大鳍鳠卵子发生的研究
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摘要
1988年6月至9月及2001年9月至2002年5月用组织学、组织化学方法研究大鳍鳢性分化及卵细胞发育过程的形态结构变化、卵黄物质的积累、滤泡细胞的发育、精孔细胞的生成、卵膜的形成等。结果表明:
     大鳍鳠仔鱼出膜后3天,可见一个PGCs位于肠的背方、肾管下的中胚层中,其大小为15×17.5μm,核径7.5μm,具一核仁,核膜清晰,核中基质着色淡,胞质略透明,细胞一端有稍微变形的突起,与其周围的中胚层细胞有明显的区别。
     出膜后5天,肾管下腹膜中胚层细胞聚集形成一对嗜碱性细胞纵行隆起,为生殖嵴,个别PGCs已迁入其中,与生殖嵴共同组成生殖腺。生殖腺继而深入体腔,悬挂于体腔系膜上,并逐步向精巢或卵巢分化。组成生殖嵴的体细胞没有明显的二重起源。PGCs来源于胚胎发育早期非生殖嵴以外的胚胎细胞,经过迁移而进入生殖嵴。早期生殖腺的纵切面中可见PGCs呈“链状”分布。
     出膜后19天,生殖腺明显分支,在某些区域两分支的游离端亦有融合的趋势,且在一个横切面上可见有多个PGCs,可能为生殖腺向卵巢分化的早期特征。
     大鳍鳠卵子的发育过程分为7个时相:原始生殖细胞时相、卵原细胞时相、卵母细胞单层滤泡时相、卵黄泡出现时相、卵黄充满时相、卵母细胞成熟和退化时相。刚迁移入生殖嵴的PGCs,呈卵原形,核质比例较大,胞径(7-13×7.5-15μm),核径(5.2-6.25×6.0-7μm),细胞质微嗜酸性,核膜清晰,核中基质着色淡,具1-2个明显的嗜碱性大核仁。PGCs进而增殖、分化为精原细胞或卵原细胞。卵原细胞呈卵圆形至椭圆形,核比例较大,位于细胞中央,核膜内缘有分
    
     理学硕士学位论文:大幻鲍卵于发生的研究
     散的核质,卵径*q,核径610pm。胞质少,在各种染色中均相对淡
     染,细胞核膜清晰,有卜 个核仁。卵原细胞排列杂乱无章,往往成堆存在于
     生殖上皮内,除了在生殖季节期间处于比较静止外,其它时间都十分活跃,均
     在进行频繁的有丝分裂,并进一步发育为初级卵母细胞。处于卵母细胞单层滤
     泡时相时,卵母细胞大小增加,其周围开始出现一薄层结缔组织样膜,随后其
     内缘分化出2-3’个滤泡细胞(不成层,细胞小,长梭形),胞质嗜碱性增强,
     各种染色中比卵原细胞染色深,其中可见有“外周环”,晚期时可见卵黄核。
     卵母细胞卵黄泡出现时相的主要特征有:滤泡细胞2-3层,内层细胞进一步发
     育为立方形颗粒细胞,卵黄膜出现并逐渐增厚,胞质中出现卵黄泡,核膜波曲
     明显,核仁物质外排,卵母细胞体积增大明显。卵黄充满时相的主要特征有:
     卵黄膜停止增长并逐渐变薄,颗粒细胞柱状、胞质中充满颗粒物,卵母细胞中
     充满了圆形、椭圆形或不规则的卵黄颗粒,大小不一,直径1.2*5Pm。生理
     成熟的卵母细胞与滤泡细胞层分离,并落入卵巢腔中,不能达到生理成熟的卵
     母细胞则逐步退化,最终被卵巢自身吸收而消失。
     大鳍鳍卵母细胞的卵黄发生有内源性和外源性两种方式,早期主要为内源
     性的卵黄发生,晚期主要为外源性的卵黄发生。卵黄的主要成分为多糖类碳水
     化合物、脂类和蛋白质,最先形成的是多糖类碳水化合物卵黄,之后出现脂肪
     卵黄,蛋白卵黄最后形成。卵黄积累时由胞质外周向内逐渐进行。
     大鳍鳍卵巢滤泡细胞来源于卵巢基质细胞,由围在卵细胞外围的卵泡膜细
     胞层和其内侧的颗粒细胞层组成。滤泡细胞从发生到退化的全过程分为7个时
     期:零散卵泡膜细胞期、单层卵泡膜细胞期、多层卵泡膜细胞期、立方形颗粒
     细胞期、柱状颗粒细胞期、颗粒细胞分泌期和颗粒细胞退化期。滤泡细胞在卵
     母细胞的发育、成熟、退化中对卵母细胞有保护、供给营养、分化出精孔细
     胞、形成次级卵膜、破坏并吞噬卵母细胞的功能。同时还具有合成卵黄半成品
     和分泌类固醇激素及协助排卵等功能。
     大鳍鳍与大多数硬骨鱼类一样仅有一受精孔具单一精孔细胞。精孔细胞由
     一滤泡细胞发育而成。
     “大鳍鳍的成熟卵外由初级卵膜(卵黄膜)和次级卵膜(胶质膜)包裹,二
     者分别由卵母细胞和晚期滤泡细胞分泌形成。
From June to September, 1988 and from October, 2001 to May, 2002, gonadal sex differentiation, changes on the structure of oocytes, formation of yolk granules, development of follicle cells as well as formation of the micropylar cell and the egg membrane in Mystus macroptems were studied by means of histological and histochemical methods. The research results are as follows:
    Three days after hatching (dan), a primordial germ cell (PGC) was discovered at the mesoblast below the segmental duct. PGC is distinct from the neighboring cells for its own specific characters: size of 15 X 17.5urn, nuclear diameter of 7.Sum, distinct nuclear membrane, one nucleolus, faint staining nuclear matrix, transparent cytoplasm as well as protuberances on cytomembrane.
    In 5 dah fish, two distinct genital ridges appeared as the outgrowth of basophilic cells from the peritoneal mesoblasts. The genital ridge and several PGCs migrated into it form the primitive gonad which stick up toward the celom and suspend on the membrane of celom. The primitive gonad will differentiate into testis or ovary gradually in the later stage. Somatic cells in genital ridge were derived from the peritoneal wall showing no recognizable signal of dual origion as amphibians before or during the sex-differentiation period. Primordial germ cells are not derived from the epithelia of genital ridge but from those cells which belong to the early embryo and migrate into the genital ridge. PGCs were seen as beads on a chain on the longitudinal section of early stage of gonad.
    The genital ridges begins to laterally elongate to form the distal and proximal sides of the gonadal folds in 12 dah fish, and the two lateral folds are almost fused in 19 dph fish. Several PGCs could be seen on a transverse section. These maybe the early signals for ovary differentiation.
    The oogenesis process in Mystus macroptems can be divided into seven phase: primordial germ cell, oogonium, oocyte with single layered follicle cells, oocyte with yolk vacuole, oocyte with full of yolk granules, mature ovum and retrogressive oocyte. The PGCs which has newly migrated into genital ridges are egg-like with a high nuclear-cytoplasmic ratio in terms of its diameter of 7-13nmX 7.5-1 Sum and diameter of nucleolus of 5.2-6.25|amX6.0-7um. When stained with H.E, acidophillic cytoplasm show light red compared with a faint stain in nuclear matrix which often contain 1-2 big basophilic nucleolus. Then PGCs grow and differentiate into spermatogonia or oogonia. Oogonia are ovate or elliptic with a high nuclear-cytoplasmic ratio and its
    
    
    nuclear matrix disperse around the inner layer of the nuclear membrane. The diameter of cell and nucleus ranges from 1 l-18um and 6-10um respectively. The oogonia, with clear nuclear membrane and 1-2 nucleolus, show faint staining when stained with various staining methods. Oogonia were seen in the genital ridge irregularly, or stacked around one area. They maintain an active state (in active mitosis or developing into primary oocyte) through all the time except in the reproductive season. During the phase of oocyte with single layered follicle cells, oocyte increase in size and become more basophillic, staining is thicker with various staining reagent compared with oogonia and an "outer ring" appeare around the outer area of the cytoplasm. Later on, yolk nucleus manifest clearly. Characteristics of oocyte with yolk vacuole were as follows: size of the body increases; follicle cells develop into 2-3 layer and the inner layer develop into cuboidal granule cells; vitelline envelope appear and thicken; yolk vac
    uole emerge in the ooplasm; Wave like-nuclear membrane become evident; components of nucleolus were released.
    Also, the characteristics during the phase of oocyte with full of yolk granules are as follows: vitelline envelope growth halt and become thinner; granule cells become columniation-shaped with its cytoplasm full of granule particles; oocyte is teemed with yolk granule which shaped round, elliptical or randomly with a diameter ranging from 1.2 urn to 4.
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