青山羊生后发育过程中GTH分泌与输卵管中GTH受体表达规律的研究
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摘要
卵泡刺激素(Follicle-stimulating hormone, FSH)和黄体生成素(Luteinizing hormone,LH)是由垂体前叶分泌的一类糖蛋白激素,FSH和LH是通过其各自的特异性受体FSHR和LHR的介导来发挥其促进动物性腺发育和调控生殖的生理学作用。FSH的生物学作用主要是促进卵巢和子宫内膜生长,刺激卵泡生长、发育,同时刺激卵泡细胞上L H受体产生;L H的生物学作用主要是促进卵泡的成熟和排卵,刺激卵泡内膜细胞产生雌激素。传统观点认为FSH和L H只对性腺有作用,但是近年来的研究发现,在性腺外的生殖组织上也存在这两种激素受体,提示FSH和L H在性腺外组织也可能起作用,而在生后发育过程中这两种激素分泌的变化和在输卵管中两者的受体的分布情况及其基因的差异表达国内外鲜有报道。本试验以山东省优良山羊品种济宁青山羊为研究对象,采用实时荧光定量RT-PCR方法,研究在生后发育过程中在mRNA水平上输卵管中FSHR和LHR基因的表达量;运用免疫组化方法研究生后发育过程中FSHR和LHR的表达及分布情况;运用放射免疫分析法对不同月龄阶段外周血中促卵泡素、促黄体生成素的含量进行测定,主要结果如下:
1.输卵管各个部位在生后发育的整个过程中都有FSHR mRNA存在,其表达的最高峰出现在0月龄,与其它各月龄之间的差异极显著(P<0.01)。生后发育过程中FSHR基因在输卵管中表达量分别为:0月龄13.914±0.75820;1月龄0.5485±0.07874;2月龄0.6109±0.04134;3月龄0.3577±0.04700;4月龄0.5722±0.3268;5月龄7.5944±0.63868;6月龄4.5153±0.72269。
2.输卵管各个部位在生后发育的整个过程中都有LHR mRNA的存在,其表达的最高峰出现在5月龄,与其它各月龄之间的差异极显著(P<0.01),生后发育过程中LHR基因在输卵管中表达量分别为:0月龄2.2441±0.24477;1月龄1.3824±0.14876;2月龄1.3990±0.14066;3月龄0.4235±0.05103;4月龄0.2560±0.3788;5月龄7.5934±0.53422;6月龄0.9044±0.8944。
3.0月龄山羊输卵管中并没有发现FSHR和LHR免疫阳性细胞,1—6月龄,输卵管中均能见到FSHR和LHR免疫阳性细胞。
4.外周血中FSH的含量各月龄之间差异不显著(P>0.05),1-2月龄时逐渐升高,至3月龄含量最多(1.58±0.52),而后4-6月龄逐渐下降,6月龄含量最低(1.28±0.27)。试验结果显示2、4、5月龄含量基本一致,略高于0、1、6月龄。
5.外周血中LH的含量在刚出生时最多(2.49±0.10),此后即迅速下降,在2月龄时最低(1.19±0.06),3月龄时出现一个峰值(2.32±0.37)。5月龄2.23±0.06略高于4、6月的含量。除2月龄外,各月龄之间差异不显著(P>0.05)。
Follicle-stimulating hormone (FSH) and Luteinizing hormone (LH) are two very important glycoprotein hormone secreted by the anterior pituitary, which play an important role in the gonadal development and reproductive modulation through respective specific receptor FSHR and LHR. The biological effects of FSH is mainly to promote the growth of ovarian and endometrial, stimulate follicle growing、developing, and stimulate LH receptors on the follicle cells; the biological effects of LH is mainly to promote follicular maturation and ovulation, stimulate thecal cells produce estrogen. Traditional view is that FSH, LH play a role on the gonad only, but recent studies found that, in extragonadal reproductive tissues, there are also two hormone receptors, suggesting that FSH, LH may also play a role outside the gonads organization, both the hormone’s secretion and their receptor’s distribution and the expression in the oviduct during the course of postnatal changes are rarely reported at home and abroad. In this experiment, the Jining Grey goats in Shandong Province as the research object, using the method of real time quantitative PCR to study FSHR and LHR gene expression in the oviduct during postnatal development in the mRNA level ;Using immunohistochemistry to test expression and distribution of FSHR and LHR during postnatal development;Using radioimmunoassay to measure follicle stimulating hormone, luteinizing hormone levels in peripheral serum of different age groups, the main results are as follows:
1. The presence of FSHR mRNA can be detected in all parts of fallopian tube after birth and during postnatal development priod, the expression peak appeared at 0 months of age, and it is very significant from the other goups(P <0.01). During postnatal development , the FSHR gene expression in the fallopian tube ratio is as follows: 0-month old group 13.914±0.75820, 1-month old group0.5485±0.07874, 2-month old group 0.6109±0.04134, 3-month old group 0.3577±0.04700, 4-month old group 0.5722±0.3268, 5-month old group 7.5944±0.63868, 6-month old group 4.5153±0.72269.
2. The presence of LHR mRNA can be detected in all parts of fallopian tube after birth and during postnatal development priod, the expression peak appeared at 5 months of age, and it is very significant from the other goups(P <0.01). During postnatal development , the FSHR gene expression in the fallopian tube ratio is as follows: 0-month old group 2.2441±0.24477, 1-month old group 1.3824±0.14876, 2-month old group 1.3990±0.14066, 3-month old group 0.4235±0.05103, 4-month old group 0.2560±0.3788, 5-month old group 7.5934±0.53422, 6-month old group 0.9044±0.8944.
3. There are no FSHR and LHR-positive cells in the oviduct in 0 month-old goat, the expression of FSHR and LHR begin from 1 month.
4. The difference of FSH levels in plasma between the the different groups is not significant, 1-2 months groups increased gradually , to 3-month group up to 1.58±0.52 whoes content is highest, and then during 4-6 months the content decreased gradually, in 6 months the content was the lowest 1.28±0.27.
5. LH levels in plasma is highest at birth which is up to 2.49±0.10, and decreased rapidly after that, to 3 months old, it have a peak 2.32±0.37,at the age of 2 months ,the content is minimum 1.19±0.06,to 5 months the content is slightly higher than 4,6 months2.23±0.06. In addition to 2 months group, the difference between the the other groups are not significant.
1.输卵管各个部位在生后发育的整个过程中都有FSHR mRNA存在,其表达的最高峰出现在0月龄,与其它各月龄之间的差异极显著(P<0.01)。生后发育过程中FSHR基因在输卵管中表达量分别为:0月龄13.914±0.75820;1月龄0.5485±0.07874;2月龄0.6109±0.04134;3月龄0.3577±0.04700;4月龄0.5722±0.3268;5月龄7.5944±0.63868;6月龄4.5153±0.72269。
2.输卵管各个部位在生后发育的整个过程中都有LHR mRNA的存在,其表达的最高峰出现在5月龄,与其它各月龄之间的差异极显著(P<0.01),生后发育过程中LHR基因在输卵管中表达量分别为:0月龄2.2441±0.24477;1月龄1.3824±0.14876;2月龄1.3990±0.14066;3月龄0.4235±0.05103;4月龄0.2560±0.3788;5月龄7.5934±0.53422;6月龄0.9044±0.8944。
3.0月龄山羊输卵管中并没有发现FSHR和LHR免疫阳性细胞,1—6月龄,输卵管中均能见到FSHR和LHR免疫阳性细胞。
4.外周血中FSH的含量各月龄之间差异不显著(P>0.05),1-2月龄时逐渐升高,至3月龄含量最多(1.58±0.52),而后4-6月龄逐渐下降,6月龄含量最低(1.28±0.27)。试验结果显示2、4、5月龄含量基本一致,略高于0、1、6月龄。
5.外周血中LH的含量在刚出生时最多(2.49±0.10),此后即迅速下降,在2月龄时最低(1.19±0.06),3月龄时出现一个峰值(2.32±0.37)。5月龄2.23±0.06略高于4、6月的含量。除2月龄外,各月龄之间差异不显著(P>0.05)。
Follicle-stimulating hormone (FSH) and Luteinizing hormone (LH) are two very important glycoprotein hormone secreted by the anterior pituitary, which play an important role in the gonadal development and reproductive modulation through respective specific receptor FSHR and LHR. The biological effects of FSH is mainly to promote the growth of ovarian and endometrial, stimulate follicle growing、developing, and stimulate LH receptors on the follicle cells; the biological effects of LH is mainly to promote follicular maturation and ovulation, stimulate thecal cells produce estrogen. Traditional view is that FSH, LH play a role on the gonad only, but recent studies found that, in extragonadal reproductive tissues, there are also two hormone receptors, suggesting that FSH, LH may also play a role outside the gonads organization, both the hormone’s secretion and their receptor’s distribution and the expression in the oviduct during the course of postnatal changes are rarely reported at home and abroad. In this experiment, the Jining Grey goats in Shandong Province as the research object, using the method of real time quantitative PCR to study FSHR and LHR gene expression in the oviduct during postnatal development in the mRNA level ;Using immunohistochemistry to test expression and distribution of FSHR and LHR during postnatal development;Using radioimmunoassay to measure follicle stimulating hormone, luteinizing hormone levels in peripheral serum of different age groups, the main results are as follows:
1. The presence of FSHR mRNA can be detected in all parts of fallopian tube after birth and during postnatal development priod, the expression peak appeared at 0 months of age, and it is very significant from the other goups(P <0.01). During postnatal development , the FSHR gene expression in the fallopian tube ratio is as follows: 0-month old group 13.914±0.75820, 1-month old group0.5485±0.07874, 2-month old group 0.6109±0.04134, 3-month old group 0.3577±0.04700, 4-month old group 0.5722±0.3268, 5-month old group 7.5944±0.63868, 6-month old group 4.5153±0.72269.
2. The presence of LHR mRNA can be detected in all parts of fallopian tube after birth and during postnatal development priod, the expression peak appeared at 5 months of age, and it is very significant from the other goups(P <0.01). During postnatal development , the FSHR gene expression in the fallopian tube ratio is as follows: 0-month old group 2.2441±0.24477, 1-month old group 1.3824±0.14876, 2-month old group 1.3990±0.14066, 3-month old group 0.4235±0.05103, 4-month old group 0.2560±0.3788, 5-month old group 7.5934±0.53422, 6-month old group 0.9044±0.8944.
3. There are no FSHR and LHR-positive cells in the oviduct in 0 month-old goat, the expression of FSHR and LHR begin from 1 month.
4. The difference of FSH levels in plasma between the the different groups is not significant, 1-2 months groups increased gradually , to 3-month group up to 1.58±0.52 whoes content is highest, and then during 4-6 months the content decreased gradually, in 6 months the content was the lowest 1.28±0.27.
5. LH levels in plasma is highest at birth which is up to 2.49±0.10, and decreased rapidly after that, to 3 months old, it have a peak 2.32±0.37,at the age of 2 months ,the content is minimum 1.19±0.06,to 5 months the content is slightly higher than 4,6 months2.23±0.06. In addition to 2 months group, the difference between the the other groups are not significant.
引文
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