On the Relation to Genetical Factors of Strain-Difference of Anesthetic Sensitivities and Brain-distribution of Neurotransmitters in ddN and C57BL mic

ddN和C57BL麦克风麻醉敏感性应变差异和神经递质脑分布与遗传因素的关系

• 批准号: 06454445
• 负责人: OGLI Kenji
• 金额: $ 4.16万
• 依托单位: Kagawa Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454445/
• 关键词: anesthetic sensitivity; loss of righting reflex; multifactorial inheritance; serotonin; metencephalin; neuropeptide Y; dopamine; glutamate receptor; 麻酔薬抵抗性; 遺伝形式; 神経伝達物質; ドーパミン分泌細胞; 正向反射; 母性因子; x染色体上因子; ニューロペプチドY

1) On the genetical analysis of anesthetic sensitivities : ED50s of Isoflurane (ISO) and Enflurane (ENF) were estimated as an index of loss of righting-reflex in ddN and C57BL mice, mutual hybrid mice and their backcross progenies. The results are as follows ;(1) There is the difference of ENF and ISO sensitivities in ddN and C57BL mice. C57BL mice is more sensitive than ddN mice. This difference decreases into 1.18 - 1.26 times compared with the value of 1.38 times in 1990. A genetical factors related to anesthetic resistance in ddN mice may be being lost by means of the continuation of inbreeding.(2) Multivariate regression analysis of parents' and F1 hybrids'data revealed that, while maternal factors and factors on autosomes were related to both anesthetics, the factors on the X chromosome were ENF specific. A wide variation i anesthetic requirements among backcross siblings progeny suggested multifacorial inheritance. The regression equations pbtained did not always predict anesthe … More tic requirement in backcross siblings progeny. These discrepancies may be due to the epistatic interaction of related genes. We conclude that multiple but different genetic factors are involved in determining ENF and ISO anesthetic requirements in ddN and C57BL mice.2) Immunohistochemical analysis of brain neurotransmitters and receptors in ddN,C57BL and their hybrid (F1) mice.(1) Serotonin : The density of serotonin-immuno reactive nerve fibers is ddN < C57BL in hippocampus CA1, CA2 and CA3, ddN > C57BL in dentate gyrus of hippocampus.(2) Metencephalin : The immunoreactivity of mossy fiber of hippocampus is ddN > F1'> F1 > C57BL.(3) Neuropeptide Y : The immunoreactive nerve cell is ddN < C57BL in parietotemmporal cortex and the immunoreactivity is 2 x ddN,F1, F1' < C57BL in suprachiasmatic nucleus, F1, F1' < ddN < C57BL in paraventricular nucleus, F1 < F1' and ddN < C57BL in spinal trigeminal nucleus, ddN < C57BL in cervical dorsal nucleus.(4) Dopamine : The nigrostriatal dopaminergic pathway is more abundant in ddN than C57BL.The number of the dopaminergic neurons in the origin (substantia nigra) of ddN is about 30% larger than that of C57BL.The area of the terminal field (striatum) of ddN is larger than C57BL,and the staining intensity of tyrosine-hydroxylase immunoreactive structure, which can be interpreted as the density of the terminal, is higher in ddN than that of C57BL.(5) Glutamate receptor : The localization of AMPA-selective gluamate receptor were examined using the antiserum made against the subunits : GluR1, GluR2/3, and GluR4. In the hippocampal CA1 and dentate gyrus, the distribution of GluR1 and GluR2/3 of C57BL are more dense than that of ddN.The staining intensity against the GluR2/3 antiserum in the parietal cerebral cortex is higher in C57BL than ddN.(The image analysis is in progress.) Less

1)麻醉敏感性的遗传分析：估计异氟醚（ISO）和恩氟醚（ENF）的ed50作为ddN和C57BL小鼠、互杂交小鼠及其回交后代右转反射丧失的指标。研究结果如下：(1) ddN和C57BL小鼠ENF和ISO敏感性存在差异。C57BL小鼠比ddN小鼠更敏感。这一差距从1990年的1.38倍下降到1.18 ~ 1.26倍。一种与ddN小鼠麻醉抗性相关的遗传因子可能会随着近亲繁殖的继续而丢失。(2)对亲本和F1杂交的数据进行多元回归分析发现，母体因素和常染色体上的因素与两种麻醉药均相关，而X染色体上的因素则是ENF特异性的。在回交的兄弟姐妹后代中，麻醉需求的广泛差异表明多重遗传。所建立的回归方程并不总能预测回交兄弟后代的需要量。这些差异可能是由于相关基因的上位性相互作用。我们得出结论，多种但不同的遗传因素参与了ddN和C57BL小鼠ENF和ISO麻醉需求的决定。2) ddN、C57BL及其杂交（F1）小鼠脑神经递质和受体的免疫组化分析。(1)血清素：海马CA1、CA2、CA3区血清素免疫反应性神经纤维密度ddN < C57BL，海马齿状回ddN > C57BL。(2) Metencephalin：海马苔藓纤维的免疫反应性为ddN > F1'> F1 > C57BL。(3)神经肽Y：顶叶颞叶皮层免疫反应性神经细胞为ddN < C57BL，视交叉上核为2倍ddN、F1、F1′< C57BL，室旁核为F1、F1′< ddN < C57BL，脊髓三叉核为F1 < F1′、ddN < C57BL，颈背核为ddN < C57BL。(4)多巴胺：ddN中黑质纹状体多巴胺能通路比C57BL更丰富。ddN起源（黑质）的多巴胺能神经元数量比C57BL大30%左右。ddN的末端场（纹状体）面积比C57BL大，酪氨酸-羟化酶免疫反应结构的染色强度也比C57BL高，这可以解释为末端密度。(5)谷氨酸受体：用针对GluR1、GluR2/3、GluR4亚基的抗血清检测ampa选择性谷氨酸受体的定位。在海马CA1和齿状回中，C57BL的GluR1和GluR2/3的分布比ddN更密集。C57BL对顶叶皮层GluR2/3抗血清的染色强度高于ddN。（图像分析正在进行中。）少

项目成果

S.Gamo, K.Morioka and K.Dodo: "Studies on ether anesthesia mutations in Drosophila melanogaster" Progress in Anesthetic Mechanism. 3(suppl.). 120-124 (1995)

S.Gamo、K.Morioka 和 K.Dodo：“黑腹果蝇乙醚麻醉突变的研究”麻醉机制进展。

小栗顕二: "“麻酔薬の作用機序"の論文をどう読むか(その2)" 臨床麻酔. 19(5). 677-684 (1995)

Kenji Oguri：“如何阅读麻醉剂作用机制的论文（第 2 部分）”《临床麻醉》19(5)。

J.Nogaya,H.Komatsu,N.Kuratani,K.Ogli and M.Kawata: "Circadian rhythmicity of open field activity and the neuropeptide Y in the suprachiasmatic nucleus." Abstract of Fourth IBRO World Congress of Neuroscience. D11. 46 (1995)

J.Nogaya、H.Komatsu、N.Kuratani、K.Ogli 和 M.Kawata：“开放场活动的昼夜节律和视交叉上核中的神经肽 Y”。

H.komatsu, J.Nogaya and K.Ogli: "Volatile Anaesthetics as Central Nervous System Excitants" Ann Acad Med Singapore. 23(Suppl). 130-138 (1994)

H.komatsu、J.Nogaya 和 K.Ogli：“挥发性麻醉剂作为中枢神经系统兴奋剂”新加坡 Ann Acad Med。

H.Komatsu, J.Nogaya, M.Ueki, S.Yokono and K.Ogli: "Possible Participation of NMDA and Glycine Receptors but not GABA-A Receptors in Enflurane-induced Opisthotonus in Mice" Clin Exp Pharmacol Physiol. 21. 495-499 (1994)

H.Komatsu、J.Nogaya、M.Ueki、S.Yokono 和 K.Ogli：“NMDA 和甘氨酸受体而非 GABA-A 受体可能参与安氟烷诱导的小鼠角弓反张”Clin Exp Pharmacol Physiol。