目的:探讨胰岛素样生长因子Ⅱm RNA结合蛋白3(IMP3)及胰岛素样生长因子-2(IGF2)在不同类型胶质瘤中的表达及意义。方法:采用免疫组化S-P法观察98例胶质瘤标本及20例非瘤脑组织中IMP3和IGF2蛋白的表达情况,分析二者的表达与不同临床病理...目的:探讨胰岛素样生长因子Ⅱm RNA结合蛋白3(IMP3)及胰岛素样生长因子-2(IGF2)在不同类型胶质瘤中的表达及意义。方法:采用免疫组化S-P法观察98例胶质瘤标本及20例非瘤脑组织中IMP3和IGF2蛋白的表达情况,分析二者的表达与不同临床病理特征的关系;并随机选取IMP3及IGF2不同程度阳性表达病例行IMP3和IGF2 m RNA原位杂交检测。结果:在不同类型胶质瘤组织中IMP3蛋白、IGF2蛋白及IMP3 m RNA均呈高表达,除少突胶质细胞瘤外,二者在Ⅲ级肿瘤组织中的表达水平均明显高于Ⅱ级(P<0.05);在胶质母细胞瘤中二者的表达水平最高。IGF2 m RNA在不同类胶质瘤中均有表达,但在Ⅱ级、Ⅲ级肿瘤组织及胶质母细胞瘤之间表达差异无统计学意义(P>0.05)。IMP3、IGF2蛋白的阳性表达水平与患者年龄、肿瘤病理级别及术后生存时间关系密切(P<0.05)。结论:IMP3和IGF2对胶质瘤的恶性进展、患者预后存在一定影响,两者可能成为脑胶质瘤诊断和治疗的特异靶点。展开更多
We have identified DNA polymorphisms in the gene of insulin-like growth factor 2 by PCR-SSCP in a resource population, which was generated by Silky reciprocally crossing to Broilers. A C→G mutation was detected in th...We have identified DNA polymorphisms in the gene of insulin-like growth factor 2 by PCR-SSCP in a resource population, which was generated by Silky reciprocally crossing to Broilers. A C→G mutation was detected in the exon 2 (at position 71) by sequencing. This single nucleotide polymorphism (SNP) was found to be associated with production traits. Chicken with BB genotype showed more chest angle width but less 3 week body weight and glandular stom- ach weight than chicken with AA genotype (P<0.05); while the heterozygote (AB genotype) chicken had more abdominal fat weight, eviscerated yield with giblet than AA homozygote chicken. Further analysis showed that there were different genetic effects on some traits between heterozygote AB (paternal allele given first) and heterozygote BA: chickens with genotype BA had more birth weight and breast weight but less abdominal fat weight than chickens with geno- type AB (P<0.05), which could be hypothetically contributed by genome imprinting. Therefore, Silky chickens were selected for production of heterozygotes to confirm whether IGF2 locus was imprinting. Progeny from heterozygote × homozygote reciprocal cross was assayed for ex- pression after the genotype was determined. The transcription of IGF2 was detected by RT-PCR-SSCP. IGF2 gene was expressed bialleleically in 1-day-old neonatal liver and 90-day-old liver, kidney, heart, and muscle of both heterozygote AB and BA chickens. Therefore, IGF2 was not an imprinting gene in chicken. The different genetic effects between the het- erozygote AB and BA remain to be elucidated.展开更多
Background Chinese indigenous pigs are popular with consumers for their juiciness,flavour and meat quality,but they have lower meat production.Insulin-like growth factor 2(IGF2) is a maternally imprinted growth factor...Background Chinese indigenous pigs are popular with consumers for their juiciness,flavour and meat quality,but they have lower meat production.Insulin-like growth factor 2(IGF2) is a maternally imprinted growth factor that promotes skeletal muscle growth by regulating cell proliferation and differentiation.A single nucleotide polymorphism(SNP) within intron 3 of porcine IGF2 disrupts a binding site for the repressor,zinc finger BED-type containing 6(ZBED6),leading to up-regulation of IGF2 and causing major effects on muscle growth,heart size,and backfat thickness.This favorable mutation is common in Western commercial pig populations,but absent in most Chinese indigenous pig breeds.To improve meat production of Chinese indigenous pigs,we used cytosine base editor 3(CBE3)to introduce IGF2 intron3-C3071T mutation into porcine embryonic fibroblasts(PEFs) isolated from a male Liang Guang Small Spotted pig(LGSS),and single-cell clones harboring the desired mutation were selected for somatic cell nuclear transfer(SCNT) to generate the founder line of IGF2^(T/T) pigs.Results We found the heterozygous progeny IGF2^(C/T) pigs exhibited enhanced expression of IGF2,increased lean meat by 18%-36%,enlarged loin muscle area by 3%-17%,improved intramuscular fat(IMF) content by 18%-39%,marbling score by 0.75-1,meat color score by 0.53-1.25,and reduced backfat thickness by 5%-16%.The enhanced accumulation of intramuscular fat in IGF2^(C/T) pigs was identified to be regulated by the PI3K-AKT/AMPK pathway,which activated SREBP1 to promote adipogenesis.Conclusions We demonstrated the introduction of IGF2-intron3-C3071T in Chinese LGSS can improve both meat production and quality,and first identified the regulation of IMF deposition by IGF2 through SREBP1 via the PI3KAKT/AMPK signaling pathways.Our study provides a further understanding of the biological functions of IGF2and an example for improving porcine economic traits through precise base editing.展开更多
文摘目的:探讨胰岛素样生长因子Ⅱm RNA结合蛋白3(IMP3)及胰岛素样生长因子-2(IGF2)在不同类型胶质瘤中的表达及意义。方法:采用免疫组化S-P法观察98例胶质瘤标本及20例非瘤脑组织中IMP3和IGF2蛋白的表达情况,分析二者的表达与不同临床病理特征的关系;并随机选取IMP3及IGF2不同程度阳性表达病例行IMP3和IGF2 m RNA原位杂交检测。结果:在不同类型胶质瘤组织中IMP3蛋白、IGF2蛋白及IMP3 m RNA均呈高表达,除少突胶质细胞瘤外,二者在Ⅲ级肿瘤组织中的表达水平均明显高于Ⅱ级(P<0.05);在胶质母细胞瘤中二者的表达水平最高。IGF2 m RNA在不同类胶质瘤中均有表达,但在Ⅱ级、Ⅲ级肿瘤组织及胶质母细胞瘤之间表达差异无统计学意义(P>0.05)。IMP3、IGF2蛋白的阳性表达水平与患者年龄、肿瘤病理级别及术后生存时间关系密切(P<0.05)。结论:IMP3和IGF2对胶质瘤的恶性进展、患者预后存在一定影响,两者可能成为脑胶质瘤诊断和治疗的特异靶点。
基金This work was supported by the NationalMajor Basic Research Development Program of China(G20000161)National Natural Science Foundation of China(Grant No.39725022).
文摘We have identified DNA polymorphisms in the gene of insulin-like growth factor 2 by PCR-SSCP in a resource population, which was generated by Silky reciprocally crossing to Broilers. A C→G mutation was detected in the exon 2 (at position 71) by sequencing. This single nucleotide polymorphism (SNP) was found to be associated with production traits. Chicken with BB genotype showed more chest angle width but less 3 week body weight and glandular stom- ach weight than chicken with AA genotype (P<0.05); while the heterozygote (AB genotype) chicken had more abdominal fat weight, eviscerated yield with giblet than AA homozygote chicken. Further analysis showed that there were different genetic effects on some traits between heterozygote AB (paternal allele given first) and heterozygote BA: chickens with genotype BA had more birth weight and breast weight but less abdominal fat weight than chickens with geno- type AB (P<0.05), which could be hypothetically contributed by genome imprinting. Therefore, Silky chickens were selected for production of heterozygotes to confirm whether IGF2 locus was imprinting. Progeny from heterozygote × homozygote reciprocal cross was assayed for ex- pression after the genotype was determined. The transcription of IGF2 was detected by RT-PCR-SSCP. IGF2 gene was expressed bialleleically in 1-day-old neonatal liver and 90-day-old liver, kidney, heart, and muscle of both heterozygote AB and BA chickens. Therefore, IGF2 was not an imprinting gene in chicken. The different genetic effects between the het- erozygote AB and BA remain to be elucidated.
基金supported by the National Natural Science Foundation of China (3207269732030102)+2 种基金CARS-PIG-35R&D Programmes of Guangdong Province (2018B020203003)Laboratory of Lingnan Modern Agriculture Project (NZ2021006)。
文摘Background Chinese indigenous pigs are popular with consumers for their juiciness,flavour and meat quality,but they have lower meat production.Insulin-like growth factor 2(IGF2) is a maternally imprinted growth factor that promotes skeletal muscle growth by regulating cell proliferation and differentiation.A single nucleotide polymorphism(SNP) within intron 3 of porcine IGF2 disrupts a binding site for the repressor,zinc finger BED-type containing 6(ZBED6),leading to up-regulation of IGF2 and causing major effects on muscle growth,heart size,and backfat thickness.This favorable mutation is common in Western commercial pig populations,but absent in most Chinese indigenous pig breeds.To improve meat production of Chinese indigenous pigs,we used cytosine base editor 3(CBE3)to introduce IGF2 intron3-C3071T mutation into porcine embryonic fibroblasts(PEFs) isolated from a male Liang Guang Small Spotted pig(LGSS),and single-cell clones harboring the desired mutation were selected for somatic cell nuclear transfer(SCNT) to generate the founder line of IGF2^(T/T) pigs.Results We found the heterozygous progeny IGF2^(C/T) pigs exhibited enhanced expression of IGF2,increased lean meat by 18%-36%,enlarged loin muscle area by 3%-17%,improved intramuscular fat(IMF) content by 18%-39%,marbling score by 0.75-1,meat color score by 0.53-1.25,and reduced backfat thickness by 5%-16%.The enhanced accumulation of intramuscular fat in IGF2^(C/T) pigs was identified to be regulated by the PI3K-AKT/AMPK pathway,which activated SREBP1 to promote adipogenesis.Conclusions We demonstrated the introduction of IGF2-intron3-C3071T in Chinese LGSS can improve both meat production and quality,and first identified the regulation of IMF deposition by IGF2 through SREBP1 via the PI3KAKT/AMPK signaling pathways.Our study provides a further understanding of the biological functions of IGF2and an example for improving porcine economic traits through precise base editing.
