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Expression of insulin-like growth factors systems in cloned cattle dead within hours after birth
Shijie Li1, Yanxin Li, Shuyang Yu
1The State Key Laboratory for Agrobiotechnology in Livestock and Poultry, China Agricultural University, Beijing, China.
Abstract:
Cloning by somatic nuclear transfer is an inefficient process in which many of the cloned animals die shortly after birth and display organ abnormalities. In an effort to determine the possible roles IGFs played in neonatal death and organ abnormalities, we have examined expression patterns of eight genes in insulin-like growth factor (IGF) systems (IGF1, IGF2, IGF1R, IGF2R, IGFBP-1, IGFBP-2, IGFBP-3, and IGFBP-4) in six organs (heart, liver, spleen, lung, kidney, and brain) of both neonatal death cloned bovines (n = 9) and normal control calves (n = 3) produced by artificial insemination (AI) using real-time quantitative RT-PCR. The effect of the age of the fibroblast donor cell on the gene expression profiles was also investigated. Aberrant expressions of six genes (IGF2, IGF1R, IGF2R, IGFBP-2, IGFBP-3, and IGFBP-4) were found in some studied tissues, but the expression of two genes (IGF1 and IGFBP-1) had similar levels with the normal controls. For the studied genes, kidney was the organ that was most affected (five genes) by gene downregulation, whereas spleen was the organ that was not affected. The two upregulation genes were in brain, but both of downregulation and upregulation were found in the heart, liver, and lung. The expression of three genes (IGF2R, IGFBP-4, and IGF2) in some tissues showed significant differences between AF cell-derived and FF cell-derived clones. Our results suggest that aberrations in gene expression within IGF systems were found in most cloned bovine tissues of neonatal death. Because IGF systems play an important role in embryo development and organogenesis, the aberrant transcription patterns detected in these clones may contribute to the defects of organs reported in neonatal death of clones.
Insights
Cloned calves experiencing neonatal death show abnormal gene expression in insulin-like growth factor (IGF) systems, particularly in organs like the kidney. These IGF gene expression changes may explain organ defects in cloned animals.
Area of Science:
- Animal Cloning
- Genetics
- Developmental Biology
Background:
- Somatic nuclear transfer cloning is inefficient, leading to neonatal death and organ abnormalities in cloned animals.
- Insulin-like growth factors (IGFs) are crucial for embryonic development and organogenesis.
Purpose of the Study:
- To investigate the role of insulin-like growth factor (IGF) systems in neonatal death and organ abnormalities in cloned bovines.
- To examine the expression patterns of eight IGF-related genes in various organs of cloned and control calves.
Main Methods:
- Real-time quantitative RT-PCR was used to analyze the expression of eight IGF system genes (IGF1, IGF2, IGF1R, IGF2R, IGFBP-1, IGFBP-2, IGFBP-3, IGFBP-4).
- Gene expression was compared between cloned bovines that died neonatally (n=9) and normal control calves (n=3).
- The influence of donor cell type (adult fibroblast vs. fetal fibroblast) on gene expression was also assessed.
Main Results:
- Aberrant expression of six IGF genes (IGF2, IGF1R, IGF2R, IGFBP-2, IGFBP-3, IGFBP-4) was observed in various tissues of cloned calves.
- Kidney showed the most downregulation (five genes), while spleen was unaffected.
- Brain exhibited two upregulated genes; heart, liver, and lung displayed both up- and downregulation.
- Differences in IGF2R, IGFBP-4, and IGF2 expression were noted between clones derived from adult and fetal fibroblasts.
Conclusions:
- Aberrations in IGF gene expression are prevalent in the tissues of cloned bovines that experienced neonatal death.
- These transcriptional changes in IGF systems likely contribute to the observed organ defects and mortality in cloned animals.
- Further research into IGF system regulation is warranted to improve cloning efficiency and animal health.
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