Study Of Tbx1 Gene Expression And Interacting Proteins In Kidney Tissues From Mouse Embryos

AbstractTBX1 (GeneID:6899) gene has been mapped to chromosomal region 22q11.21. Formally known as T-boxl, it belongs to the T-box transcription factor family, which is conserved through phylogeny and evolution. These genes are named for their similar DNA binding motif, T-box. TBX1 can impact development of ear, heart, parathyroid, thymus, teeth and other tissues and organs, and plays an important role in the development of schizophrenia.Recent research has indicated that microdeletion in 22q11.2 is strongly associated with genitourinary malformations Specific anomalies include renal malformations, metanephric duct/urinary bladder and hypospadia. Among the carriers, the incidence of above defects is as high as 36%, which is significant higher than that of the general population. TBX1 has been mapped to the critical region of the deletion and is known as an important transcription factor during the development. Research has suggested that there may be a common genetic basis for the development of various tissues and organs including the kidneys, heart, ear, thymus and parathyroid glands. TBX1 may serve in dose-dependent manner as one of the important genes in SHH, FGF, WNT and BMP signaling pathways in the early development of such tissues and organs via a common regulatory mechanism. Animal experiment also confirmed that, Tbxl knockout or haploinsufficiency can lead to phenotypes similar to those seen in human 22q11.2 deletion syndrome.So far, research of the TBX1 gene has been focused on its relationship with heart malformations, and there has been no report with regard to its connection with the development of genitourinary system. In the present study, we have analyzed the expression of Tbxl gene during the kidney development. In addition, by identifying its interacting protein, we also explored their role in the kidney development. This may help with delineation of the regulatory genetics networks for the embryonic development of the kidneysMaterial and Methods1,To analyze the expression of Tbxl gene in kidney tissue from mouse embryos with RT-PCR;2,To identify its interacting protein with immunoprecipitation (IP) and mass spectrometry (MALDI-TOF-MS). Subsequently, the protein was verified with Western Blotting. Expression of the interacting genes in mouse embryonic kidneys was also studied with RT-PCR;3,To explore the relationship between Tbx1 and Hoxd10 proteins through RNA interference.Results1,The expression of Tbxl gene reaches the first peak at the E15.5d in embryonic mouse kidneys, then decreases gradually, and reaches the second peak just after birth. In addition, the gene also expresses in adult mouse kidney at a low level;2,Hoxd10 and Tbxl proteins may interact during embryonic development of mouse kidneys;3,In myoblasts C2C12 cell line, interference of either Hoxd10 or Tbxl gene can downregulates the expression of both genesConclusion1,The expression of Tbxl gene changes along with the development of mouse kidney, indicating that it may play an important role. The gene may also play certain roles in the adult kidney;2,Tbxl may interact with Hoxd10 in embryonic kidneys of mice to regulate its development and function;3,Tbxl and Hoxd10 may interact the protein level, and feedback to each other by expression.