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Impaired hematopoiesis and delayed thrombopoietic recovery following sublethal irradiation in SRC‑3 knockout mice.

Jin J, Wang Y, Wang J, Xu Y, Chen SL, Wang JP, Su YP - Mol Med Rep (2014)

Bottom Line: Furthermore, peripheral blood cell counts, BM cellularity and colony-forming unit (CFU) assays were performed following irradiation.The results showed that peripheral blood cells were significantly lower in number and recovered less rapidly in irradiated SRC-3-/‑ mice as compared with control animals.In conclusion, the present study demonstrated that the hematopoietic ability in SRC-3 knockout mice is severely impaired following a sublethal dose of irradiation.

View Article: PubMed Central - PubMed

Affiliation: Institute of Combined Injury, State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing Engineering Research Center for Nanomedicine, College of Preventive Medicine, Third Military Medical University, Chongqing 400038, P.R. China.

ABSTRACT
The objective of the present study was to investigate the role of the steroid receptor coactivator-3 (SRC-3) in hematopoiesis of mouse bone marrow (BM) following total body irradiation (TBI). SRC-3-/‑ mice and wild-type (WT) mice were exposed to 4.5 Gy γ rays. Immunoblotting analysis revealed that the SRC-3 protein (p160) levels in normal BM-nucleated cells in WT were higher than in SRC-3-/‑ mice. Furthermore, peripheral blood cell counts, BM cellularity and colony-forming unit (CFU) assays were performed following irradiation. The results showed that peripheral blood cells were significantly lower in number and recovered less rapidly in irradiated SRC-3-/‑ mice as compared with control animals. BM-nucleated cell and CFU counts were significantly decreased in SRC-3-/‑ mice on the 7th and 14th day. Of note, the recovery of platelet (PLT) and megakaryocytic lineage were more depressed than the granulocytic and erythroid lineage in SRC-3-/‑ mice. In conclusion, the present study demonstrated that the hematopoietic ability in SRC-3 knockout mice is severely impaired following a sublethal dose of irradiation.

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Expression of SRC-3 protein in BM-nucleated murine cells. (A) Confirmation of genotype in SRC-3 mutant mice. Lane M, DNA molecular size marker; lane 1, tail DNA of SRC-3+/− mice with mixture of three primers and bands at 450 and 230 bp indicate the heterozygote; lane 2, tail DNA of SRC-3−/− mice with a mixture of primers 1 and 3, with the 230 bp band indicating the knockout (SRC-3−/−) mice; lane 3, tail DNA of SRC-3+/+ mice with a mixture of primer 1 plus primer 2, with the 450 bp band indicating wild-type (SRC-3+/+) mice. (B) Differential expression of SRC-3 protein in BM-nuclear cells of SRC-3+/+ and SRC-3−/− mice. SRC-3, steroid receptor coactivator-3; BM, bone marrow.
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f1-mmr-09-05-1629: Expression of SRC-3 protein in BM-nucleated murine cells. (A) Confirmation of genotype in SRC-3 mutant mice. Lane M, DNA molecular size marker; lane 1, tail DNA of SRC-3+/− mice with mixture of three primers and bands at 450 and 230 bp indicate the heterozygote; lane 2, tail DNA of SRC-3−/− mice with a mixture of primers 1 and 3, with the 230 bp band indicating the knockout (SRC-3−/−) mice; lane 3, tail DNA of SRC-3+/+ mice with a mixture of primer 1 plus primer 2, with the 450 bp band indicating wild-type (SRC-3+/+) mice. (B) Differential expression of SRC-3 protein in BM-nuclear cells of SRC-3+/+ and SRC-3−/− mice. SRC-3, steroid receptor coactivator-3; BM, bone marrow.

Mentions: SRC-3−/− mice were kindly provided by Professor Jianming Xu (Molecular and Cellular Biology Laboratory, Baylor College of Medicine, Houston, USA). The SRC-3 mutant colony was maintained by interbreeding heterozygous pairs. The mice had a mixed 129/SvEvxC57BL/6J genetic background. Female SRC-3−/− mice and wild-type (WT) counterparts (age, 8–10 weeks) were used in this experiment. Mice were provided with sterilized water and food ad libitum in a pathogen-free animal facility. Experimental protocols were approved by the Animal Care Committees of the Third Military Medical University. Genotypes were determined by PCR using tail DNA (Fig. 1A) (7). For PCR analysis, the WT (SRC-3+/+) allele was detected using primer 1: 5′-GATGAGTGGACTAGGCGAAAGCTCT-3′ and primer 2: 5′-GCTGAGATTTGCAGAGATGAGCTC-3′. This primer pair amplified a 450-bp fragment from the SRC-3+/+ mice. DNA was also amplified using primers 1 and 3: 5′-GGCGATTAAGTTGGGTAACGCCAG-3′, which is located in the LacZ indicator to detect the mutant of the SRC-3 allele. This primer pair amplified a 230-bp fragment from SRC-3−/− mice. The three mixed primers amplified the of 450 and 230 bp fragments to detect the heterozygote.


Impaired hematopoiesis and delayed thrombopoietic recovery following sublethal irradiation in SRC‑3 knockout mice.

Jin J, Wang Y, Wang J, Xu Y, Chen SL, Wang JP, Su YP - Mol Med Rep (2014)

Expression of SRC-3 protein in BM-nucleated murine cells. (A) Confirmation of genotype in SRC-3 mutant mice. Lane M, DNA molecular size marker; lane 1, tail DNA of SRC-3+/− mice with mixture of three primers and bands at 450 and 230 bp indicate the heterozygote; lane 2, tail DNA of SRC-3−/− mice with a mixture of primers 1 and 3, with the 230 bp band indicating the knockout (SRC-3−/−) mice; lane 3, tail DNA of SRC-3+/+ mice with a mixture of primer 1 plus primer 2, with the 450 bp band indicating wild-type (SRC-3+/+) mice. (B) Differential expression of SRC-3 protein in BM-nuclear cells of SRC-3+/+ and SRC-3−/− mice. SRC-3, steroid receptor coactivator-3; BM, bone marrow.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4020484&req=5

f1-mmr-09-05-1629: Expression of SRC-3 protein in BM-nucleated murine cells. (A) Confirmation of genotype in SRC-3 mutant mice. Lane M, DNA molecular size marker; lane 1, tail DNA of SRC-3+/− mice with mixture of three primers and bands at 450 and 230 bp indicate the heterozygote; lane 2, tail DNA of SRC-3−/− mice with a mixture of primers 1 and 3, with the 230 bp band indicating the knockout (SRC-3−/−) mice; lane 3, tail DNA of SRC-3+/+ mice with a mixture of primer 1 plus primer 2, with the 450 bp band indicating wild-type (SRC-3+/+) mice. (B) Differential expression of SRC-3 protein in BM-nuclear cells of SRC-3+/+ and SRC-3−/− mice. SRC-3, steroid receptor coactivator-3; BM, bone marrow.
Mentions: SRC-3−/− mice were kindly provided by Professor Jianming Xu (Molecular and Cellular Biology Laboratory, Baylor College of Medicine, Houston, USA). The SRC-3 mutant colony was maintained by interbreeding heterozygous pairs. The mice had a mixed 129/SvEvxC57BL/6J genetic background. Female SRC-3−/− mice and wild-type (WT) counterparts (age, 8–10 weeks) were used in this experiment. Mice were provided with sterilized water and food ad libitum in a pathogen-free animal facility. Experimental protocols were approved by the Animal Care Committees of the Third Military Medical University. Genotypes were determined by PCR using tail DNA (Fig. 1A) (7). For PCR analysis, the WT (SRC-3+/+) allele was detected using primer 1: 5′-GATGAGTGGACTAGGCGAAAGCTCT-3′ and primer 2: 5′-GCTGAGATTTGCAGAGATGAGCTC-3′. This primer pair amplified a 450-bp fragment from the SRC-3+/+ mice. DNA was also amplified using primers 1 and 3: 5′-GGCGATTAAGTTGGGTAACGCCAG-3′, which is located in the LacZ indicator to detect the mutant of the SRC-3 allele. This primer pair amplified a 230-bp fragment from SRC-3−/− mice. The three mixed primers amplified the of 450 and 230 bp fragments to detect the heterozygote.

Bottom Line: Furthermore, peripheral blood cell counts, BM cellularity and colony-forming unit (CFU) assays were performed following irradiation.The results showed that peripheral blood cells were significantly lower in number and recovered less rapidly in irradiated SRC-3-/‑ mice as compared with control animals.In conclusion, the present study demonstrated that the hematopoietic ability in SRC-3 knockout mice is severely impaired following a sublethal dose of irradiation.

View Article: PubMed Central - PubMed

Affiliation: Institute of Combined Injury, State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing Engineering Research Center for Nanomedicine, College of Preventive Medicine, Third Military Medical University, Chongqing 400038, P.R. China.

ABSTRACT
The objective of the present study was to investigate the role of the steroid receptor coactivator-3 (SRC-3) in hematopoiesis of mouse bone marrow (BM) following total body irradiation (TBI). SRC-3-/‑ mice and wild-type (WT) mice were exposed to 4.5 Gy γ rays. Immunoblotting analysis revealed that the SRC-3 protein (p160) levels in normal BM-nucleated cells in WT were higher than in SRC-3-/‑ mice. Furthermore, peripheral blood cell counts, BM cellularity and colony-forming unit (CFU) assays were performed following irradiation. The results showed that peripheral blood cells were significantly lower in number and recovered less rapidly in irradiated SRC-3-/‑ mice as compared with control animals. BM-nucleated cell and CFU counts were significantly decreased in SRC-3-/‑ mice on the 7th and 14th day. Of note, the recovery of platelet (PLT) and megakaryocytic lineage were more depressed than the granulocytic and erythroid lineage in SRC-3-/‑ mice. In conclusion, the present study demonstrated that the hematopoietic ability in SRC-3 knockout mice is severely impaired following a sublethal dose of irradiation.

Show MeSH
Related in: MedlinePlus