Ion transport across the early chick embryo: I. Electrical measurements, ionic fluxes and regional heterogeneity

Kučera, Pavel; Abriel, Hugues; Katz, Uri

doi:10.1007/bf00238248

Cited by 8 publications

(3 citation statements)

References 30 publications

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“…Supporting this hypothesis are reports of unidirectional movement of sodium ions from the yolk sac across the blastoderm and early embryonic tissue (Abriel et al, 1994;Kučera et al, 1994;Stern & MacKenzie, 1983). Sodium transport by the blastoderm is electrogenic, sensitive to ouabain applied ventrally, and independent of extracellular chloride (Deeming et al, 1987;Kučera et al, 1994). The active sodium transport drives glucose and fluid into the intraembryonic space, across and around the blastoderm which, in the absence of blood circulation, could allow the renewal of extracellular fluid and disposal of wastes and thus maintain cell homeostasis (Abriel et al, 1994).…”

Section: Discussionmentioning

confidence: 90%

“…Chloride ions and water move down the electroosmotic gradient and the saline solution floats to the top of the yolk because of its low density (Elias, 1964). Supporting this hypothesis are reports of unidirectional movement of sodium ions from the yolk sac across the blastoderm and early embryonic tissue (Abriel et al, 1994;Kučera et al, 1994;Stern & MacKenzie, 1983). Sodium transport by the blastoderm is electrogenic, sensitive to ouabain applied ventrally, and independent of extracellular chloride (Deeming et al, 1987;Kučera et al, 1994).…”

Section: Discussionmentioning

confidence: 97%

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Shell‐less culture system for chick embryos from the blastoderm stage to hatching

Dunn

2023

J Exp Zool Pt A

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Since 2014, methods have been described to hatch chick embryos from shell‐less culture after egg contents are first incubated within shells for 55–70 h. The present report describes for the first time a shell‐less culture system for chick embryos from the blastoderm stage to hatching. For the first 69–70 h, egg contents suspended in polymethylpentene kitchen wrap (F.O.R. Wrap, Riken Fabro, Tokyo, Japan) supported in 6.35 or 6.67 cm inside diameter tripods and covered with a disc of immobilized Milli‐Wrap, were rotated back and forth through 90° at 16 or 22 cycles per minute (CPM). Subsequently, the Milli‐Wrap disc was removed and culture tripods were transferred to environmental chambers, which were rocked ±20° through incubation day 8.5 (E8.5). From E9, environmental chambers were maintained in the horizontal position through to hatching with controlled O2 and CO2. To provide supplemental calcium, an aqueous solution containing 100 mg/mL of calcium l‐lactate hydrate was injected through the plastic wrap into the albumen at E9 (2.5 mL) and at E13 (1.0 mL) or E15 (1.0 mL). After incubation for 69–70 h at 16 or 22 CPM, 80%–83% of previously unincubated egg contents yielded apparently normal embryos. Hatch rate of normal embryos resulting from turntable incubation at 16 or 22 CPM was approximately 43%. Of note, egg contents remained in the same culture tripod from blastoderm stage to hatching. This technique may find use as an educational tool and in basic investigations of early embryogenesis, teratogenesis, and gene transfer experiments.

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Section: Discussionmentioning

confidence: 90%

Section: Discussionmentioning

confidence: 97%

Shell‐less culture system for chick embryos from the blastoderm stage to hatching

Dunn

2023

J Exp Zool Pt A

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“…The mucosae were microdissected from the underlying tissue and cut into 7ϫ7 mm fragments, which were mounted in a transparent culture chamber designed for epithelia. 37,38,36 The mucosa divided the chamber into apical ͑epithelial͒ and basal ͑submucosal͒ compartments. The chamber was fixed onto the plate of an epifluorescence microscope ͑Leitz Orthoplan͒ and thermostabilized at 37Ϯ0.5°C.…”

Section: Preparation Of Pig Bladder and Sheep Paranasal Mucosamentioning

confidence: 99%

Routine experimental system for defining conditions used in photodynamic therapy and fluorescence photodetection of (non-) neoplastic epithelia

et al. 2001

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A common method to induce enhanced short-term endogenous porphyrin synthesis and accumulation in cell is the topical, systemic application of 5-aminolevulinic acid or one of its derivatives. This circumvents the intravenous administration of photosensitizers normally used for photodynamic therapy (PDT) of fluorescence photodetection. However, in the majority of potential medical indications, optimal conditions with respect to the porphyrin precursor or its pharmaceutical formulation have not yet been found. Due to ethical restrictions and animal right directives, the number of available test objects is limited. Hence, definition and use of nonanimal test methods are needed. Tissue and organ cultures are a promising approach in replacing cost intensive animal models in early stages of drug development. In this paper, we present a tissue culture, which can among others be used routinely to answer specific questions emerging in the field of photodynamic therapy and fluorescence photodetection. This technique uses mucosae excised from sheep paranasal sinuses or pig bladder, which is cultured under controlled conditions. It allows quasiquantitative testing of different protoporphyrin IX precursors with respect to dose-response curves and pharmacokinetics, as well as the evaluation of different incubation conditions and/or different drug formulations. Furthermore, this approach, when combined with the use of electron microscopy and fluorescence-based methods, can be used to quantitatively determine the therapeutic outcome following protoporphyrin IX-mediated PDT.

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Ion transport across the early chick embryo: II. Characterization and pH sensitivity of the transembryonic short-circuit current

1994

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The ectoderm of the one-day chick embryo generates dorsoventrally oriented short-circuit current (Isc) entirely dependent on extracellular sodium. At the dorsal cell membrane, the Isc was modified reversibly and in a concentration-dependent manner by: amiloride (60% decrease at 1 mM, with 2 apparent IC50S: 0.13 and 48 microM), phlorizin (0.1 mM) or removal of glucose (30% decrease, additive to that of amiloride), SITS (1 mM, 13% decrease). Acidification of alkalinization of the dorsal (but not ventral) superfusate produced, respectively, decrease or increase of Isc with a pH50 of 7.64. Ba2+ (0.1-1 mM) from either side of the ectoderm decreased the Isc by 30%. Anthracene-9-carboxylic acid, furosemide and inducers of cAMP had no effect on electrophysiological properties of the blastoderm. The chick ectoderm is therefore a highly polarized epithelium containing, at the dorsal membrane, the high and low affinity amiloride-sensitive Na+ channels, Na(+)-glucose cotransporter, K+ channels and pH sensitivity, and, at the ventral membrane, the Na+, K(+)-ATPase and K+ channels. The Na+ transport reacts to pH, but lacks the cAMP regulatory system, well known in many epithelia. The active Na+ transport drives glucose and fluid into the intraembryonic space, across and around the blastoderm which, in the absence of blood circulation, could secure renewal of extracellular fluid and disposal of wastes and thus maintain the cell homeostasis.

show abstract

Ion transport across the early chick embryo: I. Electrical measurements, ionic fluxes and regional heterogeneity

Cited by 8 publications

References 30 publications

Shell‐less culture system for chick embryos from the blastoderm stage to hatching

Shell‐less culture system for chick embryos from the blastoderm stage to hatching

Routine experimental system for defining conditions used in photodynamic therapy and fluorescence photodetection of (non-) neoplastic epithelia

Ion transport across the early chick embryo: II. Characterization and pH sensitivity of the transembryonic short-circuit current

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