日本沼虾（Macrobrachium nipponense）卵黄蛋白及切除眼柄对其积累影响的研究
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摘要
卵黄蛋白是甲壳动物胚胎和幼体生长发育所需的能量和营养来源，因此，它的合成、积累、多寡等等对于胚胎和幼体来说是至关重要的。本文研究了卵黄蛋白的主要成分——卵黄磷蛋白的分离纯化、生化性质等，为最终揭示卵黄蛋白的合成机理奠定基础。同时研究了切除单侧眼柄对于卵黄蛋白积累的影响，为探讨切除单侧眼柄对卵质的影响提供基础资料。
1．日本沼虾卵黄磷蛋白的研究
利用聚丙烯酰胺凝胶电泳，采用不同的染色方法，从成熟日本沼虾卵巢中鉴定出一种卵黄磷蛋白，用电泳洗脱的方法对其进行了纯化，并对其部分生化性质进行了分析。
研究发现，日本沼虾卵黄磷蛋白总分子量为458Ku，SDS—聚丙烯酰胺凝胶电泳分析它由三个多肽亚基组成，分子量分别为110Ku，96Ku，89Ku。同时发现，三亚基的过碘酸-Schiff试剂染色反应均呈阳性，说明三亚基上均有糖辅基的结合。样品缓冲液中β-巯基乙醇的有无并不影响亚基的分离，推测亚基间不存在二硫键。
采用常规免疫方法，以纯化的卵黄磷蛋白作为抗原免疫家兔，制备卵黄磷蛋白的特异性抗血清。利用此抗血清，并结合双向免疫扩散和组织免疫学方法对日本沼虾的雌性特异蛋白进行了免疫学研究，发现：日本沼虾的卵黄磷蛋白为雌性特异蛋白；卵黄磷蛋白和卵黄蛋白原有相似的免疫原性；肝胰腺可能是其外源卵黄蛋白的合成场所。日本沼虾雌性特异蛋白与其他物种(罗氏沼虾、中国明对虾、凡纳滨对虾、中华绒螯蟹、斑马鱼)的雌性特异蛋白间没有免疫交叉反应。
2．切除单侧眼柄对卵黄蛋白积累的影响
采用剪烫法去除日本沼虾单侧眼柄，对其卵巢发育、蛋白积累进行跟踪。发现切除眼柄可以加快卵巢发育，使其提早成熟。SDS—聚丙烯酰胺凝胶电泳比较不同时期切除眼柄的实验组与正常未切眼柄的对照组卵巢中可溶性蛋白组分
摘要
...................................曰................口..................................甲....................................
的异同，发现切除眼柄三周后，实验组的蛋白组分发生了一些变化，比对照组
多出一分子量约为93Ku的蛋白条带，推测切除眼柄对亲虾卵黄蛋白的积累有一
定的影响。利用考马斯亮蓝法(Bradford法)比较实验组与对照组卵巢中总蛋
白含量，利用免疫火箭电泳比较成熟卵巢中卵黄磷蛋白含量，发现成熟卵巢中
无论是总蛋白含量还是卵黄磷蛋白含量，实验组的都要低于对照组的。可见，
切除眼柄虽然可以加快卵巢发育，缩短产卵时间，但是成熟卵巢中总蛋白含量
和卵黄磷蛋白含量降低，从而可能影响卵子质量以及幼体的成活率。
Yolk proteins are the nutritious source in the developmental stage of crustaceans embryos and nauplii. Therefore, the synthesis, the accumulation and the quantity of them is very crucial to the embryos and nauplii. Vitellin is the major component of yolk proteins. In this dissertation, isolation, purification and biochemical characterization of the vitellin from the mature ovary of the freshwater shrimp, Macrobrachium nipponense was studied. This research established the theoretical basis on the synthesis mechanism of yolk proteins. The paper also focused on the effect to the accumulation of yolk proteins as the unilateral eyestalk is ablated. The study of it provided basic data in the further analysis of the effect of unilateral eyestalk ablation to the quality of eggs.
1. The research on the vitellin of M. nipponense
With the PAGE, still applying various staining methods, our study proved one form of vitellin in the mature ovary of M nipponense. According to this paper, the vitellin was purified by electric-elution. And an analysis was carried to its partial biochemical characterization.
During the research, the molecular weight of the vitellin was 458Ku in a gradient native PAGE. Three subunits (110, 96, and 89 Ku) in purified vitellin were detected with SDS-PAGE. In the mean time, we found these three subunits all contained carbohydrate moieties based on the staining with periodic acid-Schiff s reagent. Three subunits of vitellin were also observed in SDS-PAGE whether the vitellin was reduced with j3-mercaptoethanol or not. So, we supposed that the disulfide bonds was no-existence among the three polypeptie subunits.
By applying traditional immunity way and taking the purified vitellin as antigen to inject rabbit, the vitellin's specific antiserum was obtained. With this specific antiserum and applying immunology method, the vitellin of M nipponense were studied . The results showed: the vitellin of M nipponense was the female-specific protein ; both vitellin and vitellogenin had the same immunogenicity ; hepatopancreas might be the extraovarian site of vitellogenin synthesis. There was no immunological cross-reaction between the vitellin of M. nipponense and the vitellin or vitellogenin of other species , for example , M. rosenbergii , Fenneropenaeus chinensis , Litopenaeus vannamei , Eriocheir sinensis , and Brachydanio rerio.
2. Effects on the accumulation of yolk proteins by unilateral eyestalk ablation
Unilateral eyestalk ablation was performed with fine scissors, and the cut stump was cauterized. Our research proved that eyestalk ablation could accelerate ovary's growth speed, so as to pre-mature the shrimp. With the help of SDS-PAGE, we compared the elements of soluble proteins in the ovary between the shrimps of the experiment group (eyestalk ablation) and that of the control group(without ablation). Three weeks later, the proteins component in the experiment group had a little change. In contrast with the control group, the proteins component in the experiment group had a more protein which molecular weight was 93 Ku. We also compared the total proteins concentration between the experiment group and the control group by using Bradford, and compared vitellin in mature ovaries by rocket immunoelectrophoresis. In the mature ovaries, we found that both the proteins concentration and vitellin concentration in the experiment group were lower than that in the control group. It was obvious that, although eyest
alk ablation could quicken the development of ovary, shorten the period of laying eggs, it could reduce the concentration of the total proteins and vitellin. As a result, it may effect on the quality of eggs and the survival rate of nauplii.
1．日本沼虾卵黄磷蛋白的研究
利用聚丙烯酰胺凝胶电泳，采用不同的染色方法，从成熟日本沼虾卵巢中鉴定出一种卵黄磷蛋白，用电泳洗脱的方法对其进行了纯化，并对其部分生化性质进行了分析。
研究发现，日本沼虾卵黄磷蛋白总分子量为458Ku，SDS—聚丙烯酰胺凝胶电泳分析它由三个多肽亚基组成，分子量分别为110Ku，96Ku，89Ku。同时发现，三亚基的过碘酸-Schiff试剂染色反应均呈阳性，说明三亚基上均有糖辅基的结合。样品缓冲液中β-巯基乙醇的有无并不影响亚基的分离，推测亚基间不存在二硫键。
采用常规免疫方法，以纯化的卵黄磷蛋白作为抗原免疫家兔，制备卵黄磷蛋白的特异性抗血清。利用此抗血清，并结合双向免疫扩散和组织免疫学方法对日本沼虾的雌性特异蛋白进行了免疫学研究，发现：日本沼虾的卵黄磷蛋白为雌性特异蛋白；卵黄磷蛋白和卵黄蛋白原有相似的免疫原性；肝胰腺可能是其外源卵黄蛋白的合成场所。日本沼虾雌性特异蛋白与其他物种(罗氏沼虾、中国明对虾、凡纳滨对虾、中华绒螯蟹、斑马鱼)的雌性特异蛋白间没有免疫交叉反应。
2．切除单侧眼柄对卵黄蛋白积累的影响
采用剪烫法去除日本沼虾单侧眼柄，对其卵巢发育、蛋白积累进行跟踪。发现切除眼柄可以加快卵巢发育，使其提早成熟。SDS—聚丙烯酰胺凝胶电泳比较不同时期切除眼柄的实验组与正常未切眼柄的对照组卵巢中可溶性蛋白组分
摘要
...................................曰................口..................................甲....................................
的异同，发现切除眼柄三周后，实验组的蛋白组分发生了一些变化，比对照组
多出一分子量约为93Ku的蛋白条带，推测切除眼柄对亲虾卵黄蛋白的积累有一
定的影响。利用考马斯亮蓝法(Bradford法)比较实验组与对照组卵巢中总蛋
白含量，利用免疫火箭电泳比较成熟卵巢中卵黄磷蛋白含量，发现成熟卵巢中
无论是总蛋白含量还是卵黄磷蛋白含量，实验组的都要低于对照组的。可见，
切除眼柄虽然可以加快卵巢发育，缩短产卵时间，但是成熟卵巢中总蛋白含量
和卵黄磷蛋白含量降低，从而可能影响卵子质量以及幼体的成活率。
Yolk proteins are the nutritious source in the developmental stage of crustaceans embryos and nauplii. Therefore, the synthesis, the accumulation and the quantity of them is very crucial to the embryos and nauplii. Vitellin is the major component of yolk proteins. In this dissertation, isolation, purification and biochemical characterization of the vitellin from the mature ovary of the freshwater shrimp, Macrobrachium nipponense was studied. This research established the theoretical basis on the synthesis mechanism of yolk proteins. The paper also focused on the effect to the accumulation of yolk proteins as the unilateral eyestalk is ablated. The study of it provided basic data in the further analysis of the effect of unilateral eyestalk ablation to the quality of eggs.
1. The research on the vitellin of M. nipponense
With the PAGE, still applying various staining methods, our study proved one form of vitellin in the mature ovary of M nipponense. According to this paper, the vitellin was purified by electric-elution. And an analysis was carried to its partial biochemical characterization.
During the research, the molecular weight of the vitellin was 458Ku in a gradient native PAGE. Three subunits (110, 96, and 89 Ku) in purified vitellin were detected with SDS-PAGE. In the mean time, we found these three subunits all contained carbohydrate moieties based on the staining with periodic acid-Schiff s reagent. Three subunits of vitellin were also observed in SDS-PAGE whether the vitellin was reduced with j3-mercaptoethanol or not. So, we supposed that the disulfide bonds was no-existence among the three polypeptie subunits.
By applying traditional immunity way and taking the purified vitellin as antigen to inject rabbit, the vitellin's specific antiserum was obtained. With this specific antiserum and applying immunology method, the vitellin of M nipponense were studied . The results showed: the vitellin of M nipponense was the female-specific protein ; both vitellin and vitellogenin had the same immunogenicity ; hepatopancreas might be the extraovarian site of vitellogenin synthesis. There was no immunological cross-reaction between the vitellin of M. nipponense and the vitellin or vitellogenin of other species , for example , M. rosenbergii , Fenneropenaeus chinensis , Litopenaeus vannamei , Eriocheir sinensis , and Brachydanio rerio.
2. Effects on the accumulation of yolk proteins by unilateral eyestalk ablation
Unilateral eyestalk ablation was performed with fine scissors, and the cut stump was cauterized. Our research proved that eyestalk ablation could accelerate ovary's growth speed, so as to pre-mature the shrimp. With the help of SDS-PAGE, we compared the elements of soluble proteins in the ovary between the shrimps of the experiment group (eyestalk ablation) and that of the control group(without ablation). Three weeks later, the proteins component in the experiment group had a little change. In contrast with the control group, the proteins component in the experiment group had a more protein which molecular weight was 93 Ku. We also compared the total proteins concentration between the experiment group and the control group by using Bradford, and compared vitellin in mature ovaries by rocket immunoelectrophoresis. In the mature ovaries, we found that both the proteins concentration and vitellin concentration in the experiment group were lower than that in the control group. It was obvious that, although eyest
alk ablation could quicken the development of ovary, shorten the period of laying eggs, it could reduce the concentration of the total proteins and vitellin. As a result, it may effect on the quality of eggs and the survival rate of nauplii.
引文
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Lee F. Y., Shih T. W., Chang C. F.. Isolation and characterization of the female-specific protein (Vitellogenin) in mature female hemolymph of the freshwater prawn Macrobrachium rosenbergii: comparison with ovarian vitellin. General and Comparative Endocrinology, 1997, 108: 406-415.
Lee R. F., Walker A.. Lipovitellin and lipid droplet accumulation in oocytes during ovarian maturation in the blue crab, Callinects sapidus. J. Exp. Zool., 1995, 271(5): 401-412.
Liu C. W., O' Connor J D.. Biosynthesis of lipovitellin by the crustacean ovary-Ⅱ. Characterization of an in vitro incorporation of amino acid into the purified subunits. J. Exp. Zool., 1976, 195: 41-52.
Liu C. W., O' Connor J. D.. Biosynthesis of crustacean lipovitellin-Ⅲ the incorporation of lablled amino acid into the purified lipovitellin of the crab Pachygrapsus crassipes. J. Exp. Zool., 1977, 199: 105-108.
Longyant S., Sithigomgul P., Thammapalerd N., et al. Characterization of vitellin and vitellogenin of giant tiger prawn Penaeus monodon using monoclonal antibodies specific to viteltin subunits. Invertebr. Reprod. Dev., 2000, 37(3): 211-221.
Masaharu K., Seiichi A.. Isolation of crustacean egg yolk lipoprotein by differential densitygradient ultracentrifugation. Comp. Biochem. Physiol., 1992, 103B: 363-368.
Meusy J. J.. Vitellogenin, the extraovarian precusor of the yolk protein in crustacea, aview. Reprod Nutr Det, 1980, 20: 1-20.
Millamena O. M., Quinitio E.. The effects of diets on reproductive performance of eyestalk ablated and intact mud crab Scylla serrata. Aquaculture, 2000, 181: 81-90.
Moshe T., Menachem G., Michael O.. Localization of the vitellin and its possible precursors in various organs of Parapenaeus longirostis (Crustacea: Decapoda: Penaeidae). Intenational Journal of Invertebrate Reproduction and Development, 12(1987): 1-12.
Moshe T., Milton F., Timothy K., et al. A comparative study of the ovarian proteins from two penaeid shrimps, Penaeus semisulcatus de haan and Penaeus monodon(Boone). Comp. Biochem. Physiol., 1992, 102B(3): 483-490.
Oberdrster E., Rice C. D, Irwin L. K.. Purification of vitellin form grass shrimp Palaemonetes pugio, generation of monoclonal antibodies, and validation for the detection of lipovitellin in Crustacea. Comp. Biochem. Physiol. Part C, 2000, 127: 199-207.
Pateraki L. E, Stratakis E.. Synthesis and organization of vitellogenin and vitellin molecules form the land crab Potamon potamios. Comp. Biochem. Physiol. Part B, 2000, 125: 53-61.
Picaud J. L.. Vitellogenin synthesis by the fat body of Porcellio dilatatus Brand (Crustacea: Isopoda). Invert. Reprod. Dev., 1980, 2: 341-349.
Qiu Y. W, NG T. B and Ghu K. H.. Purification and characterization of vitellin from the ovaries of the shrimp Metapenaeus ensis(Crustacea: Decapoda: Penaeidae). Invertebr. Reprod. Dev., 1997, 31(1-3): 217-223.
Quackenbush L. S.. Vitellogenesis in the shrimp, Penaeus Vannamei: in vitro studies of the isolated hepatopancreas and ovary. Comp. Biochem. Physiol., 1989, 94B(2): 253-261.
Quackenbush L. S.. Yolk protein production in the marine shrimp Penaeus vannamei. J. Crust. Biol., 1989, 9: 509-516.
Quackenbush L. S.. Yolk synthesis in the marine shrimp, Penaeus vannamei. Comp. Biochem. Physiol., 1992, 103A(4): 711-714.
Quinitio E. T, Hara A., Yamauchi K., et al. Identification and characterization of vitellin in ahermaphrodite shrimp, Pandalus Kessleri. Comp. Biochem. Physiol. 1989, 94A(3): 445-452.
Quinitio E. T, Hara A., Yamauchi K., et al. Isolation and characterization of vitellin from the ovary of Penaeus monodon. Invertebr. Reprod. Dev., 1990, 17(3): 221-228.
Rankin S. M, Bradfield J. Y, Keeley L. L.. Ovarian protein synthesis in the South American white shrimp, Penaeus vannamei, during the reproductive cycle. Invertebr. Reprod. Dev. 1989, (15): 27-33.
Reley L. G, Tsukimura B.. Yolk protein synthesis in the rice land tadpole shrimp, Triops longicaudatus, measured by in vitro incorporation of ~3H-leucine. J. Exp. Zool., 1998, 281(3): 238-247.
Serrano-Pinto V., Vazquez-Boucard C., Villarreal-Colmenares H.. Yolk proteins during ovary and egg development of mature female freshwater crayfish (Cherax Quadricarinatus). Comp. Biochem. Physiol., Part A, 2003, 134: 33-43.
Shih J. T.. Sex steroid-like substances in the ovaries、hepatopancreases and body fluid of female Mictyris brevidactylus. Zool. Stud., 1997, 36(2). 136-145.
Shimizu K., Saikaw W.. Identification and characterization of vitellin from the barnacle, Balanus amphitrist. Comp. Biochem. Physiol., 1996, 115B(1): 114-119.
Soroka Y., Milner Y., Sagi A.. The hepatopancreas as a site of yolk protein synthesis in the prawn Macrobrachium rosenbergii. Invertebr. Reprod. Dev., 2000, 37(1): 61-68.
Spaziani E. P., Hinsch G. W.. Variation in selected unsaturated fatty acids during vitellogenesis in the florida fresh water crayfish Procambarus paeninsulanus. Invertebr. Reprod. Dev., 1997, 32(1): 21-25.
Tom M., Fingerman M., Hayes T. K., et al. A copatative study of the ovarian
proteins from two Penaeus shrimp, Penaeus semisulcatus de Hann and Penaeus vannamei(Boone). Comp. Biochem. Physiol., 1992, 102B: 483-490.
Tom M., Goren M., Ovadia M.. Purification and partical characterization of vitellin from the ovaries of Parapenaeus longirostris (Crustacea: Decapoda: Penaeidae). Comp. Biochem. Physiol., 1987, 87B: 17-23.
Tsukimura B., Bender J. S., Linder C. J.. Development of an anti- vitellin ELISA for the assessment of reproduction in the ridgeback shrimp, Sicyonia ingentis. Comp. Biochem. Physiol., Part A, 2000, 127: 215-224.
Vania S. P, Celia V. B, Humberto V. C.. Yolk proteins during ovary and egg development of mature female freshwater crayfish (Cherax quadricarinatus). Comp. Biochem. Physiol., PartA, 2003: 33-43.
Vázquez-Boucard C. G., Levy P., Ceccaldi H. J., et al. Developmental changes in concentrations of vitellin, vitellogenin, and lipids in hemolymph, hepatopancreas, and ovaries from different ovarian stages of Indian white prawn Fenneropenaeus indicus. J. Exp. Mar. Biol. Ecol., 2002, 281(1-2): 63-75.
Wilder M. N., Okumura T., Suzuki Y., et al. Vitellogenin prodution induced by eyestalk ablation in juvenile giant freshwater prawn Macrobramchium rosenbergii and trial methyl farneaoate administration. Zool. Sci., 1994, 11: 45-53.
Yano I., Chinzei Y.. Ovary if the site of vitellogenins synthesis in Kuruma Prawn, Penaeus japonicus. Comp. Biochem. Physiol, 1987, 86B: 213-218.
Yehezkel G., Chayoth R., Abdu U., et al. High-density lipoprotein associated with secondary vitellogenesis in the hemolymph of the crayfish Cherax quadricarinatus. Comp. Biochem. Physiol. Part B, 2000, 127: 411-421.
Zagalsky P. F.. A study if the astactanthin lipovitellin ovoverdin isolated from the ovaries of the lobster Homarus ammarus. Comp. Biochem. Physiol., 1985, 80B: 589-598.
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Eastman-Reks S., Fingrman M.. In vitro synthesis of vitellin by the ovary of the fiddler crab, Uca pugilator. Exp. Zool., 1985, 233: 111-116.
Fainzilber M., Tom M., Shafir S., et al. Is there extraovarian synthesis of vitellogenin in penaeid shrimp?. Biol. Bull, 1992, 183: 233-241.
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García-Orozco, Karina D., Francisco V. A., et al. Molecular characterization of vitellin from the ovaries of the white shrimp Penaeus (Litopenaeus) vannamei. Comp. Biochem. Physiol., Part B, 2002, 133(3): 361-369.
Han C. H., Bae H. H.. Purification of the yolk protein, and identification of the synthetic site of its prucursor in Ericheirjaponicus (Decapoda: Brachiura). Bull Korean FisSoc, 1992, 25(2): 432-442.
Jo Q. T., Laufer H., Biggers W. J., et al. Methyl farnesoate induced ovarian maturation in the spider crab, Libinia emarginata. Invertebr. Reprod. Dev., 1999, 36(1-3): 79-85.
Kawazoe I., Jasmani S., Shih T. W., et al. Purification and characterization of vitellin from the ovary of kuruma prawn, Penaeus japonicus. Fish Sci., 2000; 66: 390-396.
Komatsu M., Ando S.. Isolation. of crustacean egg yolk lipoproteins by differential
density gradient ultracentrifugation. Comp. Biochem. Physiol. 1992, 103B(2): 363-368.
Lee C. Y., Watson R. D.. In vitro study of vitellogenesis in the blue crab (Callinectes sapidus): site and control of vitellin synthesis. J. Exp. Zool., 1995, 271(5): 364-372.
Lee F. Y., Chang C. F.. The concentrations of vitellogenin (vitellin) and protein in hemolymph ovary and hepatopancreas in different ovarian stages of the freshwater prawn Macrobrachium rosenbergii. Comp. Biochem. Physiol., 1997, 117A(4): 433-439.
Lee F. Y., Shih T. W., Chang C. F.. Isolation and characterization of the female-specific protein (Vitellogenin) in mature female hemolymph of the freshwater prawn Macrobrachium rosenbergii: comparison with ovarian vitellin. General and Comparative Endocrinology, 1997, 108: 406-415.
Lee R. F., Walker A.. Lipovitellin and lipid droplet accumulation in oocytes during ovarian maturation in the blue crab, Callinects sapidus. J. Exp. Zool., 1995, 271(5): 401-412.
Liu C. W., O' Connor J D.. Biosynthesis of lipovitellin by the crustacean ovary-Ⅱ. Characterization of an in vitro incorporation of amino acid into the purified subunits. J. Exp. Zool., 1976, 195: 41-52.
Liu C. W., O' Connor J. D.. Biosynthesis of crustacean lipovitellin-Ⅲ the incorporation of lablled amino acid into the purified lipovitellin of the crab Pachygrapsus crassipes. J. Exp. Zool., 1977, 199: 105-108.
Longyant S., Sithigomgul P., Thammapalerd N., et al. Characterization of vitellin and vitellogenin of giant tiger prawn Penaeus monodon using monoclonal antibodies specific to viteltin subunits. Invertebr. Reprod. Dev., 2000, 37(3): 211-221.
Masaharu K., Seiichi A.. Isolation of crustacean egg yolk lipoprotein by differential densitygradient ultracentrifugation. Comp. Biochem. Physiol., 1992, 103B: 363-368.
Meusy J. J.. Vitellogenin, the extraovarian precusor of the yolk protein in crustacea, aview. Reprod Nutr Det, 1980, 20: 1-20.
Millamena O. M., Quinitio E.. The effects of diets on reproductive performance of eyestalk ablated and intact mud crab Scylla serrata. Aquaculture, 2000, 181: 81-90.
Moshe T., Menachem G., Michael O.. Localization of the vitellin and its possible precursors in various organs of Parapenaeus longirostis (Crustacea: Decapoda: Penaeidae). Intenational Journal of Invertebrate Reproduction and Development, 12(1987): 1-12.
Moshe T., Milton F., Timothy K., et al. A comparative study of the ovarian proteins from two penaeid shrimps, Penaeus semisulcatus de haan and Penaeus monodon(Boone). Comp. Biochem. Physiol., 1992, 102B(3): 483-490.
Oberdrster E., Rice C. D, Irwin L. K.. Purification of vitellin form grass shrimp Palaemonetes pugio, generation of monoclonal antibodies, and validation for the detection of lipovitellin in Crustacea. Comp. Biochem. Physiol. Part C, 2000, 127: 199-207.
Pateraki L. E, Stratakis E.. Synthesis and organization of vitellogenin and vitellin molecules form the land crab Potamon potamios. Comp. Biochem. Physiol. Part B, 2000, 125: 53-61.
Picaud J. L.. Vitellogenin synthesis by the fat body of Porcellio dilatatus Brand (Crustacea: Isopoda). Invert. Reprod. Dev., 1980, 2: 341-349.
Qiu Y. W, NG T. B and Ghu K. H.. Purification and characterization of vitellin from the ovaries of the shrimp Metapenaeus ensis(Crustacea: Decapoda: Penaeidae). Invertebr. Reprod. Dev., 1997, 31(1-3): 217-223.
Quackenbush L. S.. Vitellogenesis in the shrimp, Penaeus Vannamei: in vitro studies of the isolated hepatopancreas and ovary. Comp. Biochem. Physiol., 1989, 94B(2): 253-261.
Quackenbush L. S.. Yolk protein production in the marine shrimp Penaeus vannamei. J. Crust. Biol., 1989, 9: 509-516.
Quackenbush L. S.. Yolk synthesis in the marine shrimp, Penaeus vannamei. Comp. Biochem. Physiol., 1992, 103A(4): 711-714.
Quinitio E. T, Hara A., Yamauchi K., et al. Identification and characterization of vitellin in ahermaphrodite shrimp, Pandalus Kessleri. Comp. Biochem. Physiol. 1989, 94A(3): 445-452.
Quinitio E. T, Hara A., Yamauchi K., et al. Isolation and characterization of vitellin from the ovary of Penaeus monodon. Invertebr. Reprod. Dev., 1990, 17(3): 221-228.
Rankin S. M, Bradfield J. Y, Keeley L. L.. Ovarian protein synthesis in the South American white shrimp, Penaeus vannamei, during the reproductive cycle. Invertebr. Reprod. Dev. 1989, (15): 27-33.
Reley L. G, Tsukimura B.. Yolk protein synthesis in the rice land tadpole shrimp, Triops longicaudatus, measured by in vitro incorporation of ~3H-leucine. J. Exp. Zool., 1998, 281(3): 238-247.
Serrano-Pinto V., Vazquez-Boucard C., Villarreal-Colmenares H.. Yolk proteins during ovary and egg development of mature female freshwater crayfish (Cherax Quadricarinatus). Comp. Biochem. Physiol., Part A, 2003, 134: 33-43.
Shih J. T.. Sex steroid-like substances in the ovaries、hepatopancreases and body fluid of female Mictyris brevidactylus. Zool. Stud., 1997, 36(2). 136-145.
Shimizu K., Saikaw W.. Identification and characterization of vitellin from the barnacle, Balanus amphitrist. Comp. Biochem. Physiol., 1996, 115B(1): 114-119.
Soroka Y., Milner Y., Sagi A.. The hepatopancreas as a site of yolk protein synthesis in the prawn Macrobrachium rosenbergii. Invertebr. Reprod. Dev., 2000, 37(1): 61-68.
Spaziani E. P., Hinsch G. W.. Variation in selected unsaturated fatty acids during vitellogenesis in the florida fresh water crayfish Procambarus paeninsulanus. Invertebr. Reprod. Dev., 1997, 32(1): 21-25.
Tom M., Fingerman M., Hayes T. K., et al. A copatative study of the ovarian
proteins from two Penaeus shrimp, Penaeus semisulcatus de Hann and Penaeus vannamei(Boone). Comp. Biochem. Physiol., 1992, 102B: 483-490.
Tom M., Goren M., Ovadia M.. Purification and partical characterization of vitellin from the ovaries of Parapenaeus longirostris (Crustacea: Decapoda: Penaeidae). Comp. Biochem. Physiol., 1987, 87B: 17-23.
Tsukimura B., Bender J. S., Linder C. J.. Development of an anti- vitellin ELISA for the assessment of reproduction in the ridgeback shrimp, Sicyonia ingentis. Comp. Biochem. Physiol., Part A, 2000, 127: 215-224.
Vania S. P, Celia V. B, Humberto V. C.. Yolk proteins during ovary and egg development of mature female freshwater crayfish (Cherax quadricarinatus). Comp. Biochem. Physiol., PartA, 2003: 33-43.
Vázquez-Boucard C. G., Levy P., Ceccaldi H. J., et al. Developmental changes in concentrations of vitellin, vitellogenin, and lipids in hemolymph, hepatopancreas, and ovaries from different ovarian stages of Indian white prawn Fenneropenaeus indicus. J. Exp. Mar. Biol. Ecol., 2002, 281(1-2): 63-75.
Wilder M. N., Okumura T., Suzuki Y., et al. Vitellogenin prodution induced by eyestalk ablation in juvenile giant freshwater prawn Macrobramchium rosenbergii and trial methyl farneaoate administration. Zool. Sci., 1994, 11: 45-53.
Yano I., Chinzei Y.. Ovary if the site of vitellogenins synthesis in Kuruma Prawn, Penaeus japonicus. Comp. Biochem. Physiol, 1987, 86B: 213-218.
Yehezkel G., Chayoth R., Abdu U., et al. High-density lipoprotein associated with secondary vitellogenesis in the hemolymph of the crayfish Cherax quadricarinatus. Comp. Biochem. Physiol. Part B, 2000, 127: 411-421.
Zagalsky P. F.. A study if the astactanthin lipovitellin ovoverdin isolated from the ovaries of the lobster Homarus ammarus. Comp. Biochem. Physiol., 1985, 80B: 589-598.