Aim: To extend the analysis of the proteome of human spermatozoa and establish a 2-D gel electrophoresis (2-DE) reference map of human spermatozoal proteins in a pH range of 3.5-9.0. Methods: In order to reveal mo...Aim: To extend the analysis of the proteome of human spermatozoa and establish a 2-D gel electrophoresis (2-DE) reference map of human spermatozoal proteins in a pH range of 3.5-9.0. Methods: In order to reveal more protein spots, immobilized pH gradient strips (24 cm) of broad range of pH 3-10 and the narrower range of pH 6-9, as well as different overlapping narrow range pH immobilized pH gradient (IPG) strips, including 3.5-4.5, 4.0-5.0, 4.5-5.5, 5.0-6.0 and 5.5-6.7, were used. After 2-DE, several visually identical spots between the different pH range 2-D gel pairs were cut from the gels and confirmed by mass spectrometry and used as landmarks for computer analysis. Results: The 2-D reference map with pH value from 3.5 to 9.0 was synthesized by using the ImageMaster analysis software. The overlapping spots were excluded, so that every spot was counted only once. A total of 3 872 different protein spots were identified from the reference map, an approximately 3-fold increase compared to the broad range pH 3-10 IPG strip (1 306 spots). Conclusion: The present 2-D pattern is a high resolution 2-D reference map for human fertile spermatozoal protein spots. A comprehensive knowledge of the protein composition of human spermatozoa is very meaningful in studying dysregulation of male fertility.展开更多
To better understand the spatial structure of Amur tigers(Panthera tigris altaica)at the southern edge of their range we fitted 14 tigers(6♀♀and 8♂♂)with 15 GPS-Argos collars between 2008 and 2011 in 2 study sites...To better understand the spatial structure of Amur tigers(Panthera tigris altaica)at the southern edge of their range we fitted 14 tigers(6♀♀and 8♂♂)with 15 GPS-Argos collars between 2008 and 2011 in 2 study sites:the Ussuriskii Reserve of southern Sikhote-Alin and the Land of the Leopard National Park in southwest Primorye,Russian Far East.Fixed kernel estimates of male home ranges were larger than those of female home ranges(P<0.05[mean 95%fixed kernel♀=401±205 km2;mean 95%fixed kernel♂=778±267 km2]).The home range size of females varied greatly,but on average was similar to estimates derived from earlier work further north.Low overlap of adjacent home ranges suggested that females retained exclusive territories.Real core areas of females overlapped only slightly,and remained stable over multiple years.The home ranges of adult males were smaller than those of males to the north,and in contrast to previous studies,high overlap among males indicated the absence of territoriality.Nonetheless,real core areas of males did not overlap,suggesting some spatial separation.In comparison to other tiger populations and other areas of the Russian Far East,the sex ratio in our 2 study areas was highly skewed towards males.We believe this skewed sex ratio resulted in the dissolution of territoriality of males due to an inability to defend individual females,with males resorting to scramble competition for mates.Continued monitoring of these sites to determine whether shifts in the sex ratio might result in a return to male territoriality would provide confirmation of our tentative hypothesis.展开更多
基金This work was supported by two grants from the National Natural Science Foundation of China (No. 30170480 and No. 30470884). We would like to thank Professor Guang-Yin Lu for enthusiastic reading and amending of the manuscript. We would like to thank Mr Ji-Xian Xiong, College of Life Science at the Hunan Normal University for enthusiastic support of technology and theory of MS. We are also very grateful to collaborators in our laboratories for help and valuable discussions and suggestions during the course of this work.
文摘Aim: To extend the analysis of the proteome of human spermatozoa and establish a 2-D gel electrophoresis (2-DE) reference map of human spermatozoal proteins in a pH range of 3.5-9.0. Methods: In order to reveal more protein spots, immobilized pH gradient strips (24 cm) of broad range of pH 3-10 and the narrower range of pH 6-9, as well as different overlapping narrow range pH immobilized pH gradient (IPG) strips, including 3.5-4.5, 4.0-5.0, 4.5-5.5, 5.0-6.0 and 5.5-6.7, were used. After 2-DE, several visually identical spots between the different pH range 2-D gel pairs were cut from the gels and confirmed by mass spectrometry and used as landmarks for computer analysis. Results: The 2-D reference map with pH value from 3.5 to 9.0 was synthesized by using the ImageMaster analysis software. The overlapping spots were excluded, so that every spot was counted only once. A total of 3 872 different protein spots were identified from the reference map, an approximately 3-fold increase compared to the broad range pH 3-10 IPG strip (1 306 spots). Conclusion: The present 2-D pattern is a high resolution 2-D reference map for human fertile spermatozoal protein spots. A comprehensive knowledge of the protein composition of human spermatozoa is very meaningful in studying dysregulation of male fertility.
基金This study was part of the long-term“Program of Studying the Amur Tiger in the Russian Far East”within the framework of the Permanent Expedition of RAS for the Study of Animals Listed of the Red Data Book of the Russian Federation and Other Highly Important Animals of the Russian Fauna as well as within the Siberian Tiger Project of the Wildlife Conservation Society(WCS-Russia).The authors are grateful to the staff of the V.L.Komarov“Ussuriskii”State Nature Reserve,FEB RAS and the Land of the Leopard National Park for fruitful support in data collection,to Dr Andrey D.Poyarkov from the A.N.Severtsov Institute of Ecology and Evolution,RAS and to Mr Alexander L.Salman for discussion and technical support.This research was made within the framework of the Program for the Study of the Amur Tiger in the Russian Far East and was supported by the Russian Geography Society。
文摘To better understand the spatial structure of Amur tigers(Panthera tigris altaica)at the southern edge of their range we fitted 14 tigers(6♀♀and 8♂♂)with 15 GPS-Argos collars between 2008 and 2011 in 2 study sites:the Ussuriskii Reserve of southern Sikhote-Alin and the Land of the Leopard National Park in southwest Primorye,Russian Far East.Fixed kernel estimates of male home ranges were larger than those of female home ranges(P<0.05[mean 95%fixed kernel♀=401±205 km2;mean 95%fixed kernel♂=778±267 km2]).The home range size of females varied greatly,but on average was similar to estimates derived from earlier work further north.Low overlap of adjacent home ranges suggested that females retained exclusive territories.Real core areas of females overlapped only slightly,and remained stable over multiple years.The home ranges of adult males were smaller than those of males to the north,and in contrast to previous studies,high overlap among males indicated the absence of territoriality.Nonetheless,real core areas of males did not overlap,suggesting some spatial separation.In comparison to other tiger populations and other areas of the Russian Far East,the sex ratio in our 2 study areas was highly skewed towards males.We believe this skewed sex ratio resulted in the dissolution of territoriality of males due to an inability to defend individual females,with males resorting to scramble competition for mates.Continued monitoring of these sites to determine whether shifts in the sex ratio might result in a return to male territoriality would provide confirmation of our tentative hypothesis.