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Experimental Research on Development and Senescence of C57/BL6 Mouse Hippocampal Formation

Author: ZhangJingKun
Tutor: GuoMin
School: Liaoning Medical
Course: Human Anatomy and Embryology
Keywords: Mice Hippocampal Development Senescence Proliferation Apoptosis
CLC: R329.1
Type: Master's thesis
Year: 2011
Downloads: 54
Quote: 0
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Abstract


Objective To investigate the the C57/BL6 mice hippocampal formation, development and the aging process morphological changes of cell proliferation and apoptosis variation. Methods light microscopy serial sectioning and stereological methods on the, 1,3,5,7,14,21,28 day the embryo age 10,12,14,16,18,20 day and after birth pups after birth hippocampus of adult rats and aged rats, 3,6,12,15,18 months the structure CA area and DG region and its layers of volume and body density systematic application of immunohistochemistry morphological observation and quantitative analysis; immunofluorescence technique, transmission electron microscopy and western blotting their respective time points cell proliferation and apoptosis and apoptosis-related protein Bcl-2, Bax and Caspase-3 expression in hippocampal structure qualitative and quantitative analysis. One, C57/BL6 mouse hippocampal structure, development and aging volume change (a) light microscopy observations the E12d appear hippocampal structure primordia. E18d CA pyramidal layer \Health, CA gradually mature. P7d DG granular layer shell formation. Inner and outer arms of the P21d DG granular layer thickness considerable subgrain appears layer until 18M still exist. Hippocampal formation (b) stereological measurements, CA, DG, CA layers volume and DG layers volume P7d of the former growth is slow, P7 ~ P14d rapid growth the P14d growth slowing, 3M stabilize. Of C57/BL6 mouse hippocampal structure, cell proliferation and apoptosis in the process of development and aging (a) shows a large number of positive cells DCX expression E12d hippocampal structure primordia. The E18d diffuse distribution in the various layers of the hippocampal formation district. In the P1 ~~ P14d, CA pyramidal layer outer periphery of positive cells are arranged in a tight zone, its P1d widest to P14d disappeared. P1 ~ P7d positive cells dispersed in the DG layers. P14d is mainly distributed in the particle layer within 1/2. P21d ~ 18M mainly distributed in Asia granule cell layer. The positive cell Vv value of CA pyramidal layer P1d ~ P7d downward trend. DG granular layer Vv value P1d ~ P7d upward trend, P14d ~ 18M downward trend. CA pyramidal layer V P1d to the P3d upward trend, P7d, reduce. DG granular layer V P1d to the P14d increase; P21d ~ 18M downward trend. The CA District pyramidal cell layer of positive cells Vv value P1dCA3 downward trend in the CA1 region upward trend districts are reducing P7d. E18d ~ E20d, CA3 gt; CA1, P1d ~ P7d, CA3 lt; CA1. Western blotting showed: E18 d to P3 d, DCX expression gradually increased; P3d ~ 18M downward trend. (B) Hoechst33258 staining results E12d ~ E16d apoptotic cells gradually increased. E18d apoptotic cells dispersed, mainly located in CA and DG polymorphic layer and molecular layer. P1 ~ P7d gradually reduce the number of apoptotic cells. P14d ~ 2M mainly in the dentate back hippocampal sulcus and hippocampus slot visible white matter distribution of a small number of apoptotic cells. 3M ~ 18M only see sporadic distribution in the granule cell layer of the dentate gyrus Asia. CA and DG polymorphic layer and molecular layer cell apoptosis rate P1d to P14d decreased P14d obvious; P21d ~ 2M only see sporadic distribution; 3M after distribution. CA and DG the master cell layer apoptosis rate P1d highest, and then gradually reduce, the pyramidal layer P14d after distribution only see sporadic distribution of the granular layer 3M. (C) semi-thin and ultrathin sections observations hippocampal formation in mice alive after birth, there are cell proliferation, and gradually reduced. The E12d mouse hippocampal formation of apoptotic cells with or without, P1d reduced, P7d. , C57/BL6 mouse hippocampal structure, the expression of apoptosis-related factors in the process of development and aging (a) Bcl-2 and Bax the E16d hippocampal structure can be seen a small amount of positive cells. E18d pyramidal layer and more door positive cells, and CA3 region is the most densely pyramidal layer CA1 region most sparse. The P1 ~ P7d, hippocampal formation district two positive cells were gradually increased. The P14 ~ P21d CA area two positive cells were gradually reduced; DG of Bcl-2 positive cells gradually decreased, P21d mainly located in Asia granule cell layer continues to grow; DG of Bax positive cells P14d, P21d reduce P28d stabilized. Districts expression of Bcl-2 and Bax positive cells AO value E18 d to P21 d first increases and then decreases, P7 d is the maximum stabilized P28 d. District the Bcl-2/Bax addition P1d plunged, no significant difference between the rest of the time points. Stereology and Western Blotting similar results with this. (B) Caspase-3 expression in the E12d hippocampal structure primordia can be seen a small amount of positive expression. The E16d positive cells increased, mainly located in the hippocampus lamellar edge layer. E18d ~ P3d positive expression continues to grow, the floors are distributed mostly CA pyramidal layer and DG particle layer. P5 ~ P14d distribution gradually reduced. The P21d ~ 18M districts only see a small number of positive cells. Western Blotting: The results showed that the increase in caspase-3 E18 P3d expression; the P5 ~ P21 d reduced expression; P21d stabilized. Conclusion of C57/BL6 mouse hippocampal structure the E12d occurred 3M basic developmental maturity. Aged mice hippocampal formation volume did not change significantly. Main cell layer of C57/BL6 mice hippocampal structure alive proliferation and apoptosis were gradually increased, a week after birth, although the downward trend, but remain at a relatively high level, and the same level; a week later at a lower level, but cell proliferation in comparative advantage; the P14d both scarce or no. 3, Bcl-2, Bax and Caspase-3 in apoptosis during early development in the the C57/BL6 mouse hippocampus structure play an important role.

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CLC: > Medicine, health > Basic Medical > Human morphology > Human histology > Human Histology and Embryology ( Schools)
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