The Molecular Mechanism And Relationship With The Heat Shock Protein 70 (HSP70) Of The Cow Suffered From The Alcohol Positive Milk

In order to study the etiopathogenisis and pathogenesy of the cow Alcohol Positive Milk (APM) further, On the basis of our early study and our investigation of the APM. We analyzed the difference of the heat shock protein 70 (HSP70) gene expression quantity difference in health cow and the cow which have APM. We also researched the localization of the HSP70 in the mammary gland by immunohistochemistry. The result of this experiment will reveal the relationship between the APM and the stress of the cow.HSPs are a family of highly conserved cellular proteins that have been observed in all organisms. HSPs could be induced in cells when exposed to stress, and are considered to be involved in a general mechanism that helps cells to survive many lethal stresses. Extensive studies on model species have revealed several HSP can be the act as an index to detect the heat stress of the animal. We research the HSP70 by molecular biology and immunohistochemistry methods. Which provide the theory and practice foundation in preventing the occurrence of the cow APM.Experiment 1 Immunohistochemistry research of HSP70 in the mammary gland of the cow.We collected the mammary gland tissue of the cow which suffering from alcohol positive milk cattle. we used the immunohistochemical method to identify the localization of the HSP70 in the bovine mammary tissue. the result suggests that staining of the HSP70 is more obvious in the APM cow. The positive signals were mainly found in the mammary epithelial cells of mammary gland alveolus. Indicating that heat stress made damage to the epithelial cells and resulting in the occurrence of the alcohol positive milk. Experiment 2 Different expression of HSP70 mRNA in the mammary gland of the APM cow and healthy cow.We extracted the total RNA from the cow mammary gland, Usingβ–actin as an inner control, an optimal semi-quantitative RT-PCR method was established to analyze HSP70 mRNA expression levels in the mammary gland of APM cow. The results showed that: HSP70 mRNA in bovine mammary gland in APM cow with strong expression(0.805±0.05). The expression in the APM cow was significantly higher than that in the healthy cow (P<0.05). We also compared the fragments which obtained for the mammary gland tissue of HSP70 mRNA sequence with the HSP70 mRNA (sequence number BtU09861) sequence in the GenBank and the homology is of up to 100%.