scholarly journals Review of research on haploid production in cucumber and other cucurbits

2013 ◽  
Vol 25 (1) ◽  
pp. 67-78 ◽  
Author(s):  
Joanna Gałązka ◽  
Katarzyna Niemirowicz-Szczytt
Keyword(s):  
Citrullus Lanatus ◽  
Microspore Culture ◽  
Ovary Culture ◽  
Irradiated Pollen ◽  
Haploid Production ◽  
Treatment Conditions ◽  
Pre Treatment ◽  
Haploid Embryos ◽  
The Impact

ABSTRACT This review provides a summary of haploid induction methods and factors affecting the efficacy of specific methodologies as applied to cucumber (Cucumis sativus L.), melon (Cucumis melo L.), watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai), winter squash (Cucurbita maxima Duch. ex Lam.), summer squash (Cucurbita pepo L.) and other cucurbits. This report is focused on studies that were carried out during the last 20 years. The main objective of the research on the production of haploid cucurbit plants is to accelerate breeding programs through the use of homozygous double haploid lines (DHL) and to facilitate the selection of desired (e.g. disease-resistant) genotypes for breeding. Unfortunately, currently used protocols result in a low number of double haploids (DH). The most common and best-known method of obtaining haploid cucurbit plants is via pollination with irradiated pollen, which induces parthenogenetic development of haploid embryos in planta. The embryos are extracted from immature seeds and cultured in vitro to facilitate the maturation and development of plants. The studies described below were primarily aimed at the determination of an appropriate dose of radiation and the evaluation of the impact of the genotype and the time of year on the number of haploid embryos and plants obtained. A less popular method of haploid production - ovule and ovary culture - is based on in vitro gynogenesis. The studies related to this method concentrated on optimising the composition of the medium and pre-treatment conditions (primarily temperature) to which the flower buds were subjected. Recently, increasing attention has been paid to anther and microspore culture. As in the case of in vitro ovule and ovary culture, the medium composition and flower bud pre-treatment conditions were optimised. The most recent studies suggest that anther culture is comparable in effectiveness to the irradiated pollen technique.

scholarly journals Induction of Parthenogenetic Haploid Embryos after Pollination by Irradiated Pollen in Watermelon

HortScience ◽  
1994 ◽  
Vol 29 (10) ◽  
pp. 1189-1190 ◽  
Author(s):  
N. Sari ◽  
K. Abak ◽  
M. Pitrat ◽  
J.C. Rode ◽  
R. Dumas de Vaulx
Keyword(s):  
In Vitro Culture ◽  
Doubled Haploid ◽  
Chromosome Doubling ◽  
Citrullus Lanatus ◽  
Doubled Haploid Lines ◽  
Irradiated Pollen ◽  
Haploid Embryos ◽  
Haploid Plants

Parthenogenetic haploid embryos of `Crimson Sweet', `Halep Karasi', `Sugar Baby' and `Panonia F1' watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai] were obtained after pollination with γ-irradiated (200 or 300 Gy) pollen. Some globular and heart-shaped embryos were observed in fruit harvested 2 to 5 weeks after pollination. The number of embryos per 100 seeds was highest for `Halep Karasi'. After in vitro culture, 17 haploid plants were obtained and doubled haploid lines were generated after chromosome doubling using colchicine.

Interferon-Gamma Impairs Expansion and Hematopoietic Support of Bone Marrow Mesenchymal Stromal Cells

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 3884-3884
Author(s):  
Marieke Goedhart ◽  
Anne Cornelissen ◽  
Carlijn Kuijk ◽  
Sulima Geerman ◽  
Fernanda Pascutti ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Stromal Compartment ◽  
Self Renewal ◽  
Hematopoietic Stem ◽  
Ifn Γ ◽  
Immunomodulatory Properties ◽  
Pre Treatment ◽  
The Impact

Abstract Maintenance of hematopoietic stem cells (HSCs) and regulation of their quiescence and self-renewal is critical for maintaining a lifelong supply of blood cells. The ability of HSCs to stay quiescent is thought to depend on their specific niche in the bone marrow (BM). Mesenchymal stromal cells (MSC) in the BM are multipotent stem cells that form part of the vascular HSC niche and provide micro-environmental support to HSCs both in vivo and upon expansion ex vivo. Culture-expanded MSCs also exhibit immunomodulatory properties that can be enhanced by pre-treatment with interferon-gamma (IFN-γ). BM MSC are thus attractive candidates for cellular therapy after hematopoietic stem cell transplantation, for promoting rapid hematopoietic recovery and reducing the incidence or severity of graft versus host disease. Although IFN-γ pre-treatment can improve the immunomodulatory properties of MSCs, elevated IFN-γ levels have also been associated with anemia and BM failure in multiple chronic inflammatory diseases. While the impact of IFN-γ on HSC has been elucidated in recent years, it remains largely unknown whether IFN-γ can also influence hematopoietic support by BM stromal cells. In this study, we aim to elucidate the impact of IFN-γ on hematopoietic support of BM MSC. We show that in vitro expansion of primary BM MSC cultures from healthy donors was significantly reduced in the presence of IFN-γ, and this effect could be reproduced in the BM stromal cell line MS-5. Concurrently, IFN-γ diminished the clonal capacity of BM MSC, as measured by CFU-F assays. In addition, BM MSC that were pre-stimulated with IFN-γ produced significantly lower levels of CXCL12, suggesting a loss of hematopoietic support potential. Indeed, support of CD34+ hematopoietic stem and progenitor cells (HSPC) in a co-culture assay was greatly reduced in when MSC were pre-treated with IFN-γ. To determine the impact of IFN-γ on BM MSC in vivo, we investigated the BM stromal compartment of IFN-γ AU-rich element deleted (ARE-Del) mice, which constitutively express IFN-γ in steady state conditions. FACS analysis revealed a remodeling of the BM stromal compartment in ARE-Del mice compared to littermate controls, with significantly fewer MSCs, identified as CD45-Ter119-CD31-CD51+PDGFRa+ cells. Numbers of other stromal cell subsets, such as osteoblasts and fibroblasts, were not altered. The reduction of BM MSC in ARE-Del mice coincided with a loss of quiescence in HSCs; only 35% of long term HSC (LT-HSC) in ARE-Del mice were quiescent, compared to 70% in WT mice, as determined by Ki-67 staining. Loss of quiescence in LT-HSC did not lead to increased self-renewal, but rather induced increased differentiation towards short-term HSC and multi-potent progenitors. We then sorted LT-HSC from WT and ARE-Del mice and performed in vitro HSC culture assays in the absence of IFN-γ. Absolute numbers of LT-HSC were rapidly decreased in ARE-Del compared to WT cultures after 3 and 7 days of HSC culture, while numbers of more differentiated progenitors were increased. These data indicate that an IFN-γ-mediated loss of BM MSC in ARE-Del mice leads to loss of quiescent LT-HSCs and induces a tendency towards HSC differentiation over self-renewal. In conclusion, we have shown that IFN-γ has a negative impact on expansion and hematopoietic support of BM MSC in vitro and in vivo across species. Although IFN-γ treatment enhances the immunomodulatory function of MSCs in a clinical setting, it is obvious from our data that IFN-γ impairs their HSC supporting function. These data also provide more insight in the underlying mechanism by which IFN-γ contributes to the pathogenesis of anemia and BM failure. Disclosures No relevant conflicts of interest to declare.

Effects of different genotypes and gamma ray doses on haploidization with irradiated pollen technique in watermelon (Citrullus lanatus L.)

2013 ◽  
Vol 93 (6) ◽  
pp. 1165-1168 ◽  
Author(s):  
Hatıra Taşkın ◽  
Namık Kemal Yücel ◽  
Gökhan Baktemur ◽  
Songül Çömlekçioğlu ◽  
Saadet Büyükalaca
Keyword(s):  
Gamma Rays ◽  
Irradiation Dose ◽  
Gamma Ray ◽  
Citrullus Lanatus ◽  
Embryo Rescue ◽  
Genotype 1 ◽  
Irradiated Pollen ◽  
Glass Tubes ◽  
Male Flowers ◽  
Haploid Embryos

Taşkın, H., Yücel, N. K., Baktemur, G., Çömlekçioğlu, S. and Büyükalaca, S. 2013. Effects of different genotypes and gamma ray doses on haploidization with irradiated pollen technique in watermelon ( Citrullus lanatus L.). Can. J. Plant Sci. 93: 1165–1168. Two watermelon genotypes, one commercial watermelon variety (Ustun F1) and five different doses of gamma rays coming from Co60 were tested to develop useful haploidization procedures in watermelon. For this purpose, male flowers collected a day before anthesis were irradiated with 50, 150, 200, 275 and 300 Gy doses of gamma rays, and female flowers were pollinated with irradiated pollen the next day. Seeds extracted from fruits harvested 25 d later were opened individually in a laminar flow hood. Embryos obtained via embryo rescue technique were placed in glass tubes containing CP medium with 30 g L−1 sucrose, 8 g L−1 agar, 0.08 mg L−1 B12, and 0.02 mg L−1 IAA. Sixty haploid embryos were obtained from 43 watermelon fruits in this study. Genotype 1 was found to be the most successful genotype with 3.57 haploid embryos per 100 seeds. Among tested irradiation doses, 275 Gy was better than other doses, with 5.26 haploid embryos per 100 seeds. Considered together with irradiation dose and genotypes, the maximum number of haploid embryos was obtained from Genotype 1 pollinated with 275 Gy irradiation dose, with 6.25 haploid embryos per 100 seeds.

PROGESTERONE TREATMENT INCREASES PITUITARY OESTRADIOL RETENTION IN THE OVARIECTOMIZED NORMAL FEMALE RAT BUT NOT IN THE MALE NOR IN THE ANDROGEN- OR OESTROGEN-STERILIZED FEMALE RAT

1977 ◽  
Vol 84 (2) ◽  
pp. 268-280 ◽  
Author(s):  
Robert D. Lisk ◽  
Lawrence A. Reuter
Keyword(s):  
Testosterone Propionate ◽  
Ovariectomized Rats ◽  
Normal Female ◽  
Nuclear Fraction ◽  
Female Rat ◽  
Treatment Conditions ◽  
Oestradiol Benzoate ◽  
Pre Treatment

ABSTRACT Pituitary retention of [3H]oestradiol in ovariectomized rats was measured following in vivo progesterone pre-treatment and found to be significantly increased after 48, 72, 96 and 120 h of pre-treatment. Increased [3H]oestradiol retention was also observed for at least up to 72 h after removal of the progesterone pre-treatment source. This retention was measured as dpm per mg dry tissue weight. [3H]Oestradiol retention was also measured in the nuclear fraction of tissues incubated with [3H]oestradiol in vitro. Following 72 h of in vivo progesterone pre-treatment, the nuclear fraction from the pituitary was found to retain significantly more [3H]oestradiol than corresponding fractions from non-treated animals. In contrast to ovariectomized females, no increase in [3H]oestradiol retention was found in the pituitary of orchidectomized males pre-treated with progesterone for 72 h. [3H]Oestradiol retention by pituitaries of ovariectomized rats injected on the day of birth with 200 μg oestradiol benzoate (OeB) or 500 μg testosterone propionate (TP) was significantly decreased in comparison to control animals. When the rats were pre-treated in vivo with oestradiol for 6 or 72 h and [3H]oestradiol retention was measured 6 or 24 h after this pre-treatment, the OeB and TP treated animals retained significantly less [3H]oestradiol under most treatment conditions. Progesterone pretreatment for 24 or 72 h in vivo followed by measurement of [3H]oestradiol retention immediately or 6 or 24 h later resulted in a significant increase in [3H]oestradiol retention for the control animals. In contrast, the neonatally OeB or TP treated animals differed significantly by not showing increased retention. When [3H]oestradiol retention of the pituitary was measured in vitro following homogenization at 0°C and incubation at 37°C for 1 h, the nuclear fraction from both OeB and TP treated animals was found to retain less hormone per unit DNA; however, this decrease was significant only for the TP animals. Thus, males and androgen- or oestrogensterilized females have an altered and reduced augmentation of pituitary oestradiol retention in response to both oestrogen and progesterone pretreatments.

scholarly journals Modulating the Heat Sensitivity of Prostate Cancer Cell Lines In Vitro: A New Impact for Focal Therapies

Biomedicines ◽  
2020 ◽  
Vol 8 (12) ◽  
pp. 585
Author(s):  
Oliver Hahn ◽  
Franziska M. Heining ◽  
Jörn Janzen ◽  
Johanna C. R. Becker ◽  
Marina Bertlich ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Lines ◽  
Therapeutic Option ◽  
Neuroendocrine Differentiation ◽  
High Intensity Focused Ultrasound ◽  
Heat Sensitivity ◽  
Heat Response ◽  
Pre Treatment ◽  
The Impact

Focal therapies such as high-intensity focused ultrasound (HiFU) are an emerging therapeutic option for prostate cancer (PCA). Thermal or mechanical effects mediate most therapies. Moreover, locally administered drugs such as bicalutamide or docetaxel are new focal therapeutic options. We assessed the impact of such focal medical treatments on cell viability and heat sensitivity by pre-treating PCA cell lines and then gradually exposing them to heat. The individual heat response of the cell lines tested differed largely. Vertebral-Cancer of the Prostate (VCaP) cells showed an increase in metabolic activity at 40–50 °C. Androgen receptor (AR)-negative PC3 cells showed an increase at 51.3 °C and were overall more resistant to higher temperatures. Pre-treatment of VCaP cells with testosterone (VCaPrev) leads to a more PC3-like kinetic of the heat response. Pre-treatment with finasteride and bicalutamide did not cause changes in heat sensitivity in any cell line. Mitoxantrone treatment, however, shifted heat-induced proliferation loss to lower temperature in VCaP cells. Further analysis via RNAseq identified a possible correlation of heat resistance with H3K27me3-dependent gene regulation, which could be related to an increase in the histone methyltransferase EZH2 and a possible neuroendocrine differentiation. Pre-treatment with mitoxantrone might be a perspective for HiFU treatment. Further studies are needed to evaluate possible combinations with Hsp90 or EZH2 inhibitors.

Tumor Necrosis Factor-α and Interleukin-1β Treatment of Endometrial Stromal Cells Does not Promote Their Adhesion to Peritoneal Mesothelial Cells in an in Vitro Model of Early-Stage Endometriosis

2011 ◽  
Vol 3 (2) ◽  
pp. 67-72
Author(s):  
Jean-Christophe Lousse ◽  
Sylvie Defrere ◽  
Sébastien Colette ◽  
Anne Van Langendonckt ◽  
Jacques Donnez
Keyword(s):  
Cell Adhesion ◽  
In Vitro Model ◽  
Early Stage ◽  
Mesothelial Cells ◽  
Endometrial Stromal Cells ◽  
Peritoneal Mesothelial Cells ◽  
Pre Treatment ◽  
Vitro Model ◽  
The Impact

In this study, we developed an original and reproducible quantitative in vitro model of endometrial cell adhesion to peritoneal mesothelial cells in order to better assess the impact of pro-inflammatory cytokines on early-stage endometriosis development. We demonstrated that pre-treatment with TNF-α and IL-1β does not promote endometrial stromal cell adhesion to peritoneal mesothelial cells.

scholarly journals Potential Neuroprotective and Anti-Apoptotic Properties of a Long-Lasting Stable Analog of Ghrelin: an In Vitro Study Using SH-SY5Y Cells

2018 ◽  
pp. 339-346 ◽  
Author(s):  
A. POPELOVÁ ◽  
A. KÁKONOVÁ ◽  
L. HRUBÁ ◽  
J. KUNEŠ ◽  
L. MALETÍNSKÁ ◽  
...  
Keyword(s):  
Neurodegenerative Disorders ◽  
Neuroblastoma Cells ◽  
In Vitro Study ◽  
Ghrelin Receptor ◽  
Positive Effects ◽  
Ldh Assay ◽  
Pre Treatment ◽  
The Impact ◽  
Viability Markers

Neurodegenerative disorders, such as Alzheimer’s disease (AD) and Parkinson’s disease (PD), are increasing in prevalence. Currently, there are no effective and specific treatments for these disorders. Recently, positive effects of the orexigenic hormone ghrelin on memory and learning were demonstrated in mouse models of AD and PD. In this study, we tested the potential neuroprotective properties of a stable and long-lasting ghrelin analog, Dpr3ghrelin (Dpr3ghr), in SH-SY5Y neuroblastoma cells stressed with 1.2 mM methylglyoxal (MG), a toxic endogenous by-product of glycolysis, and we examined the impact of Dpr3ghr on apoptosis. Pre-treatment with both 10-5 and 10-7 M Dpr3ghr resulted in increased viability in SH-SY5Y cells (determined by MTT staining), as well as reduced cytotoxicity of MG in these cells (determined by LDH assay). Dpr3ghr increased viability by altering pro-apoptotic and viability markers: Bax was decreased, Bcl-2 was increased, and the Bax/Bcl-2 ratio was attenuated. The ghrelin receptor GHS-R1 and Dpr3ghr-induced activation of PBK/Akt were immuno-detected in SH-SY5Y cells to demonstrate the presence of GHS-R1 and GHS-R1 activation, respectively. We demonstrated that Dpr3ghr protected SH-SY5Y cells against MG-induced neurotoxicity and apoptosis. Our data suggest that stable ghrelin analogs may be candidates for the effective treatment of neurodegenerative disorders.

scholarly journals Efficient Heat Shock Response Affects Hyperthermia-Induced Radiosensitization in a Tumor Spheroid Control Probability Assay

Cancers ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 3168
Author(s):  
Oleg Chen ◽  
Soňa Michlíková ◽  
Lisa Eckhardt ◽  
Marit Wondrak ◽  
Adriana M. De Mendoza ◽  
...  
Keyword(s):  
Stress Response ◽  
Heat Shock ◽  
Heat Shock Response ◽  
Cellular Stress Response ◽  
Proteotoxic Stress ◽  
Shock Response ◽  
Pre Treatment ◽  
Shock Proteins ◽  
The Impact

Hyperthermia (HT) combined with irradiation is a well-known concept to improve the curative potential of radiotherapy. Technological progress has opened new avenues for thermoradiotherapy, even for recurrent head and neck squamous cell carcinomas (HNSCC). Preclinical evaluation of the curative radiosensitizing potential of various HT regimens remains ethically, economically, and technically challenging. One key objective of our study was to refine an advanced 3-D assay setup for HT + RT research and treatment testing. For the first time, HT-induced radiosensitization was systematically examined in two differently radioresponsive HNSCC spheroid models using the unique in vitro “curative” analytical endpoint of spheroid control probability. We further investigated the cellular stress response mechanisms underlying the HT-related radiosensitization process with the aim to unravel the impact of HT-induced proteotoxic stress on the overall radioresponse. HT disrupted the proteome’s thermal stability, causing severe proteotoxic stress. It strongly enhanced radiation efficacy and affected paramount survival and stress response signaling networks. Transcriptomics, q-PCR, and western blotting data revealed that HT + RT co-treatment critically triggers the heat shock response (HSR). Pre-treatment with chemical chaperones intensified the radiosensitizing effect, thereby suppressing HT-induced Hsp27 expression. Our data suggest that HT-induced radiosensitization is adversely affected by the proteotoxic stress response. Hence, we propose the inhibition of particular heat shock proteins as a targeting strategy to improve the outcome of combinatorial HT + RT.

INHIBITION OF THYROGLOBULIN BIOSYNTHESIS AND DEGRADATION BY EXCESS IODIDE. SYNERGISM WITH LITHIUM

1976 ◽  
Vol 81 (2) ◽  
pp. 495-506 ◽  
Author(s):  
A. Radvila ◽  
R. Roost ◽  
H. Bürgi ◽  
H. Kohler ◽  
H. Studer
Keyword(s):  
Protein Synthesis ◽  
Thyroid Gland ◽  
Inhibitory Action ◽  
Inhibit Protein Synthesis ◽  
Thyrotrophic Hormone ◽  
Thyroid Glands ◽  
Pre Treatment ◽  
Excess Iodide ◽  
Kidney Liver

ABSTRACT Lithium and excess iodide inhibit the release of thyroid hormone from preformed stores. We thus tested the hypothesis that this was due to an inhibition of thyroglobulin breakdown. Rats were pre-treated with propylthiouracil (PTU) for 3 weeks in order to deplete their thyroids of thyroglobulin. While the PTU was continued, lithium chloride (0.25 mEq./100 g weight) or potassium iodide (3 mg per rat) were injected every 12 h for 3 days. Thereafter the thyroglobulin content in thyroid gland homogenates was measured. PTU pre-treatment lowered the thyroglobulin content from 4.21 to 0.22 mg/100 mg gland. Lithium caused a marked re-accumulation of thyroglobulin to 0.60 mg/100 mg within 3 days. While iodide alone had only a borderline effect, it markedly potentiated the action of lithium and a combination of the two drugs increased the thyroglobulin content to 1.04 mg/100 mg. Thyroxine was injected into similarly pre-treated animals to suppress secretion of thyrotrophic hormone. This markedly inhibited the proteolysis of thyroglobulin and 1.3 mg/100 mg gland accumulated after 3 days. Excess iodide, given in addition to thyroxine, decreased the amount of thyroglobulin accumulated to 0.75 mg/100 mg gland. To study whether this could be explained by an inhibitory action of iodide on thyroglobulin biosynthesis, thyroid glands from animals treated with excess iodide were incubated in vitro in the presence of 0.2 mm iodide for 3 h. Iodide decreased the incorporation of radioactive leucine into total thyroidal protein and into thyroglobulin by 25 and 35 % respectively. Iodide did not inhibit protein synthesis in the kidney, liver or muscle tissue. Thus, large doses of iodide selectively inhibit thyroglobulin biosynthesis.

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