Molecular cloning of cDNA sequences for avian malic enzyme. Nutritional and hormonal regulation of malic enzyme mRNA levels in avian liver cells in vivo and in culture.

ABSTRACT A cloned cDNA has been isolated by probing a sheep blastocyst cDNA library using a synthetic oligonucleotide representing the N-terminal amino acid sequence of the antiluteolytic protein, ovine trophoblast protein-1. Sequence analysis of the cDNA confirms the 70% homology between the antiluteolysin and the interferon-α family of proteins; however, the sequence reported here differs at several points from previously reported amino acid and cDNA sequences for the antiluteolysin. In-vitro translation of day-16 poly(A)+ RNA indicated that antiluteolysin mRNA is a major constituent of total mRNA at this stage of blastocyst development, and Northern blotting confirmed that antiluteolysin mRNA production occurred between days 13 and 22 after oestrus. This is consistent with the stage at which embryonic extracts are antiluteolytic on administration in vivo. These and other data confirm that the ovine trophoblast antiluteolysin is an interferon, and suggest that at least five isoforms of this protein may exist.

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Molecular cloning and analysis of in vivo expression of murine P- selectin

Blood ◽

10.1182/blood.v80.3.795.795 ◽

1992 ◽

Vol 80 (3) ◽

pp. 795-800 ◽

Cited By ~ 104

Author(s):

WE Sanders ◽

RW Wilson ◽

CM Ballantyne ◽

AL Beaudet

Keyword(s):

Molecular Cloning ◽

Adhesion Molecule ◽

Cellular Response ◽

Tissue Injury ◽

Amino Acid Identity ◽

Mrna Levels ◽

Surface Adhesion ◽

New Synthesis

Abstract P-selectin (CD62) is a rapidly inducible cell surface adhesion molecule that is expressed on platelets and endothelial cells and mediates their interaction with leukocytes. In vitro studies have suggested that this receptor may play an important role in hemostasis and in inflammatory response to tissue injury. We report the molecular cloning and sequencing of murine cDNA for P-selectin. The lectin, epidermal growth factor (EGF)-like, transmembrane, and cytoplasmic domains are highly conserved between mouse and human, with an overall amino acid identity of 79%. To further investigate the biology of this adhesion molecule in vivo, we analyzed mRNA levels for P-selectin in mice after injection with endotoxin. Northern blot data indicate that the cellular response in vivo includes a rapid increase in the level of mRNA, presumably for new synthesis of P-selectin. The increase in mRNA is maximal at 4 hours, and turnover is relatively rapid, with levels of RNA having decreased substantially by 6 hours following stimulation with endotoxin. After administration of endotoxin, the highest levels of mRNA expression were detected in liver, lung, kidney, and heart.

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Regulation of the epithelial Ca2+ channels in small intestine as studied by quantitative mRNA detection

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00036.2003 ◽

2003 ◽

Vol 285 (1) ◽

pp. G78-G85 ◽

Cited By ~ 127

Author(s):

Monique van Abel ◽

Joost G. J. Hoenderop ◽

Annemiete W. C. M. van der Kemp ◽

Johannes P. T. M. van Leeuwen ◽

René J. M. Bindels

Keyword(s):

Knockout Mice ◽

Hormonal Regulation ◽

Transport Proteins ◽

Ovariectomized Rats ◽

Mrna Levels ◽

Hydroxyvitamin D ◽

Dihydroxyvitamin D ◽

Genes Encoding ◽

Calbindin D

The epithelial Ca2+ channels TRPV5 and TRPV6 are localized to the brush border membrane of intestinal cells and constitute the postulated rate-limiting entry step of active Ca2+ absorption. The aim of the present study was to investigate the hormonal regulation of these channels. To this end, the effect of 17β-estradiol (17β-E2), 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], and dietary Ca2+ on the expression of the duodenal Ca2+ transport proteins was investigated in vivo and analyzed using realtime quantitative PCR. Supplementation with 17β-E2 increased duodenal gene expression of TRPV5 and TRPV6 but also calbindin-D9K and plasma membrane Ca2+-ATPase (PMCA1b) in ovariectomized rats. 25-Hydroxyvitamin D3-1α-hydroxylase (1α-OHase) knockout mice are characterized by hyperparathyroidism, rickets, hypocalcemia, and undetectable levels of 1,25(OH)2D3 and were used to study the 1,25(OH)2D3-dependency of the stimulatory effects of 17β-E2. Treatment with 17β-E2 upregulated mRNA levels of duodenal TRPV6 in these 1α-OHase knockout mice, which was accompanied by increased serum Ca2+ concentrations from 1.69 ± 0.10 to 2.03 ± 0.12 mM ( P < 0.05). In addition, high dietary Ca2+ intake normalized serum Ca2+ in these mice and upregulated expression of genes encoding the duodenal Ca2+ transport proteins except for PMCA1b. Supplementation with 1,25(OH)2D3 resulted in increased expression of TRPV6, calbindin-D9K, and PMCA1b and normalization of serum Ca2+. Expression levels of duodenal TRPV5 mRNA are below detection limits in these 1α-OHase knockout mice, but supplementation with 1,25(OH)2D3 upregulated the expression to significant levels. In conclusion, TRPV5 and TRPV6 are regulated by 17β-E2 and 1,25(OH)2D3, whereas dietary Ca2+ is positively involved in the regulation of TRPV6 only.

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Glucocorticoids regulate mRNA levels for subunits of the 19 S regulatory complex of the 26 S proteasome in fast-twitch skeletal muscles

Biochemical Journal ◽

10.1042/bj20031660 ◽

2004 ◽

Vol 378 (1) ◽

pp. 239-246 ◽

Cited By ~ 52

Author(s):

Lydie COMBARET ◽

Daniel TAILLANDIER ◽

Dominique DARDEVET ◽

Daniel BÉCHET ◽

Cécile RALLIÈRE ◽

...

Keyword(s):

Hormonal Regulation ◽

Skeletal Muscles ◽

Conclusive Evidence ◽

Mrna Levels ◽

Glucocorticoid Treatment ◽

Regulatory Subunits ◽

Regulatory Complex ◽

Insulin Dependent ◽

Fast Twitch

Circulating levels of glucocorticoids are increased in many traumatic and muscle-wasting conditions that include insulin-dependent diabetes, acidosis, infection, and starvation. On the basis of indirect findings, it appeared that these catabolic hormones are required to stimulate Ub (ubiquitin)–proteasome-dependent proteolysis in skeletal muscles in such conditions. The present studies were performed to provide conclusive evidence for an activation of Ub-proteasome-dependent proteolysis after glucocorticoid treatment. In atrophying fast-twitch muscles from rats treated with dexamethasone for 6 days, compared with pair-fed controls, we found (i) increased MG132-inhibitable proteasome-dependent proteolysis, (ii) an enhanced rate of substrate ubiquitination, (iii) increased chymotrypsin-like proteasomal activity of the proteasome, and (iv) a co-ordinate increase in the mRNA expression of several ATPase (S4, S6, S7 and S8) and non-ATPase (S1, S5a and S14) subunits of the 19 S regulatory complex, which regulates the peptidase and the proteolytic activities of the 26 S proteasome. These studies provide conclusive evidence that glucocorticoids activate Ub-proteasome-dependent proteolysis and the first in vivo evidence for a hormonal regulation of the expression of subunits of the 19 S complex. The results suggest that adaptations in gene expression of regulatory subunits of the 19 S complex by glucocorticoids are crucial in the regulation of the 26 S muscle proteasome.

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Hormonal regulation of aquaporin 3: opposing actions of prolactin and cortisol in tilapia gill

Journal of Endocrinology ◽

10.1530/joe-16-0162 ◽

2016 ◽

Vol 230 (3) ◽

pp. 325-337 ◽

Cited By ~ 18

Author(s):

Jason P Breves ◽

Mayu Inokuchi ◽

Yoko Yamaguchi ◽

Andre P Seale ◽

Bethany L Hunt ◽

...

Keyword(s):

Hormonal Regulation ◽

Cell Membranes ◽

Mrna Levels ◽

Gill Epithelium ◽

Aquaporin 3 ◽

Gill Filaments ◽

Branchial Epithelium ◽

Hydromineral Balance

Aquaporins (Aqps) are expressed within key osmoregulatory tissues where they mediate the movement of water and selected solutes across cell membranes. We leveraged the functional plasticity of Mozambique tilapia (Oreochromis mossambicus) gill epithelium to examine how Aqp3, an aquaglyceroporin, is regulated in response to osmoregulatory demands. Particular attention was paid to the actions of critical osmoregulatory hormones, namely, prolactin (Prl), growth hormone and cortisol. Branchial aqp3 mRNA levels were modulated following changes in environmental salinity, with enhanced aqp3 mRNA expression upon transfer from seawater to freshwater (FW). Accordingly, extensive Aqp3 immunoreactivity was localized to cell membranes of branchial epithelium in FW-acclimated animals. Upon transferring hypophysectomized tilapia to FW, we identified that a pituitary factor(s) is required for Aqp3 expression in FW. Replacement with ovine Prl (oPrl) was sufficient to stimulate Aqp3 expression in hypophysectomized animals held in FW, an effect blocked by coinjection with cortisol. Both oPrl and native tilapia Prls (tPrl177 and tPrl188) stimulated aqp3 in incubated gill filaments in a concentration-related manner. Consistent with in vivo responses, coincubation with cortisol blocked oPrl-stimulated aqp3 expression in vitro. Our data indicate that Prl and cortisol act directly upon branchial epithelium to regulate Aqp3 in tilapia. Thus, within the context of the diverse actions of Prl on hydromineral balance in vertebrates, we define a new role for Prl as a regulator of Aqp expression.

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Molecular cloning of a cDNA sequence for rat malic enzyme. Direct evidence for induction in vivo of rat liver malic enzyme mRNA by thyroid hormone.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(17)44234-7 ◽

1983 ◽

Vol 258 (20) ◽

pp. 12712-12717 ◽

Cited By ~ 13

Author(s):

M A Magnuson ◽

V M Nikodem

Keyword(s):

Thyroid Hormone ◽

Molecular Cloning ◽

Rat Liver ◽

Malic Enzyme ◽

Cdna Sequence ◽

Direct Evidence

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Estrogen-induced inhibition of spermatogenesis in zebrafish is largely reversed by androgen

Journal of Molecular Endocrinology ◽

10.1530/jme-17-0177 ◽

2018 ◽

Vol 60 (4) ◽

pp. 273-284 ◽

Cited By ~ 11

Author(s):

Luiz Henrique de Castro Assis ◽

Rafael Henrique de Nóbrega ◽

Nuria Esther Gómez-González ◽

Jan Bogerd ◽

Rüdiger Winfried Schulz

Keyword(s):

Growth Factors ◽

Germ Cell ◽

Hormonal Regulation ◽

Mrna Levels ◽

Transcript Levels ◽

Gonadotropin Release ◽

Androgen Treatment ◽

Germ Cell Differentiation ◽

Estrogen Exposure

The hormonal regulation of spermatogenesis involves both gonadotropins and steroid hormones. Long-term in vivo exposure of adult zebrafish to estrogen impaired spermatogenesis associated with an androgen insufficiency, possibly induced by inhibiting gonadotropin release. Using this experimental model, we investigated if androgen treatment could enhance spermatogenesis, while maintaining the inhibition of gonadotropin release through continued estrogen exposure. Moreover, we also exposed animals to androgen alone, in order to examine androgen effects in the absence of estrogen-induced gonadotropin inhibition. Estrogen exposure depleted type B spermatogonia, meiotic and postmeiotic germ cells from the adult testis, but promoted the proliferation of type A undifferentiated spermatogonia, which accumulated in the testis. This change in germ cell composition was accompanied by reduced mRNA levels of those growth factors (e.g. insl3 and igf3) expressed by testicular somatic cells and known to stimulate spermatogonial differentiation in zebrafish. Additional androgen (11-ketoandrostenedione, which is converted to 11-ketotestosterone) treatment in vivo reversed most of the effects of estrogen exposure on spermatogenesis while insl3 and igf3 transcript levels remained suppressed. When androgen treatment was given alone, it promoted the production of haploid cells at the expense of spermatogonia, and increased transcript levels of some growth factor and hormone receptor genes, but not those of insl3 or igf3. We conclude that estrogen exposure efficiently inhibits spermatogenesis because it induces androgen insufficiency and suppresses gonadotropin-regulated growth factors known to stimulate germ cell differentiation. Moreover, our results suggest that androgens and the growth factors Insl3 and Igf3 stimulate spermatogenesis via independent pathways.

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Molecular cloning and analysis of in vivo expression of murine P- selectin

Blood ◽

10.1182/blood.v80.3.795.bloodjournal803795 ◽

1992 ◽

Vol 80 (3) ◽

pp. 795-800 ◽

Cited By ~ 4

Author(s):

WE Sanders ◽

RW Wilson ◽

CM Ballantyne ◽

AL Beaudet

Keyword(s):

Molecular Cloning ◽

Adhesion Molecule ◽

Cellular Response ◽

Tissue Injury ◽

Amino Acid Identity ◽

Mrna Levels ◽

Surface Adhesion ◽

New Synthesis

P-selectin (CD62) is a rapidly inducible cell surface adhesion molecule that is expressed on platelets and endothelial cells and mediates their interaction with leukocytes. In vitro studies have suggested that this receptor may play an important role in hemostasis and in inflammatory response to tissue injury. We report the molecular cloning and sequencing of murine cDNA for P-selectin. The lectin, epidermal growth factor (EGF)-like, transmembrane, and cytoplasmic domains are highly conserved between mouse and human, with an overall amino acid identity of 79%. To further investigate the biology of this adhesion molecule in vivo, we analyzed mRNA levels for P-selectin in mice after injection with endotoxin. Northern blot data indicate that the cellular response in vivo includes a rapid increase in the level of mRNA, presumably for new synthesis of P-selectin. The increase in mRNA is maximal at 4 hours, and turnover is relatively rapid, with levels of RNA having decreased substantially by 6 hours following stimulation with endotoxin. After administration of endotoxin, the highest levels of mRNA expression were detected in liver, lung, kidney, and heart.

Download Full-text