Relationship between thymus ontogeny and DNA ligase expression in the chicken

Jotereau, Francine; Maniey, Dominique; David, Jean‐Claude

doi:10.1002/eji.1830170115

Cited by 3 publications

(1 citation statement)

References 18 publications

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“…Expression of this enzyme correlates with an altered incidence of N insertion in coding joints (Landau et al 1987) of neonatal versus adult mouse lymphocytes (Lafaille et al 1989;Feeney 1990;Gu et al 1990). It is also of note that two distinct ligase activities are sequentially expressed in successive developmental waves of avian thymocytes (Jotereau et al 1987), although it is not clear whether this has significance for VDJ recombination.…”

Section: Basis For Limited Tcr 3 Gene Recombination and General Absenmentioning

confidence: 99%

T-lymphocyte development in scid mice is arrested shortly after the initiation of T-cell receptor delta gene recombination.

Carroll

Bosma²

1991

Genes Dev.

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Scid mice lack functional lymphocytes because they carry a mutation that impairs rearrangement of immunoglobulin and T-cell receptor (TCR) genes. Rearrangement of TCR ~, but not ~, and 13 genes, was routinely observed in DNA of scid thymocytes and thymocyte hybridomas. TCR 8 gene rearrangements appeared to involve DS1, D82, and JSl elements only; rearrangement of elements upstream of D~I (e.g., VS1) was not observed, and transcripts corresponding to fully assembled TCR 8 genes (VDJ8 or VDDJS) were not detected in RNA from scid thymocytes. These findings suggest that DS1, D82, and JS1 may be among the first TCR gene elements to undergo recombination and that scid T-lineage cells are developmentally arrested during or shortly after this stage of differentiation. One class of TCR 8 recombination fragments (D82-JS1) was amplified by the polymerase chain reaction (PCR) and cloned, and the recombination junctions were sequenced. Most fragments showed normal coding joints. Interestingly, five of seven coding joints that lacked N insertions showed evidence of recombination between short stretches (2-3 bp) of homologous sequence. As discussed, the general absence of VS-, J~/-, and ll3-associated rearrangements, despite the occurrence of normal D82-JS1 rearrangements, raises the possibility that the scid mutation may cause premature cessation of TCR gene recombination and thereby arrest early T-cell development.[Key Words: scid mice; T-cell receptor; T-cell development; polymerase chain reaction] Received January 14, 1991; revised version accepted May 17, 1991.Mice homozygous for the autosomal recessive mutation, scid (scid mice) are severely deficient in mature lymphocytes (Bosma et al. 1983). Early B-and T-lineage-committed cells are clearly present in scid lymphopoietic tissues but are unable to differentiate into functional B and T lymphocytes. This developmental arrest is believed to reflect a defect in the system responsible for the recombination of V (variable), D (diversity), and J (joining) gene elements that code for immunoglobulin and T-cell receptor (TCR) variable regions. Evidence of this defect is seen in rearranged immunoglobulin heavy-chain (IgH) genes of transformed scid pre-B cell lines (Schuler et al. 1986;Hendrickson et al. 1988;Kim et al. 1988;Malynn et al. 1988;Okazaki et al. 1988;Blackwell et al. 1989) and rearranged TCR ~/and genes of scid thymic lymphomas (Schuler et al. 1986(Schuler et al. , 1990. Most such rearranged genes show abnormal deletions of D-and J-or V-and J-coding elements, frequently extending 1 kb or more. In addition, transfection of transformed scid lymphocytes with extrachromosomal (Lieber et al. 1988) or retroviral recombination substrates (Ferrier et al. 1990;Hendrickson et al. 1990) has revealed a defect in the recombinase activity associated with VDJ 1Present address: Albany Medical College, Department of Microbiology and Immunology, Albany, New York 12208 USA. recombination: Although this activity can recognize, cleave, and join the recombination signal sequences that flank...

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Section: Basis For Limited Tcr 3 Gene Recombination and General Absenmentioning

confidence: 99%

T-lymphocyte development in scid mice is arrested shortly after the initiation of T-cell receptor delta gene recombination.

Carroll

Bosma²

1991

Genes Dev.

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Purification and characterization of DNA ligase II from Drosophila melanogaster

Takahashi¹,

Tomizawa²

1990

European Journal of Biochemistry

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Drosophila melanogaster contains DNA ligases I and 11. The activity of DNA ligase I is especially high during early embryonic periods, but decreases rapidly afterwards. Although the activity of DNA ligase I1 is low, it persists throughout all developmental stages. The specific activity of DNA ligase I1 is high in embryos, but the total activity per body mass was highest in pupae. To characterize the properties of DNA ligase I1 further and to clarify its differences from DNA ligase I, DNA ligase I1 was prepared from pupae of D . melanogaster. The enzyme was purified about 3200-fold by ammonium sulfate fractionation (40 -70% saturation), phosphocellulose (P1 1) and Ultrogel column chromatography. Some of the properties have been reported previously. The isoelectric point of DNA ligase I1 was 6.4 while those of DNA ligase I were 4.9 and 5.8. The optimum pH of DNA ligase I1 was 7.8-8.1 but 8.0-8.5 for DNA ligase I.The molecular masses of DNA ligase I1 adducts with AMP were determined as 90 and 70 kDa. These adducts were degraded to 42 and 14.4 kDa by trypsin digestion. For preparation of monoclonal antibodies, a mouse was immunized with the purified enzyme. Two clones, 10-6 and 3-3 IgM, were obtained and purified from mouse ascites.These antibodies showed both binding and neutralizing activities toward DNA ligase I1 from D. melanoguster, but did not react with DNA ligase I from the same origin.These results showed clearly that DNA ligases I and I1 have different properties and suggest they have different roles during the developmental stages of D. melanoguster.DNA ligase catalyzes the formation of phosphodiester linkages between DNA chains. In eukaryotes, two forms of DNA ligase have been identified: I and 11. Because DNA ligase I is most abundant in proliferating cells, it is thought to be involved primarily in DNA replication. Likewise, the presence of DNA ligase I1 in resting cells suggests that it is involved in DNA repair. DNA ligases I and I1 are also distinguishable by their affinity for ATP, optimum ionic strength and substrate specificity [l, 21. We are interested in studying the biochemical role of the two DNA ligases in developmental stages of eukaryotes. Drosophila melanogaster was chosen for this purpose because this organism is well characterized biochemically and genetically. In a previous paper [3], we reported the detection of two DNA ligases which were separated by phosphocellulose chromatography. DNA ligase I eluted at 0.2 M KC1 from phosphocellulose and was most abundant in embryos from early stages of development. DNA ligase I1 eluted at 0.6 M KCl from phosphocellulose, was less abundant than DNA ligase I, and was present in relatively similar quantities throughout development. These enzymes appear to be similar to mammalian DNA ligases I and 11. Rabin et al. [4, 51 have also purified and characterized a DNA ligase from embryos of D . melunogaster which seems to correspond to DNA ligase 1.This paper describes the further purification and characterization of DNA ligase 11 from pupae. A comparis...

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Effect of embryonic thermal manipulation on heat shock protein 70 expression and immune system development in Pekin duck embryos

2018

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Two experiments were conducted to study the effects of thermal manipulation on heat shock protein 70 (HSP70) transcription and immune cell characteristics' in Pekin duck embryos. In experiments I and II, fertile duck eggs were incubated at either a standard (S; 37°C) or high (H; 38.0°C) temperature from embryonic day (ED) 1 to 10. At ED11, half the eggs incubated at 37.5°C or 38.0°C were either kept at 37.5°C (SS) or 38.0°C (HH) respectively, or moved to the opposite incubator/temperature of either 38.0°C (SH) or 37.5°C (HS). Beginning on ED 21, all eggs (embryos) were incubated at 37.5°C. In experiments I and II, the mean egg shell temperature (EST) of eggs incubated at 38.0°C was higher than that from 37.5°C at ED10 (P < 0.01). In both experiments, the yolk free wet embryo body weight (YFWEBW) of embryos from the HS treatment was increased while that in the HH treatments was decreased compared with the SS treatment at ED25 (P < 0.01). In experiment II, embryos from the HH treatment had increased HSP70 mRNA compared with the SS treatment in the liver, thymus, and bursa (P < 0.05). The thymus and bursa from embryos in the SH treatment had increased MHCI mRNA when compared with the SS treatment (P < 0.05). The thymus from HH and SH treatments had increased MHCII mRNA while the bursa from the HH and SH treatments had decreased MHCII mRNA when compared with the SS treatment (P < 0.05). The spleen and bursa from the HH treatment had increased IL-6, IFNγ, and IL-10 mRNA when compared with the SS treatment (P < 0.05). The thymus and spleen from embryos in the HH treatment had increased CD8+/CD4+ ratios and CD4+CD25+ cell percentages compared with the SS treatment (P < 0.05). In conclusion, embryos incubated at 0.5°C higher than the standard incubation temperature from ED1 to 21 had increased HSP70 mRNA, MHCI, and inflammatory cytokines, and the immune response was skewed toward cell-mediated immunity.

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Relationship between thymus ontogeny and DNA ligase expression in the chicken

Cited by 3 publications

References 18 publications

T-lymphocyte development in scid mice is arrested shortly after the initiation of T-cell receptor delta gene recombination.

T-lymphocyte development in scid mice is arrested shortly after the initiation of T-cell receptor delta gene recombination.

Purification and characterization of DNA ligase II from Drosophila melanogaster

Effect of embryonic thermal manipulation on heat shock protein 70 expression and immune system development in Pekin duck embryos

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