Limits...
Bcl11a is essential for lymphoid development and negatively regulates p53.

Yu Y, Wang J, Khaled W, Burke S, Li P, Chen X, Yang W, Jenkins NA, Copeland NG, Zhang S, Liu P - J. Exp. Med. (2012)

Bottom Line: Myeloid development, in contrast, is not obviously affected by the loss of Bcl11a.Bcl11a regulates expression of Bcl2, Bcl2-xL, and Mdm2, which inhibits p53 activities.Bcl11a is therefore essential for lymphopoiesis and negatively regulates p53 activities.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China.

ABSTRACT
Transcription factors play important roles in lymphopoiesis. We have previously demonstrated that Bcl11a is essential for normal lymphocyte development in the mouse embryo. We report here that, in the adult mouse, Bcl11a is expressed in most hematopoietic cells and is highly enriched in B cells, early T cell progenitors, common lymphoid progenitors (CLPs), and hematopoietic stem cells (HSCs). In the adult mouse, Bcl11a deletion causes apoptosis in early B cells and CLPs and completely abolishes the lymphoid development potential of HSCs to B, T, and NK cells. Myeloid development, in contrast, is not obviously affected by the loss of Bcl11a. Bcl11a regulates expression of Bcl2, Bcl2-xL, and Mdm2, which inhibits p53 activities. Overexpression of Bcl2 and Mdm2, or p53 deficiency, rescues both lethality and proliferative defects in Bcl11a-deficient early B cells and enables the mutant CLPs to differentiate to lymphocytes. Bcl11a is therefore essential for lymphopoiesis and negatively regulates p53 activities. Deletion of Bcl11a may represent a new approach for generating a mouse model that completely lacks an adaptive immune system.

Show MeSH

Related in: MedlinePlus

Dynamic expression patterns of Bcl11a in hematopoiesis. (A) Schematic diagram of the Bcl11a-eGFP reporter allele. The eGFP reporter cassette flanked by two F3 sites is introduced to the 3′UTR region of Bcl11a, 8 bp after the stop codon TAG. (B) Flow cytometry tracking Bcl11a expression using the Bcl11aeGFP/eGFP reporter mice. Cell surface markers for defining these cells are described in Table S1. (C) Expression of Bcl11a (eGFP+) and Bcl11b (Tdtomato+) in double negative (DN) thymocytes was measured by flow cytometry. DN thymocytes are identified as described in Fig. S1 D. (D) qRT-PCR analysis of Bcl11a expression in sorted hematopoietic populations. Data represent mean values of three independent biological replicates and all values are normalized to the expression of the Gapdh gene. Error bars indicate the SD. Thy, thymus; PB, peripheral blood; M, macrophages; G, granulocytes; Mk, megakaryocytes; B, BM CD19+ B cells; T, spleen CD3+T cells. In all flow cytometry assays, at least four mice were analyzed for each cell type in independent experiments.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC3526365&req=5

fig1: Dynamic expression patterns of Bcl11a in hematopoiesis. (A) Schematic diagram of the Bcl11a-eGFP reporter allele. The eGFP reporter cassette flanked by two F3 sites is introduced to the 3′UTR region of Bcl11a, 8 bp after the stop codon TAG. (B) Flow cytometry tracking Bcl11a expression using the Bcl11aeGFP/eGFP reporter mice. Cell surface markers for defining these cells are described in Table S1. (C) Expression of Bcl11a (eGFP+) and Bcl11b (Tdtomato+) in double negative (DN) thymocytes was measured by flow cytometry. DN thymocytes are identified as described in Fig. S1 D. (D) qRT-PCR analysis of Bcl11a expression in sorted hematopoietic populations. Data represent mean values of three independent biological replicates and all values are normalized to the expression of the Gapdh gene. Error bars indicate the SD. Thy, thymus; PB, peripheral blood; M, macrophages; G, granulocytes; Mk, megakaryocytes; B, BM CD19+ B cells; T, spleen CD3+T cells. In all flow cytometry assays, at least four mice were analyzed for each cell type in independent experiments.

Mentions: We determined Bcl11a expression at the single-cell level by making and analyzing an eGFP reporter mouse where an IRES-eGFP-FRT-Neo-FRT cassette was targeted to the 3′UTR of the Bcl11a locus (Fig. 1 A). The homozygous Bcl11aeGFP/eGFP mice had normal hematopoiesis and were used for detection of Bcl11a expression (GFP+) by flow cytometry.


Bcl11a is essential for lymphoid development and negatively regulates p53.

Yu Y, Wang J, Khaled W, Burke S, Li P, Chen X, Yang W, Jenkins NA, Copeland NG, Zhang S, Liu P - J. Exp. Med. (2012)

Dynamic expression patterns of Bcl11a in hematopoiesis. (A) Schematic diagram of the Bcl11a-eGFP reporter allele. The eGFP reporter cassette flanked by two F3 sites is introduced to the 3′UTR region of Bcl11a, 8 bp after the stop codon TAG. (B) Flow cytometry tracking Bcl11a expression using the Bcl11aeGFP/eGFP reporter mice. Cell surface markers for defining these cells are described in Table S1. (C) Expression of Bcl11a (eGFP+) and Bcl11b (Tdtomato+) in double negative (DN) thymocytes was measured by flow cytometry. DN thymocytes are identified as described in Fig. S1 D. (D) qRT-PCR analysis of Bcl11a expression in sorted hematopoietic populations. Data represent mean values of three independent biological replicates and all values are normalized to the expression of the Gapdh gene. Error bars indicate the SD. Thy, thymus; PB, peripheral blood; M, macrophages; G, granulocytes; Mk, megakaryocytes; B, BM CD19+ B cells; T, spleen CD3+T cells. In all flow cytometry assays, at least four mice were analyzed for each cell type in independent experiments.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3526365&req=5

fig1: Dynamic expression patterns of Bcl11a in hematopoiesis. (A) Schematic diagram of the Bcl11a-eGFP reporter allele. The eGFP reporter cassette flanked by two F3 sites is introduced to the 3′UTR region of Bcl11a, 8 bp after the stop codon TAG. (B) Flow cytometry tracking Bcl11a expression using the Bcl11aeGFP/eGFP reporter mice. Cell surface markers for defining these cells are described in Table S1. (C) Expression of Bcl11a (eGFP+) and Bcl11b (Tdtomato+) in double negative (DN) thymocytes was measured by flow cytometry. DN thymocytes are identified as described in Fig. S1 D. (D) qRT-PCR analysis of Bcl11a expression in sorted hematopoietic populations. Data represent mean values of three independent biological replicates and all values are normalized to the expression of the Gapdh gene. Error bars indicate the SD. Thy, thymus; PB, peripheral blood; M, macrophages; G, granulocytes; Mk, megakaryocytes; B, BM CD19+ B cells; T, spleen CD3+T cells. In all flow cytometry assays, at least four mice were analyzed for each cell type in independent experiments.
Mentions: We determined Bcl11a expression at the single-cell level by making and analyzing an eGFP reporter mouse where an IRES-eGFP-FRT-Neo-FRT cassette was targeted to the 3′UTR of the Bcl11a locus (Fig. 1 A). The homozygous Bcl11aeGFP/eGFP mice had normal hematopoiesis and were used for detection of Bcl11a expression (GFP+) by flow cytometry.

Bottom Line: Myeloid development, in contrast, is not obviously affected by the loss of Bcl11a.Bcl11a regulates expression of Bcl2, Bcl2-xL, and Mdm2, which inhibits p53 activities.Bcl11a is therefore essential for lymphopoiesis and negatively regulates p53 activities.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China.

ABSTRACT
Transcription factors play important roles in lymphopoiesis. We have previously demonstrated that Bcl11a is essential for normal lymphocyte development in the mouse embryo. We report here that, in the adult mouse, Bcl11a is expressed in most hematopoietic cells and is highly enriched in B cells, early T cell progenitors, common lymphoid progenitors (CLPs), and hematopoietic stem cells (HSCs). In the adult mouse, Bcl11a deletion causes apoptosis in early B cells and CLPs and completely abolishes the lymphoid development potential of HSCs to B, T, and NK cells. Myeloid development, in contrast, is not obviously affected by the loss of Bcl11a. Bcl11a regulates expression of Bcl2, Bcl2-xL, and Mdm2, which inhibits p53 activities. Overexpression of Bcl2 and Mdm2, or p53 deficiency, rescues both lethality and proliferative defects in Bcl11a-deficient early B cells and enables the mutant CLPs to differentiate to lymphocytes. Bcl11a is therefore essential for lymphopoiesis and negatively regulates p53 activities. Deletion of Bcl11a may represent a new approach for generating a mouse model that completely lacks an adaptive immune system.

Show MeSH
Related in: MedlinePlus