| Objective: To observe the morphology and distributional characteristics of Sox2 positive cells in subfornical organ(SFO) of SD rats in different stages and to Observe the expression.Methods: Healthy male SD rats were divided into two groups: the young group and the adult group with 24 rats in each group. The rats were in clean grade without further process.1 sample preparationNormal sample preparation:animals were anesthetized by Chloral hydrate(0.3ml/100g),primed by 4% paraformaldehyde(0.1mol/L,PH7.2)through the ascending aorta,fixed,got the brain,then fixed.Choose the tissue samples with organum subfornicale after dehydration,transparention,embedding.Use Paraffin slicing machine to get the coronal slice plane with the thickness of 5μm and make them paraffin sections.The disposition conform to the requirements of animal ethics to the experimental animals.2 StainingImmunohistochemical staining:The Paraffin sections were stained by Sox2 and were observed under mocroscope and photographed after cemented by glycerol.3 Statistical analysisChoose the same cross-section(organum subfornicale) and use the multimedia image pathology graphic analysis system to count Sox2 positive cells and the total number of cells in organum subfornicale at the same magnification(20 times). Then calculate the percentage of the positive cells which adopt ±s measurement data.1 The distribution of Sox2 positive cells in SFO of the rats at different developmental stages.In the SFO of adult group rats, the ventral part ependymal Sox2 positive cells are arranged orderly and densely. Ependymal positive cells are mostly double layers and some are monolayer in the central part and nd three layers or even multi-layers in the lateral part.Sparse Sox2 weakly positive cells scattered distributed in the backsides. Strong positive cells distrubuted densely in the great vessels around the outer portions which mostly located in the ventral sides or even between the vessels. Negtive cells were distrubuted in the back sides of the great vessels. The central department has intensive Sox2 immune positive cells. Small blood vessels were seen around the central with Sox2 negative cells around.2 The morphological characteristics of Sox2 positive cells in SFO of different developmental stages rats.In the SFO of the rats, strong positive cells were seen in or under the ependymal cells.The cell nucleus is dark brown,round or ovoid,3.7μm ~5.3μm*2.6μm~5.4μm;the strong positive cell nucleus between the vessels are oblong or elongated, 3.3μm~9.4μm*2.5μm~3.3μm;weakly positive cell nucleus in central partis beige, round or ovoid, 6.1μm~7.5μm*3.8μm~7.1μm; weakly positive cells with size of11.1μm~12.7μm *8.3μm~9.2μm were rarely seen.3 The comparison of Sox2 positive cells distribution at different developmental stages.The percentage of Sox2 positive cell in SFO of SD rats of the young group is 65.58%+4.23% and 52.13%+6.56% of the adult group.The Data comparison of the two groups had statistically significant difference(P < 0.05).4 Sox2 positive cell mitosis was found near the SFO later dorsal parts. In its lower left, subependymal zone, the same types of nerve cells were seen that connected into a chain.5 Sox2 strong positive cell clusters were seen in the ventral sideResults: ependyma of SFO which were consist of 4 strong positive cells. In the back side of it were three cell nucleus which with same morphological characters, 4.0μm~5.6μm*3.0μm~1.7μm, the ventral nucleus is elongated, 5.5μm*1.7μm.Conclusion:1 The expression of quantity in Sox2 positive cells in the SFO of adult SD rats are more than that in the adult group.2 The Sox2 positive expression cells are sparse in the back side and dense in the ventral side in both groups.3 Dense Sox2 positive cells are around the great vessels and negative cells are around central small vessels of adult SD rats. There are no dense Sox2 strong positive cells around the great vessels of SFO of SD rats in the adult group but negative cells around the central small blood vessels. |
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