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Objective To observe the calcium-binding protein D28k (Calbindin-D28k) Expression and positioning in the developing mouse kidney, Calbindin-D28k mouse kidney development occurs. Methods weighing 25 ~ 30g healthy adult Kunming mice, check vaginal suppository shedding, determine its conception, select the days of embryonic age (E) 10,12,14,16,18 fetal and postnatal days of age (N days) 1,3,7,14,21,40 offspring, each group of eight mice. Each embryo-old mice (day) caesarean section to remove the kidneys, routinely paraffin sections. Application of light microscopy (LM) were observed; different developmental stages of the mouse kidney using immunohistochemistry (IHC) staining (SABC) and immunofluorescence technique (IFT) of each group of mice kidneys Calbindin-D28k expression qualitative observation ; apply image analysis methods (CLAS-1000-type cell image analysis system) for each group of mice kidneys Calbindin-D28k expression quantitative detection; Western blot (Western blotting) of different developmental stages of the mouse kidney Calbindin-D28k expression quantitative determination. The results a light mirror results: embryonic age 12 days, the ureteric bud around postnatal renal tissue around the ureteric bud. Embryonic age 14 days, the kidney area students around the ureteric bud branching vesicles, comma bodies, S bodies. Embryonic age 16 days, III, IV renal corpuscles. Embryonic age 18 days, early medulla during scattered in a large number of interstitial packed outside the area, the equivalent of the adult medulla. One day after birth, the kidney capsule swells reduced kidney area, visible renal papilla. Seven days after birth, the cortex of kidney area disappear, renal development is basically mature, ureteric bud disappears marrow actinomycetes appears, showing a nearly straight tubule, the far straight tubule and collecting duct the renal papillary visible area within the medulla packed. 14 days after birth, cortical thickening, medulla can be divided into the medulla packed inside and outside the district. 21 days after birth, the cortical thickness was significantly increased renal papillary seen inside the medulla with an increase in the number of collecting duct. 40 days after the birth and development of the kidney and renal reach adult levels, the medulla packed well within the band clearly with adult rats identical. 2 immunohistochemical staining results: embryonic age 10 days, Calbindin-D28k in fetal rat kidneys no expression. 12 days of embryonic age, Calbindin-D28k ureteric bud expressed relatively weak intensity of expression, no expression in renal tissue after birth. 14 days of embryonic age, Calbindin-D28k expression in ureteric bud branching enhancement, no expression in the kidney area. Embryonic age 16 days, Calbindin-D28k expression in the ureteric bud ampulla strong, the tubular and S bodies no expression in S, however, expression and the ureteric bud ampulla connection at. 3 days embryonic age 18 days after birth, the renal capsule, proximal tubule, medullary no expression, enhanced expression of the distal nephron. Seven days after birth, Calbindin-D28k located in the kidney distal convoluted tubule, connecting tubule, and cortical collecting duct. After birth, 14 days, 21 days, 40 days Calbindin-D28k in the kidney of mice were expressed its expression intensity and stabilize. Calbindin-D28k expression in mouse kidney parts of the cytoplasm. 3 CLAS-1000-type cell image analysis system detects gray value: kidney development in different periods Calbindin-D28k expression there are significant differences: 12 days of embryonic age, Calbindin-D28k in the expression of the highest gray value, Calbindin-D28k certain expression. Calbindin-D28k in the expression of the gray values ??from embryonic age 12 days to 7 days after birth, reduce the amount of expression gradually increased and reached the peak at 7 days after birth. From 14 days to 40 days after birth, after birth, Calbindin-D28k expression reduced and stabilized, the difference was not significant. 4 immunofluorescence results: Calbindin-D28k no expression in fetal rats at embryonic age 10 days, 12 days of embryonic age expression in the ureteric bud trace embryonic age 14 days in the Y-shaped ureteric bud branching expression, enhanced expression of embryonic age of 16 days in ureteric bud ampulla express strongly, and a small amount of nephron and ureteric bud ampulla connection the Ministry of expression. Embryonic age 18 days to 7 days after birth, Calbindin-D28k expression increased, and located in the distal convoluted tubule, connecting tubule, and cortical collecting duct. 7 days after birth 40 days after birth, Calbindin-D28k in the distal convoluted tubule, connecting tubule, cortical collecting duct rich decreased expression in the medullary collecting duct. Calbindin-D28k expression sites in ureteric bud or distal convoluted tubule, connecting tubule, and cortical collecting duct epithelial cells. 5 Western blotting results: embryonic age 12 days, Calbindin-D28k expression level is very low, can not be detected. Embryonic age 14 days to 16 days of embryonic age Calbindin-D28k in the expression increased, the difference was statistically significant (12.0994 ± 0.6911) (13.2383 ± .1729), P <0.01]. Embryonic age 16 days to 18 days of embryonic age Calbindin-D28k in the expression of no significant difference. Embryonic age 18 days to 7 days after birth, Calbindin-D28k in the expression amount was significantly increased, differences have significant [(13.8447 ± 0.2385), (15.2718 ± .2019), (16.3208 ± .2839), (19.3383 ± 0.4044), P < 0.01]. 7 days to 14 days after birth, after birth, Calbindin-D28k expression levels decreased, the difference was statistically significant (19.3383 ± 0.4044) (15.8915 ± 1.0185), P <0.01]. 14 days to 40 days after birth, after birth, Calbindin-D28k expression was no significant difference. Conclusion 1. In the mouse kidney development, Calbindin-D28k in fetal rats at embryonic age 12 days ureteric bud weakly expressed, with the embryo (days) Age increased its expression level gradually increased, reached the maximum level 7 days after birth, and is located in the distal convoluted tubule, connecting tubule and cortical collecting duct, after which the expression is reduced and stabilized. 2. Calbindin-D28k expression sites in ureteric bud or distal convoluted tubule, connecting tubule, and cortical collecting duct epithelial cells. 3. Calbindin-D28k expression pattern in kidney development and positioning with renal calcium reabsorption process of intracellular calcium transport is closely related to, and may play an important role in calcium ion transport regulator.
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