Preparation Of Antibody Against GAS7 And Expression Of GAS7 In Hippocampus Of The OVX Rats

Growth arrest-specific (gas) genes are expressed preferentially in cells that enter a quiescent state. gas7, which we identified in serum-starved murine fibroblasts, is reported to be expressed in vivo selectively in the system of nervous, and expression has been observed during the growth arrest that accompanies terminal differentiation of cells. Gas7 is necessary for the formation of neurite in cultured cerebellar preneurons. Whereas antisense inhibition of Gas7 expression impedes neurite outgrowth from maturing Purkinje cells in culture, overexpression of the Gas7 protein promotes neurite outgrowth from undifferentiated PC12 cells. Therefore, Gas7 is regard as the marker od matured nervous cells and a tumor suppressor gene. Few antibodies are available now,which are not commercial yet. For further research on gas7,we prepared polyclonal antibodies , the methods, such as western blot, immunohistochemistry and RT-PCR were used to examine the expression and distribution ,and we select SD rats and establish a model of ovariectomized rats, followed by estrogen replacement therapy, observe the change of expression of gas7 in hippocampus of the OVX rats.Objective:1. to prepare anti-gas7 polyclonal antibodies2. to observe the expression and distribution of gas7 isoforms in the system of nervous of rats3. to investigate the change of expression of gas7 isoforms in hippocampus of the OVX rats. observeMethods:1. the GST fusion protein which rat carboxy-group terminals were fused with glutathione S-transferase (GST) separately with gene engineering technology as immunogen is injected into rabbit for production of polyclonal antibodies2. implicate western blot, immunohistochemistry and RT-PCR to examine the expression and distribution of gas7 isoforms3. implicate western blot to observe the change of expression of gas7 isoforms in hippocampus of the OVX rats .Results:1. prepare successfully anti-gas7 polyclonal antibodies.2. gas7 express in vivo selectively in the system of nervous, and different gas7 isoforms express in different region. 3. estrogen can up regulate the expression of gas7b and down regulate the expression of gas7a.conclusion:1. different gas7 isoforms may participate in the different functions of the system of nervous2. estrogen may contribute to anti-aging in female animal by regulate the expression of gas7 isoforms in hippocampus of the OVX rats.