Nesfatin-1在犬生殖軸上的表達
發(fā)布時間：2018-11-05 10:20
【摘要】：Nesfatin-1是一種多肽,它參與多種生理活動。目前國內(nèi)外學者對Nesfatin-1的研究僅限于鼠、魚,在犬上的報道較少。本試驗以幼兒期(1.5M)、初情前期(4M)、初情期(7.5M)以及成年期(24M)犬為研究對象,觀察Nesfatin-1在犬生殖軸上的分布及母犬不同生理發(fā)育階段生殖軸上NUCB2 mRNA的變化規(guī)律,初步闡述Nesfatin-1在犬生殖軸上的分布及其基因的時間表達規(guī)律,為Nesfatin-1調(diào)控犬的生殖提供科學的理論依據(jù)。成年公犬、母犬各3只,放血處死,取其下丘腦、垂體、卵巢及睪丸置入4%多聚甲醛中固定24~48 h后包埋,常規(guī)石蠟切片切片,厚度為5μm,再進行蘇木精免疫組化染色顯微鏡下觀察并拍照。另外,取不同生理發(fā)育階段的母犬(1.5M,4 M,7.5M,24M)各4只。處死后取下丘腦、垂體、卵巢放入事先加入TRIzol的凍存管內(nèi),-80℃保存。研磨組織后提取RNA,熒光定量PCR測定各發(fā)育階段不同組織中NUCB2mRNA的表達。結(jié)果表明:Nesfatin-1免疫陽性細胞主要分布于下丘腦的視上核(Supraoptic nucleus,SON)、下丘腦外側(cè)區(qū)(Lateral hypothalamic area,LHA)區(qū)域,神經(jīng)垂體、腺垂體,卵巢中的卵母細胞及間質(zhì)中,睪丸中未見其分布的生精細胞中。不同發(fā)育階段的母犬卵巢和垂體中NUCB2 mRNA在幼兒期最低(P0.01),隨著發(fā)育至初情前期、初情期NUCB2 mRNA的含量不斷上升,在初情期的表達達到最高(P0.01),成年期NUCB2 mRNA的表達則下降(P0.01)。下丘腦中NUCB2 mRNA的水平自幼兒期開始至成年期呈逐漸上升的趨勢(P0.01)。在生殖軸器官中,卵巢中NUCB2mRNA的水平在四個發(fā)育階段中皆是最高(P0.05)。根據(jù)以上的結(jié)果可以得出結(jié)論:Nesfatin-1及其基因存在于犬的生殖軸。NUCB2mRNA水平在初情期最高,提示Nesfatin-1可能調(diào)控犬的初情期啟動。在生殖器官中卵巢中NUCB2 mRNA水平最高,表明Nesfatin-1可能直接調(diào)控卵巢的發(fā)育及其功能。
[Abstract]:Nesfatin-1 is a polypeptide involved in many physiological activities. At present, the research on Nesfatin-1 is limited to rats and fish, and few reports are made on dogs. The subjects of this study were young children (1.5m), early estrus (4M), early estrus (7.5M) and adult dogs (24M). To observe the distribution of Nesfatin-1 on the reproductive axis of dogs and the changes of NUCB2 mRNA in the reproductive axis of female dogs at different physiological stages, and to explain the distribution of Nesfatin-1 on the reproductive axis of dogs and the temporal expression of genes. To provide a scientific theoretical basis for Nesfatin-1 to regulate the reproduction of dogs. Three adult male dogs and three female dogs were killed by bloodletting. The hypothalamus, pituitary, ovary and testis were fixed in 4% paraformaldehyde for 24 hours, then embedded in paraffin section with thickness of 5 渭 m. Then the hematoxylin immunohistochemical staining microscope was observed and photographed. In addition, four female dogs with different physiological development stages (1.5 MN 4 M7. 5 MU 24 M) were selected. After death, hypothalamus, pituitary and ovary were taken and stored at-80 鈩，
本文編號：2311761
[Abstract]:Nesfatin-1 is a polypeptide involved in many physiological activities. At present, the research on Nesfatin-1 is limited to rats and fish, and few reports are made on dogs. The subjects of this study were young children (1.5m), early estrus (4M), early estrus (7.5M) and adult dogs (24M). To observe the distribution of Nesfatin-1 on the reproductive axis of dogs and the changes of NUCB2 mRNA in the reproductive axis of female dogs at different physiological stages, and to explain the distribution of Nesfatin-1 on the reproductive axis of dogs and the temporal expression of genes. To provide a scientific theoretical basis for Nesfatin-1 to regulate the reproduction of dogs. Three adult male dogs and three female dogs were killed by bloodletting. The hypothalamus, pituitary, ovary and testis were fixed in 4% paraformaldehyde for 24 hours, then embedded in paraffin section with thickness of 5 渭 m. Then the hematoxylin immunohistochemical staining microscope was observed and photographed. In addition, four female dogs with different physiological development stages (1.5 MN 4 M7. 5 MU 24 M) were selected. After death, hypothalamus, pituitary and ovary were taken and stored at-80 鈩，
本文編號：2311761
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