Development of vertebral column and appendicular skeleton in larvae and juveniles of Platax teira
-
摘要: 掌握尖翅燕鱼 (Platax teira) 仔稚鱼脊柱及附肢骨骼系统的发育时序特征,可为其苗种繁育、环境适应及分类鉴定提供理论依据。采用软骨-硬骨双染色法对尖翅燕鱼仔稚鱼 [0~30日龄 (dah)] 脊柱、胸鳍、腹鳍、背鳍、臀鳍以及尾鳍进行染色观察与分析。结果表明,尖翅燕鱼有24枚脊椎骨,尾部骨骼由5枚尾下骨和2枚尾上骨构成,附肢支鳍骨的发育顺序为胸鳍、腹鳍、尾鳍、臀鳍和背鳍。尖翅燕鱼在初孵仔鱼已具备胸鳍支鳍骨原基和脊索,4 dah仔鱼胸鳍和腹鳍最先发育;7 dah仔鱼髓弓、脉弓开始出现;13 dah仔鱼骨骼形态和数量基本稳定;26 dah左右稚鱼开始骨化,脊柱和脊椎最先完成骨化;30 dah进入幼鱼阶段,其外表形态已经固定,与成鱼无异,典型特征为头部、躯干部和尾部各有3条黑色条带,身体呈雪白银色,受到外界刺激时体色瞬间变为黑色。尖翅燕鱼脊柱及附肢骨的发育顺序与其他海水鱼基本一致,但骨骼发育时间节点和数量有所不同。Abstract: Understanding the developmental time sequence characteristics of the spine and appendage skeletal system of Platax teiracan provide a theoretical basis for its seedling breeding, environmental adaptation and classification. We stained and analyzed the spine, pectoral fin, pelvic fin, anal fin and caudal fin ofP. teira larvae (0–30 dah) by the method of cartilaginous and bony staining. The results show that there were 24 vertebrae, and the tail bone was composed of five subcaudal bones and two upper caudal bones. The development sequence of appendage branch fin bones was pectoral fin, abdominal fin, caudal fin, anal fin and dorsal fin. The first hatched fish already had pectoral branch fin bone primordium and notochord. At 4 dah, the pectoral fin and pelvic fin developed first; at 7 dah, medullary arch and vein arch appeared; at 13 dah, the morphology and number of larval bones were basically stable; at about 26 dah, ossification began in juvenile fish, and the spine and spine were the first to complete ossification; at 30 dah, the larvae entered the juvenile stage, and their appearance has been fixed. The typical characteristics were three black bands on the head, trunk and tail, and the body was white silver. When being stimulated or stressed by external stimulation, the body color changed to black instantly, and the shape had no difference from that of the juvenile. The developmental sequence of the spine and appendage bones was basically the same with that of other sea fish, but the time nodes and number of the development of bone were different.
-
Key words:
- Platax teira /
- Bone staining /
- Larval and juvenile development /
- Spine /
- Appendicular skeleton
-
表 1 阿尔新蓝-茜素红S染色法
Table 1. Alcian blue and alizarin red S staining technique
染色步骤
Staining step实验过程
Experimental process样本处理
Sample treatment将仔稚鱼麻醉,测量体长和全长,注意不要采集外形畸形的样品,用10%的甲醛溶液浸泡样本,4 ℃条件下浸泡超过24 h,完成样品的固定。 脱水
Dehydration取出固定样品后,用纱布包裹样品,蒸馏水浸泡至少24 h (中间多换几次蒸馏水),或用自来水流水冲洗过夜,以清除固定剂。再用不同浓度的乙醇梯度脱水,乙醇溶液浓度依次为:50%、75%、90%和100%,脱水时间均为6 h。 软骨染色
Cartilage staining从脱水液中取出的样品,直接浸入软骨染色液 (将无水乙醇和冰醋酸1∶1混合,加入 0.3 g阿利新蓝粉末,配置成1 000 mL溶液) 中,染色24 h,期间可以更换染液。 漂洗
Rinsing在光线较强的环境下进行操作,观察到样品染色好后,从软骨染色液中取出样品放入漂洗液 [将30%的过氧化氢 (H2O2) 和蒸馏水按照1∶5混合配制成1 000 mL溶液,并加入8 g氢氧化钾 (KOH)] 中,不间断用玻璃棒搅拌,直到样品表面颜色变淡。 中和
Neutralize将漂洗好的样品放入染色中和液 [蒸馏水中加入过量硼砂 (Na2B4O7),得饱和硼砂溶液即可] 中进行中和,时间为20~24 h。 硬骨染色
Bone staining将样品放入硬骨染色液 (1 000 mL蒸馏水中加入8 g 氢氧化钾 (KOH) 完全溶解后,再加入0.3~0.5 g茜素红粉末) 中,每隔12 h取出样品观察染色情况,直至染色完成。 酶解
Zymolysis将样本浸泡在酶溶液 (将1.25 g重金属螯合剂EDTA-Na2和4.5 g胰蛋白酶溶解于1 000 mL饱和硼砂溶液) 中,根据酶解程度调整消化时间,直至稍见骨骼即可,中间最好更换酶解液。 保存
Preservation用蒸馏水清洗样本,然后依次移入体积比为3∶1、1∶1、1∶3的1% KOH-甘油合剂中,最后保存于纯甘油中,并加入3~4 粒麝香草酚保存。 -
[1] 周胜杰, 马婷, 胡静, 等. 尖吻鲈仔鱼骨骼发育观察[J]. 南方农业学报, 2018, 49(3): 592-598. [2] 韩明洋, 周胜杰, 马振华. 鱼类仔稚鱼骨骼及其畸形发生研究进展[J]. 水产学杂志, 2021, 34(2): 94-100. [3] 邓平平, 施永海, 徐嘉波, 等. 金钱鱼仔稚鱼脊柱及附肢骨骼系统早期发育研究[J]. 水产科学, 2021, 40(4): 596-602. [4] 郑攀龙. 卵形鲳鲹仔稚鱼骨骼发育及骨骼畸形研究[D]. 上海: 上海海洋大学, 2015: 8-9. [5] 吕雪娇, 王雨浓, 刘清华, 等. 鞍带石斑鱼仔稚幼鱼骨骼发育与生长特性研究[J]. 海洋科学, 2018, 42(5): 116-121. doi: 10.11759/hykx20180308002 [6] BILECENOGLU M, KAYA M. A new alien fish in the Mediterranean Sea: Platax teira (Forsskal, 1775) (Osteichthyes: Ephippidae)[J]. Aquat Invasions, 2006, 1(2): 80-83. doi: 10.3391/ai.2006.1.2.5 [7] ZENG C, LIU B S, ZHU K C, et al. Comparison of body height, body length, full length and body weight growth curves of the longfin batfish, Platax teira[J]. Pak J Zool, 2021, 53(2): 1-7. [8] ZENG C, LIU B S, ZHU K C, et al. Histological study on development of the digestive system of longfin batfish (Platax teira)[J]. Isr J Aquac-Bamidgeh, 2020, 72: 9. [9] LIU B, GUO H Y, ZHU K C, et al. Nutritional compositions in different parts of muscle in the longfin batfish, Platax teira (Forsskal, 1775)[J]. J Appl Anim Res, 2019, 47(1): 403-407. doi: 10.1080/09712119.2019.1649680 [10] 郑攀龙, 马振华, 郭华阳, 等. 卵形鲳鲹尾部骨骼胚后发育研究[J]. 南方水产科学, 2014, 10(5): 45-50. [11] 张伟, 刘莹, 官曙光, 等. 大泷六线鱼仔稚鱼头部骨骼发育观察[J]. 水生生物学报, 2021, 45(4): 898-905. [12] 郑珂, 岳昊, 郑攀龙, 等. 海水养殖鱼类仔、稚鱼骨骼发育与畸形发生[J]. 中国水产科学, 2016, 23(1): 250-261. [13] 王永梅, 唐文乔. 中国鲤形目鱼类的脊椎骨数及其生态适应性[J]. 动物学杂志, 2014, 49(1): 1-12. [14] 王秋荣, 毕建功, 林利民, 等. 青石斑鱼骨骼发育异常的形态特征[J]. 大连海洋大学学报, 2012, 27(5): 417-421. [15] 李仲辉, 杨太有. 大口黑鲈和尖吻鲈骨骼系统的比较研究[J]. 动物学报, 2001, 47(S1): 110-115. [16] 崔科. 红鳍笛鲷胚后发育的条件优化[D]. 上海: 上海海洋大学, 2018: 8-22. [17] KOUMOUNDOUROS G, GAGLIARDI F, DIVANACH P, et al. Normal and abnormal osteological development of caudal fin in Sparus aurata L. fry[J]. Aquaculture, 1997, 149(3/4): 215-226. [18] 邓平平, 施永海, 徐嘉波, 等. 美洲鲥仔稚鱼脊柱及附肢骨骼系统的早期发育[J]. 中国水产科学, 2017, 24(1): 73-81. [19] 于超勇, 官曙光, 于道德, 等. 大泷六线鱼仔稚鱼脊柱及附肢骨骼系统的发育观察[J]. 大连海洋大学学报, 2020, 35(1): 47-55. [20] GAVAIA P J, DINIS M T, CANCELA M L. Osteological development and abnormalities of the vertebral column and caudal skeleton in larval and juvenile stages of hatchery-reared Senegal sole (Solea senegalensis)[J]. Aquaculture, 2002, 211(1/2/3/4): 305-323. [21] 王秋荣, 倪玥莹, 林利民, 等. 大黄鱼仔稚鱼脊柱、胸鳍及尾鳍骨骼系统的发育观察[J]. 水生生物学报, 2010, 34(3): 467-472. [22] BAI L R, ZHAO Z Y. Study on embryonic development, larva, juvenile and young fish of Epinephelus fuscoguttatus[J]. Agric Sci Technol, 2016, 17(6): 1305-1314,1354. [23] 周胜杰, 胡静, 杨其彬, 等. 尖吻鲈胚胎发育及胚后发育观察[J]. 水产研究, 2020, 7(1): 58-68. [24] 黄贤克, 单乐州, 闫茂仓, 等. 黄姑鱼胚胎发育及其与温度和盐度的关系[J]. 海洋科学, 2017, 41(7): 44-50. doi: 10.11759/hykx20160920003 [25] 韩明洋. 温度胁迫下卵形鲳鲹的转录响应及其脊椎的组织病理和分子表征研究[D]. 上海: 上海海洋大学, 2021: 25-32. [26] 吕雪娇. 大菱鲆 (Scophthalmus maximus) 和鞍带石斑鱼 (Epinephelus lanceolatus) 生长特性以及骨骼发育与畸形研究[D]. 青岛: 中国科学院大学 (中国科学院海洋研究所), 2018: 70-79. [27] 邓平平, 严银龙, 施永海. 褐菖鲉仔稚鱼脊柱及附肢骨骼系统的早期发育[J]. 浙江大学学报 (农业与生命科学版), 2018, 44(6): 735-742. [28] 张宗锋, 施永海, 张根玉, 等. 刀鲚脊柱及附肢骨骼早期发育研究[J]. 水产科技情报, 2015, 42(4): 175-178. [29] LANGILLE R M, HALL B K. Development of the head skeleton of the Japanese medaka, Oryzias latipes (Teleostei)[J]. J Morphol, 1987, 193(2): 135-158. doi: 10.1002/jmor.1051930203 -