Gene expression during mammalian oogenesis and early embryogenesis: quantification of three messenger RNAs abundant in fully grown mouse oocytes

Development ◽  
1989 ◽  
Vol 106 (2) ◽  
pp. 251-261 ◽  
Author(s):  
R.J. Roller ◽  
R.A. Kinloch ◽  
B.Y. Hiraoka ◽  
S.S. Li ◽  
P.M. Wassarman
Keyword(s):  
Gene Expression ◽  
Steady State ◽  
Early Embryogenesis ◽  
Messenger Rnas ◽  
Oocyte Growth ◽  
Mouse Oocytes ◽  
Mouse Tissues ◽  
Specific Mrnas ◽  
Steady State Levels ◽  
Ribonuclease Protection

Ribonuclease protection assays have been used to quantitatively assess changes in steady-state levels of specific mRNAs during oogenesis and early embryogenesis in mice. The mRNAs encode ZP3 (a glycoprotein that serves as a sperm receptor), LDH-B (heart-type lactate dehydrogenase), and MOM-1 (a protein of unknown function). MOM-1 and LDH-B are expressed in a variety of adult mouse tissues and midgestation embryos, whereas ZP3 expression is restricted completely to oocytes. All three mRNAs are expressed by growing mouse oocytes and accumulate to unusually high levels in fully grown oocytes as compared to somatic cells; 240,000, 200,000 and 74,000 copies mRNA per fully grown oocyte for ZP3, LDH-B and MOM-1, respectively. Steady-state levels of LDH-B and MOM-1 mRNA undergo a modest decline (approximately 20–40%) during ovulation when fully grown oocytes become unfertilized eggs and, in general, mirror the reported change in poly(A)+RNA levels during this period of development. On the other hand, the level of ZP3 mRNA declines dramatically (approximately 98%) during ovulation, from approximately 240,000 copies per oocyte to approximately 5000 copies per unfertilized egg, and ZP3 mRNA is undetectable in fertilized eggs (less than 1000 copies per fertilized egg). MOM-1 mRNA is expressed at relatively low levels in morulae (approximately 2000 copies per embryo) and blastocysts (approximately 5000 copies per embryo), whereas ZP3 mRNA remains undetectable (less than 1000 copies per embryo) at these stages of preimplantation development. These findings are discussed in the context of overall gene expression during oocyte growth, meiotic maturation and early embryogenesis in mice.

Regulation of prostatic genes: role of androgens and zinc in gene expression

1986 ◽  
Vol 64 (6) ◽  
pp. 601-607 ◽  
Author(s):  
R. J. Matusik ◽  
C. Kreis ◽  
P. McNicol ◽  
R. Sweetland ◽  
C. Mullin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Signal Peptide ◽  
Prostate Gland ◽  
Male Rats ◽  
Castration Resistant ◽  
Gene Transcripts ◽  
Specific Mrnas ◽  
Steady State Levels ◽  
Cloned Cdna

Gene expression in the rat dorsolateral prostate gland has been studied using cloned cDNA probes to the most abundant expressed mRNAs. One cDNA clone (pM-40) corresponds to two closely homologous mRNAs of about 880 nucleotides which code for two proteins of 23 and 21 kilodaltons (kDa). At least the 23-kDa protein contains a signal peptide. Another clone (pRWB) corresponds to a 1550-nucleotide mRNA which codes for a 52-kDa protein which also contains a signal peptide. The steady-state levels of these specific mRNAs increase in the dorsolateral prostate with sexual maturation. In castrated mature male rats, the M-40 mRNAs are inducible either by androgens or zinc, while the RWB mRNA is only responsive to androgens. In situ cDNA–mRNA hybridization histochemistry has been used to study the localization of the M-40 and RWB gene transcripts. Both M-40 and RWB mRNAs are most abundant in the epithelium of the lateral tip of the dorsolateral prostate. Following castration, the RWB mRNA decreases, while the M-40 mRNAs continue to be expressed in isolated areas of the epithelium. These castration-resistant cells maintain normal morphology in the absence of androgens.

Drosophila melanogaster homologs of the raf oncogene

1987 ◽  
Vol 7 (6) ◽  
pp. 2134-2140
Author(s):  
G E Mark ◽  
R J MacIntyre ◽  
M E Digan ◽  
L Ambrosio ◽  
N Perrimon
Keyword(s):  
Drosophila Melanogaster ◽  
In Situ Hybridization ◽  
Steady State ◽  
Kinase Domain ◽  
Early Embryogenesis ◽  
The Other ◽  
Chromosome 2 ◽  
Related Gene ◽  
Steady State Levels

A murine v-raf probe, representing the kinase domain, was used to identify two unique loci in Drosophila melanogaster DNA. The most closely related to v-raf was mapped by in situ hybridization to position 2F5-6 (Draf-1) on the X chromosome, whereas the other raf-related gene (Draf-2) was found at position 43A2-5 on chromosome 2. The nucleotide and amino acid homologies of Draf-1 to the kinase domain of v-raf are 61 and 65%, respectively. The large amount of a 3.2-kilobase Draf-1 transcript detected in eggs as a maternal message decreases during embryonic development, and significant steady-state levels are observed throughout the remainder of morphogenesis. We speculate that the Draf-1 locus plays an important role in early embryogenesis.

scholarly journals Dopaminergic regulation of avian prolactin gene transcription

2003 ◽  
Vol 31 (1) ◽  
pp. 185-196 ◽  
Author(s):  
A Al Kahtane ◽  
Y Chaiseha ◽  
M El Halawani
Keyword(s):  
Gene Expression ◽  
Steady State ◽  
Gene Transcription ◽  
Mrna Stability ◽  
Anterior Pituitary ◽  
Good Evidence ◽  
Pituitary Cells ◽  
Anterior Pituitary Cells ◽  
Steady State Levels ◽  
Prl Gene

It is well documented that prolactin (PRL) release and PRL gene expression in birds are controlled by the tonic stimulation of hypothalamic vasoactive intestinal peptide (VIP). However, there is good evidence that dopamine (DA) exerts both stimulatory (at the hypothalamic level) and inhibitory (at the pituitary level) effects on PRL secretion. The interactions between VIP and DA in the regulation of PRL gene transcription are not known. This study was designed to examine the effects of a D(2) DA receptor agonist (D(2)AG; R(-)-propylnorapomorphine HCl) on basal and VIP-stimulated PRL gene transcription rate, PRL mRNA steady-state levels, PRL mRNA stability and PRL release from cultured turkey anterior pituitary cells. The D(2)AG (10(-)(10) M) completely inhibited the stimulatory effect of VIP (10(-)(7) M) upon nascent PRL mRNA as determined utilizing a nuclear run-on transcription assay. To examine further the effect of the D(2)AG on PRL mRNA post-transcriptional events, anterior pituitary cells were treated with different concentrations of D(2)AG (10(-)(12)-10(-)(4) M). Semi-quantitative RT-PCR and RIA were performed to determine the levels of PRL mRNA and PRL content in the medium respectively. The results show that D(2)AG inhibited VIP-stimulated PRL mRNA steady-state levels as well as basal and VIP-stimulated PRL release, effects which were diminished by the D(2) DA receptor antagonist, S(-)-eticlopride HCl (10(-)(10) M). Actinomycin D (5 microg/ml), an inhibitor of mRNA synthesis, was used to assess the effect of D(2)AG on PRL mRNA stability in response to VIP. The stimulatory effect of VIP on PRL mRNA stability was completely negated by the D(2)AG (from a half-life of 53.0+/-2.3 h in VIP-treated cells to 25.5+/-1.6 h in D(2)AG+VIP-treated cells, P<or=0.05). These results support the hypothesis that VIP and DA play a major role in the regulation of PRL gene expression in avian species, at both the transcriptional and post-transcriptional levels. In addition, these findings suggest that the DAergic system inhibits PRL release and synthesis by antagonizing VIP at the pituitary level via D(2) DA receptors.

Steady-state levels of 7-methylguanine increase in nuclear DNA of postmitotic mouse tissues during aging

1990 ◽  
Vol 237 (5-6) ◽  
pp. 229-238 ◽  
Author(s):  
Boen H. Tan ◽  
F.Aladar Bencsath ◽  
James W. Gaubatz
Keyword(s):  
Steady State ◽  
Nuclear Dna ◽  
Mouse Tissues ◽  
Steady State Levels

Induction of gene expression during involution of the lactating mammary gland of the rat

1994 ◽  
Vol 12 (1) ◽  
pp. 47-60 ◽  
Author(s):  
R S Guenette ◽  
H B Corbeil ◽  
J Léger ◽  
K Wong ◽  
V Mézl ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mammary Gland ◽  
Steady State ◽  
Epithelial Cells ◽  
Tissue Transglutaminase ◽  
State Level ◽  
Ultrastructural Changes ◽  
Steady State Level ◽  
Hsp 27 ◽  
Steady State Levels

ABSTRACT After weaning, the mammary gland ceases lactation and involutes. The wet weight of the gland decreases by 70% within 4 days of weaning. This involves significant tissue remodelling as the ducts regress and return to the resting state. The presence of apoptotic bodies in the luminal epithelial compartment 2 to 3 days after weaning provides clear evidence that a substantial proportion of the regression is attributable to the induction of active cell death (ACD) of the epithelial cells. These changes in the architecture of the gland were found to be mirrored by changes in gene expression. The steady-state level of β-casein mRNA decreased rapidly after weaning from the high levels seen during lactation to undetectable levels by 8 days after weaning. The steady-state levels of expression of a number of genes associated with ACD, including TRPM-2, tissue transglutaminase (TGase) and poly(ADP-ribose) polymerase (PARP), increased transiently during this time-frame. The steady-state level of TRPM-2 mRNA increased 2 days after weaning, reaching a peak on day 4, and decreasing to undetectable levels by day 8 after weaning. The steady-state levels of two other mRNAs, TGase and PARP, showed very similar kinetics. In contrast, the mRNA for Hsp 27, which has been shown to be induced during prostate regression, was not significantly induced in the regressing mammary gland. In-situ hybridization demonstrated that the TRPM-2, TGase and PARP genes were expressed predominantly in the luminal epithelial cells of the ducts. These cells expressed β-casein mRNA during lactation, and underwent ACD after weaning. While the ultrastructural changes in the mammary gland after weaning, and the induction of TRPM-2, TGase and PARP mRNAs, are reminiscent of apoptosis in the prostate, several features of the process are different. Most notably, the disruption of the secretory processes and the lack of increased expression of Hsp 27 in the regressing mammary gland suggest that there may be a number of important events in ACD that are not common to all cells.

scholarly journals A novel negative cis-regulatory element on the hepatitis B virus S-(+)-strand

1999 ◽  
Vol 80 (10) ◽  
pp. 2673-2683 ◽  
Author(s):  
Markus Wagner ◽  
Michael Alt ◽  
Peter Hans Hofschneider ◽  
Matthias Renner
Keyword(s):  
Gene Expression ◽  
Steady State ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Regulatory Element ◽  
Regulatory Elements ◽  
Luciferase Gene ◽  
B Virus ◽  
Luciferase Mrna ◽  
Steady State Levels

Hepatitis B virus (HBV) has a double-stranded DNA genome. The minus-strand contains coding regions for all known HBV proteins and most of the cis-regulatory elements. Little is known about transcription from the S-(+)-strand and its regulation. Thus, the presence of regulatory elements located on the S-(+)-strand was investigated by inserting nt 1038–1783 of HBV in both orientations between the human cytomegalovirus (HCMV) promoter and a luciferase gene. Transfection experiments revealed that the plasmid containing this HBV DNA fragment in an orientation allowing expression from the S-(+)-strand (antisense) led to inhibition of luciferase gene expression compared to the plasmid containing this sequence in an orientation that allows gene expression from the L-(−)-strand (sense). Deletion analyses delimit the sequence essential for the inhibitory effect to a 150 bp region that also carries part of the enhancerII/core promoter complex. However, the possible influence of this regulatory element has been excluded in various experiments. The repressing HBV sequence acts in an orientation- and position-dependent manner; no inhibition was observed when this DNA element was inserted upstream of the HCMV promoter or downstream of the luciferase gene. Northern blot analyses revealed reduced luciferase mRNA steady-state levels in cells transfected with constructs containing the essential HBV sequence in antisense orientation compared to plasmids containing this sequence in sense orientation. Since nuclear run-on experiments showed similar transcription initiation rates with these plasmids, the diminished luciferase mRNA steady-state levels must be due to altered stabilities, suggesting that nt 1783–1638 of HBV encode an RNA-destabilizing element.

Sign in / Sign up

Export Citation Format

Share Document