根據過去的研究已知陽氯離子共同運輸蛋白中的sodium-potassium-chloride co-transporter 1(NKCC1)和potassium-chloride co-transporter 2(KCC2)，會造成氯離子內外濃度分布不同，使得成熟大腦的GABA神經傳導為抑制性(inhibition)的反應，呈現過極化(hyperpolarization)的現象；而尚未發展成熟的大腦，則為去極化(depolarizes)現象，導致GABA傳導為興奮性(excitation)。且過去研究已證實，調控此兩共同運輸蛋白的上游因子可能為STE20 (sterile 20)/SPS1-related proline/alanine-rich kinase (SPAK)。此外，亦有文獻指出在思覺失調症(Schizophrenia)患者的海馬迴相較於正常人，可發現NKCC1的表現較KCC2高，因此，本研究欲探討SPAK對於精神疾病的行為及NKCC1和KCC2表現的影響。實驗方法乃利用孤立養育模式(Isolation rearing)建立類精神疾病之動物模式，並以相關的行為試驗測定，最後再採取前皮質額葉(Prefrontal cortex)及海馬迴(Hippocampus)進行生化分析，藉此與一般社交養育模式(Social rearing)的小鼠作一對照。結果發現不論是否經過孤立養育的過程，SPAK-/-小鼠在感覺運動門閥功能、新奇事物認知中，皆表現得比WT好，且較不焦慮；而在生化分析部分，社交養育的SPAK-/-小鼠在前皮質額葉的KCC2 表現高於NKCC1，而WT小鼠則無顯著改變。因此，根據上述的結果可推論SPAK基因，對於精神疾病所產生的異常行為表現及NKCC1和KCC2的蛋白質表現，具有影響力。

SPAK (Sterile 20/SPS1-related proline/alanine-rich kinase) is a protein kinase belonging to the superfamily of mitogen-activated protein kinase (MAPK) and has been found extensively distributed across the body. The SPAK associated variations of NKCC1 and KCC2 in the central nervous system are important in the interpretation of developmental mental dysfunctions. The present study aimed to elucidate the role of SPAK-NKCC1/KCC2 in schizophrenia with a rodent schizophrenia-like model in which the mouse paradigm of isolation rearing (IR) was employed as it simulates the sensorimotor gating abnormalities of schizophrenia. SPAK transgenic mice were used and divided to four groups: social-wild type, social-SPAK-/-, isolation-wild type, isolation-SPAK-/-. Prepulse inhibition (PPI) following an acoustic startle and novel object recognition test (NORT) were performed to index the schizophrenia-associated dysfunctions of gating ability and the novelty recognition. Finally, protein expressions of NKCC1/KCC2 in prefrontal cortex and hippocampus were detected to collaborate with the behavioral data. Our results demonstrated that SPAK KO mice had a superior PPI and novelty recognition than the wild type controls, with a concomitant increase of KCC2 in the prefrontal cortex. IR sabotaged the performances in PPI and NORT with associated increases of NKCC1 and KCC2. Further, the effects of SPAK KO on PPI and NORT were distinctively modulated by IR, as it preserved the genetic effects on NORT but nullified those effects on PPI, and reversed the SPAK KO-increased KCC2 in prefrontal cortex. Our data supported the gene-environment hypothesis and demonstrated that the manipulation of SPAK gene might be useful in schizophrenia study.

目錄 I
圖表目錄 III
中文摘要 IV
ABSTRACT V
中英文簡寫對照表 VII

第一章 序言 1
第一節 孤立養育動物模式 1
第二節 STE20 (STERILE 20)/SPS1-RELATED PROLINE/ALANINE-RICH KINASE , SPAK 4
第三節 陽氯離子(CATION CHLORIDE COTRANSPTOR, CCCS) 5
第四節 GABA對神經生理的影響 7

第二章 研究目的與實驗設計 9

第三章 材料與方法 11
第一節 實驗動物 11
第二節 基因篩選(GENOTYPING) 11
第三節 西方點墨分析法(WESTERN BLOT) 13
第四節 動向行為測試（LOCOMOTOR ACTIVITY） 15
第五節 前脈衝抑制(PREPULSE INHIBITION)及驚嚇反應(STARTLE RESPONSE)測試 15
第六節 高腳T型迷宮( ELEVATED T-MAZE, ETM) 17
第七節 新奇事物認知測試(NOVEL OBJECT RECOGNITION TEST, NORT) 18
第八節 統計方法 19

第四章 實驗結果 21
第一節 由SPAK+/-小鼠配對所繁衍出的後代基因型表現 21
第二節 感覺運動門閥功能(SENSORIMOTOR GATING FUNCTION)的相關測定 21
第三節 新奇事物認知記憶及好奇心相關測試 23
第四節 似焦慮表現相關測定 24
第五節 活動動向力測試(LOCOMOTOR ACTIVITY) 26
第六節 NKCC1和KCC2在前額葉皮質 (PFC)蛋白質的表現 26

第五章 討論 27
第一節 感覺運動門閥功能(SENSORIMOTOR GATING FUNCTION)的相關測定 27
第二節 新奇事物認知記憶及好奇心相關測試 32
第三節 焦慮表現相關測定 35
第四節 活動動向力測試(LOCOMOTOR ACTIVITY) 38
第五節 NKCC1和KCC2在前額葉皮質 (PFC)蛋白質的表現 39

第六章 結論 41

附圖 42
參考文獻 53

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