杂交石斑鱼Epinephelus fuscoguttatus♀×E. lanceolatus♂血细胞的类型及发生
作者简介:周晖(1978—), 男, 广西玉林市人, 讲师, 博士, 研究方向为鱼类生理生态学。E-mail: 346005548@qq.com
收稿日期: 2017-01-09
要求修回日期: 2017-04-23
网络出版日期: 2017-09-22
基金资助
国家海洋局海洋公益行业科研专项 (201205028)
广东省级科技计划项目(2016B020201009)
广东省科技厅省部产学研结合项目(2013B090700010)
广东普通高校创新团队项目(2015KCXTD021)
广东省高校重点实验室专项(2013CXZDA019)
广东省海洋经济创新发展区域示范专项(GD2012-A02-003)
Blood cell types and production of hybrid grouper E. fuscoguttatus ♀×E. lanceolatus ♂
Received date: 2017-01-09
Request revised date: 2017-04-23
Online published: 2017-09-22
Supported by
Public Science and Technology Research Funds Projects of Ocean (201205028)
Science and Technology Plan Projects of Guangdong Province (2016B020201009)
Special Project on the Integration of Industry, Education and Research of Guangdong Province (2013B090700010)
The Innovation Team Building Project of Guangdong Province Ordinary University (2015KCXTD021)
Special Project on Key Laboratory of Guangdong Higher Education (2013CXZDA019)
Regional Demonstration of Marine Economic Innovation and Development in Guangdong Province (GD2012-A02-003)
Copyright
为查明杂交石斑鱼Epinephelus fuscoguttatus♀ × Epinephelus lanceolatus♂血细胞形态分类及血细胞发生特点, 文章通过血液和组织涂(印)片、细胞染色和显微观察, 研究了体重263.18±12.31g的杂交石斑鱼外周血液白细胞的分类组成, 以及头肾、脾脏、体肾和肝脏等4种组织中各类血细胞的发生情况, 并将结果与其母本进行比较。结果表明, 杂交石斑鱼外周血液白细胞由淋巴细胞[(55.46±12.38)%]、血栓细胞[(21.11±5.38)%]、嗜中性粒细胞[(19.44±12.02)%]、单核细胞[(1.66±2.13)%]和嗜酸性粒细胞[(2.33±1.91)%]组成; 4种组织印迹片中, 头肾中未成熟的细胞总数比例最高[(16.12±0.30)%]], 体肾次之[(7.90±0.35)%], 脾脏[(2.56±0.21)%]和肝脏[(0.97±0.18)%]未成熟细胞比例较低; 由此推断杂交石斑鱼主要造血组织是头肾, 其次是体肾。在杂交石斑鱼4种组织中均观察到浆细胞, 在头肾、体肾和脾脏印迹片中观察到嗜碱性粒细胞和巨噬细胞, 在体肾印迹片中观察到巨大原单核细胞。杂交石斑鱼与其母本相比, 外周血液白细胞中淋巴细胞比例接近(+2.16%), 血栓细胞较少(-14.58%), 嗜中性粒细胞(+9.10%)、嗜酸性粒细胞(+2.18%)和单核细胞(+1.38%)较多; 主要造血组织都是头肾, 但淋巴细胞的主要发生组织不同(分别是体肾和头肾), 次要造血组织也不同(分别是体肾和脾脏); 此外杂交石斑鱼各组织中未成熟细胞比例明显高于母本, 显示出活跃的造血活动。本研究结果能为杂交石斑鱼的血液生理学、病理学研究及石斑鱼杂交选育提供基础资料。
周晖 , 陈刚 , 邓崇新 . 杂交石斑鱼Epinephelus fuscoguttatus♀×E. lanceolatus♂血细胞的类型及发生[J]. 热带海洋学报, 2017 , 36(5) : 49 -58 . DOI: 10.11978/2017006
To study the blood cell morphological classification and production characteristics of hybrid grouper Epinephelus fuscoguttatus♀ × Epinephelus lanceolatus♂, peripheral blood smears and tissues stamps (pronephros, kidney, spleen and liver) were made; cell staining and microscopic observation, differential leucocyte counts (DLC) of peripheral blood and observation of various types of blood cell development in the four types of tissues were conducted, and the results were compared with its female parent. The results showed that lymphocytes [(55.46±12.38)%], thrombcytes [(21.11±5.38)%], neutrophils [(19.44±12.02)%], monocytes [(1.66±2.13)%] and eosinophils [(2.33±1.91)%] were observed in peripheral blood. The pronephros showed the highest immature cells percentage [(16.12±0.30)%], followed by kidney [(7.90±0.35)%], spleen [(2.56±0.21)%] and liver [(0.97±0.18)%]. The results indicated that the major hematopoietic tissue of hybrid grouper was pronephros, followed by kidney. Besides, plasma cells were observed in the four types of tissues, basophils and macrophages were observed in stamps of pronephros, kidney and spleen, and huge primitive monocytes were observed in kidney stamps. Compared with the female parent, the ratio of lymphocytes in peripheral blood leukocytes was similar (+2.16%), with less thrombotic cells (-14.58%), and more neutrophils (+9.10%), eosinophils (+2.18%) and monocytes (+1.38%). The main hematopoietic tissues of hybrid grouper and its female parent were both pronephros, but the major tissues for lymphocytes production were different (kidney and pronephros, respectively); and the secondary hematopoietic tissues were different ((kidney and spleen). The proportion of immature cells in the tissues of the hybrid grouper was significantly higher than that of its female parent, showing active hematopoietic activity. The results of this study provide basic data of morphology classification and blood cell production for studying blood physiology, pathology of grouper and hybrid breeding of grouper.
Tab.1 Differential leucocyte counts (DLC) in peripheral blood of hybrid grouper表1 杂交石斑鱼外周血液中主要白细胞分类计数(DLC) |
细胞类型 | 个数 | 百分比/% |
---|---|---|
淋巴细胞 | 119.00±30.18 | 55.46±12.38 |
血栓细胞 | 45.29±9.01 | 21.11±5.38 |
嗜中性粒细胞 | 41.71±22.13 | 19.44±12.02 |
嗜酸性粒细胞 | 5.51±5.00 | 2.33±1.91 |
单核细胞 | 3.57±3.99 | 1.66±2.13 |
注: 表中数据为8尾实验鱼数据的平均值±标准差。 |
Tab.2 Leucocyte size in peripheral blood of hybrid grouper表2 杂交石斑鱼外周血液中各类细胞大小 |
细胞类型 | 细胞大小 | 核大小 | ||
---|---|---|---|---|
长径/µm | 短径/µm | 长径/µm | 短径/µm | |
红细胞 | 12.35±1.45 | 6.21±0.51 | 4.53±0.32 | 3.41±0.18 |
大淋巴细胞 | 7.18±1.49 | 5.36±0.93 | 5.08±1.00 | 3.77±0.53 |
小淋巴细胞 | 5.74±0.62 | 4.53±0.39 | 4.86±0.64 | 3.73±0.49 |
血栓细胞 | 10.47±1.03 | 3.69±0.95 | 7.58±0.99 | 2.97±0.82 |
嗜中性粒细胞 | 9.82±1.43 | 7.50±0.92 | 6.10±0.87 | 4.61±0.61 |
嗜酸性粒细胞 | 7.42±1.05 | 6.32±0.98 | 4.85±0.50 | 4.24±0.61 |
单核细胞 | 13.21±1.12 | 11.87±1.32 | 8.98±0.68 | 7.44±1.12 |
注: 表中数据为8尾实验鱼数据的平均值±标准差。 |
Fig. 1 Blood cells of hybrid grouper. The blood cells in the plate were enlarged 1000 times; the scaleplate in the plate indicated 10 μm. (a-e) blood smear; (f-g) liver stamp; (h) kidney stamp; (i) spleen stamp; (j-n) pronephros stamp图1 杂交石斑鱼血细胞 |
Tab. 3 Percentage of different blood cell lines in four tissues of hybrid grouper表3 杂交石斑鱼4种组织中各类血细胞的分类计数 |
细胞类型 | 头肾 | 脾脏 | 肝脏 | 体肾 | ||||
---|---|---|---|---|---|---|---|---|
个数 | 百分比/% | 个数 | 百分比/% | 个数 | 百分比/% | 个数 | 百分比/% | |
原红细胞PE | 26.43±10.74 | 0.81±0.39 | 0 | 0 | 0 | 0 | 5.43±1.99 | 0.21±0.10 |
幼红细胞IE | 191.43±23.98 | 5.85±0.43 | 0 | 0 | 0 | 0 | 22.43±14.35 | 0.87±0.47 |
红细胞 Er | 278.86±62.50 | 8.53±1.82 | 968.00±31.21 | 71.24±2.42 | 3402.71±163.71 | 90.28±1.85 | 1010.14±286.53 | 38.83±2.06 |
原粒细胞PMy | 30.57±17.65 | 0.93±0.26 | 1.29±1.38 | 0.09±0.04 | 0 | 0 | 7.57±4.86 | 0.29±0.24 |
早幼粒细胞Pr | 35.14±10.12 | 1.07±0.26 | 2.43±2.51 | 0.18±0.22 | 0 | 0 | 12.71±12.61 | 0.49±0.23 |
嗜中性中幼粒细胞 MIN | 19.43±13.15 | 0.59±0.32 | 0 | 0 | 0 | 0 | 7.57±4.65 | 0.29±0.24 |
嗜中性晚幼粒细胞 LIN | 60.00±18.10 | 1.83±0.21 | 0 | 0 | 0 | 0 | 9.00±6.76 | 0.35±0.23 |
嗜中性粒细胞Ne | 118.00±17.08 | 3.61±0.41 | 17.57±6.48 | 1.29±0.24 | 2.29±5.62 | 0.06±0.19 | 26.86±11.22 | 1.04±0.31 |
嗜碱性粒细胞Ba | 4.14±3.93 | 0.13±0.13 | 3.86±4.35 | 0.28±0.33 | 0 | 0 | 1.43±1.40 | 0.06±0.06 |
嗜酸性粒细胞Eo | 15.00±4.00 | 0.46±0.15 | 58.86±13.53 | 4.33±0.53 | 8.14±0.90 | 0.22±0.24 | 13.57±8.32 | 0.53±0.30 |
原淋巴细胞PL | 37.29±6.90 | 1.14±0.32 | 7.00±2.31 | 0.52±0.04 | 1.00±1.73 | 0.03±0.08 | 33.29±8.01 | 1.29±0.21 |
幼淋巴细胞IL | 39.86±13.84 | 1.22±0.45 | 14.29±1.38 | 1.05±0.12 | 1.43±2.57 | 0.04±0.07 | 71.86±10.32 | 2.78±0.74 |
大淋巴细胞LL | 25.86±9.26 | 0.79±0.14 | 5.00±1.53 | 0.37±0.14 | 1.60±4.58 | 0.04±0.29 | 8.29±2.43 | 0.32±0.21 |
小淋巴细胞SL | 1707.86±701.32 | 52.21±2.20 | 184.00±21.40 | 13.54±1.32 | 248.57±35.88 | 6.60±0.41 | 1214.71±380.44 | 46.90±2.27 |
浆细胞Pl | 92.19±4.08 | 2.82±0.13 | 10.29±1.13 | 0.76±0.10 | 6.43±1.29 | 0.17±0.28 | 86.29±9.88 | 3.34±0.37 |
原单核细胞PMo | 30.43±6.55 | 0.93±0.25 | 4.86±3.71 | 0.36±0.28 | 2.57±3.15 | 0.07±0.07 | 7.71±6.21 | 0.30±0.20 |
幼单核细胞Imo | 56.57±19.89 | 1.73±0.25 | 4.86±4.77 | 0.36±0.36 | 31.29±6.68 | 0.83±0.47 | 26.71±11.56 | 1.03±0.72 |
单核细胞Mo | 15.14±4.56 | 0.46±0.2. | 6.86±1.19 | 0.50±0.17 | 25.57±8.98 | 0.68±0.82 | 14.14±3.76 | 0.55±0.25 |
血栓细胞Th | 486.71±30.21 | 14.88±0.97 | 69.57±13.70 | 5.12±0.97 | 37.29±8.74 | 0.99±0.23 | 15.43±3.91 | 0.60±0.34 |
注: 表中数据为8尾实验鱼数据的平均值±标准差。 |
Tab. 4 Percentage of immature blood cells in four tissues of hybrid grouper (%)表4 杂交石斑鱼4种组织中各类未成熟细胞占总细胞数的百分比(%) |
组织 | 红细胞 | 淋巴细胞 | 粒细胞 | 单核细胞 | 总数 |
---|---|---|---|---|---|
头肾 | 6.66±0.40 | 2.36±0.34 | 4.44±0.24 | 2.66±0.23 | 16.12±0.30 |
体肾 | 1.08±0.29 | 4.07±0.44 | 1.42±0.23 | 1.33±0.46 | 7.90±0.35 |
脾脏 | 0 | 1.57±0.14 | 0.27±0.13 | 0.72±0.33 | 2.56±0.21 |
肝脏 | 0 | 0.07±0.07 | 0 | 0.90±0.31 | 0.97±0.18 |
注: 表中数据为8尾实验鱼数据的平均值±标准差。 |
Tab. 5 Size of different blood cell lines at different stages in four tissues of hybrid grouper表5 杂交石斑鱼4种组织中各阶段不同类型血细胞大小 |
细胞系 | 细胞大小 | 细胞核大小 | ||
---|---|---|---|---|
长径/µm | 短径/µm | 长径/µm | 短径/µm | |
原红细胞 PE | 9.31±0.74 | 7.65±1.28 | 6.99±1.06 | 5.61±0.93 |
早幼红细胞 EIE | 7.79±1.26 | 6.60±0.71 | 5.21±0.49 | 4.72±0.64 |
中幼红细胞 MIE | 8.15±0.65 | 6.60±0.94 | 5.15±0.55 | 4.46±0.48 |
晚幼红细胞 LIE | 8.62±0.92 | 5.57±0.91 | 5.06±0.56 | 3.89±0.47 |
红细胞 Er | 12.26±1.10 | 6.38±0.48 | 4.70±0.38 | 3.29±0.21 |
原粒细胞PMy | 10.32±1.40 | 8.75±1.43 | 7.41±0.68 | 6.46±1.06 |
早幼粒细胞Pr | 9.54±0.73 | 8.86±0.55 | 7.08±0.53 | 5.95±0.51 |
嗜中性中幼粒细胞MIN | 10.34±1.23 | 9.01±1.28 | 6.81±0.74 | 5.61±0.74 |
嗜中性晚幼粒细胞LIN | 9.15±0.81 | 8.40±0.72 | 6.13±0.32 | 5.36±0.54 |
嗜中性粒细胞Ne | 10.39±0.70 | 9.78±0.69 | 5.84±0.48 | 5.16±0.37 |
嗜碱性粒细胞Ba | 8.91±0.41 | 8.01±0.60 | 6.11±0.65 | 5.19±0.59 |
嗜酸性粒细胞Eo | 7.72±0.66 | 6.77±0.69 | 5.53±0.47 | 4.77±0.38 |
原淋巴细胞PL | 7.76±0.99 | 6.31±0.76 | 6.24±0.84 | 5.33±0.75 |
幼淋巴细胞IL | 7.08±0.64 | 5.64±0.39 | 5.92±0.52 | 5.19±0.40 |
大淋巴细胞LL | 6.86±1.37 | 5.97±0.93 | 5.20±1.16 | 4.74±0.80 |
小淋巴细胞SL | 6.12±0.78 | 5.32±0.73 | 5.03±0.57 | 4.16±0.50 |
浆细胞Pl | 10.04±1.47 | 8.68±0.94 | 6.09±0.69 | 4.91±0.74 |
原单核细胞PMo | 9.37±1.16 | 8.44±1.65 | 7.62±0.78 | 6.08±1.08 |
幼单核细胞Imo | 11.72±0.99 | 10.92±0.67 | 8.53±1.17 | 5.75±0.87 |
单核细胞Mo | 13.20±1.57 | 10.37±1.25 | 8.15±1.34 | 6.46±0.96 |
血栓细胞Th | 4.47±0.21 | 3.75±0.28 | 4.24±0.16 | 3.72±0.31 |
巨噬细胞 Ma | 15.19±1.93 | 11.29±1.73 | 8.14±0.48 | 6.87±0.69 |
注: 表中数据为8尾实验鱼数据的平均值±标准差。 |
The authors have declared that no competing interests exist.
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