RESUMO
Lung branching morphogenesis relies on intricate epithelial-mesenchymal interactions and signaling networks. Still, the interplay between signaling and energy metabolism in shaping embryonic lung development remains unexplored. Retinoic acid (RA) signaling influences lung proximal-distal patterning and branching morphogenesis, but its role as a metabolic modulator is unknown. Hence, this study investigates how RA signaling affects the metabolic profile of lung branching. We performed ex vivo lung explant culture of embryonic chicken lungs treated with DMSO, 1 µM RA, or 10 µM BMS493. Extracellular metabolite consumption/production was evaluated by using 1H-NMR spectroscopy. Mitochondrial respiration and biogenesis were also analyzed. Proliferation was assessed using an EdU-based assay. The expression of crucial metabolic/signaling components was examined through Western blot, qPCR, and in situ hybridization. RA signaling stimulation redirects glucose towards pyruvate and succinate production rather than to alanine or lactate. Inhibition of RA signaling reduces lung branching, resulting in a cystic-like phenotype while promoting mitochondrial function. Here, RA signaling emerges as a regulator of tissue proliferation and lactate dehydrogenase expression. Furthermore, RA governs fatty acid metabolism through an AMPK-dependent mechanism. These findings underscore RA's pivotal role in shaping lung metabolism during branching morphogenesis, contributing to our understanding of lung development and cystic-related lung disorders.
Assuntos
Metabolismo Energético , Pulmão , Morfogênese , Transdução de Sinais , Tretinoína , Animais , Tretinoína/metabolismo , Tretinoína/farmacologia , Pulmão/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/embriologia , Metabolismo Energético/efeitos dos fármacos , Morfogênese/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Embrião de Galinha , Proliferação de Células/efeitos dos fármacos , Mitocôndrias/metabolismo , Mitocôndrias/efeitos dos fármacos , GalinhasRESUMO
Linoleic acid (LA), an n-6 polyunsaturated fatty acid (PUFA), is obtained from the maternal diet during pregnancy, and is essential for normal fetal growth and development. A maternal high-LA (HLA) diet alters maternal and offspring fatty acids, maternal leptin and male/female ratio at embryonic (E) day 20 (E20). We investigated the effects of an HLA diet on embryonic offspring renal branching morphogenesis, leptin signalling, megalin signalling and angiogenesis gene expression. Female Wistar Kyoto rats were fed low-LA (LLA; 1.44% energy from LA) or high-LA (HLA; 6.21% energy from LA) diets during pregnancy and gestation/lactation. Offspring were sacrificed and mRNA from kidneys was analysed by real-time PCR. Maternal HLA decreased the targets involved in branching morphogenesis Ret and Gdnf in offspring, independent of sex. Furthermore, downstream targets of megalin, namely mTOR, Akt3 and Prkab2, were reduced in offspring from mothers consuming an HLA diet, independent of sex. There was a trend of an increase in the branching morphogenesis target Gfra1 in females (p = 0.0517). These findings suggest that an HLA diet during pregnancy may lead to altered renal function in offspring. Future research should investigate the effects an HLA diet has on offspring kidney function in adolescence and adulthood.
Assuntos
Rim , Ácido Linoleico , Morfogênese , Proteínas Proto-Oncogênicas c-akt , Transdução de Sinais , Serina-Treonina Quinases TOR , Animais , Feminino , Gravidez , Serina-Treonina Quinases TOR/metabolismo , Rim/metabolismo , Rim/efeitos dos fármacos , Ratos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Morfogênese/efeitos dos fármacos , Morfogênese/genética , Ácido Linoleico/metabolismo , Masculino , Ratos Endogâmicos WKY , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Feto/metabolismo , Feto/efeitos dos fármacosRESUMO
Coordinated morphogenic adaptation of growing plants is critical for their survival and propagation under fluctuating environments. Plant morphogenic responses to light and warm temperatures, termed photomorphogenesis and thermomorphogenesis, respectively, have been extensively studied in recent decades. During photomorphogenesis, plants actively reshape their growth and developmental patterns to cope with changes in light regimes. Accordingly, photomorphogenesis is closely associated with diverse growth hormonal cues. Notably, accumulating evidence indicates that light-directed morphogenesis is profoundly affected by two recently identified phytochemicals, karrikins (KARs) and strigolactones (SLs). KARs and SLs are structurally related butenolides acting as signaling molecules during a variety of developmental steps, including seed germination. Their receptors and signaling mediators have been identified, and associated working mechanisms have been explored using gene-deficient mutants in various plant species. Of particular interest is that the KAR and SL signaling pathways play important roles in environmental responses, among which their linkages with photomorphogenesis are most comprehensively studied during seedling establishment. In this review, we focus on how the phytochemical and light signals converge on the optimization of morphogenic fitness. We also discuss molecular mechanisms underlying the signaling crosstalks with an aim of developing potential ways to improve crop productivity under climate changes.
Assuntos
Lactonas , Transdução de Sinais , Lactonas/metabolismo , Luz , Piranos/metabolismo , Piranos/farmacologia , Furanos/metabolismo , Furanos/farmacologia , Desenvolvimento Vegetal/efeitos da radiação , Desenvolvimento Vegetal/efeitos dos fármacos , Morfogênese/efeitos da radiação , Morfogênese/efeitos dos fármacos , Adaptação Fisiológica/genéticaRESUMO
The integration of nanoparticles (NPs) holds promising potential to bring substantial advancements to plant cryopreservation, a crucial technique in biodiversity conservation. To date, little attention has been focused on using nanoparticles in cryobiology research. This study aimed to assess the effectiveness of NPs in enhancing the efficiency of plant cryopreservation. In-vitro-derived shoot tips of bleeding heart (Lamprocapnos spectabilis (L.) Fukuhara) 'Gold Heart' and 'Valentine' were used as the plant material. The encapsulation-vitrification cryopreservation protocol included preculture, encapsulation, dehydration, storage in liquid nitrogen, rewarming, and recovery steps. Gold (AuNPs), silver (AgNPs), or zinc oxide (ZnONPs) nanoparticles were added at various concentrations either into the preculture medium or the protective bead matrix during encapsulation. The explant survival and further morphogenic and biochemical events were studied. Results showed that the impact of NPs on cryopreservation outcomes was cultivar-specific. In the 'Valentine' cultivar, incorporating 5 ppm AgNPs within the alginate bead matrix significantly improved cryopreservation efficiency by up to 12%. On the other hand, the 'Gold Heart' cultivar benefited from alginate supplementation with 5 ppm AgNPs and 5-15 ppm ZnONPs, leading to an over 28% increase in the survival rate of shoot tips. Interestingly, adding NPs to the preculture medium was less effective and sometimes counterproductive, despite promoting greater shoot proliferation and elongation in 'Valentine' explants compared to the control. Moreover, nanoparticles often induced oxidative stress (and enhanced the activity of APX, GPOX, and SOD enzymes), which in turn affected the biosynthesis of plant primary and secondary metabolites. It was found that supplementation of preculture medium with higher concentration (15 ppm) of gold, silver and zinc oxide nanoparticles stimulated the production of plant pigments, but in a cultivar-dependent matter. Our study confirmed the beneficial action of nanoparticles during cryopreservation of plant tissues.
Assuntos
Criopreservação , Ouro , Nanopartículas Metálicas , Criopreservação/métodos , Nanopartículas Metálicas/química , Ouro/química , Ouro/farmacologia , Prata/química , Prata/farmacologia , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Morfogênese/efeitos dos fármacos , VitrificaçãoRESUMO
BACKGROUND: Gymnema sylvestre is a medicinal woody perennial vine known for its sweetening properties and antidiabetic therapeutic uses in the modern and traditional medicines. Its over-exploitation for the therapeutic uses and to meet the demand of pharmaceutical industry in raw materials supply for the production of anti-diabetic drugs has led to considerable decline in its natural population. RESULTS: An efficient system of shoot bud sprouting from nodal segment explants and indirect plant regeneration from apical meristem-induced callus cultures of G. sylvestre have been developed on Murashige and Skoog (MS) medium amended with concentrations of cytokinins. Of the three growth regulators tested, N6-benzylaminopurine (BAP) was the most efficient and 2.0 mg L-1 gave the best shoot formation efficiency. This was followed by thidiazuron (TDZ) and kinetin (Kin) but, most of the TDZ-induced micro shoots showed stunted growth. Multiple shoot formation was observed on medium amended with BAP or TDZ at higher concentrations. The produced micro shoots were rooted on half strength MS medium amended with auxins and rooted plantlets acclimatized with 87% survival of the regenerates. CONCLUSIONS: The developed regeneration system can be exploited for genetic transformation studies, particularly when aimed at producing its high yielding cell lines for the anti-diabetic phytochemicals. It also offers opportunities for exploring the expression of totipotency in the anti-diabetic perennial vine.
Assuntos
Reguladores de Crescimento de Plantas/farmacologia , Regeneração/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Gymnema sylvestre/crescimento & desenvolvimento , Morfogênese/efeitos dos fármacos , Compostos de Fenilureia/farmacologia , Purinas/farmacologia , Tiadiazóis/farmacologia , Compostos de Benzil/farmacologia , Brotos de Planta/efeitos dos fármacos , Gymnema sylvestre/efeitos dos fármacos , Cinetina/farmacologiaRESUMO
Aspectos concernentes à morfogênese de Gallus gallus domesticus viabilizam estudos relacionados à ação da insulina sobre estruturas e topografia embrionárias. Na temperatura de 37,5ºC, ovos foram incubados por 24 h, injetados com 5 ml de insulina de suínos em 3 concentrações e reincubados por mais 72 h. As características morfológicas de 80 embriões foram avaliadas e, de acordo com a organização apresentada, classificados em 5 níveis de morfogênese. Em 21 embriões submetidos aos testes com a insulina registrou-se dismorfismo generalizado (4o nível), enquanto nos de experimento de controle foi verificado morfogênese-padrão (1o nível) e dismorfismo localizado (3o nível). Aqueles espécimes mostraram corpo com dimensões reduzidas, caracterizado por limites ântero-dorsal organizados em uma projeção cefálica e regiões posterior-ventral alteradas, evidenciando um padrão de anormalidades na determinação do eixo ântero-posterior, que indica a ação específica da insulina na morfogênese embrionária no período de 96 horas de incubação.
Assuntos
Animais , Embrião de Galinha/fisiologia , Insulina , Embrião de Galinha/efeitos dos fármacos , Resumo em Inglês , Insulina , Morfogênese/efeitos dos fármacosRESUMO
Amphibian gastrulation was used as a model system to study the action of the nucleoside 1-beta-D-arabinofuranosylcytosine (Ara-C) on the early events of amphibian morphogenesis. Ara-C inhibits both glycoprotein and glycolipid synthesis and interferes with DNA synthesis. Thus, it is useful to investigate the importance of the cell surface and the nucleous during Bufo arenarum morphogenesis. Living embryos were incubated with Ara-C at blastula and gastrula stages. Treated-embryos undergo abnormal gastrulation, most of the embryos exogastrulate, although some do not gastrulate at all. This antimetabolite did not interfere with neural induction, as partial exogastrulae developed a small neural tube. We have proven that Area-C disturbs the typical intercellular organization and inhibits the radial intercalation of the blastocoelic roof. The mesodermal migration is the most affected morphogenetic process. The results described in this paper demonstrate that the timing of gastrulation movements strongly involves the participation of surface and extracellular molecules in cell recognition and cell interaction but does not involve a significant increase in cell division rate and can also occur in the absence of the cell division.