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1 44 2 Vol.44, No OCEANOLOGIA ET LIMNOLOGIA SINICA Mar., 2013 (Acrossocheilus fasciatus) * 1, 2 1, ( ; ; ; ), (Acrossocheilus fasciatus),, 4 5, ; 6 18,, ; 19 20,,, ; 21 22,, ; 23 24,, ; 25, 6 7, 9, 12,, 4 ; ; ; S917,,, 鲃, (Tor brevifilis)(, 1998) (Varicorhinus macrolepis)(, 2005) 鲃 (Spinibarbus caldwelli)(, 2004;, 2008) 鲃 (Spinibarbus hollamdi)(, 2004) (Acrossocheilus hemispinus)(, 2010) 1) (Acrossocheilus fasciatus) (, 1989), 鲃,,,, (, 2010);, (, 2012) (, 2009) (, 2012) ;,,,, ; 50 80m 2 ;, 5 26 ; /m 2,, *, 2011GA ;, ,, realjiangjianhu@126.com; :,,, daihaiping2005@163.com 1), :, :,, zhujunquan@nbu.edu.cn : , :

2 2期姜建湖等: 养殖光唇鱼(Acrossocheilus fasciatus)卵巢发育的组织学观察 349 保持池水中溶解氧在 4mg/L 以上, ph ; 投喂粗蛋白含量为 35%的配合饲料; 养殖期间定期清污, 保持池中整洁 2010 年 11 月 2011 年 10 月, 每月从养殖的及 2010 世代光唇鱼中各随机取样 20 尾; 2011 年 8 10 月, 增加 2011 世代光唇鱼的取样, 每月 20 尾共获样品鱼 780 尾(其中雄鱼 365 尾, 雌鱼 415 尾) 实验用鱼用丁香酚麻醉后进行解剖, 观察卵巢的形态并拍照, 同时测定体长体质量卵巢质量等; 计算卵巢成熟系数=卵巢质量/体质量 100% 取部分卵巢样品用 Bouin 氏液固定后进行石蜡切片制样, 切片经 H.E 染色后于 Olympus BX51 显微镜下观察, 用 Image-Pro Plus 6.0 图像分析软件拍照及测量生殖细胞大小卵巢发育分期参照刘筠(1993)和楼允东(1999)的方法图1 光唇鱼卵巢发育的解剖学观察 Fig.1 Anatomical observation on ovary development of A. fasciatus a.Ⅰ期卵巢 b. 第一次性周期内的Ⅱ期卵巢 c. 产后恢复的Ⅱ期卵巢 d. Ⅲ期卵巢 e. Ⅳ 期卵巢 f. V 期卵巢 g. Ⅵ Ⅳ期卵巢 h. Ⅵ期卵巢 2 结果 2.1 卵巢发育的形态及组织学结构特征光唇鱼卵巢一对, 位于体腔中鳔腹面的两侧, 外育的Ⅱ期卵巢结实而富有弹性, 颜色偏黄(图 1c) 切被卵巢膜, 属被卵巢根据解剖及组织学切片观察结 2c); 根据第Ⅱ时相卵母细胞的大小及形态结构变化, 果, 将光唇鱼卵巢发育划分为Ⅰ Ⅵ时期将其分为早中晚三期早期: 卵母细胞呈多角圆 Ⅰ期扁塌, 半透明略显浅粉色(图 1b); 性成熟产后重复发片显示, 该期卵巢主要由第Ⅱ时相卵母细胞组成(图卵巢呈透明细条状(图 1a) 切片显示, 形或椭圆形, 卵径 μm, 细胞质强嗜碱性, 呈内部主要由第Ⅰ时相卵细胞即卵原细胞及由卵原深蓝色; 细胞核大, 中位, 核径 47 53μm, 核质稀, 细胞刚转化而来的早期初级卵母细胞组成卵原细胞呈浅红色; 核仁个, 大小不等, 沿核膜内缘分布是由原始生殖细胞分化产生, 多成团分布, 胞体近圆 (图 2d) 中期: 卵母细胞体积增大变圆, 卵径 138 形, 卵径 8 12μm; 细胞核大, 呈圆形, 核径 5 7μm, 165μm, 胞质嗜碱性减弱; 细胞核增大, 核径 65 中央大核仁明显可见; 核外为一层薄细胞质(图 2a) 70μm; 核仁个, 多数沿核膜分布, 少数分散早期初级卵母细胞多呈圆形或近圆形, 卵径 17 于核质中; 细胞外周开始出现滤泡细胞, 滤泡细胞核 23μm; 细胞核圆形, 核径 11 13μm, 在核膜内侧出扁平, 嗜碱性, 呈蓝色(图 2e) 晚期: 卵径 172 现一些小核仁; 细胞质增多(图 2b) Ⅰ期卵巢仅在第 208μm, 胞质呈弱嗜碱性; 椭圆形卵黄核在细胞质中一次性周期内的 4 5 月龄鱼中出现(表 1) 出现; 近核膜处的个别核仁被囊泡包裹, 似要排入细 Ⅱ期卵巢呈扁带状, 微血管丰富, 肉眼尚看胞质中; 卵母细胞质膜外的滤泡细胞数量不断增多, 不清卵粒第一次性周期内初次发育的Ⅱ期卵巢松软最终形成单层滤泡膜(图 2f) 第一次性周期内 6 18

3 表月龄光唇鱼的卵巢发育 Tab.1 Ovary development of A. fasciatus of 4 to 27 month-old after hatching (. ) (cm) (g) ± ± ± ± ± ± ± ± ± ± ± ±7.15 1) ± ± ± ± ± ±8.91 2) ± ± ± ±8.72, ± ±9.54, ± ± ± ±9.24 : 1) 2 ; 2) 2 19, ( 1),, ;, ( 1d),, 35% 67%( 2g), :, μm, μm; 44 56, ;, ; ( 2h, 2i) :, μm;,, ; ;, 2.1μm( 2j, 2k) :, μm;, ( 2l), ( 1),,, ( 1e),, ; 55% 86%( 3a), :, μm;, μm,, ;, ;, ; 5μm( 3b, 3c) : μm,,,, ( 3d) : μm, ;, 11μm( 3e, 3f), ( 1), ( 1f),,,,, 8% 17%, 3% 14%( 3g),, μm( 3h, 3i),, ( 1) 7, ( 1g),,,,,

4 2期姜建湖等: 养殖光唇鱼(Acrossocheilus fasciatus)卵巢发育的组织学观察 351 Ⅳ卵巢; 8 月份的大部分雌鱼产后卵巢松弛萎缩, 呈时相卵母细胞和空滤泡组成(图 3i) 第一次性周期内, 橘黄色(图 1h), 切片显示, 卵巢中主要由第Ⅱ 第Ⅲ 24 月龄少部分鱼及 25 月龄鱼卵巢处于该期(表 1) 图2 光唇鱼卵巢发育的组织学观察(1) Fig.2 Histological observation on ovary development of A. fasciatus a. Ⅰ期卵巢, 示卵原细胞及早期初级卵母细胞 b. Ⅰ期卵巢, 示早期初级卵母细胞 c. Ⅱ期卵巢, 示第Ⅰ 第Ⅱ时相卵母细胞 d. 第Ⅱ时相早期卵母细胞, 示核仁 e. 第Ⅱ时相中期卵母细胞, 示核仁及滤泡细胞 f. 第Ⅱ时相晚期卵母细胞, 示核仁卵黄核及滤泡膜 g. Ⅲ期卵巢, 示第Ⅱ 第Ⅲ时相卵母细胞 h. 第Ⅲ时相早期卵母细胞, 示液泡 i. 第Ⅲ时相早期卵母细胞, 示鞘膜细胞及液泡 j. 第Ⅲ时相中期卵母细胞, 示卵黄粒及液泡 k. 第Ⅲ时相中期卵母细胞, 示卵黄粒液泡放射带及滤泡膜 l. 第Ⅲ时相晚期卵母细胞, 示液泡 EPO. 早期初级卵母细胞; FC. 滤泡细胞; FM. 滤泡膜; NU. 核仁; OG. 卵原细胞; TC. 鞘膜细胞; VA. 液泡; YG. 卵黄粒; YN. 卵黄核; ZR. 放射带

5 海 352 图3 洋与湖沼 44 卷光唇鱼卵巢发育的组织学观察(2) Fig.3 Histological observation on ovary development of A. fasciatus a. Ⅳ期卵巢, 示第Ⅱ 第Ⅲ及第Ⅳ时相卵母细胞 b. 第Ⅳ时相早期卵母细胞, 示细胞核及卵黄粒 c. 第Ⅳ时相早期卵母细胞, 示放射带 d. 第Ⅳ时相中期卵母细胞, 示细胞核及卵黄粒 e. 第Ⅳ时相晚期卵母细胞, 示卵黄粒 f. 第Ⅳ时相晚期卵母细胞, 示放射带 g. Ⅴ期卵巢, 示第Ⅲ 第Ⅳ及第Ⅴ时相卵母细胞 h. 第Ⅴ时相卵母细胞, 示放射带 i. Ⅵ期卵巢, 示第Ⅱ 第Ⅲ时相卵母细胞及空滤泡 EF. 空滤泡; N. 细胞核 2.2 性成熟后卵巢发育周年变化 2010 年 11 月 2011 年 10 月, 对 2008 世代月龄光唇鱼产后卵巢发育及月平均成熟系数变化情况的观察分析显示(图 4), 11 月卵巢发育处Ⅱ期, 平均成熟系数为 1.81%; 12 月卵巢发育进入第Ⅲ期进行越冬, 12 3 月平均成熟系数为 2.66% 3.74%; 翌年 4 月, 卵巢发育达Ⅳ期, 进入产卵前期, 平均成熟系数达 8.23%; 5 月, 平均成熟系数达 12.56%, 部分雌鱼卵巢发育至第 V 期, 可进行产卵; 6 月, 所有雌鱼卵巢发育达第 V 期, 平均成熟系数达最高值 25.9%, 进入产卵盛期; 7 月, 平均成熟系数为 15.14%, 产后雌图4 Fig 世代光唇鱼卵巢成熟系数的周年变化 Annual change of ovary gonadosomatic index of A. fasciatus of generation 2008

6 2 : (Acrossocheilus fasciatus) 353,,, 8, 5.15%, 9,,, 10, % 1.71% , 22 23, 2 3, (, 1998) 鲃 (Spinibarbus sinensis)(, 2003) (Varicorhinus simus)(, 2008) ,,, (, 1998) (, 2005) (, 2010) 1) ; 4 ; 6 7,,, (, 2010) 3.2, V,, ;, ; V V, 25%, 7,,,,,, (, 2005), (4 5 6 ), (6 7 ),, 3.3,,, ;,, ;,, 11μm, (, 2005) (Tridentiger trigonocephalus)(, 2009) (Ilisha elongata)(, 2001),, ; (Boleophthalmus pectinirostris)(, 2010),,,, ( 瑔, 1988; Jiang et al, 2010; Ma et al, 2012),,,, ;, (Brandao et al, 2003; Suzuki et al, 2000),,,,,, 鲃., 50(2): ,,, ( ). :, , :, 20 41,,, 鲃., 38(1): ), :, 17 40

7 354 44,,, ( ), 28(4): ,, ( ), 29(3): ,,, , 21(4): ,,, , 41(6): 瑔, , 12(3): ,,, (Acrossocheilus fasciatus)., 43(2): ,, , 25(4): ,,, 鲃., 39(4): 7 10,,, , 34(2): ,,, , 33(2): ,, ( ), 36(3): ,, , 2(2): , :, ,,, 鲃., 33(3): Brandao C A, Valentim M D, Caramaschi E P, Ovary maturation stages and oocyte features in three species of the neotropical fish Hemiodus (M ller, 1842). Brazilian Archives of Biology and Technology, 46(3): Jiang Y Q, Zhang T T, Yang W X, Formation of zona radiata and ultrastructural analysis of egg envelope during oogenesis of Chinese perch Siniperca chuatsi. Micron, 41(1): 7 14 Ma X X, Zhu J Q, Zhou H et al, The formation of zona radiata in Pseudosciaena crocea revealed by light and transmission electron microscopy. Micron, 43(2 3): Suzuki H I, Agostinho A A, Winemiller K O, Relationship between oocyte morphology and reproductive strategy in loricariid catfishes of the Paran River, Brazil. Journal of Fish Biology, 57(3): HISTOLOGICAL OBSERVATION ON THE OVARY DEVELOPMENT OF CULTIVATED ACROSSOCHEILUS FASCIATUS JIANG Jian-Hu 1, 2, DAI Hai-Ping 1,4, ZHU Jun-Quan 1, ZHANG Yu-Ming 3, LEI Shi-Yong 1 (1. Key Laboratory of Applied Marine Biotechnology, Ministry of Education, Ningbo University, Ningbo, ; 2. Zhejiang Institute of Freshwater Fisheries, Huzhou, ; 3. Xinchang Bureau of Water Conservancy, Xinchang, ; 4. Fishery Administration and Technical Service Station of Yinzhou, Ningbo, ) Abstract Morphological and histological features of ovary development of Acrossocheilus fasciatus were studied using dissecting and tissue sectioning techniques. The results show that the ovary development in the first sexual cycle contained I IV continuous stages. Four to five months after hatching (MAH), the ovary was at stage I and contained stage I oocytes. Six to 18 MAH, the ovary was at stage II, and contained stage II oocytes, during which unilaminar follicle membrane formed. Nineteen to 20 MAH, the ovary was at stage III, in which stage III oocytes were contained, the number of layers in follicle membrane changed from one to two, the zona radiate formed between the plasma membrane and follicle membrane. Furthermore, the vacuole and yolk granule appeared in the cytoplasm. Twenty-one to 22 MAH, the ovary was at stage IV, stage IV oocytes were contained, the oocyte cytoplasm was full of yolk granule. Twenty-three to 24 MAH, the ovary was at stage V, stage V oocytes were contained, and the follicle membrane at this stage began to desquamate. Twenty-five MAH, the ovary was at stage VI, empty follicle and oocytes at stage II and III could be found. The spawning season of this species was from June to July, the ovary changed back to stage II in September; in December it changed back to stage III, and over all the winter the ovary remained at this stage. In April, the ovary continued to develop and entered into stage IV. According the morphological and histological characteristics of ovary development, A. fasciatus should be classified into the group of short-term batch-releasing patterned fish. Key words Acrossocheilus fasciatus; ovary development; oocyte; histology

2 : 419 ( -3) 第 2 时相 (1) :, ;,,,,,,, 5 7 ( -4) (2) :, 0.2 0.5 µm ( -5),, (3) :,,,,,,, ( -6) ( -7) 第 3 时相 (1) :,,, ( -8) ( -9), 2.0 3.0 µm,, (2) : 2

2 : 419 ( -3) 第 2 时相 (1) :, ;,,,,,,, 5 7 ( -4) (2) :, 0.2 0.5 µm ( -5),, (3) :,,,,,,, ( -6) ( -7) 第 3 时相 (1) :,,, ( -8) ( -9), 2.0 3.0 µm,, (2) : 2 34 2 Vol. 34, No.2 2010 3 ACTA HYDROBIOLOGICA SINICA Mar., 2 0 1 0 DOI: 10.3724/SP.J.1035.2010.00418 曹伏君罗杰刘楚吾 (, 524025) :, : 1, 3, 4 6, 7 8 10, 6 :,, 2, 5 GSI, 0.70%,, 7 9 GSI 11 2 GSI 0.08% 0.20%,,