全國中小學科展

動物學

探討中型金蛛(Argiope aetheroides)與長圓金蛛(Argiope aemula)隱帶功能之研究

本次研究主要探討中型及長圓金蛛蛛網隱帶的結構不同,所造成的振動模式差異。蜘蛛網面的結構會直接影響到蛛網結構的振動方式,以及能量散布的情形。為了了解蜘蛛網有無隱帶結構對於獵物衝擊網面的影響,使用釣魚線及蠶絲進行模擬蛛網。實驗結果顯示,不論長圓或是中型金蛛其隱帶的長度與體長成正相關。金蛛隱帶的長度與環境溫度、照度和風速的相關性不高。而在複式顯微鏡下發現金蛛隱帶的構造有疏密之分。仿生金蛛網不論是何種類型隱帶,皆無法減少受模擬風吹吹拂網面的振幅,僅有蠶絲密十字型隱帶能夠有效減少受模擬獵物撞擊網面的振幅。此外,蠶絲仿生網的振動幅度較釣魚線小,穩固性較佳。

初探渦蟲RNAi的作用機制與限制

核糖核酸干擾 (RNA interference, RNAi) 被廣泛應用在以渦蟲為模式生物的相關研究,其中一種進行RNAi方法是利用餵食渦蟲雙股RNA (dsRNA) 的方式達到 RNAi 的目的,但所餵食的dsRNA只在渦蟲腸道中存在餵食後的4天內。dsRNA究竟如何從渦蟲腸道進入組織中以及RNAi於渦蟲的詳細作用機轉尚不明確。 tgs-1的表現限定於渦蟲成體幹細胞 (pluripotent stem cells, PSCs) 中的前驅幹細胞— cNeoblasts中,可利用 RT-qPCR (Reverse Transcription-Quantitative Polymerase Chain Reaction) 來定量檢測渦蟲tgs-1基因的表現差異。tgs-1基因的產物是一種膜蛋白,為cNeoblasts的標記 (marker) 之一,因此選擇其作為探討RNAi基因緘默 (gene silencing) 效率的目標基因,未來可以應用於渦蟲幹細胞與再生相關研究。 本研究利用RT-qPCR檢測tgs-1的表現量,探討餵食dsRNA在東亞渦蟲Dugesia japonica的RNAi作用機制與效率。實驗發現,六次的dsRNA餵食比餵食2次或4次有更好的效率、餵食不同長度的dsRNA對RNAi 效率沒有顯著影響。tgs-1的基因緘默作用也會影響其他基因的表現,包括少量增加Ago2的表現量,明顯提升gata的表現量,然而piwi-1表現量明顯降低。除此之外,透過胺基酸序列比對發現渦蟲與線蟲SID-1同源基因具有相當高的相似度。 由實驗結果結論多次dsRNA餵食可提高基因緘默的效率和dsRNA 的 3' 序列在RNAi中的關鍵角色。tgs-1 RNAi也影響Ago2、gata 和piwi-1的表現,這些基因與RNAi的機制和dsRNA的攝入量有關。未來將進一步探討dsRNA是如何通過渦蟲腸道、是否透過SID-1蛋白攜帶進入體腔中、dsRNA在渦蟲體內的切割與作用位置和胞吞作用在RNAi所扮演的角色等問題,相關實驗仍在進行中。 本研究結果將有助於了解餵食渦蟲dsRNA經由渦蟲腸道進入體內細胞引起RNAi現象的機制,除了能比較扁形動物RNAi系統與其他物種,例如哺乳類細胞、果蠅、以及線蟲RNAi的作用差異之外,更能有效應用於於探討基因參與渦蟲再生與體軸形成的發育的研究上。

高基氏體如何在細胞中偏安一隅?

高基氏體為細胞內重要的胞器,可加工修飾蛋白質並運輸胞囊。我們利用螢光染色法觀察三種不同物種的細胞,高基氏體顯著地聚集在細胞一側。細胞分裂時如何平均分配高基氏體給子細胞?實驗利用基因重組轉殖螢光蛋白標記中心體、微管、內質網、粒線體,發現只有中心體與其延伸的微管會與高基氏體顯著地聚集在同一側,而破壞掉中心體延伸出的微管會使高基氏體無法聚集,由此可推測細胞利用中心體延伸的微管來聚集高基氏體。我們還發現細胞分裂時,高基氏體會先分散到細胞質各處,細胞分裂後再聚集到中心體周遭。在拍攝活細胞細胞分裂的實驗中,顯示中心體會藉由微管聚集高基氏體,以重新排列微管的方式來牽動平均分配高基氏體至兩側。本實驗確認高基氏體的位置與微管、中心體移動有高度相關。高基氏體移動並聚集一側有助於建立細胞極性,這與物質運送分泌、細胞爬行皆有關聯。

建立線蚓 (Enchytraeus sp.) RNAi實驗模式並探究Hedgehog基因對其生長與再生之功能

線蚓(Enchytraeus sp.)與蚯蚓同屬於環節動物門,具有很強的再生能力。Hedgehog(Hh)基因在兩側對稱動物中對體節發育有重要的調控作用,而先前研究中已發現線蚓體內的Hh基因序列,所以本研究希望利用調控線蚓體內Hh基因的表現,探討Hh基因在環節動物生長與再生中的功能。 本實驗首次嘗試將RNAi技術應用在線蚓上。將線蚓浸泡在含有Enc-Hh的dsRNA溶液中以操作RNAi。利用免疫螢光染色法發現Hh基因受到抑制後,線蚓新生體節神經的連接不完整,顯示Hh基因與神經發育有相關性。由於線蚓快速的再生速度、飼養容易、構造簡單及身體透明且方便染色觀察,透過線蚓RNAi技術的建立,除了能進一步探討Hh基因功能外,也希望提供再生研究時的另一種選擇方式。

蚊幼蟲與搖蚊幼蟲在水中分布與其血紅蛋白基因表現之關係

蚊蟲呼吸透過氣管系統,本研究假設底棲孑孓與搖蚊幼蟲具備血紅蛋白系統以擷取氧氣。戶外採集8種孑孓和1種搖蚊幼蟲,錄影觀察白腹叢蚊、竹生翠蚊和鹽埕搖蚊幼蟲長時間停留底層;家蚊、斑蚊則反覆上下至水面呼吸。顯微鏡下發現白腹叢蚊幼蟲體壁內側充滿紅點,色澤隨著富氧或缺氧而變化;然二種斑蚊則不明顯。PCR增殖6種蚊蟲和1種搖蚊血紅蛋白基因片段並建構親緣關係樹。定量RT-PCR顯示,白腹叢蚊與鹽埕搖蚊在缺氧條件下,血紅蛋白相對表現量分別增加5倍和150倍,上層埃及斑蚊和白線斑蚊則增加3.9倍和1.5倍。西方墨點法證實孑孓缺氧條件下,17-kDa血紅蛋白皆有大量表現。本研究首次確定6種蚊蟲具血紅蛋白基因,證明孑孓血紅蛋白基因在缺氧下的表現量增加,包括mRNA與蛋白質。此等反應氣候變遷,暖化導致水溫升高與低溶氧條件下,蚊蟲可能有因應環境改變的呼吸機制。未來將進一步標幟血紅蛋白在蚊幼蟲體內組織之表現與其他環境變因之影響。

The Reproduction success of the Cyprinidae and a Claridae fish species and its impact on small- scale fisheries

To investigate the reproduction success and natural recruitment of several Cyprinidae fish and Claridae fish species in the Allemanskraal Dam. The purpose of the project included investigating whether each individual fish species studied has a successful 2020/2021 spawning season in comparison with each other. Sections of the seine net were measured along with a distance of 10 along the shoreline. The ends of the seine nets were attached to one foot and the top of the net was held by hand. Both volunteers moved in unison while covering the 10m. The volunteer in the “deep end” moved towards the shoreline creating a semi-circle while the other volunteer remained stationary. The two ends of the net were then pulled onto the shore and the fish were collected. The results found that the Labeo Umbratus and Cyprinus carpio had the most successful spawning seasons with the highest recorded numbers. These high numbers of the Labeo Umbratus can be due to the fact that the species lays a large number of eggs. The high numbers of the Cyprinus carpio is due to the lower numbers of the other fish species as previous studies have shown that the species negatively impacts the environment which could in turn negatively impact the other fish species. The Claridae gariepinus and Labeobarbus aeneusas were the lowest. The low numbers of the Labeobarbus aneusas may be due to their slow growth and late maturity rate. The Labeo capensis had an average number relative to the other species and this is due to the fact that during the sampling period the dam was at 100% capacity as this is essential for the survival of the juvenile fish. The hypothesis was accepted as the Labeo Umbratus, Cyprinus carpio and Labeo capensis all have a successful spawning season. However, due to the size of the Cyprinus carpio, they would be most suited for a small scale fishery.

The Population Structure of the Orange River mudfish (Labeo capensis) in Allemanskraal Dam and Its potential as a Fishery Species

The aim of this research was to investigate whether the ecology and biology of the Orange River mudfish Labeo capensis were suitable for the species to be used in fisheries. Three fleets of the gill nets were set, parallel to the shore. One fleet was lifted, and the fish were collected by hand. The two remaining fleets were lifted the next day. The seine net was pulled for 10 metres within the littoral zone. The net was then pulled towards the shore of the dam and the procedure was repeated four times. The four fyke nets were set parallel to the shore and were left for two nettings nights and then lifted. All fish caught were collected by hand and placed into buckets. The majority (82.93%) of the fish caught were within the 0-100 mm size class. The 101-200mm and 201-300mm size classes contain similar numbers of fish, while no fish were caught in the 301-400mm size class. The hypothesis was accepted. Allemanskraal Dam, as of the study period, has a very small juvenile fish population of L. capensis, as only 7 out of 41 fish individuals caught were within the 101- 300mm fork length size class. These results show that the population of L. capensis is not established as of yet, as the research did was right after their breeding season. Historical research has shown that sexually mature individuals of the L. capensis species tend to be a minimum of 300mm SL, 4-6 years after hatching. The population was largely young-of-the-year and may develop into an established population in 3-4 years (after sexual maturity). The L. capensis population in Allemanskraal Dam has the potential to be a fishery species if suitable conditions are maintained. Establishing this species’ potential will therefore allow economically viable fisheries to utilise them sustainably and to their full economic potential.

The Reproduction success of the Cyprinidae and a Claridae fish species and its impact on small- scale fisheries

To investigate the reproduction success and natural recruitment of several Cyprinidae fish and Claridae fish species in the Allemanskraal Dam. The purpose of the project included investigating whether each individual fish species studied has a successful 2020/2021 spawning season in comparison with each other. Sections of the seine net were measured along with a distance of 10 along the shoreline. The ends of the seine nets were attached to one foot and the top of the net was held by hand. Both volunteers moved in unison while covering the 10m. The volunteer in the “deep end” moved towards the shoreline creating a semi-circle while the other volunteer remained stationary. The two ends of the net were then pulled onto the shore and the fish were collected. The results found that the Labeo Umbratus and Cyprinus carpio had the most successful spawning seasons with the highest recorded numbers. These high numbers of the Labeo Umbratus can be due to the fact that the species lays a large number of eggs. The high numbers of the Cyprinus carpio is due to the lower numbers of the other fish species as previous studies have shown that the species negatively impacts the environment which could in turn negatively impact the other fish species. The Claridae gariepinus and Labeobarbus aeneusas were the lowest. The low numbers of the Labeobarbus aneusas may be due to their slow growth and late maturity rate. The Labeo capensis had an average number relative to the other species and this is due to the fact that during the sampling period the dam was at 100% capacity as this is essential for the survival of the juvenile fish. The hypothesis was accepted as the Labeo Umbratus, Cyprinus carpio and Labeo capensis all have a successful spawning season. However, due to the size of the Cyprinus carpio, they would be most suited for a small scale fishery.

The Population Structure of the Orange River mudfish (Labeo capensis) in Allemanskraal Dam and Its potential as a Fishery Species

The aim of this research was to investigate whether the ecology and biology of the Orange River mudfish Labeo capensis were suitable for the species to be used in fisheries. Three fleets of the gill nets were set, parallel to the shore. One fleet was lifted, and the fish were collected by hand. The two remaining fleets were lifted the next day. The seine net was pulled for 10 metres within the littoral zone. The net was then pulled towards the shore of the dam and the procedure was repeated four times. The four fyke nets were set parallel to the shore and were left for two nettings nights and then lifted. All fish caught were collected by hand and placed into buckets. The majority (82.93%) of the fish caught were within the 0-100 mm size class. The 101-200mm and 201-300mm size classes contain similar numbers of fish, while no fish were caught in the 301-400mm size class. The hypothesis was accepted. Allemanskraal Dam, as of the study period, has a very small juvenile fish population of L. capensis, as only 7 out of 41 fish individuals caught were within the 101- 300mm fork length size class. These results show that the population of L. capensis is not established as of yet, as the research did was right after their breeding season. Historical research has shown that sexually mature individuals of the L. capensis species tend to be a minimum of 300mm SL, 4-6 years after hatching. The population was largely young-of-the-year and may develop into an established population in 3-4 years (after sexual maturity). The L. capensis population in Allemanskraal Dam has the potential to be a fishery species if suitable conditions are maintained. Establishing this species’ potential will therefore allow economically viable fisheries to utilise them sustainably and to their full economic potential.

在壓力下更要堅強,探討ACE2/Ang(1-7)軸與韌力的關係

背景:韌力 (resilience)指個體遭受壓力時快速調適及復原的能力,是許多疾病的保護因子但生理機轉仍不明。Angiotensin-converting enzyme 2/Angiotensin(1-7) [ACE2/Ang(1-7)]軸具有保護心血管、抗焦慮及保護神經的潛力。目前仍無ACE2/Ang(1-7)軸與韌力相關的研究。另外ACE2是SARS-CoV-2病毒進入人體的受體,相關研究在疫情下更顯重要。 目的:本研究中我們以習得無助行為模式篩選具韌力行為的大鼠(resilient, R),探討韌力鼠與非韌力鼠(non-R) ACE2/Ang(1-7)軸表現的差異。 假說:韌力鼠的ACE2/Ang(1-7)軸活性較非韌力鼠高。 結果:韌力鼠在壓力下其corticosterone (CORT)濃度與控制組無差異。韌力鼠的Ang(1-7)濃度較非韌力鼠及控制組高,暗示較高的Ang(1-7)濃度與韌性表現相關。以Ang(1-7)/Angiotensin II (Ang II)比率作為反映ACE2 轉化Ang(1-7) 功能的指標,發現韌力鼠的ACE2濃度及功能均與未主動逃跑而受電擊的時間相關,暗示其ACE2活性能因應壓力調控上升。韌力鼠的ACE2濃度雖較非韌力鼠低,但其ACE2功能則較其他兩組高。並且,Ang (1-7)濃度及ACE2功能僅在韌力鼠與ACE2濃度呈現正相關,但在非韌力鼠則無相關。暗示在壓力下韌力鼠的ACE2仍維持有效率的功能但非韌力鼠的ACE2功能則受影響。另一方面,Ang (1-7)濃度僅在非韌力鼠與CORT濃度負相關,暗示非韌力鼠個體的ACE2/Ang (1-7)]軸活性與海馬迴-下視丘-腦下垂體(HPA)軸的活性可能相關。 結論:韌力個體的ACE2/Ang (1-7)軸活性較非韌力鼠及控制組佳且可能因應壓力而調節增加。相反的,非韌力鼠個體的ACE2/Ang (1-7)軸功能較差且可能與HPA軸的活性相關。我們的研究顯示ACE2/Ang (1-7)軸扮演調節韌力的重要角色。ACE2/Ang (1-7)軸活性是否可以做為個體韌力的指標或具有增進韌力的臨床價值需進一步研究。