In vitro translation and estradiol-17beta induction of Xenopus laevis vitellogenin messenger RNA.

Shapiro, David J.; Baker, Henry J.; Stitt, David T.

doi:10.1016/s0021-9258(17)33504-4

Cited by 72 publications

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References 35 publications

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“…In parallel cultures maintained over the same time period in medium containing insulin and 0.1% propylene glycol (estradiol solvent), <1% of secreted protein was immunoprecipitable on each day analyzed . Overall, the duration of the lag and the magnitude of the response to estradiol in the cultured hepatocytes closely resemble those found in vivo (18,23).…”

Section: Induction Of Vitellogenin Synthesis By Estradiolsupporting

confidence: 63%

“…At peak hormonal response, vitellogenin is the predominant protein synthesized by the liver (12,18,23) . The commitment of such a large part of the liver's protein synthetic capacity to the production of a single protein along with the availability of large amounts of tissue for biochemical analysis makes this a generally attractive system for studies on the hormonal regulation of gene expression .…”

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Primary induction of vitellogenin synthesis in monolayer cultures of amphibian hepatocytes.

Stanchfield

Yager

1980

The Journal of Cell Biology

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Direct induction of vitellogenin production in cultured male amphibian hepatocytes by estradiol-l7ß has been accomplished. Liver cells were isolated from adult male bullfrogs by collagenase perfusion and maintained as monolayers in serumfree medium containing insulin and estradiol . Vitellogenin production was measured by direct immunoprecipitation from radioactively labeled secreted protein with a specific antiserum against vitellogenin . Significant quantities of vitellogenin were detected in the exported protein on the second day of hormone treatment . Vitellogenin production increased with duration of culture in the presence of estradiol until by the eighth day -90% of secreted protein was vitellogenin . This response is largely comparable to that obtainable in vivo .Indirect immunofluorescence microscopy was used to identify cells synthesizing vitellogenin in response to estradiol . An increase in cytoplasmic fluorescence could be seen in cells throughout the cultures, with increasing time in the presence of estradiol . By the sixth day of treatment, the majority of cells showed significant fluorescence labeling . The results suggest that studies on the mechanisms underlying the primary activation of the vitellogenin gene may now be conducted under well defined conditions in a monolayer liver cell culture system .

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Section: Induction Of Vitellogenin Synthesis By Estradiolsupporting

confidence: 63%

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confidence: 99%

Primary induction of vitellogenin synthesis in monolayer cultures of amphibian hepatocytes.

Stanchfield

Yager

1980

The Journal of Cell Biology

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“…Γιά νά απαντήσουν σ'αυτήν την ερώτη ση, διάψορες ομάδες εξέτασαν τήν παρουσία του mRNA της βιτελλογενίνης,-μεταφράζοντας in vitro τό κυτταροπλασματικό RNA. Τά αποτελέσματα πού πήραν γιά τόν Xenopus (Shapiro et al, 1976* Skipper καί Hamilton, 1977' Farmer et al, 1978 καί γιά τήν κόττα (Mullinix et al, 1976), άπέδει£αν τήν υπαρ.ξη μεταφραστικά ένεργοϋ mRNA στό κυτταρόπλασμα, με τά τή χορήγηση οιστρογόνου. 'Αλλά σ'αύτά τά πειράματα, τό ποσό τοϋ mRNA της βιτελλογενίνης μετρήθηκε μέ βάση τή με ταφραστική του ένεργότητα καί ως εκ τούτου, δέν είναι δυ νατόν ν'αποκλεισθεί ή παρουσία ενός ποσοστού mRNA της βιτελλογενίνης πού είναι ανενεργό γιά in vitro μετάφραση.…”

Section: ορμονικη ρυθμιση της βιτελλογενεσηςunclassified

Δομη Χρωματινης Και Ρυθμιση Της Γονιδιακης Εκφρασης

Δημητριαδησ¹

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Τό αρχικό μέρος της διατριβής αυτής έγινε στό 'Εθνικό 'Ινστιτούτο 'Ιατρικής "Ερευνας (N.I.M.R.) της Μ.Βρεττανίας. ένώ τό τελικό μέρος καί ή θεωρητική επεξεργασία των αποτε λεσμάτων στό 'Εθνικό "Ιδρυμα 'Ερευνών. Ευχαριστώ τόν δι ευθυντή του τμήματος 'Αναπτυξιακής Βιολογίας του N.I.M.R. Dr J.R.Tata γιά τή φιλοξενία του καθώς καί τόν διευθυντή τοϋ Κέντρου Βιολογικών 'Ερευνών Καθηγητή κ. Κ.Σέκερη τόσο γιά τήν ηθική συμπαράσταση του δσο καί για τήν κριτική με λέτη των χειρογράφων αυτής της διατριβής. 'Επίσης θέ λω νά ευχαριστήσω γιά τή διόρθωση τών κειμένων τίς Drs Βά σω Ζόγκζα-Δημητριάδη καί Μερόπη Πατρινου-Γεωργούλα καθώς καί τήν δ. 'Ελένη Μιχαλοπούλου γιά τή δακτυλογράφηση τών κειμένων. V 1.3.7. Μετατροπές ίστονών και ενεργή χρωματίνη. Σελίδα 1.3.8. Μηχανισμοί άποσυγκρότησης των νουκλεοσωμάτων στην ενεργή χρωματίνη " 2. ΕΛΕΓΧΟΣ ΤΗΣ ΓΟΝΙΔΙΛΚΗΣ ΕΚΦΡΑΣΗΣ ΚΑΤΑ ΤΗΝ ΔΙΑ-2.1. ΘΕΩΡΙΕΣ ΤΗΣ ΔΙΑΦΟΡΟΠΟΙΗΣΗΣ....«,. ....... 2.1.1. θεωρία της κατευθυνόμενης σωματικής μεταλ λαγής ο 2.1.1.1. Πληροφοριακό περιεχόμενο των γονιδιωμάτων των διαφοροποιημένων κυττάρων 2.1.

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Use of the protein A–gold immunocytochemical and enzyme‐gold cytochemical techniques in studies of vitellogenesis

Herbener

1989

Am. J. Anat.

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Vitellogenesis in the frog hepatocyte was investigated by applying the protein A-gold immunocytochemical and RNase-gold cytochemical techniques in conjunction with morphometric and biochemical analyses. The morphometric studies demonstrated that the surface density of rough endoplasmic reticulum (RER) and nucleolar size increased more than fourfold and 1.25-fold, respectively, while the nuclear size and the mitochondrial compartment size remained constant following estrogen treatment. Concurrently, liver RNA concentration increased 2.5-fold while protein and DNA concentrations did not change. In addition, total plasma protein more than doubled, with vitellogenin accounting for 40% of the final volume. The secretory proteins vitellogenin and protein-RcX (a nonvitellogenin, estrogen-induced plasma protein of unknown function, found in the plasma of Rana catesbeiana) were detected immunocytochemically in the RER, Golgi apparatus, and secretory granules in hepatocytes only of estrogen-treated frogs. Lysosomes also were labeled. These observations established that protein-RcX was synthesized and secreted by the hepatocyte in parallel with vitellogenin and that both of these export proteins were confined to the secretory pathway and lysosomes. Quantitation of labeling density indicated that the concentration of vitellogenin increased as it progressed along the secretory vector. Albumin was detected immunocytochemically also within these same hepatocyte entities from both untreated and treated animals. In the untreated animals, albumin concentration also increased progressively along the secretory vector. A marked alteration of albumin processing was observed following estrogen treatment. While albumin concentration in the RER was unchanged, its concentrations within the Golgi apparatus and secretory granules were lower than those observed in the RER or in counterpart compartments under control conditions. RNase-gold cytochemistry for total RNA demonstrated a 1.5-fold increase in labeling density over the nucleolus but no change in RER labeling following estrogen treatment. These labeling data, in combination with the morphometric data, suggest an increase of approximately 80% in the total amount of RNA in the nucleolus and 430% in the RER in response to estrogen. This review thus illustrates the significant contributions which can be made by gold-probe techniques, alone or in combination with morphometric and biochemical techniques, to investigations of the intracellular processing of secretory proteins.

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In vitro translation and estradiol-17beta induction of Xenopus laevis vitellogenin messenger RNA.

Cited by 72 publications

References 35 publications

Primary induction of vitellogenin synthesis in monolayer cultures of amphibian hepatocytes.

Primary induction of vitellogenin synthesis in monolayer cultures of amphibian hepatocytes.

Δομη Χρωματινης Και Ρυθμιση Της Γονιδιακης Εκφρασης

Use of the protein A–gold immunocytochemical and enzyme‐gold cytochemical techniques in studies of vitellogenesis

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