ABSTRACT
OBJECTIVE@#To explore whether the Ad5-atoh1/EGFP could transdifferent the supporting cells into the new hair cells in young adult guinea pigs cochlea in vivo.@*METHOD@#Twelve healthy pigmented guinea pigs weighted 200-250 g were included in this experiment. 5 ul of Ad5-E1/E3 defected-atoh1/EGFP were infused into the scala media through a hole made on the lateral wall of the cochlea. Six of the 12 animal were killed 2 weeks after the infusion operation. The others were killed 4 weeks after the operation. The whole mount of the basal membranes were directly observed under the fluorescence microscope for the expression of the EGFP (enhance green fluorescent protein) or for the expression of the hair cellspecific marker and nuclear after staining with myosin VIIa rabbit polyclonal antibody and Dapi dye.@*RESULT@#New cells with big nuclear, ellipse body and expressed with EGFP were found in the region near to the outmost row of the outer hair cells in 2 animal 2 weeks after the infusion. Moreover there were 3 animals with specific morphologic new cells in the location where ever been located by the outer hair cells and the region as 2 weeks animals 4 weeks after the infusion. Those cells were stained by myosin VIIa antibody.@*CONCLUSION@#Atoh1 gene could transdifferent some supporting cells in the basal membrane into hair cell like cells in young adult guinea pigs in vivo. These supporting cells locate in the region of outer hair cells and the basal membrane which do not belong to the region of outer hair cells.
Subject(s)
Animals , Basic Helix-Loop-Helix Transcription Factors , Genetics , Metabolism , Cell Transdifferentiation , Genetics , Cochlea , Cell Biology , Ear, Inner , Green Fluorescent Proteins , Genetics , Metabolism , Guinea Pigs , Hair Cells, Auditory , Cell Biology , Labyrinth Supporting Cells , Cell BiologyABSTRACT
Objective To modify the method of gavage administration in guinea pigs. Methods Fourty awake guinea pigs were kept rearing on the hind legs and leaning on a vertical fixture to avoid their escaping forward. A 1 mL injector was inserted into the mouth to the depth when the molar teeth were passed. Another fourty guinea pigs under general anesthesia were reversed at trendelenburg position and a children suction tube with an outer diameter of 2 mm was inserted into the stomach. Results All of the 80 guinea pigs were administered by modified gavage smoothly for seven consecutive days by one operator each time. None endured much pain or digestive tract injury, or died from air way perfusion by mistake. Conclusions We successfully modified the gavage method in guinea pigs, which would definitely take guinea pigs involved in intragastical pharmacal experiments besides the routine of rats and mice.