【摘要】：小型哺乳動物季節(jié)性繁殖與能量代謝水平之間的相互關(guān)系一直是近年來動物生態(tài)學(xué)領(lǐng)域的研究熱點。野生哺乳動物在自然環(huán)境下如何對季節(jié)性繁殖節(jié)律進行調(diào)控,能量代謝又是如何作用于繁殖節(jié)律的,這些依然是尚未解決的問題。瘦素(Leptin)作為機體能量代謝平衡的調(diào)定點信號,能將機體各器官及組織脂肪含量的多少相互聯(lián)系起來,通過下丘腦的瘦素受體發(fā)揮作用,協(xié)調(diào)各器官的能耗及代謝。促性腺激素釋腦Gn RH)是影響季節(jié)性繁殖的重要因子,在下丘腦分泌并釋丘過下丘腦-腺垂體-性腺軸的作用,調(diào)控生殖。瘦素對Gn RH在下丘腦處的表達是否有影響?二者表達模式是否相關(guān)?瘦素是否參與小型哺乳動物季節(jié)性繁殖節(jié)律的調(diào)控?為探究以上問題,選擇山東省的野生黑線倉鼠(Cricetulus barabensis)種群為實驗用動物,通過形態(tài)測量、靜止代謝率的測定、組織學(xué)、免疫組織化學(xué)和實時熒光定量PCR方法,研究了不同年齡、不同性別黑線倉鼠的生理多樣性,檢測了Leptin及受體、Gn RH及受體的蛋白表達以及m RNA表達量的季節(jié)性規(guī)律,探究了科學(xué)評價小型哺乳動物能量代謝及繁殖狀況的體質(zhì)指標。實驗內(nèi)容涉及黑線倉鼠生理多樣性的性別、年齡和季節(jié)差異,體質(zhì)指標的優(yōu)化,瘦素及其受體蛋白、Gn RH及其受體蛋白在大腦和下丘腦內(nèi)的免疫組化定位及相對定量,外周脂肪組織中OB m RNA的相對表達量的季節(jié)性動態(tài)等。研究結(jié)果表明:①Gn RH陽性細胞在雌雄黑線倉鼠下丘腦中的數(shù)量均呈顯著的季節(jié)性變化,一般在高繁殖季節(jié)(春秋季)顯著高于低繁殖季節(jié)(夏冬季),Gn RH受體陽性細胞數(shù)量的則相反。結(jié)果提示,下丘腦的Gn RH主要調(diào)控繁殖功能,而下丘腦的Gn RH受體可能主要參與Gn RH的負反饋調(diào)節(jié)作用。②Leptin陽性細胞數(shù)量在季節(jié)間差異顯著,且雌雄規(guī)律并非完全相同。雄性個體表現(xiàn)為春秋季夏季冬季;雌性個體,表現(xiàn)為秋季春夏季冬季。Leptin受體陽性細胞在季節(jié)間差異顯著,且雌雄表達規(guī)律相近,均表現(xiàn)為高繁殖季節(jié)(春秋季)低繁殖季節(jié)(夏冬季)。瘦素基因(OB)m RNA在外周脂肪組織中的表達也有顯著性的季節(jié)差異,且雌雄表達模式相近,即高繁殖季節(jié)(春秋季)低繁殖季節(jié)(夏冬季)。結(jié)果提示,瘦素在中樞神經(jīng)系統(tǒng)可能作為啟動繁殖的一種允許因子而非啟動因子而存在,并在一定程度上抑制其繁殖節(jié)律。瘦素受體在低繁殖季節(jié)的高表達可以介導(dǎo)瘦素作用,進一步抑制機體的繁殖時的能量支出,而增加能量的貯存。③黑線倉鼠靜止代謝率表現(xiàn)顯著的性別、年齡和季節(jié)性差異,其中雄性顯著低于雌性,且隨年齡和溫度的增加而呈降低的趨勢。在體質(zhì)指標方面,肥滿度K值和胴體重兩個指標則不存在性別間顯著性差異;重長比KWL值存在顯著的性別間差異,并表現(xiàn)為隨年齡的增加而增加,同時雌雄黑線倉鼠的KWL值均表現(xiàn)為季節(jié)間差異顯著。結(jié)果提示,在衡量小型哺乳動物體質(zhì)狀況時,建議使用重長比作為體質(zhì)指標較為方便有意義。黑線倉鼠作為北方地區(qū)農(nóng)田鼠害優(yōu)勢種,繁殖力強,種群A原因。因此,控制鼠害的綜合防控任重而道遠。研究結(jié)果有助于在分子水平上理解瘦素對黑線倉鼠能量代謝及繁殖水平方面的影響,初步解釋黑線倉鼠繁殖的季節(jié)性調(diào)控與能量代謝的關(guān)系,同時有助于更準確的評價小哺乳動物的體質(zhì)狀況,并為揭示黑線倉鼠種群暴發(fā)和季節(jié)性波動的分子機制提供借鑒。
[Abstract]:The relationship between the seasonal reproduction of small mammals and the level of energy metabolism has been a hot topic in the field of animal ecology in recent years. How wild mammals regulate the seasonal reproduction rhythm in the natural environment, and how energy metabolism works on the reproduction rhythm is still a problem that has not been solved yet. Leptin, as a point-to-point signal of the balance of energy metabolism in the body, can link the levels of fat in various organs and tissues of the body, and coordinate the energy consumption and metabolism of each organ through the role of the leptin receptor in the hypothalamus. Gonadotropin-releasing (Gn RH) is an important factor that affects the seasonal reproduction and regulates the reproduction of the hypothalamus-pituitary-gonad axis in the hypothalamus. Does leptin have an effect on the expression of Gn RH at the hypothalamus? Is the expression pattern of the two related? Is leptin involved in the regulation of the seasonal reproduction rhythm of small mammals? In order to explore the above problems, the wild black-line hamsters (Cricetulus barbadensis) in Shandong Province were selected as experimental animals, and different ages were studied by means of morphometry, measurement of resting metabolic rate, histology, immunohistochemistry and real-time fluorescence quantitative PCR. The physiological diversity of different sex black-line hamsters, the protein expression of Leptin and receptor, Gn RH and the receptor, and the seasonal regulation of the expression of m-RNA were examined, and the physical indexes of energy metabolism and reproduction of small mammals were studied. the experimental contents relate to the sex, the age and the seasonal difference of the physiological diversity of the black line hamsters, the optimization of the physical index, the immunohistochemical localization and relative quantification of the leptin and its receptor protein, Gn RH and its receptor protein in the brain and the hypothalamus, Seasonal dynamics of the relative expression of OB m RNA in peripheral adipose tissue. The results of the study show that the number of positive cells of Gn RH in the hypothalamus of the male and female black-line hamsters has a significant seasonal variation, and the number of positive cells in the Gn RH receptor is in the opposite direction during the high reproduction season (spring fall) significantly higher than that of the low reproduction season (summer winter). It is suggested that the Gn RH in the hypothalamus can regulate and control the reproductive function, while the Gn RH receptor in the hypothalamus may be involved in the negative feedback regulation of Gn RH. The number of Leptin positive cells was significant in the season, and the male and female law was not the same. The male was presented in the spring and autumn and winter in the summer, and the female was in the winter of the spring and summer in the autumn. Leptin receptor positive cells were significantly different in the season, and the male and female expression patterns were similar, all of which were in the low reproduction season (summer and winter) in the high reproduction season (spring fall). The expression of the leptin gene (OB) m RNA in the peripheral adipose tissue is also significant, and the male and female expression patterns are similar, that is, the high propagation season (spring fall) and the low reproduction season (summer winter). The results suggest that leptin may be present in the central nervous system as an allowable factor and a non-priming factor for the initiation and reproduction of the central nervous system, and to a certain extent inhibit its reproduction rhythm. The high expression of the leptin receptor in the low reproduction season can mediate the action of the leptin, further inhibit the energy expenditure during the reproduction of the body, and increase the storage of the energy. Hamster's resting metabolic rate showed a significant gender, age, and seasonal difference in which males were significantly lower than females and decreased with age and temperature. in that aspect of body constitution, there was no significant difference in the sex between the K value and the carcass weight of the body weight, and there was a significant difference in the gender between the weight length and the KWL value, and the KWL value of both male and female black-line hamsters was significantly different from that of the male and female black-line hamsters. The results suggest that the use of the weight-length ratio is more convenient to measure the physical condition of the small mammals. The black-line hamsters are the dominant species in the northern area, and the breeding force is strong and the cause of the population A. Therefore, the comprehensive prevention and control of the control of the mouse harm is a long way to go. The results of the study help to understand the effect of leptin on the energy metabolism and reproductive level of the black-line hamsters at the molecular level, and to explain the relationship between the seasonal regulation and energy metabolism of the black-line hamsters, and to help to evaluate the physical condition of the small mammals more accurately. and provides a reference for revealing the molecular mechanism of the outbreak and the seasonal fluctuation of the black-line hamsters.
【學(xué)位授予單位】：曲阜師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S852.21;S814

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