Calcium-transport function of the chick embryonic chorioallantoic membrane. I. In vivo and in vitro characterization

1986 ◽  
Vol 82 (1) ◽  
pp. 73-84
Author(s):  
R.S. Tuan ◽  
M.J. Carson ◽  
J.A. Jozefiak ◽  
K.A. Knowles ◽  
B.A. Shotwell
Keyword(s):  
Calcium Transport ◽  
Calcium Uptake ◽  
Chorioallantoic Membrane ◽  
Transport Function ◽  
In Ovo ◽  
In Vitro Characterization ◽  
Microsomal Membranes

During chick embryonic development, the chorioallantoic membrane (CAM) is responsible for the mobilization of shell calcium into the embryonic circulation. The calcium-transport function of the CAM was studied here by measuring CAM calcium uptake in vivo and in vitro. The in vivo technique involved the use of an uptake chamber constructed on top of the CAM in situ. The in vitro methods included two systems: CAM tissue disks and cell-free microsomal membranes isolated from the CAM. Analyses using these three assays show that calcium uptake by the CAM exhibited characteristics indicative of active transport, such as temperature dependence, saturability, energetic requirement and ion specificity. The data also show that calcium-uptake activities of the CAM increase as a function of embryonic age in a manner coincident with the increased accumulation of calcium by the developing embryo in ovo.

scholarly journals The Role of Pre-Clinical 3-Dimensional Models of Osteosarcoma

2020 ◽  
Vol 21 (15) ◽  
pp. 5499
Author(s):  
Hannah L. Smith ◽  
Stephen A. Beers ◽  
Juliet C. Gray ◽  
Janos M. Kanczler
Keyword(s):  
Three Dimensional ◽  
Chorioallantoic Membrane ◽  
3D Models ◽  
In Ovo ◽  
Future Directions ◽  
3 Dimensional ◽  
In Vivo Animal Models

Treatment for osteosarcoma (OS) has been largely unchanged for several decades, with typical therapies being a mixture of chemotherapy and surgery. Although therapeutic targets and products against cancer are being continually developed, only a limited number have proved therapeutically active in OS. Thus, the understanding of the OS microenvironment and its interactions are becoming more important in developing new therapies. Three-dimensional (3D) models are important tools in increasing our understanding of complex mechanisms and interactions, such as in OS. In this review, in vivo animal models, in vitro 3D models and in ovo chorioallantoic membrane (CAM) models, are evaluated and discussed as to their contribution in understanding the progressive nature of OS, and cancer research. We aim to provide insight and prospective future directions into the potential translation of 3D models in OS.

Nerve-dependent accumulation of myosin light chain 3 in developing limb musculature

Development ◽  
1987 ◽  
Vol 101 (4) ◽  
pp. 673-684
Author(s):  
P.A. Merrifield ◽  
I.R. Konigsberg
Keyword(s):  
Neural Tube ◽  
Light Chain ◽  
Limb Development ◽  
Myosin Light Chain ◽  
Chorioallantoic Membrane ◽  
In Ovo ◽  
Fast Myosin ◽  
Limb Musculature

Myosin alkali light chain accumulation in developing quail limb musculature has been analysed on immunoblots using a monoclonal antibody which recognizes an epitope common to fast myosin light chain 1 (MLC1f) and fast myosin light chain 3 (MLC3f). The limb muscle of early embryos (i.e. up to day 10 in ovo) has a MLC profile similar to that observed in myotubes cultured in vitro; although MLC1f is abundant, MLC3f cannot be detected. MLC3f is first detected in 11-day embryos. To determine whether this alteration in MLC3f accumulation is nerve or hormone dependent, limb buds with and without neural tube were cultured as grafts on the chorioallantoic membrane of chick hosts. Although differentiated muscle develops in both aneural and innervated grafts, innervated grafts contain approximately three times as much myosin as aneural grafts. More significantly, although aneural grafts reproducibly accumulate normal levels of MLC1f, they fail to accumulate detectable levels of MLC3f. In contrast, innervated grafts accumulate both MLC1f and MLC3f, suggesting that the presence of neural tube in the graft promotes the maturation, as well as the growth, of muscle tissue. This is the first positive demonstration that innervation is necessary for the accumulation of MLC3f that occurs during normal limb development in vivo.

scholarly journals Investigating the ocular toxicity potential and therapeutic efficiency of in situ gel nanoemulsion formulations of brinzolamide

2020 ◽  
Vol 9 (4) ◽  
pp. 578-587
Author(s):  
Sima Talaei ◽  
Mohammad Mehdi Mahboobian ◽  
Mojdeh Mohammadi
Keyword(s):  
Intraocular Pressure ◽  
Membrane Surface ◽  
Chorioallantoic Membrane ◽  
In Situ Gel ◽  
Toxicity Potential ◽  
Vessel Injury ◽  
Hen’S Egg

Abstract Glaucoma is an ocular disease i.e. more common in older adults with elevated intraocular pressure and a serious threat to vision if it is not controlled. Due to the limitations regarding the conventional form of brinzolamide (Azopt®), two optimum formulations of in situ gel nanoemulsion were developed. To ensure the safety and efficacy of developed formulations for ocular drug delivery, the current study was designed. MTT assay was carried out on the human retinal pigmentation epithelial cells. To investigate the irritation potential of the chosen formulations, hen’s egg test-chorioallantoic membrane as a borderline test between in vivo and in vitro methods has been done. The modified Draize method was utilized to evaluate eye tolerance against the selected formulations. Intraocular pressure was measured by applying the prepared formulations to the eyes of normotensive albino rabbits in order to assess the therapeutic efficacy. Based on MTT test, cell viability for NE-2 at 0.1% and NE-1 at 0.1 and 0.5% concentrations was acceptable. The results of the hen’s egg test-chorioallantoic membrane test indicated no sign of vessel injury on the chorioallantoic membrane surface for both formulations. Also, during 24 h, both formulations were well-tolerated by rabbit eyes. The pharmacodynamics effects of formulations had no difference or were even higher than that of suspension in case of adding lower concentration (0.5%) of brinzolamide to the formulations. With regard to the results of the mentioned methods, our advanced formulations were effective, safe, and well-tolerated, thus can be introduced as an appropriate vehicle for ocular delivery of brinzolamide.

scholarly journals Localization of calcium in roots and microsomal membranes of corn by direct pyroantimonate precipitation.

1987 ◽  
Vol 35 (7) ◽  
pp. 781-788 ◽  
Author(s):  
M A Vaughan ◽  
T J Mulkey ◽  
C W Goff
Keyword(s):  
Plasma Membrane ◽  
Calcium Transport ◽  
Intact Cell ◽  
Root Cells ◽  
Meristematic Cells ◽  
Calcium Sequestration ◽  
Corn Roots ◽  
Microsomal Membranes

Many cell membrane systems, including microsomal vesicles of corn, are able to regulate calcium levels both in vivo and in vitro, often in an ATP-dependent, calmodulin-stimulated fashion. The purpose of this study was to determine calcium distribution in meristematic cells of intact tissue and microsomal vesicles from corn roots using direct pyroantimonate-osmium fixation. In root cells, precipitates were localized in mitochondria, plastids, the nucleus, endoplasmic reticulum, Golgi apparatus, and along the plasma membrane. Plasma membrane-enriched microsomal vesicles isolated from corn roots incubated in media to permit calcium transport before pyroantimonate-osmium fixation show internal precipitates associated with the membrane and in the lumen of the vesicles. De-staining of the sections with 1 mM EDTA or EGTA removed precipitate from the sections, confirming the presence of calcium in the antimonate precipitates. These data support biochemical data that this same membrane preparation exhibited ATP-dependent calcium sequestration that was stimulated by calmodulin, as measured by retention of 45Ca. This provides evidence that these membranes are responsible for ATP-requiring, calmodulin-stimulated calcium transport in the intact cell.

Abstract P401: The Anti-arrhythmic Effects Of VDAC Isoforms Are Mediated By Glutamate 73 Through Regulation Of Mitochondrial Calcium Transport

2021 ◽  
Vol 129 (Suppl_1) ◽  
Author(s):  
Lauren Crisman ◽  
Hirohito Shimizu ◽  
Adam Langenbacher ◽  
Jie Huang ◽  
Kevin Wang ◽  
...  
Keyword(s):  
Calcium Transport ◽  
Calcium Uptake ◽  
Calcium Overload ◽  
Mitochondrial Calcium ◽  
Cellular Processes ◽  
Excess Calcium ◽  
Evolutionarily Conserved ◽  
Mitochondrial Calcium Uptake

Mitochondria critically regulate cellular processes such as bioenergetics, metabolism, calcium homeostasis and apoptosis. VDAC proteins are abundant proteins that control the passage of ions and metabolites across the outer mitochondrial membrane. We have previously shown that activation of VDAC2, is able to buffer excess calcium and thereby suppress calcium overload induced arrhythmogenic events in vitro and in vivo. However, the mechanism by which VDAC2 regulates calcium transport and cardiac contractions remained unclear. It is also unclear whether all three VDAC isoforms (VDAC1,2 and 3) possess similar cardioprotective activity. The zebrafish tremblor/ncx1 mutant lacks functional NCX1 in cardiomyocytes leading to calcium overload, and the manifestation of fibrillation-like phenotypes. Using the tremblor/ncx1 mutant as a model, we observed isoform-specific differences between the VDAC family members. VDAC1 and VDAC2 enhanced mitochondrial calcium trafficking and restore rhythmic contraction in tremblor mutants, whereas, VDAC3 did not. We found that the differing rescue capabilities of VDAC proteins were dependent upon residues in their N-terminal halves. Phylogenetic analysis further revealed the presence of an evolutionarily conserved glutamate at position 73 (E73) within VDAC1 and VDAC2, but a glutamine (Q73) in VDAC3. Excitingly, we showed that replacing VDAC2 E73 with Q73 ablated its calcium transporting activity. Conversely, substituting the Q73 with E73 allows VDAC3 to gain calcium trafficking and cardioprotective abilities. Overall, our study demonstrates an essential role for the evolutionarily conserved glutamate-73 in determining the anti-arrhythmic effect of VDAC isoforms through their regulation of mitochondrial calcium uptake.

scholarly journals Promoting the Calcium-Uptake Bioactivity of Casein Phosphopeptides in vitro and in vivo

2021 ◽  
Vol 8 ◽  
Author(s):  
Guo Liu ◽  
Baoyan Guo ◽  
Shengwei Sun ◽  
Minna Luo ◽  
Fei Liu ◽  
...  
Keyword(s):  
Calcium Binding ◽  
Calcium Transport ◽  
Calcium Uptake ◽  
Calcium Absorption ◽  
Serum Alkaline Phosphatase ◽  
Binding Capacity ◽  
Animal Experiments ◽  
Casein Phosphopeptides

Casein phosphopeptides have been studied widely for their ability to chelate calcium. However, systematic studies on the effects of casein phosphopeptides (CPP) on calcium absorption in vitro and in vivo are scarce. The purities of two commercially available products, CPP1 and CPP2, are 18.37 and 25.12%, respectively. Here, the in vitro calcium binding capacity of CPP2 was 142.56 ± 7.39 mg/g, which was higher than that of CPP1 (107.15 ± 6.27 mg/g). The calcium transport results in a Caco-2 monolayer model indicated that, relative to controls, CPP1 and CPP2 increased calcium transport by 21.78 and 53.68%, respectively. Subsequent animal experiments showed that the CPP2-Ca-H group (1% Ca, 0.4% CPP2) had significant increases in the femur index, serum Ca2+ and serum osteocalcin levels, and femoral Ca content. The CPP2-Ca-H animal also had decreased serum alkaline phosphatase levels, parathyroid hormone content, and urinary pyridinoline content. Overall, our results demonstrated that CPP2 had stronger effects on promoting calcium uptake than CPP1.

Raphanus Sativus Seeds OilArrested In Vivo Inflammation and Angiogenesis Through Down-Regulation of TNF-a

2021 ◽  
Vol 22 ◽  
Author(s):  
Muhammad Asif ◽  
Hafiz Muhammad Yousaf ◽  
Mohammed Saleem ◽  
Liaqat Hussain ◽  
Mahrukh ◽  
...  
Keyword(s):  
Raphanus Sativus ◽  
Molecular Mechanisms ◽  
Radical Scavenging ◽  
Chorioallantoic Membrane ◽  
In Ovo ◽  
In Vivo Inflammation ◽  
Tnf Α ◽  
Anti Inflammatory

Background: Raphanus sativus is traditionally used as an anti-inflammatory agent. Objectives: The current study was designed to explore the in vivo anti-inflammatory and antiangiogenic properties of Raphanus sativus seeds oil. Methods: Cold press method was used for the extraction of oil (RsSO) and was characterised using GC-MS techniques. Three in vitro antioxidant assays (DPPH, ABTS, and FRAP) were performed to explore antioxidant potential of RsSO. Disc diffusion methods were used to study in vitro antimicrobial properties. In vivo anti-inflammatory properties were studied in both acute and chronic inflammation models. In ovo chicken, a chorioallantoic membrane assay was performed to study antiangiogenic effects. Molecular mechanisms were identified using serum TNF-α ELISA kit and docking tools. Results: GC-MS analysis of RsSO revealed the presence of hexadecanoic and octadecanoic acid. Findings of DPPH, ABTS, and FRAP models indicated relatively moderate radical scavenging properties of RsSO. Oil showed antimicrobial activity against a variety of strains tested. Data of inflammation models showed significant (p < 0.05) anti-inflammatory effects of RsSO in both acute and chronic models. 500 mg/kg RsSO halted inflammation development significantly better (p < 0.05) as compared with lower doses. Histopathological evaluations of paws showed minimal infiltration of inflammatory cells in RsSO-treated animals. Findings of TNF-α ELSIA and docking studies showed that RsSO has the potential to downregulate the expression of TNF-α, iNOS, ROS, and NF-κB, respectively. Moreover, RsSO showed in vivo antiangiogenic effects. Conclusion: Data of the current study highlight that Raphanus sativus seeds oil has anti-inflammatory, and antiangiogenic properties and can be used as an adjunct to standard NSAIDs therapy to reduce its dose and side effects.

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