Differentiation markers in the Drosophila ovary

Development ◽  
1984 ◽  
Vol 84 (1) ◽  
pp. 275-286
Author(s):  
Robert A. H. White ◽  
Norbert Perrimon ◽  
Walter J. Gehring
Keyword(s):  
Monoclonal Antibody ◽  
Wheat Germ ◽  
Germ Line ◽  
Anterior Part ◽  
Differentiation Markers ◽  
Ovarian Cell ◽  
Adult Ovary ◽  
Preferential Binding ◽  
Tunica Propria ◽  
Drosophila Ovary

A library of monoclonal antibodies, raised against imaginal discs of Drosophila melanogaster, was screened for binding to differentiation antigens in the adult ovary by immunofluorescence. Several lectins were similarly assayed. Two antibodies, DOV 1 and DOV 2, and wheat germ agglutinin exhibited binding which was restricted to particular stages of ovarian cell differentiation. DOV 2 also showed a marked preferential binding to the cell surface of germ line cells in the ovary. A differentiation of the portion of the tunica propria covering the anterior part of the germarium was revealed by the monoclonal antibody DOV 3. Another monoclonal antibody, DOV 4, identified a molecular specialization of the chorion at the tip of the micropyle. These markers should provide tools for the molecular analysis of oogenesis.

Developmental expression of c-kit, a proto-oncogene encoded by the W locus

Development ◽  
1990 ◽  
Vol 109 (4) ◽  
pp. 911-923 ◽  
Author(s):  
A. Orr-Urtreger ◽  
A. Avivi ◽  
Y. Zimmer ◽  
D. Givol ◽  
Y. Yarden ◽  
...  
Keyword(s):  
Germ Cells ◽  
Receptor Tyrosine Kinase ◽  
Germ Line ◽  
Primordial Germ Cells ◽  
Mouse Embryos ◽  
Developmental Expression ◽  
Male Germ Line ◽  
Polypeptide Growth Factors ◽  
Adult Ovary

Developmental expression of the c-kit proto-oncogene, a receptor tyrosine kinase encoded by the W locus, was investigated by in situ hybridization in normal mouse embryos. Early after implantation transcripts were detectable only in the maternal placenta (6 1/2-7 1/2 days p.c.). Subsequently (8 1/2 days p.c.) numerous ectodermal (neural tube, sensory placodes) and endodermal (embryonic gut) derivatives expressed c-kit. Later transcripts were detected also in the blood islands of the yolk sac and in the embryonic liver, the main sites of embryonic hemopoiesis. Around midgestation, transcripts accumulated in the branchial pouches and also in primordial germ cells of the genital ridges. This complex pattern of expression remained characteristic also later in gestation, when c-kit was expressed in highly differentiated structures of the craniofacial area, in presumptive melanoblasts and in the CNS. In the adult ovary, maternal c-kit transcripts were detected. They were present in the oocytes of both immature and mature ovarian follicles, but not in the male germ line, where c-kit expression may be down regulated. Thus, c-kit activity is complex and appears in multiple tissues including those that also display defects in mutations at the W locus where c-kit is encoded. Correlation between W phenotypes and c-kit expression, as well as the regulation of the complex and multiple expression of polypeptide growth factors and receptors, is discussed.

The positional effect of presumptive primordial germ cells (pPGCs) on their differentiation into PGCs in Xenopus

Development ◽  
1988 ◽  
Vol 102 (3) ◽  
pp. 527-535
Author(s):  
K. Ikenishi ◽  
Y. Tsuzaki
Keyword(s):  
Germ Cells ◽  
Germ Plasm ◽  
Germ Line ◽  
Anterior Part ◽  
Primordial Germ Cells ◽  
Positional Effect ◽  
In Series

To determine whether the location of ‘germ plasm’-bearing cells [presumptive primordial germ cells (pPGCs)] is crucial for their differentiation into PGCs in Xenopus, [3H]thymidine-labelled pPGCs were implanted into the anterior or posterior halves of the endoderm in unlabelled host neurulae. Labelled PGCs in the genital ridges of experimental tadpoles were investigated by autoradiography. When the labelled pPGCs were implanted into posterior halves of the endoderm where host pPGCs are situated, 65 and 77% of the experimental tadpoles (designated as p-tadpoles) had the labelled PGCs in series I and II, respectively. When implanted into the anterior halves, 20 and 27% of the experimental tadpoles (a- tadpoles) had the labelled PGCs in series I and II, respectively. In p-tadpoles, the average numbers of labelled PGCs per tadpole were 8á7 in series I and 10 in series II, whereas they were 2á0 in a-tadpoles of both series. Both the proportion and the average number in p-tadpoles of both series were significantly different from those in a-tadpoles. In both series, labelled PGCs in p-tadpoles were found to be distributed throughout the genital ridges while those in a-tadpoles were localized only in the anterior part of the ridges. These facts indicate that the location of pPGCs in the endoderm affects their successful migration into the genital ridges, and that not only the presence of the germ plasm but also the proper location in endoderm are prerequisites to PGC differentiation of the germ line cells.

scholarly journals Structure of the germarium and female germ-cell clusters in Thulinius ruffoi (Bertolani, 1982) (Tardigrada: Eutardigrada: Parachela)

2019 ◽  
Author(s):  
Kamil Janelt ◽  
Marta Jezierska ◽  
Sebastian Student ◽  
Izabela Poprawa
Keyword(s):  
Germ Cell ◽  
Laser Scanning ◽  
Spatial Organization ◽  
Germ Line ◽  
Anterior Part ◽  
Confocal Laser ◽  
Freshwater Species ◽  
Cell Clusters ◽  
Scanning Transmission ◽  
Female Germ Cell

Abstract Thulinius ruffoi is a freshwater species that has the ability to reproduce via parthenogenesis. A meroistic polytrophic ovary is present in this species. Analyses of the germarium structure, and formation and organization of female germ-cell clusters were performed using light, confocal laser scanning, transmission electron and serial block-face scanning electron microscopy. The germarium is the small, anterior part of an ovary that contains putative germ-line stem cells. In the studied species, the female germ-cell clusters are large and branched. Only one cell in each cluster develops into an oocyte, while all the other cells become trophocytes. In this paper, we present the first report on the presence of F-actin as a component of the intercellular bridges that connect the cells in the germ-cell cluster of T. ruffoi. Moreover, our results show that the female germ-cell clusters are formed as the result of both synchronous and asynchronous divisions and that their organization can vary not only between individuals of the investigated species, but also that clusters developing simultaneously within the same ovary can have a different spatial organization.

Null alleles reveal novel requirements for Bic-D during Drosophila oogenesis and zygotic development

Development ◽  
1994 ◽  
Vol 120 (5) ◽  
pp. 1233-1242 ◽  
Author(s):  
B. Ran ◽  
R. Bopp ◽  
B. Suter
Keyword(s):  
Germ Line ◽  
Null Alleles ◽  
Cell Fates ◽  
New Class ◽  
Specific Accumulation ◽  
Normal Polarity ◽  
Female Germ Line ◽  
Low Levels ◽  
Drosophila Ovary ◽  
Sister Cells

In the Drosophila ovary, the Bicaudal-D (Bic-D) gene is required for the differentiation of one of 16 interconnected cystocyte sister cells into an oocyte. A new class of Bic-Dnull alleles reveals a novel requirement for Bic-D for zygotic viability. In the germ line, the null mutations show that developmental processes that take place in germarial region 1, even those that create asymmetry, are independent of Bic-D function. Bic-D is then required to establish oocyte identity in one cystocyte and is essential, not only for the oocyte-specific accumulation of all oocyte markers that we have tested so far, but also for the posterior migration of the oocyte. In addition, normal polarity amongst the nurse cells requires Bic-D, indicating that the creation of different nurse cell identities may depend on oocyte determination. Our results show that different processes in early oogenesis require different amounts of Bic-D in a process-specific way and certain later processes can proceed at low levels of Bic-D. This suggests that the patterning of the female germ line and the development of an oocyte depend on differential responses to a single activity that is capable of initiating distinct oogenesis processes and can establish different cell fates.

Abstract 630: Novel Chimeric Anti-proteoglycan Antibody Inhibits Arterial Retention of Proatherogenic Lipoproteins in a Rat Model of Insulin Resistance

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Yosdel Soto ◽  
Rabban Mangat ◽  
Ana M Vázquez ◽  
Spencer D Proctor
Keyword(s):  
Insulin Resistance ◽  
Monoclonal Antibody ◽  
Ex Vivo ◽  
Dependent Manner ◽  
Insulin Resistant ◽  
Remnant Lipoproteins ◽  
Preferential Binding ◽  
Carotid Vessels

Background: The response-to-retention hypothesis for atherosclerosis describes subendothelial retention of apolipoprotein B-containing lipoproteins mediated by proteoglycans (PG). Further we know that diabetes is also associated with both increased circulating chylomicron remnants and remodeling of proatherogenic PGs. We have recently reported antiatherogenic properties of a novel chimeric monoclonal antibody (chP3R99) that recognizes PG sulfated molecules. Hypothesis: chP3R99 monoclonal antibody may interfere with the interaction of atherogenic lipoproteins with arterial sulfated PGs during insulin resistance. Methods and Results: chP3R99 antibody recognized sulfated glycosaminoglycans by ELISA showing a preferential binding to chondroitin sulfate. Also, chP3R99 blocked the interaction of proatherogenic lipoproteins with this glycosaminoglycan in vitro in a dose-dependent manner and its intravenous injection into healthy Sprague-Dawly rats (n=6, 1 mg/animal) inhibited LDL (4 mg/kg; intraperitoneally) aortic retention. To further assess this property in an insulin resistant condition, carotid arteries from control and JCR:LA-cp rats (n=4) were perfused ex vivo with apoB48 containing remnant lipoproteins (prepared via rabbit hepatectomy procedure), with or without Cy3-LDL (150 μg/mL) for 20 minutes. Confocal microscopy analysis revealed an increased arterial retention of both remnants (3.6 fold) and LDL (2.8 fold) in carotid vessels from insulin resistant rats relative to control. However, chP3R99 pre-perfusion resulted in decreased retention of remnants (-30%) and LDL (-60%) associated arterial cholesterol. Data suggests that the chP399 antibody may interfere with the arterial attachment of both remnants and LDL in vivo, but with differential efficacy. Conclusions: Relative to LDL, remnant lipoproteins had preferential accumulation in arterial vessels from insulin resistant rats ex vivo , which could then be inhibited by acute pre-exposure to the chP3R99 antibody. These in vivo data support the concept for an innovative approach to target the retention of proatherogenic lipoproteins in a pre-clinical setting.

XY follicle cells in ovaries of XX—XY female mouse chimaeras

Development ◽  
1988 ◽  
Vol 104 (4) ◽  
pp. 683-688 ◽  
Author(s):  
P.S. Burgoyne ◽  
M. Buehr ◽  
A. McLaren
Keyword(s):  
Female Mouse ◽  
Germ Line ◽  
Follicle Cells ◽  
Cell Type ◽  
Ovarian Cell ◽  
Female Mice ◽  
Xy Female

Oocytes with adhering follicle cells were sampled from ovaries obtained from 11 GPI-1A—GPI-1B chimaeras, comprising 10 females and 1 hermaphrodite. GPI analysis of individual oocytes revealed a marked bias towards the GPI-1B component in the germ line of this chimaeric combination. GPI-1B XY oocytes were identified in the ovary from the hermaphrodite, the bias towards the GPI-1B germ line perhaps helping to counterbalance the normally severe selection against XY oocytes. GPI analysis of follicle cells revealed a much more balanced contribution of the two components to this ovarian cell type. Importantly, GPI-1A follicle cells were identified in more than half the follicles from an XX—XY female in which the GPI-1A component was XY, supporting an earlier conclusion of Ford et al. (1974) that XY cells can contribute to the follicles of XX—XY female mice. It is suggested that XY cells can be recruited to form follicle cells in XX—XY chimaeras when there is a developmental mismatch between the two components, such that an ovary-determining signal produced by the XX component pre-empts the testis-determining action of the Y.

Alkaline phosphatase isoenzymes of liver and bone origin are incompletely resolved by wheat-germ-lectin affinity chromatography.

1989 ◽  
Vol 35 (1) ◽  
pp. 29-32 ◽  
Author(s):  
D G Gonchoroff ◽  
E L Branum ◽  
J F O'Brien
Keyword(s):  
Monoclonal Antibody ◽  
Alkaline Phosphatase ◽  
Affinity Chromatography ◽  
Wheat Germ ◽  
Solid Phase ◽  
Lectin Affinity Chromatography ◽  
Serum Samples ◽  
Lectin Affinity ◽  
Alp Activity ◽  
Wheat Germ Lectin

Abstract We used wheat-germ-lectin affinity chromatography as a tool to investigate the structure of alkaline phosphatase (ALP, EC 3.1.3.1) and to obtain fractions enriched in either bone or liver ALP activity. Liver and bone isoenzymes in serum samples were incompletely resolved except that the activity in the nonretained fraction (fraction 1) always represented pure liver isoenzyme and constituted a larger percentage of total activity in pooled sera with increased liver ALP activity than in pooled sera with increased bone activity. In contrast, a more avidly retained ALP activity, presumably with high glycosylation, was found in human serum with high activity of bone ALP. Using a solid-phase immunoassay, we examined the fractions obtained from the wheat-germ-lectin-Sepharose 4B column to determine whether the isoenzyme preference of the monoclonal antibody was markedly influenced by the degree of glycosylation. Whether samples contained high proportions of liver or of bone isoenzyme activity, the nonretained fraction contained a higher percentage of liver ALP, whereas the more strongly bound fraction contained a higher percentage of bone ALP. Except for eluted fractions that either contained no detectable N-acetylglucosamine or the highest percentage of it, the avidity of the liver-isoenzyme-specific monoclonal antibody for ALP seemed to be independent of the degree of glycosylation, suggesting that the epitope for monoclonal antibody may be expressed in some structure other than the carbohydrate moieties.

scholarly journals Generation of a Neutralizing Human Monoclonal Antibody Fab Fragment to Surface Antigen 1 ofToxoplasma gondiiTachyzoites

2010 ◽  
Vol 79 (1) ◽  
pp. 512-517 ◽  
Author(s):  
Yong-Feng Fu ◽  
Meng Feng ◽  
Kenji Ohnishi ◽  
Tamon Kimura ◽  
Johbu Itoh ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Surface Antigen ◽  
Heavy Chain ◽  
Germ Line ◽  
Human Monoclonal Antibody ◽  
Passive Immunization ◽  
Positive Staining ◽  
Immunoglobulin Gene ◽  
Fab Fragment ◽  
Fab Fragments

ABSTRACTA combinatorial immunoglobulin gene library was constructed from lymphocytes in peripheral blood of a patient with toxoplasmosis and screened for production of human monoclonal antibody Fab fragments to recombinant surface antigen 1 (SAG1) ofToxoplasma gondii. Two Fab clones, Tox203 and Tox1403, which consisted of a common heavy chain and different light chains, showed positive staining on the entire surface of tachyzoites in confocal microscopy. Sequence analysis of the heavy-chain gene revealed that the closest germ line V segments were VH3-23. The germ line D segment was D1-7, and the closest germ line J segment was JH4. In the light-chain genes, the closest germ line V segment was Vκ1-17 with the Jκ1 or Jκ4 segments. The dissociation constants of these Fab fragments with recombinant SAG1 were 3.09 × 10−9M for Tox203 and 2.01 × 10−8M for Tox1403, indicating that the affinity of Tox203 was 7 times higher than that of Tox1403. Preincubation ofT. gondiitachyzoites with Tox203 significantly inhibited their attachment to cultured MDBK cells. Passive immunization of mice with Tox203 also significantly reduced mortality after challenge withT. gondiitachyzoites. This is the first report of bacterial expression of human monoclonal antibody Fab fragments to SAG1 ofT. gondii. These results also demonstrate that human Fab fragments to SAG1 might be applicable for immunoprophylaxis of toxoplasmosis.

scholarly journals A Novel Repeat-Associated Small Interfering RNA-Mediated Silencing Pathway Downregulates Complementary Sense gypsy Transcripts in Somatic Cells of the Drosophila Ovary

2006 ◽  
Vol 81 (4) ◽  
pp. 1951-1960 ◽  
Author(s):  
Alain Pélisson ◽  
Emeline Sarot ◽  
Geneviève Payen-Groschêne ◽  
Alain Bucheton
Keyword(s):  
Small Interfering Rna ◽  
Germ Line ◽  
Endogenous Retrovirus ◽  
Transcript Accumulation ◽  
Selfish Dna ◽  
Independent Manner ◽  
Somatic Tissues ◽  
Interfering Rna ◽  
Drosophila Ovary ◽  
Complementary Sense

ABSTRACT Replication of the gypsy endogenous retrovirus involves contamination of the female germ line by adjacent somatic tissues. This is prevented by flam, an as-yet-uncloned heterochromatic pericentromeric locus, at the level of transcript accumulation in these somatic ovarian tissues. We tested the effect of a presumptive RNA silencing mechanism on the accumulation of RNAs produced by constructs containing various gypsy sequences and report that the efficiency of silencing is indeed correlated with the amount of complementary RNAs, 25 to 30 nucleotides in length, in the ovary. For instance, while these RNAs were found to display a three- to fivefold excess of the antisense strands, only the transcripts that contain the complementary sense gypsy sequences could be repressed, indicating that they are targeted at the RNA, not DNA, level. Their size and asymmetry in strand polarity are typical of the novel repeat-associated small interfering RNA (rasiRNA)-mediated pathway, recently suspected to prevent the deleterious expression of selfish DNA specifically in the germ line. Unlike microRNAs (but like rasiRNAs and, surprisingly, siRNAs as well), gypsy rasiRNAs are modified at the 3′ end. The rasiRNA-associated protein Piwi (but not Aub) is required for gypsy silencing, whereas Dicer-2 (which makes siRNAs) is not. In contrast, piwi, aub, and flam do not appear to affect somatic siRNA-mediated silencing. The amount of gypsy rasiRNAs is genetically determined by the flam locus in a provirus copy number-independent manner and is triggered in the somatic tissues by some pericentromeric provirus(es), which are thereby able to protect the germ line from retroviral invasion.

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