bluuebunny/crossref_metadata_2025_split huggingface dataset:
Modulation of gene expression in the preimplantation mouse embryo by TGF‐α and TGF‐β
Authors: Babalola Gbolagade O. , Schultz Richard M. 
Published: February 2005

AbstractThe effect of growth factors on regulating gene expression in the preimplantation mouse embryo was examined, since results of previous experiments revealed a stimulatory effect of exogenously‐added growth factors on preimplantation development in vitro. Treatment of early cavitating blastocysts with either 250 pM TGF‐α or TGF‐β results in changes in the pattern of total protein synthesis as assessed by high‐resolution two‐dimensional gel electrophoresis. In some cases, the synthesis of a particular polypeptide is either up‐ or downregulated by each growth factor, whereas in other instances the synthesis of a polypeptide is modulated by one but not the other growth factor. Use of the mRNA differential display method permitted the identification of genes whose expression is either up‐ or downregulated by these growth factors. Treatment of mouse blastocysts with either TGF‐α or TGF‐β results in the increased expression of the b subunit of the F0 ATPase. TGF‐β also stimulates the expression of the DNA polymerase α. TGF‐α treatment results in the increase in expression of a gene homologous to the human HEPG2 cDNA, as well as in a decrease in expression of fibronectin. © 1995 Wiley‐Liss, Inc.
Differential constitutive expression of interferon genes in early mouse embryos
Authors: Riego Eileen , Pérez Aimeé , Martínez Rebeca , Castro Fidel O. , Lleonart Ricardo , de la Fuente José 
Published: February 2005

AbstractRecent evidence suggests that several processes during mammalian embryogenesis may be regulated by IFNs or IFN‐like molecules. With the use of MAPPing, the simultaneous presence of transcripts homologous to IFN‐α, IFN‐β, IRF‐1, and IRF‐2 was examined in mouse embryos and in embryonal carcinoma (EC) P19 cells, which are equivalent to epiblast cells of the early postimplantation blastocyst. Transcripts for IFN‐α, but not for IFN‐β, were detected as maternal transcripts in the ovulated oocyte and persisted over early embryogenesis. IRF‐1 transcripts appeared only after the first cell cleavage in the two‐cell stage embryo. IRF‐2 transcripts were analyzed only in EC P19 cells and were found in both undifferentiated (D−) and differentiated (D+) cells. The IFN‐α transcripts present in (D−) P19 cells were cloned and the partial cDNA sequences determined. Mu IFN‐αA and a new Mu IFN‐α species (Mu IFN‐α12) were isolated from (D−) P19 cells. The presence of constitutive IFN‐α transcripts in early mouse embryos suggests a role for these molecules during embryogenesis. © 1995 Wiley‐Liss, Inc.
Monoallelic expression of IGF2 at the human fetal/maternal boundary
Authors: Ekström Tomas J. , Cui Hengmi , Nyström Anders , Rutanen Eeva‐Marja , Ohlsson Rolf 
Published: February 2005

AbstractIGF2 is expressed in both placental and decidual tissues, enabling an analysis of the parental imprinting over the fetomaternal boundary. Evidence is provided that IGF2 is monoallelically expressed in both placenta and pregnant, as well as nonpregnant, endometrium. These observations suggest that the maternally derived IGF2 allele is inactivated during germline transmission. Comparison of promoter usage in decidua and placental samples shows that the P3 promoter appears to regulated independently of the others. These observations are discussed with respect to current models of IGF2 imprinting and the hypothesized conflict of parental reproductive interests which bears on the phenomenon of parental imprinting. © 1995 Wiley‐Liss, Inc.
Rabbit blastocyst: Allocation of cells to the inner cell mass and trophectoderm
Authors: Giles J. R. , Foote R. H. 
Published: February 2005

AbstractThe proportion of total cells in the blastocyst allocated to the inner cell mass (ICM) and trophectoderm (TE) is important for future development and may be a sensitive indicator to evaluate culture conditions. The number of cells and their distribution within the two primary cell lineages were determined for the rabbit embryo developing in vivo after superovulation or nonsuperovulation or embryo transfer and compared with embryos developing in vitro. Comparisons were made with cultured embryos or embryos grown in vivo until 3.5, 4.0, and 4.5 days of age. Embryos from superovulated rabbits developed in vivo for 3.5, 4.0, and 4.5 days, respectively, had 361, 758, and 902 total cells (P<0.05), and in nonsuperovulated rabbits 130, 414, and 905 total cells (P<0.05), with increasing proportions of ICM cells over time (P<0.05). One‐cell embryos recovered from superovulated females and transferred to nonsuperovulated recipients developed more slowly with 70, 299, and 550 total cells after 3.5, 4.0, and 4.5 days of culture (P<0.05), respectively. The proportion of ICM cells increased with age of the embryo. Corresponding values for one‐cell embryos cultured in vitro resulted in 70, 299, and 550 total cells (P<0.05). However, in vitro culture of morula‐stage embryos in the presence of fetal bovine serum for 24 hr did not delay growth. In addition, the proportions of ICM/total cells were 0.17, 0.25, and 0.29 for embryos developing in vitro at 3.5, 4.0, and 4.5 days, respectively, similar to those for embryos developing in vivo at each of the three recovery times. These data establish for the first time the number and proportion of cells allocated to the ICM of the rabbit embryo developing in vivo or under defined conditions in vitro. © 1995 Wiley‐Liss, Inc.
Preimplantation development of mouse embryos in KSOM: Augmentation by amino acids and analysis of gene expression
Authors: Ho Yugong , Wigglesworth Karen , Eppig John J. , Schultz Richard M. 
Published: February 2005

AbstractSimplex optimization has generated several media that have improved the development of mouse preimplantation embryos in vitro. One objective of this study was to compare the development of preimplantation mouse embryos in one of these computer‐optimized media, KSOM, with embryos that developed in vivo, in terms of the relative abundances of specific mRNAs involved in metabolism, transcription, and cell proliferation. First, however, since studies have indicated an improvement of other simple embryo culture media by addition of amino acids, the effects of the addition of amino acids to KSOM (KSOM/AA) on preimplantation development were assessed. We find that addition of both essential and non‐essential amino acids to KSOM augments development in vitro, as compared to development supported by KSOM without amino acids. This augmentation is observed starting at the blastocyst stage, and is associated with increased rate of development to the blastocyst stage, increased frequency of hatching, and increased number of cells in the blastocysts. Reverse‐transcription PCR was then used to assess the relative abundance of mRNAs for actin, glyceraldehyde‐3‐phosphate dehydrogenase, Na+, K+‐ATPase, Sp1, TATA box‐binding protein TBP, IGF‐I, IGF‐II, IGF‐I receptor, and IGF‐II receptor in embryos that developed in vivo and in vitro using KSOM/AA. Eight out of 9 of these mRNAs were present in the 8‐cell embryos and blastocysts raised in KSOM/AA in amounts that were indistinguishable from those in embryos that developed in vivo. It is concluded that KSOM/AA provides an environment in which preimplantation mouse embryos can undergo development that is quantitatively similar to that occurring in vivo. © 1995 Wiley‐Liss, Inc.
Transcriptional up‐regulation of the mouse gene for the muscle‐specific subunit of enolase during terminal differentiation of myogenic cells
Authors: Lamandé Noël , Brosset Sophie , Lucas Marguerite , Keller Angélica , Rouzeau Jean‐Denis , Johnson Thomas R. , Gros François , Ilan Joseph , Lazar Monique 
Published: February 2005

AbstractThe glycolytic enzyme enolase (EC 4.2.1.11) exists as dimers formed from three structurally related subunits α, β, and γ, encoded by separate genes. The gene encoding the β‐subunit is expressed only in striated muscles. We have previously shown that the β‐enolase gene belongs to a small subset of muscle‐specific genes showing transcriptional activity in cultured myoblasts, prior to withdrawal from the cell cycle. An increase in the level of β‐enolase mRNA occurs during terminal differentiation of myoblasts. To investigate the mechanisms underlying this increase, we have simultaneously estimated, under steady state conditions, the rate of synthesis and the stability of β‐enolase mRNA in proliferating C2.7 myoblasts as well as in differentiating myotubes. The method used is based on the isolation of newly synthesized RNA from the total RNA pool, following pulse‐labeling of intact cells in the presence of 4‐thiouridine. The results described here demonstrate a coordinate increase in newly synthesized and total β‐enolase mRNA, while the mRNA half‐life, about 4 hr, remains unchanged in the course of terminal differentiation. The expression of the gene for insulin‐like growth factor‐II (IGF‐II), a major positive regulator of myogenesis, was analyzed using the same approach.It is concluded that the up‐regulation of β‐enolase as well as IGF‐II gene expression in differentiating muscle cells reflects an increased rate of entry of newly synthesized mRNAs into the general pool of transcripts without changes in their respective half‐lives. © 1995 Wiley‐Liss, Inc.
Rat sperm galactosyl receptor: Purification and identification by polyclonal antibodies raised against multiple antigen peptides
Authors: Mertz James R. , Banda Phillip W. , Kierszenbaum Abraham L. 
Published: February 2005

AbstractWe have previously reported the purification of rat testis galactosyl receptor, an equivalent to the Ca2+‐dependent (C‐type) minor variant of rat hepatic lectin‐2/3 (RHL‐2/3). We now report the purification of galactosyl receptor from rat sperm and its immunolocalization in the intact rat testis and sperm by polyclonal antibodies prepared using multiple antigen peptides (MAP) as immunogens. Two MAP antigens (designated 27‐mer and 28‐mer), corresponding to amino acid sequences of the carbohydrate‐recognition domain (galactose) and adjacent Ca2+‐binding sites of RHL‐2/3, were used for immunization. Anti‐RHL‐2/3, anti‐p27, and anti‐p28 sera crossreacted with rat hepatocyte RHL‐2/3 and its rat testis and sperm equivalent, galactosyl receptor, purified by chromatofocusing followed by galactose‐Hydropore‐EP affinity chromatography. Neither anti‐p27 nor anti‐p28 sera crossreacted with the major hepatocyte variant, RHL‐1. A RHL‐1‐equivalent was not detected in rat testis and sperm. Immunofluorescence studies demonstrated that anti‐p27 and anti‐p28 sera recognize galactosyl receptor sites at the Sertoli cell‐spermatogenic cell interface and on the dorsal surfacae of the sperm head, overlying the acrosome. The characteristic crescent‐shaped immunoreactive pattern in sperm was lost after induction of the acrosome reaction. Further studies should determine whether antisera to MAP antigens 27‐mer and 28‐mer, corresponding to specific protein motifs, can serve as immunological probes for examining cell‐cell interaction events during spermatogenesis and at fertilization. © 1995 Wiley‐Liss, Inc.
Oviductins possess chitinase‐ and mucin‐like domains: A lead in the search for the biological function of these oviduct‐specific ZP‐associating glycoproteins
Authors: Malette Brigitte , Paquette Yves , Merlen Yannick , Bleau Gilles 
Published: February 2005

AbstractOver the last 10 years considerable progress has been made in the immunological and biochemical characterization of oviduct‐specific glycoproteins. It is now well established that a subclass of these secretory products, designated as oviductins, associate with the zona pellucida of the ovulated oocyte and with the early embryo. Recent reports on the cloning of cDNAs of oviductins from various species, including that of golden hamster (Mesocricetus auratus) oviductin by our laboratory, allowed us to compare their deduced amino acid sequences with those of other proteins. Optimal alignment analysis showed that oviductins contain regions of significant similarity with catalytically inactive mammalian members of the bacterial and microfilarial chitinase protein family. Most importantly, a close examination of the hamster and human deduced amino acid sequences revealed that both glycoproteins possess contiguous Ser/Thr rich repeated units, clustered in their carboxy‐terminal portions. These mucin‐type motifs are similar in the hamster and human glycoprotein, although hamster oviductin contains more of these complete units. This striking feature might indicate that these molecules play a similar role to mucin‐type glycoproteins, e.g., in protecting the oocyte and early embryo against attacks from their environment. We propose a model whereby oviductins are targeted to the oocyte via the interaction of their chitinase‐like domains with specific oligosaccharide moieties of the zona pellucida. Once localized to this structure, oviductin molecules would act as a protective shield around the oocyte and early embryo by virtue of their densely glycosylated mucin‐type domains. © 1995 Wiley‐Liss, Inc.
Expression of whey acidic protein (WAP) genes in tissues other than the mammary gland in normal and transgenic mice expressing mWAP/hGH fusion gene
Authors: Wen Jawyuan , Kawamata Yuji , Tojo Hideaki , Tanaka Satoshi , Tachi Chikashi 
Published: February 2005

AbstractWhey acidic protein (WAP) is a major whey protein secreted in rodents' milk. Murine WAP (mWAP) genes have been assumed to be expressed solely in the mammary gland. However, several heterologous genes fused with the mWAP promoter and artificially introduced into animal genomes as transgene were expressed not only in the mammary gland but also in other tissues as well. In the present study, we investigated, by means of the reverse transcription polymerase chain reaction (RT‐PCR), the patterns of expression of endogenous WAP genes in tissues of normal mice and in transgenic mice carrying hGH gene coupled to the mWAP promoter sequence. The results revealed that the genes driven by the mWAP promoter, regardless of whether they are endogenous genes or transgenes, were transcribed in a variety of tissues other than the mammary gland of lactating normal female mice, although the expression levels are generally low. The expression of WAP genes in the cerebrum and the liver is regulated, as in the mammary gland, according to the reproductive stages. However, the tissue distribution of endogenous WAP gene expression in mature virgin transgenic female mice was the same as that in lactating normal female mice. © 1995 Wiley‐Liss, Inc.
Localization of nucleic acids in the nucleoli of oocytes and early embryos of mouse and hamster: An autoradiographic study
Authors: Kopečný V. , Landa V. , Pavlok A. 
Published: February 2005

AbstractMouse preovulatory oocytes, zygotes, parthenogenetically activated pronuclear oocytes, and early embryos, as well as hamster zygotes, were analyzed, by autoradiography, for the distribution of either “maternal” or newly synthesized RNAs. Early mouse embryos were also examined for the distribution of newly replicated DNA. Special attention was attributed to NLBs in oocytes or to NPBs in early embryos. In mouse oocytes, [5‐3H]uridine radioactivity accumulated (after a 2‐hr pulse) in vitro, in addition to other nuclear compartments, in the central compact material of the NLBs. There was no cytoplasmic labeling. In all parthenogenetic pronuclear embryos developed from similarly labeled oocytes, this label was distinctly detectable in the central compact material of the NPBs; less intensive labeling was seen in the nucleoplasm and cytoplasm. On the contrary, the central compact part of the mouse NPB did not show labeling in DNA after a continuous culture with [6‐3H]thymidine. In mouse and hamster pronuclear zygotes, convincing evidence was obtained for a lack of any newly synthesized nucleic acids in the compact material of NPBs using 4‐ to 10‐hr culture with [8‐3H]adenosine. Based on these data, it was shown that the NLBs of oocytes or NPBs of early embryos probably contain RNAs synthesized during the last stages of antral follicle oocyte differentiation. This unique pathway of RNAs in the oocyte—embryo system may explain the specific morphology of both oocyte and early embryo “nucleoli.” © 1995 Wiley‐Liss, Inc.
Stimulation of bovine sperm motility and respiration by the triazine dye cibacron blue F3GA
Authors: Schoff Patrick K. , First Neal L. 
Published: February 2005

AbstractBovine sperm motility and respiration were stimulated by the triazine dye Cibacron Blue F3GA (CB), which may operate as a nucleotide mimic. CB stimulation of respiration was half‐maximal at about 35 μM and respiration reached maximal levels about 1.5 minutes after CB addition. Respiratory stimulation was preceded by a transient increase in cytosolic cAMP. Sperm cAMP titers were elevated from 5 to 10 pmoles/108 cells within 30 seconds of CB addition, but rapidly dropped to a stable level of about 7.5 pmoles/108 cells. CB was a potent inhibitor of sperm membrane adenylyl cyclase and inhibited respiration in permeabilized cells. Taken together, the data indicated that CB stimulation was not manifested via the cytosol. In addition, a nonpermeant blue dextran preparation synthesized with CB also stimulated sperm respiration and motility. CB inhibited sperm membrane phosphodiesterase activity, suggesting that the transient pulse of cAMP resulted from CB interaction with this enzyme in the sperm membrane. © 1995 wiley‐Liss, Inc.
Further indications of the multicomponent nature of the acrosome reaction‐inducing substance of human follicular fluid
Authors: Fehl Petra , Miska Werner , Henkel Ralf 
Published: February 2005

AbstractHuman follicular fluid (hFF), which has been treated with either unspecific proteases or dextran‐coated charcoal (DCC) to remove proteins and/or steroids, cannot successfully induce the acrosome reaction (AR). After the removal of steroids, AR‐inducing activity can be restored to hFF by supplementation with exogenous progesterone, but only in the presence of intact protein. Gel filtration experiments with 3H‐progesterone‐labelled hFF showed elution of the radioactive signal in the high molecular weight range, corresponding to bound progesterone. AR‐inducing activity was seen in exactly the same fraction. Based on these results, the acrosome reaction‐inducing substance (ARIS) appears to be a complex of progesterone and a progesterone‐binding protein, which was shown to be identical with the plasma protein corticosteroid‐binding globulin (CBG) by immunological techniques. AR induction was only observed in the presence of both CBG and progesterone, suggesting a combined effect of the two components. © 1995 wiley‐Liss, Inc.
Transforming growth factor β1 expression in the endometrium of the mare during placentation
Authors: Lennard S. N. , Stewart Francesca , Allen W. R. 
Published: February 2005

AbstractIn situ hybridization, Northern blotting, and immunohistochemical techniques were used to study the expression of transforming growth factor beta 1 (TGFβ1) in the endometrium of the mare during the first 150 days of pregnancy (term = 330–340 days).In situ hybridization using an oligonucleotide (45mer) probe, based on a homologous region within all known mammalian TGFβ1 DNA sequences, demonstrated TGFβ1 mRNA accumulation in the glandular and lumenal epithelial cells of the endometrium from day 33 onwards which corresponds to the time of implantation (day 33–45). Expression in the endometrium remained at a high level to the end of the sampling period (day 150). There was also marked expression of TGFβ1 in the mononuclear cells accumulated around the periphery of the specialized trophoblast cells of the endometrial cup within the endometrium and in the mononuclear cells accumulated in the endometrial stroma of mares carrying failing donkey‐in‐horse pregnancies created by embryo transfer. The sense (control) oligonucleotide probe exhibited no hybridization to any tissue at any stage. Northern blot analysis demonstrated that the oligonucleotide probe and a porcine TGFβ1 cDNA clone hybridized to a single 2.5 kb transcript in horse endometrial and lymphocyte RNA, thus validating the oligonucleotide probe for detection of horse TGFβ1 mRNA. Furthermore, both probes demonstrated an increased signal in the pregnant endometrium from day 33 onwards, thereby confirming the in situ hybridization results. Immunostaining with a specific anti‐bovine TGFβ1 serum also showed increasing TGFβ1 accumulation in endometrial epithelia during pregnancy, and localization of the protein in endometrial stroma and in the trophoblast layer of the placenta after day 60 of pregnancy.These results show that TGFβ1 expression increases in the maternal endometrium of the mare at the time of implantation and that it may play a role in regulating endometrial and/or trophoblast growth and differentiation during placentation in this species. It may also influence fetal development (via placental transfer) at a later stage of gestation. © 1995 wiley‐Liss, Inc.
Mouse androgen‐dependent epididymal glycoprotein crisp‐1 (DE/AEG): Isolation, biochemical characterization, and expression in recombinant form
Authors: Eberspaecher Uwe , Roosterman Dirk , Krätzschmar Jörn , Haendler Bernard , Habenicht Ursula‐Friederike , Becker Andreas , Quensel Christina , Petri Thomas , Schleuning Wolf‐Dieter , Donner Peter 
Published: February 2005

AbstractIn the rat, the secretory glycoprotein DE/AEG is one of the main constitutents of the epididymal fluid. We have recently reported the cloning of the cDNA for the related cysteine‐rich secretory protein‐1 (CRISP‐1) from murine epididymis (Haendler et al., 1993; Endocrinology 133:192–198). The protein has now been isolated from the same organ and its N‐terminal amino acid sequence has been determined. CRISP‐1 exhibited an iso‐electric point of ∼6.8. High levels of CRISP‐1 antigen were detected in the corpus and cauda of the epididymis, vas deferens, seminal vesicle, prostate, and in the salivary gland by immunohistochemistry. A quantitative analysis of the cauda epididymal fluid by sandwich ELISA revealed that CRISP‐1 represented ∼15% of the total protein. For heterologous expression, the CRISP‐1 coding sequence was introduced into the pMPSV/CMV vector before transfection of baby hamster kidney (BHK) cells and selection with puromycin and neomycin. Expression in insect cells was achieved by co‐transfection of Sf9 cells with a transfer vector and baculovirus DNA. Recombinant CRISP‐1 was isolated in quantities sufficient for structural analysis. Ethyl maleimide treatment showed that all 16 cysteines were engaged in disulfide bonds. Proteolytic digestion demonstrated that the six cysteines localized in the N‐terminal moiety formed three bonds with each other, suggesting the existence of two discrete domains in the protein. © 1995 wiley‐Liss, Inc.