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動物學

果蠅緯度相關晝夜節律特徵:穩定性、活動量分佈與演化意義 Latitude-Dependent Circadian Traits in Drosophila: Stability, Activity Peaks, and Evolutionary Implications

生物時鐘可對生物體的行為與生理造成影響,在探討晝夜節律特徵的差異時,過去研究常侷限於北美大陸的品系,缺少赤道及南半球品系的晝夜節律特徵探討。有鑒於黑腹果蠅在全球各大洲的廣泛分佈,因此我們以黑腹果蠅(近赤道與中高緯度品系)為材料,研究果蠅是否因緯度而有相異的晝夜節律特徵?結果顯示不同緯度的果蠅品系展現出相異的晝夜節律特徵。赤報品系在全暗狀態下仍維持原本光暗12小時的穩定節律,而南北半球的中高緯度品系則具有相似節律特徵,即在全暗狀態下的節律不對齊原本正常光源的穩定節律,其他如活動量、週期、及節律強度等皆有著品系間的差異。更進一步比對實驗中各個品系基因序列,研究發現per和tim在調控區段有許多SNP變異,顯示其與晝夜節律特徵的關係,有助於後續尋找更多造成晝夜節律特徵差異的可能遺傳變異並探討。

Straw manufacturing for epidemic prevention

Recently,thefeedpricesforlivestockfarmshavebeencontinuouslyincreasing, while the prices of calves have been declining, leading to many livestock and dairy farms facing financial losses. If livestock farms are affected by diseases such as foot-and-mouth disease, they suffer significant losses. This is because reducedproductivityinlivestocknotonlyaffectstheirproductionbutalsoentails substantial costs for vaccines and treatments. Therefore, there is a need to explore how to effectively prevent diseases, focusing on common diseases in cattle such as subclinical mastitis and mastitis, and alleviate the burden on farms economically and environmentally. This study aims to investigate the production of bacterial growth-inhibiting straw bedding using substances such as illite and charcoal to contribute to disease prevention.

利用粒線體UQCRC1基因缺失之巴金森氏症果蠅模式探討細胞自噬作用對於神經系統退化之影響

巴金森氏症是最常見的神經退化性疾病之一,目前尚無根治之法。研究已知,粒線體如果出現問題,會加速神經細胞退化。細胞自噬作用可協助清除功能不佳的粒線體,以維持神經細胞運作。電子傳遞鏈第三複合體核心中的UQCRC1 (Ubiquinol-Cytochrome C Reductase Core Protein 1) 基因點突變會導致巴金森氏症。然而,細胞自噬作用對於UQCRC1引起神經退化影響的仍不清楚。本研究中,在果蠅神經系統以RNA干擾方式降低UQCRC1表現量,建立巴金森氏症模式;而後利用不同藥物分別抑制或促進細胞自噬作用,觀察果蠅爬行能力;同時,利用MARCM遺傳工具,使果蠅組織細胞分為UQCRC1缺失和正常細胞兩群,觀察細胞自噬相關的蛋白表現量在兩群細胞間的差異。研究結果顯示細胞自噬對UQCRC1參與的粒線體功能缺損的神經退化有代償作用。

Whose feather is that? A cross-views between a naturalist and a molecular biologist

Identifyingthespeciesorsexofabirdbasedonafeatherfoundinnatureisoftenchallenging,evenwith the help of reference books. However, determining the presence of a rare species in a habitat using an indirectpresenceindicator,suchasafeather,canhelpinimplementingspecificmeasuresforpreserving the species. The aim of this study is to investigate whether DNAgenotyping is better than specialized books when identifying bird feathers. Toanswerthisquestion,Icollectedfeathersinthewildand,withthehelpoftwobooks,triedtoidentify theirspeciesandsex.Then,assistedbyDrGwenaëlJacob(UNIFR),Iisolatedtwogenesinnineselected feathers. The investigated genes were the CHD gene for sexing and the COI gene for species identification.Todothis,theDNAwasfirstextractedfromthefeathers,purified,andamplifiedbyPCR. Subsequently,anelectrophoresis wasperformedtosexthe samples andcheckthatthe PCRamplification hadworkedproperly.Finally,thesamplesweresequencedbytheMicrosynthlaboratory(St-Gall),and the obtained sequences were entered into the NCBI database. Acomparisonoftheresultsobtainedwitheachofthetwodifferentmethodsshowsthattheidentification with specialized books was fairly successful. 56% of the species identification made with the books were indeed confirmed by genotyping. DNAanalysis provided a different result only for feather #16. However,33%ofgeneticidentificationfailed,eitherduetogeneticmaterialqualityorlaboratoryerrors. Asitwaspossibletoidentifythesexofonlyonesample(feather#14)withthebooks,itwasnotpossible tomakeatruecomparisonofthetwoapproaches.However,asgeneticsexingworkedwell(onefailure, feather #28), it can be inferred that genetic sexing is more effective than using books. This work demonstrated that DNAis not infallible and that sometimes books are equally effective in identifyingbirdspeciesfromafeather.However,insexingbird,DNAremainsmoreefficient.Thus,one can conclude that DNAgenotyping is not superior but rather complementary to specialized books for identifying bird feathers.

短期睡眠剝奪對小鼠免疫系統的影響

現代社會中,睡眠剝奪已成為普遍問題,人們對其對免疫系統及整體健康的負面影響愈加關注。本研究使用特製的旋轉鼠籠讓小鼠連續72小時保持清醒,探討急性睡眠剝奪對小鼠免疫反應的影響。研究發現NK細胞與脾臟中的記憶CD8 T細胞比例明顯減少,顯示細胞毒性功能受損或記憶免疫反應下降。與此同時,抗炎細胞因子的表達增加,而促炎細胞因子和相關基因的表達則有顯著下調。此外,雖然觀察到B細胞比例有所增加,這可能是免疫系統在細胞免疫功能受損時,維持免疫穩態的反應。這些發現揭示了睡眠剝奪可能抑制免疫系統造成損害。本研究強調適量睡眠對維持免疫平衡的重要性,並指出睡眠不足可能促進慢性免疫問題的發展。在此基礎上,後續研究可探討短期睡眠剝奪與腫瘤及免疫系統的關聯,並延伸至長期剝奪的影響。

Non-invasive study of the electrical activity of the brain of various chordate animals

In clinical practice, EEG is used to diagnose a number of neurological diseases and to diagnose epilepsy. But at present, the question of the nature of EEG has not been completely resolved and is of great scientific interest. There have been no studies at all on the non-invasive study of the electrical activity of the brain of the shark superorder, which belongs to the class of cartilaginous fish. By studying the electrical activity of the brain of various gnathostomes, it is possible to obtain an answer to the question of the emergence of rhythms from the point of view of phylogenesis and evolution, and by comparing their EEG with the human EEG, one can identify similar patterns that help in the study of reactions to various influences. During the work, for the first time, EEG indicators of spotted cat sharks, ECG, heart rate and respiratory rate of cat sharks and toads were obtained. In the future, it is planned to assemble a smaller neuroheadset for non-invasive studies of the electrical activity of the brain of small animals (sharks, toads, monitor lizards). This data can be used for evolutionary and medical research. *No animals were harmed during or after the experiments.

短期睡眠剝奪對小鼠免疫系統的影響

現代社會中,睡眠剝奪已成為普遍問題,人們對其對免疫系統及整體健康的負面影響愈加關注。本研究使用特製的旋轉鼠籠讓小鼠連續72小時保持清醒,探討急性睡眠剝奪對小鼠免疫反應的影響。研究發現NK細胞與脾臟中的記憶CD8 T細胞比例明顯減少,顯示細胞毒性功能受損或記憶免疫反應下降。與此同時,抗炎細胞因子的表達增加,而促炎細胞因子和相關基因的表達則有顯著下調。此外,雖然觀察到B細胞比例有所增加,這可能是免疫系統在細胞免疫功能受損時,維持免疫穩態的反應。這些發現揭示了睡眠剝奪可能抑制免疫系統造成損害。本研究強調適量睡眠對維持免疫平衡的重要性,並指出睡眠不足可能促進慢性免疫問題的發展。在此基礎上,後續研究可探討短期睡眠剝奪與腫瘤及免疫系統的關聯,並延伸至長期剝奪的影響。

探討在秀麗隱桿線蟲中IFE-1經由sRNA路徑對於精子生成機制的影響

sRNA在各種物種的精子功能中起著至關重要的作用。在秀麗隱桿線中,當缺少精子相關的sRNA「ALG-3/4 26G sRNA」會導致其在25度時不孕。此外,IFE-1是人類真核轉譯起始因子EIF4E的直系同源基因,主要表達於雄性生殖細胞系統中。在先前研究中我們觀察到當「真核轉譯起始因子IFE-1有缺陷」或「精子缺少相關sRNA」時,亦會導致精子具有缺陷。由於三者的相似性,我們認為IFE-1和26G sRNA的生成路徑有關。因此我們假設IFE-1參與協助酵素NYN-3辨認並切割msd-1 mRNA模板後促進26G sRNA生成。我們使用Western Blot、IP、螢光顯微鏡等方法,探討了IFE-1和MSD-1::GFP的關係,發現在ife-1正常的情況下,高溫對於MSD-1::GFP的表現量沒有影響。並且因該蛋白只表現在公蟲精子,我們可以推論msd-1:gfp 只作用於公蟲精子。而此疑似可正向調控基因表現的26G sRNA,有望發展成有別於過往sRNA藥物抑制基因表現的一種新基因治療方法。

探討果蠅神經膠細胞核的遷移機制

細胞核是真核細胞內最大且至關重要的細胞結構之一。其具體位置在各種細胞中可能有所不同。為了深入了解細胞核位置對於細胞功能的影響,我們選擇以果蠅幼蟲眼疊為研究對象,探究細胞核在神經系統發育過程中所扮演的角色。神經系統在生物體中扮演著極為關鍵的角色,包括神經元和神經膠細胞。如果失去神經膠細胞,將導致神經退化或死亡。在我們的研究中發現,神經膠細胞核在其發育過程中會發生大規模的內部移動。為了限制神經膠細胞核的移動,我們利用了果蠅作為研究動物,並應用了果蠅常用的 GAL4-UAS系統和GrabFP技術,這使得我們能夠限制神經膠細胞內細胞核的移動。我們的實驗成功證明,限制神經膠細胞核的移動會影響神經細胞的軸突發育,但不會影響神經細胞 R1-R8 聚叢的發育。未來,我們計劃將 GrabFP 技術應用於研究不同胞器在細胞內相對位置對其功能的影響。

探討在秀麗隱桿線蟲中IFE-1經由sRNA路徑對於精子生成機制的影響

sRNA在各種物種的精子功能中起著至關重要的作用。在秀麗隱桿線中,當缺少精子相關的sRNA「ALG-3/4 26G sRNA」會導致其在25度時不孕。此外,IFE-1是人類真核轉譯起始因子EIF4E的直系同源基因,主要表達於雄性生殖細胞系統中。在先前研究中我們觀察到當「真核轉譯起始因子IFE-1有缺陷」或「精子缺少相關sRNA」時,亦會導致精子具有缺陷。由於三者的相似性,我們認為IFE-1和26G sRNA的生成路徑有關。因此我們假設IFE-1參與協助酵素NYN-3辨認並切割msd-1 mRNA模板後促進26G sRNA生成。我們使用Western Blot、IP、螢光顯微鏡等方法,探討了IFE-1和MSD-1::GFP的關係,發現在ife-1正常的情況下,高溫對於MSD-1::GFP的表現量沒有影響。並且因該蛋白只表現在公蟲精子,我們可以推論msd-1:gfp 只作用於公蟲精子。而此疑似可正向調控基因表現的26G sRNA,有望發展成有別於過往sRNA藥物抑制基因表現的一種新基因治療方法。