillage and no tillage practices. The study was performed in 2004-2005; a complete randomised block design was used with three replicates (plot dimensions 3 × 7 m). The different methodologies estimated BNF to account for 45-58% of total plant N, equivalent to 94 to 123 kg N ha-1. Depending on the methodology for estimating the BNF the soil N balance varied between –7 and 22 kg N ha-1. With an average grain yield of 1,618 kg ha-1 and a BNF accounting for approximately 50% of total plant N (i.e.,115 kg N ha-1), the soil N balance was slightly positive (14 kg ha-1) and independent of the tillage practice. The tillage systems had no effect (P < 0.05) on the mass or number of nodules, shoot biomass production at the R1 or R6 growth stages, the N content, BNF, or grain yield. Since the present results were obtained using non-commercial soybean isolines, further research is required to determine the soil N balance when high yielding soybean crops are raised

Microbiological activity and carbon mineralization in pampean soils with different agricultural use intensity

ABSTRACT 1 The processes involved in the flows of matter and energy of terrestrial ecosystems depends 2 heavily on soil biological activity, the current conventional agricultural managements could alter 3 the biological mechanisms involved in decomposition and nutrient cycling in agroecosystems. 4 The aim of this study was to compare the activity levels and soil microbial biomass between 5 different agricultural pampean soil uses and its relationship to carbon mineralization. 25 years of 6 agricultural use were compared with 25 years of ecological reserve naturalized where each 7 agroecosystem soil were collected at 61 -125 -183 -236 -302 -368 -431 -488 days for 8 measuring their moisture, organic matter, enzymatic activity, microbial biomass carbon, soil 9 respiration, metabolic quotient, microbial quotient and carbon mineralization rate. The distance 10 between agroecosystems is less than 800 m, thus assuming the same soil and climatic conditions. 11 The data were evaluated by Friedman test finding significant differences in moisture, organic 12 matter, enzymatic activity, soil respiration y microbial quotient (p< 0.01). Difference was also 13 found in the microbial mineralization rate of carbon (p< 0.1). 14 KEY WORDS 15 Microbiological activity; carbon mineralization; soil use. 16 PeerJ PrePrints | https://doi.org/10.7287/peerj.preprints.1608v1 | CC-BY 4.0 Open Access

Cotton Gossypium hirsutum L., defense in response to nitrogen fertilization and beet armyworm density.

Plants respond to insect herbivory by producing dynamic changes in an array of defense-related volatile and nonvolatile secondary metabolites. A scaled response relative to herbivory levels and nutrient availability would be adaptive, particularly under nutrient-limited conditions, in minimizing the costs of expressed defensive pathways and synthesis. In this study, we investigated effects of varying nitrogen (N) fertilization (42, 112, 196, and 280 ppm N) on levels of cotton plant (Gossypium hirsutum) phytohormones [jasmonic acid (JA) and salicylic acid (SA)], terpenoid aldehydes (hemigossypolone, heliocides H1, H2, H3, and H4), and volatile production in response to beet armyworm (Spodoptera exigua) herbivory. Additional bioassays assessed parasitoid (Cotesia marginiventris) host-searching success in response to cotton plants grown under various N fertilizer regimes. At low N input (42 ppm N), herbivore damage resulted in significant increases in local leaf tissue concentrations of JA and volatiles and in systemic accumulation of terpenoid aldehydes. However, increased N fertilization of cotton plants suppressed S. exigua-induced plant hormones and led to reduced production of various terpenoid aldehydes in damaged mature leaves and undamaged young leaves. While increased N fertilization significantly diminished herbivore-induced leaf volatile concentrations, the parasitism of S. exigua larvae by the parasitoid C. marginiventris in field cages did not differ among N treatments. This suggests that, despite significant N fertilization effects on herbivore-induced plant defenses, at short range, the parasitoids were unable to differentiate between S. exigua larvae feeding on physiologically different cotton plants that share large constitutive volatile pools releasable when damaged by herbivores