bluuebunny/crossref_metadata_2025_split huggingface dataset:
Optimization of three‐ and four‐color multiparameter DNA analysis in lymphoma specimens
Authors: Plander M. , Brockhoff G. , Barlage S. , Schwarz S. , Rothe G. , Knuechel R. 
Published: June 2003

AbstractBackgroundSimultaneous analysis of DNA and immunophenotype of lymphoma cells by flow cytometry allows the calculation of the proliferative activity and aneuploidy in even a small lymphoma population. Unfavorable DNA binding characteristics or spectral features of DNA dyes impair the accuracy of multiparameter DNA analysis and limit their clinical application. We describe here a reliable and reproducible application of both three‐ and four‐color multiparameter DNA analysis.MethodsAfter immunostaining of fresh samples of peripheral blood, bone marrow and single cell suspensions of lymph nodes from healthy and lymphoma patients, a methanol fixation for TO‐PRO‐3 and DRAQ5 staining was tested.ResultsThe red‐excitable TO‐PRO‐3 on a FACSCalibur is limited to two‐color antigen staining including fluorescein‐isothiocyanate and phycoerythrin‐labeled monoclonal antibodies due to its broad excitation spectrum. Although DRAQ5 is only applicable to flow cytometers equipped with a single argon laser emitting 488‐nm light, its emission spectrum can be easily separated from the FITC, PE, and PE/Texas‐Red emissions. DRAQ5 showed almost identical stoichiometric DNA binding characteristics as propidium iodide. Coefficient of variation produced by DRAQ5 staining is in the range of 3.5 and is adequate for detecting aneuploid amd near‐diploid cells.ConclusionsThese advantageous features of DRAQ5 make it a reliable candidate for multiparameter clinical studies. Cytometry Part A 54A:66–74, 2003. © 2003 Wiley‐Liss, Inc.
Developmental changes in neurite outgrowth responses of dorsal root and sympathetic ganglia to GDNF, neurturin, and artemin
Authors: Yan H. , Newgreen D.F. , Young H.M. 
Published: June 2003

AbstractThe ability of glial cell line–derived neurotrophic factor (GDNF), neurturin, and artemin to induce neurite outgrowth from dorsal root, superior cervical, and lumbar sympathetic ganglia from mice at a variety of development stages between embryonic day (E) 11.5 and postnatal day (P) 7 was examined by explanting ganglia onto collagen gels and growing them in the presence of agarose beads impregnated with the different GDNF family ligands. Artemin, GDNF, and neurturin were all capable of influencing neurite outgrowth from dorsal root and sympathetic ganglia, but the responses of each neuron type to the different ligands varied during development. Neurites from dorsal root ganglia responded to artemin at P0 and P7, to GDNF at E15.5 and P0, and to neurturin at E15.5, P0, and P6/7; thus, artemin, GDNF, and neurturin are all capable of influencing neurite outgrowth from dorsal root ganglion neurons. Neurites from superior cervical sympathetic ganglia responded significantly to artemin at E15.5, to GDNF at E15.5 and P0, and to neurturin at E15.5. Neurites from lumbar sympathetic ganglia responded to artemin at all stages from E11.5 to P7, to GDNF at P0 and P7 and to neurturin at E11.5 to P6/7. Combined with the data from previous studies that have examined the expression of GDNF family members, our data suggest that artemin plays a role in inducing neurite outgrowth from young sympathetic neurons in the early stages of sympathetic axon pathfinding, whereas GDNF and neurturin are likely to be important at later stages of sympathetic neuron development in inducing axons to enter particular target tissues once they are in the vicinity or to induce branching within target tissues. Superior cervical and lumbar sympathetic ganglia showed temporal differences in their responsiveness to artemin, GDNF, and neurturin, which probably partly reflects the rostrocaudal development of sympathetic ganglia and the tissues they innervate. Developmental Dynamics 227:395–401, 2003. © 2003 Wiley‐Liss, Inc.
Isolation of a novel cDNA enriched in the undifferentiated chick retina and lens
Authors: Godbout Roseline , Andison Randy , Katyal Sachin , Bisgrove Dwayne A. 
Published: June 2003

AbstractBy using a differential screening strategy, we have identified a previously uncharacterised gene that is preferentially expressed in chick retinal precursor cells as well as in the anterior epithelial cells of the lens at early stages of development. The EURL transcript has an open reading frame of 293 amino acids and a 3′ untranslated region of >850 nucleotides. The only recognizable feature in the EURL (early undifferentiated retina and lens) amino acid sequence is a putative coiled‐coil domain located at the C‐terminus of the protein. The human EURL orthologue maps to chromosome 21q21. Northern blot analysis, in situ hybridization of tissue sections, and whole‐mount hybridization indicate elevated levels of EURL mRNA in the embryonic dorsal retina from stage 14 (day 2) to stage 18 (day 3). At day 3.5, when >90% of the retinal cells are in the proliferative stage, EURL transcripts are found primarily in the peripheral dorsal retina, i.e., the most undifferentiated part of the dorsal retina. EURL transcripts are also detected in the lens at stage 18 and remain abundant in the proliferating epithelial cells of the lens until at least day 11. The distribution pattern of EURL in the developing retina and lens suggest a role before the events leading to cell determination and differentiation. Developmental Dynamics 227:409–415, 2003. © 2003 Wiley‐Liss, Inc.
Expression of Sox transcription factors in the developing mouse pancreas
Authors: Lioubinski Oleg , Müller Myriam , Wegner Michael , Sander Maike 
Published: June 2003

AbstractPrevious work has identified members of the homeodomain and basic helix‐loop‐helix families of transcription factors as critical determinants of mammalian pancreatic development. Here, we describe the identification of HMG‐box transcription factors of the Sox gene family in the mouse pancreas. We detected transcripts for Sox11, Sox4, Sox13, Sox5, Sox9, Sox8, Sox10, Sox7, Sox17, Sox18, Sox15, and Sox30 in embryonic pancreas and found Sox4, Sox9, and Sox13 in adult pancreatic islets. Expression of seven of these Sox factors was studied in more detail by in situ hybridization from the stage of early pancreatic outgrowth to birth. Expression of Sox11 was found in the mesenchyme surrounding the pancreatic buds, whereas Sox4 and Sox9 were confined to the pancreatic epithelium and later to islets. Sox13 and L‐Sox5 showed expression in most of the pancreatic epithelial cells between embryonic days 12.5 and 14.5. Sox8 and Sox10 were detected in a thin layer of cells surrounding the islets. The expression patterns of Sox genes in the embryonic pancreas suggest that they could have important and possibly redundant functions in pancreas development. Developmental Dynamics 227:402–408, 2003. © 2003 Wiley‐Liss, Inc.
AMPA‐evoked ion influx is strongest in tangential neurons of the rat neocortical intermediate zone close to the front of the migratory stream
Authors: Poluch Sylvie , Rossel Mireille , König Norbert 
Published: June 2003

AbstractIn addition to the classically described radially migrating neurons, embryonic cortical areas receive neurons originating from the basal ganglia. One of the migration routes is in the intermediate zone. The front of this migration moves toward the hippocampus synchronously with the edge of the dorsally extending cortical plate. We investigated whether cells close to the front have specific properties compared with those at less advanced positions. Activation of AMPA receptors in the presence of cobalt showed that a strong influx of divalent cations could be triggered in front cells by low agonist concentration, whereas the less advanced cells needed a higher concentration to incorporate detectable amounts of cobalt. As shown by in situ hybridization, this discrepancy was not due to differential expression of GluR‐2 (known to reduce permeability for divalent cations). In vivo, release of an endogenous agonist presumably affects more, or differently, the tangential cells close to the front. Developmental Dyanmics 227:416–421, 2003. © 2003 Wiley‐Liss, Inc.
Cathepsin L plays an active role in involution of the mouse mammary gland
Authors: Burke Michael A. , Hutter Dorothy , Reshamwala Rita P. , Knepper Janice E. 
Published: June 2003

AbstractInvolution of the mammary gland after weaning occurs in two stages. The first stage is reversible, whereas the second stage is characterized by the irreversible collapse of the alveolar structure. A differential display analysis using cDNAs from tissues obtained at various times after forced weaning of pups identified cathepsin L as up‐regulated during early involution. Levels of cathepsin L mRNA were dramatically increased within 24 hr after weaning. Cathepsin L protein detected by immunoblot was also increased during involution, reaching near maximal levels by 36 hr after weaning. In situ immunohistochemistry detected pronounced cathepsin L protein in the cytoplasm and cell periphery. Mice treated with a specific inhibitor of cathepsin L exhibited substantially reduced numbers of apoptotic cells at times up to 72 hr after weaning when compared with untreated animals. The cathepsin L inhibitor did not alter levels of cathepsin L detected in immunoblots or influence molecular weight of the cathepsin L species detected. These data suggest that cathepsin L plays a regulatory role early in the process of mammary gland involution. Developmental Dynamics 227:315–322, 2003. © 2003 Wiley‐Liss, Inc.
Expression patterns of Tgfβ1–3 associate with myocardialisation of the outflow tract and the development of the epicardium and the fibrous heart skeleton
Authors: Molin Daniël G.M. , Bartram Ulrike , Van der Heiden Kim , Van Iperen Liesbeth , Speer Christian P. , Hierck Beerend P. , Poelmann Robert E. , Gittenberger‐de‐Groot Adriana C. 
Published: June 2003

AbstractTransforming growth factor‐beta (Tgfβ) is essential for normal embryogenesis. The cardiac phenotypes obtained after knockout of each of the three mammalian isoforms suggest different roles during morphogenesis. We studied cardiovascular expression of Tgfβ1–3 in parallel tissue sections of normal mouse embryos from 9.5 to 15.5 days post coitum (dpc) by using radioactive in situ hybridisation. The Tgfβ isoforms are differentially expressed in unique and in overlapping patterns during cardiovascular development. In the vessels, Tgfβ1 is found in the intima, whereas Tgfβ2 and ‐β3 are mainly present in the media and adventitia of the great arteries. Tgfβ1 is present in the endocardium at all stages examined. The Tgfβ2 signal in the endocardium of the atrioventricular canal and outflow tract (9.5 dpc) shifts during epithelial–mesenchymal transformation toward the mesenchymal cushions (10.5–11.5 dpc) after which it exhibits a marked spatiotemporal expression pattern as the cushion differentiation progresses (11.5–15.5 dpc). The myocardium underlying the endocardial cushions and the atrial muscular septum are intensely positive for Tgfβ2 at early stages (9.5–11.5 dpc) and expression decreases at 12.5 days. In contrast to earlier reports, we find marked overlap of Tgfβ2 and ‐β3 expression. Tgfβ3 expression shows a characteristic distribution in the mesenchymal cushions, suggesting a role in cushion differentiation, possibly additional to Tgfβ2. From 14.5 dpc onward, a strong Tgfβ3 signal is found in the fibrous septum primum of the atrium and in the fibrous skeleton of the heart. Special attention was paid to the proepicardial organ and its derivatives. The proepicardial organ strongly expresses Tgfβ2 as early as 9.5 days, and all isoforms are present in the epicardium from 12.5 dpc onward. The spatiotemporal cardiovascular expression of Tgfβ1–3 supports both specific and complementary functions during cardiovascular development that might explain functional redundancy between the Tgfβ‐isoforms. The information provided favors novel roles of Tgfβ1–3 in epicardial development, of Tgfβ2 in myocardialisation, and of Tgfβ3 in differentiation of the fibrous structures of the heart. Developmental Dynamics 227:431–444, 2003. © 2003 Wiley‐Liss, Inc.
Characterization of Wnt signaling components and activation of the Wnt canonical pathway in the murine retina
Authors: Liu Hong , Mohamed Othman , Dufort Daniel , Wallace Valerie A. 
Published: June 2003

AbstractThe neuroepithelial layer of the developing eyecup contains multipotential precursor cells that give rise to all of the neurons and the one glial cell type present in the adult retina. Patterning within the retinal neuroepithelium is regulated by cell intrinsic as well as cell extrinsic mechanisms. Although the identity of some of the signaling molecules that regulate retinal development is known, the function of many others, especially members of the Wnt family, has yet to be characterized in the context of retinal development. We undertook a comprehensive in situ hybridization analysis to examine the expression of Wnt pathway components in the developing and adult mouse neural retina. Our findings confirm and extend previous expression studies in mice and other vertebrates, as we show that Wnt‐3, ‐5a, ‐5b, and ‐7b are expressed in the neural retina and that there is a dynamic pattern of Wnt receptor (Mouse frizzled [Mfz]) and Wnt antagonist (Secreted‐frizzled‐related protein [Sfrp]) gene expression in the embryonic and perinatal neural retina. Moreover, we show that Wnt‐13 is expressed in the pigment epithelium overlying the distal part of the eyecup and the ciliary margin and that Mfz‐4, ‐6, and ‐7 are expressed in different regions within the ciliary margin. To determine where activation of canonical Wnt signaling is occurring in the retina, we examined reporter gene expression in TCF/Lef‐LacZ mice and we demonstrate that the highest levels of β‐gal activity are found in the ciliary margin, adjacent to and within the Wnt‐13 expression domain, implicating Wnt‐13 signaling in the development of the ciliary margin and its derivatives. Developmental Dynamics 227:323–334, 2003. © 2003 Wiley‐Liss, Inc.
Three‐dimensional morphology of inner ear development in Xenopus laevis
Authors: Miller Bever Michele , Jean Ying Y. , Fekete Donna M. 
Published: June 2003

AbstractThe three‐dimensional morphology of the membranous labyrinth of Xenopus laevis is presented from embryonic through late tadpole development (stages 28 to 52, inclusive). This was accomplished by paint‐filling the endolymphatic spaces of Xenopus ears at a series of stages, beginning with the embryonic otic vesicle and ending with the complex ear of the late tadpole. At stage 52, the inner ear has expanded approximately 23‐fold in its anterior/posterior dimension compared with stage 28 and it is a miniature of the adult form. The paint‐filling technique illustrates the dramatic changes required to convert a simple ear vesicle into the elaborate form of the adult, including semicircular canal formation and genesis of vestibular and auditory organs, and it can serve as a basis for phenotype identification in experimentally or genetically manipulated ears. Developmental Dynamics 227:422–430, 2003. © 2003 Wiley‐Liss, Inc.
Expression of MRF4 protein in adult and in regenerating muscles in Xenopus
Authors: Becker Christel , Della Gaspera Bruno , Guyot Michèle , Donsez Evelyne , Armand Anne‐Sophie , Charbonnier Frédéric , Launay Thierry , Chanoine Christophe 
Published: June 2003

AbstractIn Xenopus, previous studies showed that the transcripts of the myogenic regulatory factor (MRF) MRF4 accumulate during skeletal muscle differentiation, but nothing is known about the accumulation of XMRF4 protein during myogenesis. In this report, an affinity‐purified polyclonal antibody against Xenopus MRF4 was developed and used to describe the pattern of expression of this myogenic factor in the adult and in regenerating muscles. From young forming myotubes, XMRF4 protein persistently accumulated in nuclei during the regeneration process and was strongly expressed in nuclei of adult muscles. No selective accumulation of XMRF4 protein was detectable at neuromuscular junctions, but XMRF4 immunoreactivity was observed in sole plate nuclei as well as in extrasynaptic myofiber nuclei. We also report that XMRF4 protein accumulated before the establishment of neuromuscular connections, showing that innervation is not necessary for the appearance of XMRF4 protein during muscle regeneration. Developmental Dynamics 227:445–449, 2003. © 2003 Wiley‐Liss, Inc.
Erk pathway and activator protein 1 play crucial roles in FGF2‐stimulated premature cranial suture closure
Authors: Kim Hyun‐Jung , Lee Mi‐Hye , Park Hyun‐Sik , Park Mi‐Hyun , Lee Sang‐Won , Kim Shin‐Yoon , Choi Je‐Yong , Shin Hong‐In , Kim Hyun‐Jung , Ryoo Hyun‐Mo 
Published: June 2003

AbstractCranial sutures are an important growth center of the cranial bones, and the suture space must be maintained to permit the cranial adjustments needed to accommodate brain growth. Craniosynostosis, characterized by premature suture closure, mainly results from mutations that generate constitutively active fibroblast growth factor (FGF) receptors. FGF signaling, thus, is responsible for the pathogenesis of craniosynostosis. Even though FGF activates many different signaling pathways, the one involved in premature suture closure has not been defined. We observed that placing FGF2‐soaked bead on the osteogenic fronts of cultured mouse calvaria accelerates cranial suture closure and strongly induces the expression of osteopontin, an early marker of differentiated osteoblasts. FGF2 treatment also induced fos and jun mRNAs and later increased the nuclear levels of activator protein 1 (AP1). FGF2 stimulates the expression of osteopontin by inducing expression of AP1, which then binds to its response element in the osteopontin promoter. Blocking of the Erk pathway by PD98059 suppressed the AP1 and osteopontin expression stimulated by FGF2. Coincidently, blocking of the Erk pathway also significantly retarded FGF2‐accelerated cranial suture closure. Thus, the Erk pathway mediates FGF/FGF receptor–stimulated cranial suture closure, probably by stimulating synthesis of AP1 that then stimulates the differentiation of osteoblasts. Developmental Dynamics 227:335–346, 2003. © 2003 Wiley‐Liss, Inc.
Ontogenic expression of TGFβ 1, 2, and 3 and its receptors in the rat gastric mucosa
Authors: de Andrade Sá Eunice Ribeiro , Jordão Luciana Renata , Takahashi Cassia Ayumi , Alvares Eliana Parisi , Gama Patrícia 
Published: June 2003

AbstractThe stomach of the rat undergoes extensive changes during the formation and maturation of gastric glands. The presence of transforming growth factor beta (TGFβ) in rat milk and in the gastrointestinal tract of pups may suggest its role in this process. The current study evaluated the in vivo dynamic expression and distribution of TGFβ1, β2, β3 and their receptors TβRI and TβRII in the gastric epithelium of 20‐day fetal rats and 1‐, 14‐, 21‐, and 30‐day‐old pups. Immunohistochemistry was used to detect the proteins, and staining was classified according to intensity and cell type. The results showed that the gastric epithelium expresses TGFβ isoforms and receptors throughout development. We found that immunoreactivity paralleled the appearance of differentiated cells, such that surface mucous cells were the first to be immunostained and chief cells were the last. The intensity of reactions followed this same pattern, showing that the expression of TGFβ isoforms spread along the gland with growth. Of interest, the highest apparent activity of TGFβ was observed from 21 days onward, a period that is concomitant with weaning and maturation of most gastric cell types. In addition, surface mucous cells were strongly labeled at the basal cytoplasm at 14 days, suggesting an interaction with the connective tissue. In conclusion, the dynamic expression of TGFβ1, β2, β3, and TβRI and TβRII through stomach development suggests significant paracrine and autocrine roles for this growth factor. We propose that temporal and spatial differences may be regulated by dietary changes, which in turn control cell proliferation and differentiation in the gastric epithelium. Developmental Dynamics 227:450–457, 2003. © 2003 Wiley‐Liss, Inc.
Differential expression of tissue inhibitor of metalloproteinases type 1 (TIMP‐1) during mouse gonad development
Authors: Guyot Romain , Magre Solange , Leduque Patrick , Le Magueresse‐Battistoni Brigitte 
Published: June 2003

AbstractIn mammals, the gene Sry initiates signaling pathways triggering the differentiation of a testis from a sexually indifferent gonad. Assuming that these morphogenetic events may alter the proteolytic balance, the expression of matrix metalloproteinases (MMPs) and inhibitors (TIMPs) was investigated in gonads from 11.5 days postcoitum (dpc) onward, when testicular organogenesis occurs. Whereas selective MMPs and TIMPs (1–3) were detected in undifferentiated gonads (11.5 dpc) and in neonatal testes, a single TIMP (TIMP‐1) was expressed in a sexually dimorphic manner from 12.5 dpc onward (i.e., after overt male gonad differentiation), demonstrated by using a semiquantitative reverse transcriptase‐polymerase chain reaction and a Western blot analysis. To gain insight into the role of TIMP‐1, the expression of gelatinases (mRNA levels and enzyme activity) was monitored. However, no sex differences could be evidenced, indicating that TIMP‐1 was not inhibiting this class of MMPs during testis organogenesis. Apart from being an inhibitor of MMPs, TIMP‐1 is known to display growth promoting activities. Of interest, testicular TIMP‐1 (but not TIMP‐2) levels were further enhanced up to 2 weeks of age, consistent with a role in the early postnatal testicular growth. We, therefore, established an organotypic culture system in which seminiferous cords may differentiate de novo and grow, depending on culture conditions. In that system and mimicking the in vivo situation, TIMP‐1 immunolocalized strongly within the male gonadal territory and weakly in female gonads, in which no organization was evident. Experiments are now under way to determine to what extent timp‐1 is a morphogenic gene involved in seminiferous cord formation and development. Developmental Dynamics 227:357–366, 2003. © 2003 Wiley‐Liss, Inc.
Hoxb1 neural crest preferentially form glia of the PNS
Authors: Arenkiel Benjamin R. , Gaufo Gary O. , Capecchi Mario R. 
Published: June 2003

AbstractThe vertebrate cranial neural crest cells give rise to many complex derivatives of the head, neck, and face, including neuronal and glial cells that act in concert for proper development of the anterior–peripheral nervous system. Several genes have been implicated in the processes of neural crest specification, migration, and differentiation; among these are the hox gene clusters. To determine the fates of hox‐expressing cranial neural crest, we describe the results of a genetic lineage analysis by using the Cre/loxP system to drive the activation of different ROSA26 reporter alleles under the regulation of the hoxb1 locus. By targeting the 3′ untranslated region of the hoxb1 gene, we have preserved endogenous gene activity and have been able to accurately follow the fates of the cells derived from the hoxb1 expression domain. Emphasis was placed on identifying the cell and tissue types that arise from the rhombomere 4‐derived neural crest. Our results demonstrate that, in addition to forming much of the cartilage, bones, and muscle of the ears and neck, a significant population of rhombomere 4‐derived neural crest is fated to generate the glial component of the seventh cranial nerve. Developmental Dynamics 227:379–386, 2003. © 2003 Wiley‐Liss, Inc.