The Mechanical Research About The Influnence Of Boron To The Hypothalamic-Pituitary-Ovary Axis Of Ostrich Chicks

African ostrich originated from African desert.The female ostrich reach sexual maturity until 2-2.5y,which is the latest one in the avian.And it's brood time lasts for 3 months.What's the reason for it? As we know the gonad must be different from others. Taking estrogen can cause serum levels of 17β-E2,copper-protein,triglyeride heightened obviously,and after taking boron,these levels heightened much more.Through the additive of boric acid to ostrich chicks,we can explore the estrogen-like mechanism of boron,especially the effect on hypothalamic- pituitary-ovarium axis.In this research, ostrich chicks were treated by additive of various doses boron acid(0mg/L,100mg/L,200mg/L,400mg/L).Using the methods of transmission electron microscopy,radioactive immunoassay,histochemical method,immunohistochemistry,terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) and western bolt,the structural characteristics of ostrich chicks' HPO and the role of boron acid and apoptosis in the HPO after additive different dose of boron acid were illustrated.The main research results are as follows:1 Changes of female ostrich chicks hypothalamus after added B45d femal ostrich chicks with adding boric acid to the hydroposia were used in the present study to investigate the effects of boric acid on hypothalamus of ostrich chicks through locating the MT1 cells in hypothalamus of ostrich chicks after making light microscope sample copy by hematoxylin-eosin staining and immunohistochemistry SABC.The results showed that the number of paraventricular hypothalamic nucleus per unit area was highest in the group of 400 mg/L,but the difference was not significant. The rate of large and middle cells was highest in group of 200 mg/L,which was significantly higher than the other three groups;The number of supraoptic hypothalamic nucleus per unit area in group of 200 mg/L was highest among the groups,which was significantly higher than that of the control group.The rate of large and middle cells was highest in group of 200 mg/L and there were mainly small size cells in group of 400 mg/L; The number of nucleus arcuatus hypothalami per unit area was highest in control group and second highest in group of 200 mg/L,the difference between the two groups was significant;it was lowest in group of 400 mg/L and the difference was extremely significant comparing with the control group.The cells in control group were most small size.The positive MTlir cells in paraventricular nucleus were most in group of 200 mg/L and fewest in group of 400 mg/L,the differences were highly significant respectively. The number of MTlir cells in supraoptic nucleus was highest in group of 400 mg/L and the cells arranged closely with the lowest average integrated optical density and morphological incompletion.The difference between group of 400 mg/L and control group was highly significant.MT1 ir cells in nuclei arciformes were most in control group and arranged intensively.The average integrated optical density was highest in group of 100 mg/L with significant difference and lowest in group of 400 mg/L with no significant difference.The results demonstrated paraventricular nucleus and supraoptic nucleus had significant auxoaction in group of 200 mg/L but possessed significant depressant effect in group of 400 mg/L.The arcuate nuclei could promote slightly in group of 100 mg/L but depress significantly in group of 400 mg/L.2 Changes of female ostrich chicks pituitary after added BThree kinds of cells and MT1 in anterior pituitary of female ostrich chicks added B were located by the means of Peacse and SABC.Peacse showed that,there were more acidophilic cells,which were triangle or triangle with clear cell boundary,in control group.Furthermore,the size of basophilic cell were unequal,and there were many chromophobe cells with small size,light stain,and unclear cell boundary.Basophilic cell population increases in the group added B at the dosage of 100mg/L,and was maximum at the dosage of 200mg/L.In the group of 400mg/L,eosinocytes and Basicytes were agglomerate,and chromophobe cells were the least.SABC showed that,MT1ir cells stained with light/dark-colored brown.Cell with irregular shape was located stacked or scattered,and positive drops were main located in nucleus.In the group of 100mg/L,both MT1ir cell population and positive cells increased.In group of 200mg/L,MT1ir cell population increased more and most cells were strong positive.In group of 400mg/L, MT1ir cell population decreased and most cells were week positive.The results indicated that,when added B at the dosage of 100mg/L,200mg/L,all basicytes,MT1ir cells and positive cells increase.On the other hand,there was an inhibition to MT1ir cells at the dosage of 400mg/L.3 Changes of female ostrich chicks ovary after added BThe histological structure of ostrich chicks ovary and the effect of boron acid on the ovary were studied by the means of histology,SABC,western blot.The result showed that the diameter of primary oocyte was increased significantly in 100mg/L group,the number of primordial follicle,primary follicle,secondary follicle increased significantly,and the MT1ir intensity in the follicular,oocyte of every follicular,but the number of primordial follicle decreased significantly in 400mg/L group.4 Changes of E₂ and P₄ in serum after added BE₂ and P₄ in serum were detected using radio-immunity technology at 15d,30d,45d after added boric acid.The results showed that the levels of E₂ and P₄ in serum were higher in 100mg/L and 200mg/L groups than that in the controls.And the disparity was not significant.The level of E₂ decreased in the 400mg/L group,and the disparity was highly significant.However,the disparity of P₄ was not significant.At the 30d after adding to boric acid,the disparities of E₂ and P₄ were not significant in 100mg/L, 200mg/L and 400mg/L groups compared to the controls.At the 45d after adding to boric acid,the level of E₂ was higher in 100mg/L and 200mg/L groups than that in the controls. And the disparity of 200mg/L group was highly significant.The level of 400mg/L group decreased,and the disparity was not significant.The level of P₄ was lower in 200mg/L and 400mg/L groups than that in the controls.And the disparity was significant.The results revealed that adding to the appropriate amount of boric acid could increase the level of E₂ in serum,but the level would decrease when the amount up to 400mg/L.The effect on the level of P₄ was slight at the 15-30d after adding to boric acid,but at the 45d, the level of P₄ in serum decreased significantly.5 The changes of HPO apoptotic after added BThe regular of Apoptosis and the effect of boron acid on apoptotic in ostrich chicks were studied by the means of Tunel method.The results showed that there are significant effect of boron acid on SON,ARC,hypophysis and ovary.The Apoptotic index was significant decreased in the 100 and 200mg/L group,but it is contra in 200mg/L group. There have no significant disparation on PVN by add boron acid.