调控小鼠肾脏功能的相关中枢核团神经解剖学研究

发布时间：2018-04-30 18:16

本文选题：肾脏 + 病毒示踪技术　；参考：《华中科技大学》2012年博士论文

【摘要】：近年来随着有生物活性的能够进行多突触传递的嗜神经示踪病毒的发现、改造与应用,有关调控内脏功能的中枢核团定位和功能认识得以深化,相关中枢核团之间的神经解剖学通路变得更加明晰。本研究运用伪狂犬病毒(pseudorabies virus, PRV)转突触示踪技术和荧光标记技术试图确定中枢神经系统参与调控小鼠肾脏功能的相关神经核团,以及这些核团之间的神经解剖学投射通路。研究方法与结果方法：将嗜神经示踪病毒PRV-614注入19只成年雄性C57BL/6J小鼠的左侧肾脏上极,按照动物的不同生存期限无痛处死后制备冰冻切片,用荧光免疫组织化学技术检测脊髓、脑干(延髓和中脑)中PRV-614感染神经元、色氨酸羟化酶(TPH)和酪氨酸羟化酶(TH)阳性神经元的表达。结果：1.PRV-614经小鼠肾脏注射后第3d,少量的PRV-614感染神经元逆行示踪标记在脊髓T9的IML区。2.注射PRV-614后第4d,少量的PRV-614感染神经元发现在脊髓T8-10的IML区、IC区和CAN区,以及延髓头端腹外侧核。3.注射PRV-614后第5d,较多的PRV-614感染神经元发现在脊髓T7-12的IML区、IC区和CAN区等,延髓头端腹内侧核、延髓头端腹外侧核、延髓中缝核、孤束核、A5区域、蓝斑核及亚蓝斑核等；较少的PRV-614感染神经元发现在中脑PPTg和LDTg区等。4.注射PRV-614后第6d,在脊髓几乎未发现PRV-614感染神经元。较多的PRV-614感染神经元发现在延髓头端腹内侧核、延髓头端腹外侧核、延髓中缝核、A5区域、蓝斑核及亚蓝斑核等,以及中脑PPTg和LDTg区。其中PRV-614感染神经元主要发现在LDTg头端和中间部位,以及PPTg中部和尾部。PRV-614/TPH双重标记神经元主要分布在LDTg头端,而PRV-614/TH神经元分散在LDTg中在3个部位。PRV-614/TPH和PRV-614/TH神经元主要位于PPTg (cPPTg)尾侧。研究总结1.从小鼠肾脏注射的PRV-614经突触逆行传递到达中枢核团具有时空表达特异性。2.主要调节肾脏交感神经活性的脊髓节段范围可能是T8-10,其中T9尤为重要。3.延髓头端腹外侧核是主要调节肾脏TH阳性交感神经活性的中枢核团,而延髓中缝核是主要调节肾脏TPH阳性交感神经活性的中枢核团。4.脑桥中A5区域、蓝斑核及亚蓝斑核是主要调节肾脏TH阳性交感神经活性的中枢核团。5. LDTg头部和PPTg尾部均参与肾脏TH/TPH阳性交感神经活性的调节。
[Abstract]:In recent years, with the discovery, modification and application of bioactive eosinophilic tracer virus capable of multi-synaptic transmission, the localization and functional understanding of central nuclei regulating visceral function have been deepened. The neuroanatomic pathways between the relevant central nuclei became clearer. In this study, pseudorabies virus (PRV) synaptic tracer technique and fluorescence labeling technique were used to identify the central nervous system involved in the regulation of renal function in mice, as well as the neuroanatomical projection pathway between these nuclei. Methods and results: a total of 19 adult male C57BL/6J mice were injected with eosinophilic tracer virus (PRV-614) into the upper pole of the left kidney. Frozen sections were prepared after painless death according to the different survival periods of the animals. The spinal cord was detected by fluorescence immunohistochemical technique. Expression of PRV-614 positive neurons, tryptophan hydroxylase and tyrosine hydroxylase in the brain stem (medulla oblongata and midbrain). Results 1. On the 3rd day after injection of PRV-614 into the kidney of mice, a small number of PRV-614 infected neurons were retrogradely labeled in the IML region of spinal cord T9. On the 4th day after PRV-614 injection, a small number of PRV-614 infected neurons were found in the IML and CAN regions of the T8-10 spinal cord, as well as in the ventrolateral nucleus ventrolateral of the medulla oblongata. On the 5th day after PRV-614 injection, more PRV-614 infected neurons were found in the IML and CAN regions of T7-12 spinal cord, the ventromedial nucleus of the medulla oblongata, the ventrolateral nucleus of the medulla oblongata, the nucleus raphe nucleus of the medulla oblongata, the A5 region of the nucleus tractus solitarius, the nucleus coeruleus and the nucleus coeruleus. Fewer PRV-614 infected neurons were found in the PPTg and LDTg regions of the midbrain. On the 6th day after PRV-614 injection, almost no PRV-614 infected neurons were found in the spinal cord. More PRV-614 infected neurons were found in the ventromedial nucleus of the medulla oblongata, the ventrolateral nucleus of the medulla oblongata, the A5 area of the nucleus raphe nucleus of the medulla oblongata, the nucleus coeruleus and the sublocus coeruleus, and the PPTg and LDTg areas of the midbrain. The PRV-614 infection neurons were mainly located in the head and middle part of LDTg, and in the middle and tail of PPTg. The double labeled neurons were mainly located in the LDTg head, while the PRV-614/TH neurons were scattered in the three sites of LDTg. The PRV-614/TH neurons were mainly located in the caudal side of PPTg cPPTg. Research summary 1. The PRV-614 injected from the kidney of mice reached the central nucleus via synaptic retrograde transmission with spatiotemporal expression specificity. 2. The range of spinal cord segments which mainly regulate renal sympathetic nerve activity may be T 8-10, of which T 9 is especially important. 3. The ventrolateral nucleus of the medulla oblongata is the central nucleus which mainly regulates the activity of the th positive sympathetic nerve in the kidney, while the nucleus raphe nucleus of the medulla oblongata is the central nucleus which mainly regulates the sympathetic activity of the kidney TPH positive. The A5 region of the pons, the nucleus coeruleus and the sublocus coeruleus nucleus are the central nuclei which mainly regulate the sympathetic activity of the renal th positive neurons. Both LDTg head and PPTg tail are involved in the regulation of renal TH/TPH positive sympathetic nerve activity.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R322

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