德国美利奴羊胎儿期骨骼肌组织学结构发育特征研究
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摘要
【目的】探究胎儿期绵羊肌纤维的形成与发育规律,揭示德国美利奴羊胎儿期骨骼肌的组织学生长发育特征。【方法】选择妊娠期第75天(D75)、第105天(D105)和第135天(D135)的胎羊,测定其体长、体高和体质量;采集背最长肌,进行组织切片和HE染色,统计不同发育阶段胎儿单位肌纤维数量、直径和密度;应用免疫组化方法研究不同发育阶段背最长肌中快/慢肌纤维的类型和密度。【结果】随着怀孕母羊妊娠时间的增加,从75~135d胎儿体质量增长近17倍,体长、体高增加近1.5倍;单位肌纤维数量总体呈先增加后减少的变化趋势,在105d时单位面积肌纤维数量最高;肌纤维直径于妊娠75~105d增长幅度较为平缓,105~135d增长速率明显变大;早中期发育主要以慢肌(Ⅰ型肌纤维)为主,后期发育主要以快肌(Ⅱ型肌纤维)为主。【结论】德国美利奴羊胎儿肌纤维数量在妊娠105d时达到峰值,且此时肌纤维数量基本恒定,是肌纤维从肌增生转为肌肥大发育的关键时期,早中期发育主要以慢肌(Ⅰ型肌纤维)为主,后期发育主要以快肌(Ⅱ型肌纤维)为主。
【Objective】The purpose of this study is to explore the formation and development pattern of muscle fiber and reveal the developmental characteristics of skeletal muscle tissue in fetal period of German Merino sheep.【Method】Three times of D75,D105 and D135 during pregnancy were selected.The body size of German Merino fetal sheep was measured.Longissimus tissue from fetal sheep was collected and subjected to tissue sections and HE staining to count for the fiber number,diameter and density at different developmental stages of fetal muscle tissue.Fiber type was determined using immunohistochemistry to clarify the number of fast/slow muscle fiber type in dfferent stages of development.【Result】At the period from D75 to D135 of pregnant,the increase of fetal weight was nearly 17 times,the body length and height is nearly 1.5 times.The numbers of muscle fibers in unit area gradually increased to D105 while reach the highest value,and the overall trend first climbed up and then declined.The growth rate of diameter of muscle fibers from D75 to D105 was relatively smooth,but that of D105-D135 obviously increased.In middle stage,the slow muscle(type Ⅰ muscle fiber)was the main type,and the latter was mainly composed offast muscle(typeⅡ muscle fiber).【Conclusion】The number of German Merino sheep fetal muscle fiber reached peak at D105,and the number of muscle fiber is almost constant.We found that the pregnant time of 105 days was a critical time window with fiber developmental transformation from muscle hyperplasia to muscle hypertrophy,and made clear the slow muscle(typeⅠ muscle fiber)was the main type during middle stage,and the latter stage was mainly composed of fast muscle(typeⅡ muscle fiber).
【Objective】The purpose of this study is to explore the formation and development pattern of muscle fiber and reveal the developmental characteristics of skeletal muscle tissue in fetal period of German Merino sheep.【Method】Three times of D75,D105 and D135 during pregnancy were selected.The body size of German Merino fetal sheep was measured.Longissimus tissue from fetal sheep was collected and subjected to tissue sections and HE staining to count for the fiber number,diameter and density at different developmental stages of fetal muscle tissue.Fiber type was determined using immunohistochemistry to clarify the number of fast/slow muscle fiber type in dfferent stages of development.【Result】At the period from D75 to D135 of pregnant,the increase of fetal weight was nearly 17 times,the body length and height is nearly 1.5 times.The numbers of muscle fibers in unit area gradually increased to D105 while reach the highest value,and the overall trend first climbed up and then declined.The growth rate of diameter of muscle fibers from D75 to D105 was relatively smooth,but that of D105-D135 obviously increased.In middle stage,the slow muscle(type Ⅰ muscle fiber)was the main type,and the latter was mainly composed offast muscle(typeⅡ muscle fiber).【Conclusion】The number of German Merino sheep fetal muscle fiber reached peak at D105,and the number of muscle fiber is almost constant.We found that the pregnant time of 105 days was a critical time window with fiber developmental transformation from muscle hyperplasia to muscle hypertrophy,and made clear the slow muscle(typeⅠ muscle fiber)was the main type during middle stage,and the latter stage was mainly composed of fast muscle(typeⅡ muscle fiber).
引文
[1]蒋小怀.新疆养羊业生产形势分析与发展对策研究[J].草食家畜,2014(2):1-5.Jiang X H.Study on the production situation and development countermeasures of Xinjiang sheep industry[J].Grass-Feeding Livestock,2014(2):1-5.
[2]赵有璋.羊生产学[M].北京:中国农业出版社,1993.Zhao Y Z.Production science of sheep and goat[M].Beijing:China Agriculture Press,1993.
[3]周金星,毕亚玲,高登惠.肌肉组织学结构及其与肉品品质的关系[J].山地农业生物学报,2004,23(5):438-441.Zhou J X,Bi Y L,Gao D H.The relationship between structure and meat quality and muscle tissue[J].Journal of Mountain Agriculture and Biology,2004,23(5):438-441.
[4]赵晓,莫德林,张悦,等.猪的骨骼肌生长发育研究进展[J].生命科学,2011,23(1):37-44.Zhao X,Mo D L,Zhang Y,et al.Research progress on skeletal muscle growth and development in pigs[J].Chinese Bulletin of Life Sciences,2011,23(1):37-44.
[5]刘居鹏,单安山,彭济昌.猪肌纤维发育及其与肉质的关系[J].中国饲料,2006(16):11-14.Liu J P,Shan A S,Peng J C.Development of porcine muscle fiber and its relationship with meat quality[J].China Feed,2006(16):11-14.
[6]郭云雁,王立贤,程笃学,等.猪肌纤维组织学特性研究进展[J].中国畜牧兽医,2007,34(8):49-52.Guo Y Y,Wang L X,Cheng D X,et al.Research progress on histological characteristics of porcine muscle fibers[J].China Animal Husbandry and Veterinary Medicine,2007,34(8):49-52.
[7]Wilson D A,Baker G J,Pijanowski G J,et al.Composition and morphologic features of the interosseous muscle in Standardbreds and Thoroughbreds[J].Am J Vet Res,1991,52(1):133-139.
[8]Ashmore C R,Addis P B,Doerr L.Development of muscle fibers in the fetal pig[J].J Anim Sci,1973,36(6):1088-1093.
[9]Wigmore P M,Stickland N C.Muscle development in large and small big fetuses[J].Journal of Anatomy,1983,40(137):235-245.
[10]Zhu M J,Ford S P,Nathanielsz P W,et al.Effect of maternal nutrient restriction in sheep on the development of fetal skeletal muscle[J].Biol Reprod,2004,71(6):1968-1973.
[11]任航行.Texel与乌珠穆沁绵羊妊娠中、后期胎儿骨骼肌基因表达谱及组织学分析[D].北京:中国农业科学院,2010.Ren H X.Texel and ujumqin sheep in late pregnancy,fetal skeletal muscle profiles of gene expression and histological analysis[D].Beijing:Chinese Academy of Agricultural Sciences,2010.
[12]陈晓涛,章焕然,陈伟德.德国美利奴羊在新疆的应用前景[J].草食家畜,2003(1):9-10.Chen X T,Zhang H R,Chen W D.The German Merino sheep application in Xinjiang[J].Grass-Feeding Livestock,2003(1):9-10.
[13]蒋烈戈,李勇忠,景亚平,等.德国美利奴羊与中国美利奴羊主要生产性能比较[J].中国草食动物科学,2013,33(6):78-79.Jiang L G,Li Y Z,Jing Y P,et al.The German Merino sheep and Chinese Merino sheep production performance comparison[J].China Herbivores Science,2013,33(6):78-79.
[14]陈璐,马恒辉,金邦茎.骨骼肌组织石蜡制片技术的改进[J].临床与实验病理学杂志,2008,24(2):233-234.Chen L,Ma H H,Jin B J.Improvement of paraffin sectioning technique for skeletal muscle tissue[J].Chinese Journal of Clinical and Experimental Pathology,2008,24(2):233-234.
[15]李艳,高友晶,张喜,等.细胞石蜡包埋切片技术的应用[J].江汉大学学报(自然科学版),2011,39(1):72-74.Li Y,Gao Y J,Zhang X,et al.Application of cell paraffin embedded section technique[J].Journal of Jianghan University(Natural Science Edition),2011,39(1):72-74.
[16]王巧.HE染色方法在临床病理诊断中的应用[J].临床合理用药杂志,2014(32):118-119.Wang Q.Application of HE staining method in clinical pathological diagnosis[J].Journal of Clinical Rational Drug Use,2014(32):118-119.
[17]张敏.HE染色在临床病理诊断中的应用[J].齐齐哈尔医学院学报,2011,35(4):552-553.Zhang M.Clinical application of HE staining in clinical pathological diagnosis[J].Journal of Qiqihar University of Medicine,2011,35(4):552-553.
[18]刘增辉.病理染色技术[M].北京:人民卫生出版社,2001:14.Liu Z H.Pathological staining technique[M].Beijing:People Health Publishing House,2001:14.
[19]吴秉铨,刘彦仿.免疫组化学病理诊断[M].北京:北京科学技术出版社,2007.Wu B Q,Liu Y F.Immunohistochemical pathological diagnosis[M].Beijing:Beijing Science and Technology Press,2007.
[20]张卫琴.免疫组化技术在病理诊断中的应用[J].安徽医药,2012,16(11):1700-1702.Zhang W Q.Application of immunohistochemical technique in pathological diagnosis[J].Anhui Medical and Pharmaceutical Journal,2012,16(11):1700-1702.
[21]Crouse H R.Palatability of individual muscles from bovine leg steaks as related to chemical and histologocal traits[J].J Food Sci,1972,37:282-287.
[22]Cornforth D P,Hecker A L,Cramger D A,et al.Maturity and its relationship to muscle characteristics of cattle[J].Journal of Animal Science,1980,50:75-80.
[23]Cassens R G,Rymkiewicz J,Szeremeta J,et al.The importance of myofiber type[J].Food Microstructure,1984,3:1-7.
[24]秦宁,徐日福,张树敏,等.长白山野猪和东北民猪骨骼肌组织学特性比较[J].吉林农业大学学报,2012,34(3):343-347.Qin N,Xu R F,Zhang S M,et al.Comparison of histological characteristics of skeletal muscle between Changbai Mountain wild boar and Northeast China pig[J].Journal of Jilin Agricultural University,2012,34(3):343-347.
[25]刘卫娟.几种肉羊肌肉组织学形状和理化性状的研究[D].河北保定:河北农业大学,2009.Liu W J.Several studies of the shape and physical-chemical properties of the muscle groups were studied[D].Baoding,Hebei:Agricultural University of Hebei,2009.
[26]Ausoni S,Gorza L,Schiaffino S,et al.Expression of myosin heavy chain isoforms in stimulated fast and slow rat muscles[J].The Journal of Neuroscience,1990,10:153-160.
[27]杨飞云.猪骨骼肌肌纤维类型分布及转化的分子机理研究[D].四川雅安:四川农业大学,2008.Yang F Y.Study on the distribution and transformation of muscle fiber types in porcine skeletal muscle[D].Ya’an,Sichuan:Sichuan Agricultural University,2008.
[28]高儒松,程志斌,杨正华.肌纤维类型与猪肉品质的关系[J].中国畜牧兽医,2009,36(4):191-195.Gao R S,Cheng Z B,Yang Z H.Relationship between muscle fiber types and pork quality[J].China Animal Husbandry and Veterinary Medicine,2009,36(4):191-195.
[29]罗军.肌肉纤维特性研究进展[J].黄牛杂志,1989,5(4):36-40.Luo J.Research progress of muscle fiber characteristics[J].Journal of Yellow Cattle Science,1989,5(4):36-40.
[30]杨飞云,陈代文,黄金秀,等.猪背最长肌肌纤维类型的发育性变化及品种与营养影响特点[J].畜牧兽医学报,2008,39(12):1701-1708.Yang F Y,Chen D W,Huang J X,et al.The developmental changes of muscle fiber types in longissimus dorsi muscle of pigs and the characteristics of the variety and nutrition[J].Chinese Journal of Animal and Veterinary Sciences,2008,39(12):1701-1708.
[31]杨晓静,赵茹茜,陈杰,等.猪背最长肌肌纤维类型的发育性变化及其品种和性别特点[J].中国兽医学报,2005,25(1):89-94.Yang X J,Zhao R Q,Chen J,et al.Developmental changes of muscle fiber types and characteristics of breed and sex of longissimus dorsi muscle in pigs[J].Chinese Journal of Veterinary Science,2005,25(1):89-94.
[32]Engel W K.The essentiality of histo and cytochemical studies in the investigation of meuromuscular disease[J].Neurology,1962,12:778-784.
[2]赵有璋.羊生产学[M].北京:中国农业出版社,1993.Zhao Y Z.Production science of sheep and goat[M].Beijing:China Agriculture Press,1993.
[3]周金星,毕亚玲,高登惠.肌肉组织学结构及其与肉品品质的关系[J].山地农业生物学报,2004,23(5):438-441.Zhou J X,Bi Y L,Gao D H.The relationship between structure and meat quality and muscle tissue[J].Journal of Mountain Agriculture and Biology,2004,23(5):438-441.
[4]赵晓,莫德林,张悦,等.猪的骨骼肌生长发育研究进展[J].生命科学,2011,23(1):37-44.Zhao X,Mo D L,Zhang Y,et al.Research progress on skeletal muscle growth and development in pigs[J].Chinese Bulletin of Life Sciences,2011,23(1):37-44.
[5]刘居鹏,单安山,彭济昌.猪肌纤维发育及其与肉质的关系[J].中国饲料,2006(16):11-14.Liu J P,Shan A S,Peng J C.Development of porcine muscle fiber and its relationship with meat quality[J].China Feed,2006(16):11-14.
[6]郭云雁,王立贤,程笃学,等.猪肌纤维组织学特性研究进展[J].中国畜牧兽医,2007,34(8):49-52.Guo Y Y,Wang L X,Cheng D X,et al.Research progress on histological characteristics of porcine muscle fibers[J].China Animal Husbandry and Veterinary Medicine,2007,34(8):49-52.
[7]Wilson D A,Baker G J,Pijanowski G J,et al.Composition and morphologic features of the interosseous muscle in Standardbreds and Thoroughbreds[J].Am J Vet Res,1991,52(1):133-139.
[8]Ashmore C R,Addis P B,Doerr L.Development of muscle fibers in the fetal pig[J].J Anim Sci,1973,36(6):1088-1093.
[9]Wigmore P M,Stickland N C.Muscle development in large and small big fetuses[J].Journal of Anatomy,1983,40(137):235-245.
[10]Zhu M J,Ford S P,Nathanielsz P W,et al.Effect of maternal nutrient restriction in sheep on the development of fetal skeletal muscle[J].Biol Reprod,2004,71(6):1968-1973.
[11]任航行.Texel与乌珠穆沁绵羊妊娠中、后期胎儿骨骼肌基因表达谱及组织学分析[D].北京:中国农业科学院,2010.Ren H X.Texel and ujumqin sheep in late pregnancy,fetal skeletal muscle profiles of gene expression and histological analysis[D].Beijing:Chinese Academy of Agricultural Sciences,2010.
[12]陈晓涛,章焕然,陈伟德.德国美利奴羊在新疆的应用前景[J].草食家畜,2003(1):9-10.Chen X T,Zhang H R,Chen W D.The German Merino sheep application in Xinjiang[J].Grass-Feeding Livestock,2003(1):9-10.
[13]蒋烈戈,李勇忠,景亚平,等.德国美利奴羊与中国美利奴羊主要生产性能比较[J].中国草食动物科学,2013,33(6):78-79.Jiang L G,Li Y Z,Jing Y P,et al.The German Merino sheep and Chinese Merino sheep production performance comparison[J].China Herbivores Science,2013,33(6):78-79.
[14]陈璐,马恒辉,金邦茎.骨骼肌组织石蜡制片技术的改进[J].临床与实验病理学杂志,2008,24(2):233-234.Chen L,Ma H H,Jin B J.Improvement of paraffin sectioning technique for skeletal muscle tissue[J].Chinese Journal of Clinical and Experimental Pathology,2008,24(2):233-234.
[15]李艳,高友晶,张喜,等.细胞石蜡包埋切片技术的应用[J].江汉大学学报(自然科学版),2011,39(1):72-74.Li Y,Gao Y J,Zhang X,et al.Application of cell paraffin embedded section technique[J].Journal of Jianghan University(Natural Science Edition),2011,39(1):72-74.
[16]王巧.HE染色方法在临床病理诊断中的应用[J].临床合理用药杂志,2014(32):118-119.Wang Q.Application of HE staining method in clinical pathological diagnosis[J].Journal of Clinical Rational Drug Use,2014(32):118-119.
[17]张敏.HE染色在临床病理诊断中的应用[J].齐齐哈尔医学院学报,2011,35(4):552-553.Zhang M.Clinical application of HE staining in clinical pathological diagnosis[J].Journal of Qiqihar University of Medicine,2011,35(4):552-553.
[18]刘增辉.病理染色技术[M].北京:人民卫生出版社,2001:14.Liu Z H.Pathological staining technique[M].Beijing:People Health Publishing House,2001:14.
[19]吴秉铨,刘彦仿.免疫组化学病理诊断[M].北京:北京科学技术出版社,2007.Wu B Q,Liu Y F.Immunohistochemical pathological diagnosis[M].Beijing:Beijing Science and Technology Press,2007.
[20]张卫琴.免疫组化技术在病理诊断中的应用[J].安徽医药,2012,16(11):1700-1702.Zhang W Q.Application of immunohistochemical technique in pathological diagnosis[J].Anhui Medical and Pharmaceutical Journal,2012,16(11):1700-1702.
[21]Crouse H R.Palatability of individual muscles from bovine leg steaks as related to chemical and histologocal traits[J].J Food Sci,1972,37:282-287.
[22]Cornforth D P,Hecker A L,Cramger D A,et al.Maturity and its relationship to muscle characteristics of cattle[J].Journal of Animal Science,1980,50:75-80.
[23]Cassens R G,Rymkiewicz J,Szeremeta J,et al.The importance of myofiber type[J].Food Microstructure,1984,3:1-7.
[24]秦宁,徐日福,张树敏,等.长白山野猪和东北民猪骨骼肌组织学特性比较[J].吉林农业大学学报,2012,34(3):343-347.Qin N,Xu R F,Zhang S M,et al.Comparison of histological characteristics of skeletal muscle between Changbai Mountain wild boar and Northeast China pig[J].Journal of Jilin Agricultural University,2012,34(3):343-347.
[25]刘卫娟.几种肉羊肌肉组织学形状和理化性状的研究[D].河北保定:河北农业大学,2009.Liu W J.Several studies of the shape and physical-chemical properties of the muscle groups were studied[D].Baoding,Hebei:Agricultural University of Hebei,2009.
[26]Ausoni S,Gorza L,Schiaffino S,et al.Expression of myosin heavy chain isoforms in stimulated fast and slow rat muscles[J].The Journal of Neuroscience,1990,10:153-160.
[27]杨飞云.猪骨骼肌肌纤维类型分布及转化的分子机理研究[D].四川雅安:四川农业大学,2008.Yang F Y.Study on the distribution and transformation of muscle fiber types in porcine skeletal muscle[D].Ya’an,Sichuan:Sichuan Agricultural University,2008.
[28]高儒松,程志斌,杨正华.肌纤维类型与猪肉品质的关系[J].中国畜牧兽医,2009,36(4):191-195.Gao R S,Cheng Z B,Yang Z H.Relationship between muscle fiber types and pork quality[J].China Animal Husbandry and Veterinary Medicine,2009,36(4):191-195.
[29]罗军.肌肉纤维特性研究进展[J].黄牛杂志,1989,5(4):36-40.Luo J.Research progress of muscle fiber characteristics[J].Journal of Yellow Cattle Science,1989,5(4):36-40.
[30]杨飞云,陈代文,黄金秀,等.猪背最长肌肌纤维类型的发育性变化及品种与营养影响特点[J].畜牧兽医学报,2008,39(12):1701-1708.Yang F Y,Chen D W,Huang J X,et al.The developmental changes of muscle fiber types in longissimus dorsi muscle of pigs and the characteristics of the variety and nutrition[J].Chinese Journal of Animal and Veterinary Sciences,2008,39(12):1701-1708.
[31]杨晓静,赵茹茜,陈杰,等.猪背最长肌肌纤维类型的发育性变化及其品种和性别特点[J].中国兽医学报,2005,25(1):89-94.Yang X J,Zhao R Q,Chen J,et al.Developmental changes of muscle fiber types and characteristics of breed and sex of longissimus dorsi muscle in pigs[J].Chinese Journal of Veterinary Science,2005,25(1):89-94.
[32]Engel W K.The essentiality of histo and cytochemical studies in the investigation of meuromuscular disease[J].Neurology,1962,12:778-784.