孵化温度对乌龟性别分化影响的研究
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摘要
许多爬行类动物的性别决定由胚胎发育期的孵化温度决定，本文以乌龟(chinemys reevesii)为研究对象，运用组织学方法研究了温度控制乌龟性别分化及性腺发育的组织学和细胞学特点。龟卵在26℃和33℃恒温和常温下孵化。33℃恒温孵化得到100％的雌性小龟，26℃恒温孵化得到100％的雄性小龟，常温孵化得到的小龟有雌性、雄性和间性。用雄性决定温度(26℃)恒温和雌性决定温度(33℃)恒温组合实验，在设计的不同时间，从26℃转移到33℃下孵化直至出壳，或从33℃转移到26℃下孵化直至出壳。研究乌龟性别分化完成的时间。33℃下第22—23天(第21期左右)卵巢分化完成，雌性定型；26℃下第34—35天(第20期)精巢分化完成，雄性定型。一旦性别分化完成，再改变孵化温度，性别不会再逆转。
33℃下孵化的小龟卵巢皮质发达，由原始生殖细胞(primordial germinal cells PGCs)产生卵原细胞，髓质退化成为卵巢腔，分化时卵巢细胞的超微结构显示线粒体和内质网的数量大大增加，它们在分化结束后解体，分化前细胞内有较多的大颗粒，在分化中大颗粒减少，小颗粒增多，在分化后小颗粒达到最多。26℃下孵化的小龟精巢皮质退化成为一层薄的白膜，髓质有许多睾丸管构成，管腔内有散落排列的精原细胞。分化时精巢细胞的超微结构显示线粒体和内质网数量也有增加，但不如分化中卵巢细胞的数量多，分化后线粒体和内质网解体，在整个分化过程中大颗粒一直存在，无小颗粒出现。作者结合国内外学者的研究分析，这种颗粒物质是类固醇激素，大颗粒应该是雄性激素，小颗粒应该是雌性激素。雌激素通过芳香化酶作用由激素合成而来。在卵巢分化中线粒体和光面内质网增多，除了适应性腺本身的发育生长外，线粒体的增多主要是为芳香化酶把雄性激素转化为雌激素的作用提供能量，光面内质网的增多是有利于类固醇激素的
合成。在精巢的分化中基本上不把雄性激素合成为雌激素，线粒体和
光面内质网的增多主要是用于性腺的发育生长，所以数目不及卵巢中
的多。
本文对于影响乌龟胚胎发育的一些因素，性腺分化时期以及胚胎
性腺的组织学和细胞学特点，孵化温度和性别决定，以及近年来国内
外关注的温度与激素，激素和性别分化，基因与性别决定等问题进行
了讨论。
In many reptiles sexual differentiation is controlled by the incubating temperature of the embryo. With histological method, this paper studies the sexual differentiation and gonadal development of Chinemys reevesii under different incubating temperature and histological and cytological characteristics of its sexual gonads. Eggs of Chinemys reevesii were incubated at 26 , 33 and normal atmopheric temperature. Incubation at 33 resulted in females in 100% of the cases; Incubation at 26 resulted in males in 100% of the cases; Incubation at normal atmospheric temperature resulted in females, males and intersexualities. This study investigates the time when sexual differentiation is finished in Chinemys reevesii, using different combinations of incubation at a male-producing temperature(26 ) and at a female-producing temperature (33 ). Incubation at 33 , the female sexual differentiation is finished after 22-23 days (around stage 19), at 26 ,the male sexual differentiation is completed after 34-35 days (aroun
d stage 20). Once suxual differentiation is finished, the sex will not be reversed when the incubating temperature is changed.
The females incubated at 33 , their ovarium has powerful cortex, the germinal epithlium produce oocyte, its medulla degenerate into ovarian cavity. During sexual differentiation, mitochondria and smooth endoplasmic reticulum in cells of ovary increase in quantity, they disintegrate after sexual differentiation. There are a lot of big grana in cells of ovary before sexual differentiation, in the course of sexual differentiation the quantity of the big grana decrease, while small grana increase and peak
after sexual differentiation. The males incubation at 26 , cortices of their testes degenerate into a thin albuginea. Medulla are formed by testicular cords in which scatter spermatogonium. During sexual differentiation, mitochondria and smooth endoplasmic reticulum in cells of testis also increase in quantity, but it is less than that in ovary, they disintegrate after sexual differentiation. The big grana always exsist in course of sexual differentiation, but small grana don't appear. The author consider that the grana are steroid hormone according to the studies of domestic and overseas scholars, the big grana should be androgens, the small grana should be estrogens. Estrogens are synthesized from androgens through the activity of aromatase. Besides fitting the own growth of gonad, the main function of the increase of mitochondria is to provide energy to the synthesis of estrogens, the main function of smooth endoplasmic reticulum is for the synthesis of the steroid hormone. But in differentiation of testi
s, estrogens are hardly synthesized from androgens, the increase of mitochondria and smooth endoplasmic reticulum is mainly used in the growth of gonad, so their quantities are less than that in ovary.
Furthermore, this paper touches other problems such as factors influencing the development of embryonic, periods of gonad in sexual differentiation, histological and cytological characteristics of the primordial gonad, the sex determination influenced by incubating temperature, and the relationship of temperature and hormone, hormone and sexual differentiation, gene and sexual differentiation what are concerned by internal-external country in recent years.
33℃下孵化的小龟卵巢皮质发达，由原始生殖细胞(primordial germinal cells PGCs)产生卵原细胞，髓质退化成为卵巢腔，分化时卵巢细胞的超微结构显示线粒体和内质网的数量大大增加，它们在分化结束后解体，分化前细胞内有较多的大颗粒，在分化中大颗粒减少，小颗粒增多，在分化后小颗粒达到最多。26℃下孵化的小龟精巢皮质退化成为一层薄的白膜，髓质有许多睾丸管构成，管腔内有散落排列的精原细胞。分化时精巢细胞的超微结构显示线粒体和内质网数量也有增加，但不如分化中卵巢细胞的数量多，分化后线粒体和内质网解体，在整个分化过程中大颗粒一直存在，无小颗粒出现。作者结合国内外学者的研究分析，这种颗粒物质是类固醇激素，大颗粒应该是雄性激素，小颗粒应该是雌性激素。雌激素通过芳香化酶作用由激素合成而来。在卵巢分化中线粒体和光面内质网增多，除了适应性腺本身的发育生长外，线粒体的增多主要是为芳香化酶把雄性激素转化为雌激素的作用提供能量，光面内质网的增多是有利于类固醇激素的
合成。在精巢的分化中基本上不把雄性激素合成为雌激素，线粒体和
光面内质网的增多主要是用于性腺的发育生长，所以数目不及卵巢中
的多。
本文对于影响乌龟胚胎发育的一些因素，性腺分化时期以及胚胎
性腺的组织学和细胞学特点，孵化温度和性别决定，以及近年来国内
外关注的温度与激素，激素和性别分化，基因与性别决定等问题进行
了讨论。
In many reptiles sexual differentiation is controlled by the incubating temperature of the embryo. With histological method, this paper studies the sexual differentiation and gonadal development of Chinemys reevesii under different incubating temperature and histological and cytological characteristics of its sexual gonads. Eggs of Chinemys reevesii were incubated at 26 , 33 and normal atmopheric temperature. Incubation at 33 resulted in females in 100% of the cases; Incubation at 26 resulted in males in 100% of the cases; Incubation at normal atmospheric temperature resulted in females, males and intersexualities. This study investigates the time when sexual differentiation is finished in Chinemys reevesii, using different combinations of incubation at a male-producing temperature(26 ) and at a female-producing temperature (33 ). Incubation at 33 , the female sexual differentiation is finished after 22-23 days (around stage 19), at 26 ,the male sexual differentiation is completed after 34-35 days (aroun
d stage 20). Once suxual differentiation is finished, the sex will not be reversed when the incubating temperature is changed.
The females incubated at 33 , their ovarium has powerful cortex, the germinal epithlium produce oocyte, its medulla degenerate into ovarian cavity. During sexual differentiation, mitochondria and smooth endoplasmic reticulum in cells of ovary increase in quantity, they disintegrate after sexual differentiation. There are a lot of big grana in cells of ovary before sexual differentiation, in the course of sexual differentiation the quantity of the big grana decrease, while small grana increase and peak
after sexual differentiation. The males incubation at 26 , cortices of their testes degenerate into a thin albuginea. Medulla are formed by testicular cords in which scatter spermatogonium. During sexual differentiation, mitochondria and smooth endoplasmic reticulum in cells of testis also increase in quantity, but it is less than that in ovary, they disintegrate after sexual differentiation. The big grana always exsist in course of sexual differentiation, but small grana don't appear. The author consider that the grana are steroid hormone according to the studies of domestic and overseas scholars, the big grana should be androgens, the small grana should be estrogens. Estrogens are synthesized from androgens through the activity of aromatase. Besides fitting the own growth of gonad, the main function of the increase of mitochondria is to provide energy to the synthesis of estrogens, the main function of smooth endoplasmic reticulum is for the synthesis of the steroid hormone. But in differentiation of testi
s, estrogens are hardly synthesized from androgens, the increase of mitochondria and smooth endoplasmic reticulum is mainly used in the growth of gonad, so their quantities are less than that in ovary.
Furthermore, this paper touches other problems such as factors influencing the development of embryonic, periods of gonad in sexual differentiation, histological and cytological characteristics of the primordial gonad, the sex determination influenced by incubating temperature, and the relationship of temperature and hormone, hormone and sexual differentiation, gene and sexual differentiation what are concerned by internal-external country in recent years.
引文
1 丁汉波《发育生物学》高等教育出版社 1986
2 芮菊生《组织切片学技术》人民出版社 1980
3 刘筠《中国养殖鱼类繁殖生理学》农业出版社 1993
4 曲漱惠《动物胚胎学》人民教育出版社 1980
5 T、M、彼尔索夫《鱼类的性别分化》
6 翟中和《细胞生物学》高等教育出版社 2000
7 刘祖洞《遗传学》高等教育出版社 1994
8 张红卫《发育生物学》高等教育出版社 2001
9 刘筠等 温度等生态因素对鳖(甲鱼)胚胎发育的影响。湖南师院学报(自然科学版)，1982，(1)：67-70
10 侯陵 孵化温度与乌龟性别。两栖爬行动物学报，1985，4(2)：130
11 陈实平，章静波 龟的性别分化．自然杂志，10(4)：294-295
12 卞伟 乌龟的生物学及养殖技术(一)．内陆水产，1997，(2)：20-21
13 刘国安等 乌龟繁殖生态的研究。水生生物学报，1988，12(3)：230-235
14 周灿 孵化温度对乌龟性别分化的影响。硕士论文，1998
15 谭立军 乌龟胚胎发育的研究。硕士论文，1988
16 沈卉君等 中华鳖的解剖研究Ⅳ：消化、呼吸、泌尿生殖系统。上海师范学院学报，1983(1)：43-50
17 叶玉珍等 稚鳖的性别分化及组织学研究。中国水产科学，1998，5(2)：11-14
18 谢万奎 不同性别鳖的生长差异及孵化温度控制鳖性别分化的初步研究。水利渔业，1993，(4)：23-24
19 罗曼等 温度对鳖卵孵化中性比影响的研究．水利渔业，1998(1)：6-7
20 任少亭等 温度对中华鳖孵育率及性别决定影响的研究。水产养殖，1998，(6)：20-21
21 阳建春等 爬行动物性别机理研究。中山大学学报论丛，1997，1：43-48
22 聂刘旺 温度依赖性别决定的分子机制。生物学杂志，1998。15(5)：35-38
23 林植华，计翔 孵化温度对北草蜥孵化卵和孵出幼体的影响．动物学研究，1998，19(6)：
439-443
24 郭超文等 三种龟类动物的细胞遗传研究．水生生物学报，1998，22(1)：17-24
25 郭超文等 中国水龟类的核型与银带带型研究。水生生物学报，1998，22(4)：314-318
26 刘丽等 加温养殖下乌龟精巢发育的研究。湛江海洋大学学报，2000，20(4)：1-4
27 侯陵 鳖胚原始生殖细胞发生的研究。两栖爬行动物学报，1987，6(2)5-9
28 刘少军 革胡子鲶原始生殖细胞的起源、迁移及性腺分化。水生生物学报，1991，15(1)：1-7
29 金岭梅等 温度对鳖卵孵化的影响．家畜生态，1998，19(4)：24-26
30 汪鸣等 中华花龟的染色体组型与Ag-NORs．安徽师范大学学报(自然科学版)，1999，22(1)：44-46
31 叶玉珍等 甲基睾丸素对中华鳖外部性征诱导作用的研究．华中农业大学学报，1999，18(1)：72-74
32 聂刘旺等 两种龟类SOX基因的PCR扩增及SSCP分析的研究．应用与环境生物学学报，1999，5(4)：378-381
33 郭超文等 中国四种龟的细胞遗传研究。遗传学报，1995，22(1)：40-45
34 张海军等 乌龟SOX gene的PCR扩增及其条件。四川动物，2001，20(2)：82-85
35 林浩然等 鳗鲡繁殖生物学研究Ⅴ．性类固醇激素诱导雌鳗鲡促性腺激素(GtH)分泌和卵巢发育的作用．水生生物学报，1994，18(3)73-78
36 毕建花 乌龟的人工繁殖．水产养殖，1997，6：9-10
37 林光华，熊敬维 革胡子鲶卵巢在第一次性周期内分化与发育的研究．动物学研究，1995，16(4)：365-372
38 邱东茹，吴振斌 环境雌激素对动物和人体的影响及其作用机制．水生生物学报，1997，21(4)：365-374
39 匡颖，翟玉梅 小鼠原生殖细胞的研究进展。动物学杂志，1999，34(1)：48-51
40 薛妹朗等 中华鳖精母细胞联会复合体的研究。遗传，1999，21(1)：6-8
41. Yntema, C.L Characteristics of gonads and oviducts in hatching and young of Chelydra serpentina resulting from three incubation temperatures.J .Morph.1981,167:297-304
42. Packard, G C et al .Influence of moisture, temperature and substrature on snapping turtle
eggs and emdryos. Ecology, 1987, 68:983-993
43 Phillips, J A et al, Influence of and temperature on eggs and emdryps of gree iguanas (Iguana iguana). Herpetologica, 1990, 46:238-245
44 Yntema, C L. Effect s of various temperature on the embryonic development of chelydra serpentina. Anat. Rec,1960,36:305-306
45 Ewert, M A. Embryology of turtles in biology of reptilia, academic press, London and New York, 1985,15:75-2667
46 Limpus, C J et al. Movement induced mortality of loggerhead eggs. Heroetologica, 1979, 35:335-338
47 Parmeter, C J. Incubation of the eggs of the gree turtle Chelonia mydas. Intorres strait Australia the effect of movement on hatchability. Aust. Wild .Res. 1980, 7:487-491
48 Pieau, C Sur la proportion sexuelle chez Ies embryous de deux cheloniens (Testudo gracea L et Emys orbicularis L.) issus d'oeufs incubes artificilellement. C. R Acad. Sci. Paris, 1971, 272D:3071-3074
49 Pieau, C. Effects de la temperature sur le developpement des glandes genitals chez les embryons de deux cheloniens, Emys orbicularis L.et testudo graeca L. C A Aca Sci. Paris. 1972, 274D: 719-722
50 Bull ,J J. Sex ditermination in reptile.Quart Rev Biol,1980,55:3-21
51 Yntema, C L. Effects of incubation temperatures on sexual defferentiation in turtle, Chelydra serpentina. J. Morph, 1976,150:453-462
52 Pieau, C. Temperature variation and sex determination in reptiles. Bioessays, 1996, 18: 19-26
53 Vogt, R C. Ecology of hatching sex ratio in map turtles. Ecology, 1984, 65(2): 582-587
54 Pieau ,C. Modalities of the action of temperature on sexual defferentiation in field-developing embryos the European pond turtle Emys orbicularis (Emydidae). J .Exp. Zool, 1982, 220:353-360
55 Georges ,A, Limpus C et al. Hatching sex in the marine turtle Caretta caretta is determined by proportion of development at a temperature, not daily duration of exposure. J.
Exp .Zool, 1994, 270: 432-444
56 Pieau, C. et al. Determination of temperature sensitive stages of sexual differentiation of the gonads in embryos of the turtle,Emys orbicularis. J .Morphol, 1981,170:373-382
57 Bull, J J ,Vogt R C. Temperature-sensitive period of sex determination in Emydid Turtles. J .Exp .Zool, 1981, 218:435-440
58 Yntema. Temperature levels and period of sex determination during incubation of eggs of Chelydra serpentina. J. Morph, 1979,159:17-28
59 Yntema,.A series of stages in the Embryonic development of Chelydra serpentina. J. Morph 1968,125:219-252
60 Crews ,D et al. Temperature-dependent sex determination in reptiles: proximate mechanisms, ultimate outcomes, and practical applications. Dev. Genet, 1994, 15: 297-312
61 Forbes, T R. Intersexuality in reptile.In intersexuality in vertebrates including Man. C.N Armstrong and A.J.Marshall, eds.Academic Press,London,1964,pp:273-283
62 Pieau C. Sur la differentiation du sexe en fonction de la temperature chez les embryons d'Emys robicularis L. (chelonien); effects des hormones sexuelles. Ann. Embryol .Morphogl, 1974, 7:365-394
63 Raynaud, A. Pieau,C.Embryonic development of the genital system.In:Biology of the Reptilia, Vol 15,Development B C.Gans and F.Billett.John Wiley and Sons. New York PP.1985:149-300
64 Pieau C. Sexual differentiation of gonads as a function of temperature in the turtli Emys orbicularis :endocrine function ,intersexuality and growth.J.Exp.Zool,1998, 281:400-408
65 Charnier, M.Action de la temperature sur la sex-ration chez I'em bryon d'Agama agama (Agamidae,Lacertlien).Soc.Biol.Af,1966,160:620-622
66 Patrick, Zaborski.et al. H-Y antigen expression in temperature sex-reverseed turtles (Emys orbicularis), differentiation ,1982,22:73-78
67 Crews,d.et al. Estrogen and sex reverseal in turtles, a dose-dependent phenomenon. Gen.Comp.Endocrinol,1991,81:357-364
68 Merchant-Larios,h. et al. Correlation among thermosensitive period, estradiol response and gonadaldifferentiation in the sea turtle Lepidochelys olivadal.Gen.Comp.Endocrinol, 1993,107:373-385
69 Dorizzi, M.et al. The ovary retains male potential after the thermosensitive period for sex determination in the turtle Emys orbicularis. Differentiation,1996,60:193-201
70 Bull, J J. et al. Sex reversal by estradiol in three reptilien orders.Gen.Comp.Endocrinol, 1988,70:425-428
71 Gutzke, W H P V. et al. Sensitive periods during embryogeny for hormonally induced sex determination in turtles.Gen.Comp.Endocrinol,1988,71:265-267
72 Smith C A. et al. Uptake of H-estradiol by embryonic crocodile gonad during the period of sexual differentiation. J.Exp.Zool,1994,270:219-224
73 Wibbels, T. et al. Synergism between temperature and estradiol : a common pathway in turtle sex determination?J.Exp.Zool,1991,260:130-134
74 Swain,A.et al.Mammalian sex determination : a molecular drama.Genes and Dev,1999,13:757-767
75 Kent, J. et al. The evolution of WT1 sequence and expression pattern in the vertebrates. Oncogene,1995,11:1781-1792
76 Westernps. Et al. Temperature-dependent sex determination: upregulation of SOX9 expression after commitment to male development. Dev.Dyn,1999,214:171-177
77 Western, P S. et al. Temperature-dependent sex determination in the American alligator. expression of SF1,WT1 and DAX1 during gonadogenesis. Gene, 2000,241:223-232
78 Thane, Wibbels.et al. Temperature-dependent sex determination in the Red-Eared Slider turtle,Trachemys scripta. J.Exp.Zool,1998,281:409-416
79 Lortta, D.et al. Sequence and expression analysis of WT1 and SOX9 in the red-eared slider turtle, Trachemys scripta. J.Exp.Zool,1998,281:417-427
80 Lynch, J P. et al. Steroidgenic factor 1, an orphan nuclear receptor, regulates the expressiong of the rat aromatase gene in gonadal tissues. Mol.Endocrinol,1993,7:776-786
81 Lala ,D S. et al. Steroidogenic factor 1, a key regulator of steroidogenic enzyme expression,
is the mouse nomologue of Fushi tarazu factor 1. Mol.Endocrinol,1992,6:1249-1258
82 Clemente, N. et al. Effect of AMH upon aromatase activity and porcine immature ovaries. Endocrine,1994,2:553-558
83 Denny, P. et al. A conserved family of genesrelated to the testis determining genes SRY. Nucl.Acid.Res,1992,20:2887-2888
84 Pieau C. Temperature-dependent sex determination and gonadal differentiation in reptiles. Genes and Mechanisms in Vertebrate Sex Determination. 2001,BIRKHAUSER Verlag Base/Switzerland: 117-141
85 Moreno-Mendoza, N. et al.Differential expression of SOX9in gonads of the sea turtle Lepidochelys olivacea at male- or female-promoting temperatures. J. Exp. Zool,1999, 284:705-710
86 Severine, N. et al. Expression of AMH, SF1, and SOX9 in gonads of genetic female chickens during sex reversal induced by an aromatase inhibitor. Dev. Dyn,2001,222:228-237
87 M oreno-Mendoza, N. et al. Temperature regulates SOXexpression in cultured gonads of Lepidochelys olivacea, a species with temperature sex determination. Dev.Biol, 2001, 229:319-326
88 Shyril O'steen. Embryonic temperature influences juvenile temperature choice and growth rate in snapping turtles Chelydra serpentina. J. Exp.Zool,1998,201:439-449
89 Kenneth, J. et al. Orientation and open-sea navigation in sea turtles. J. Exp. Biol, 1996,199:73-81
90 Bull J J. Temperature-dependent sex determination in turtles. Science,1979,206:1186-1188
91 Hugh Wallace. et al. Sex reversal of the newt Triturus cristatus reared at extreme temperature. Int J.Dev.Biol, 2000,44:807-810
92 Alice Fleming, David Crews. Estradiol and incubation temperature modulate regulation of steroidogenic factor 1 in the developing gonad of the red-eared slider turtle. Endocrinology, 2001,142(4): 1403-1411
93 Lda Torres-Maldonado. Et al. Timing of SOX9 downregulation and female sex determination in gonad of the sea turtle Lepidochelys olivacea. J.Exp.Zool,
2001,290:498-503
94 Wallace H.et al. Amphibian sex determination and sex reversal CMLS,Cell.Mol.Life Sci, 1999,55:901-909
95 Mrosovsky N, Pieau C. Transitional range of temperature, pivotal temperatures and thermosensitive stages for sex determination in reptiles. Amphibia-Reptilia, 1991, 12:169-179
96 Christian Dournon. et al. Temperature sex-reversal in amphibians and reptiles. Int.Dev.Biol, 1991,34:81-92
2 芮菊生《组织切片学技术》人民出版社 1980
3 刘筠《中国养殖鱼类繁殖生理学》农业出版社 1993
4 曲漱惠《动物胚胎学》人民教育出版社 1980
5 T、M、彼尔索夫《鱼类的性别分化》
6 翟中和《细胞生物学》高等教育出版社 2000
7 刘祖洞《遗传学》高等教育出版社 1994
8 张红卫《发育生物学》高等教育出版社 2001
9 刘筠等 温度等生态因素对鳖(甲鱼)胚胎发育的影响。湖南师院学报(自然科学版)，1982，(1)：67-70
10 侯陵 孵化温度与乌龟性别。两栖爬行动物学报，1985，4(2)：130
11 陈实平，章静波 龟的性别分化．自然杂志，10(4)：294-295
12 卞伟 乌龟的生物学及养殖技术(一)．内陆水产，1997，(2)：20-21
13 刘国安等 乌龟繁殖生态的研究。水生生物学报，1988，12(3)：230-235
14 周灿 孵化温度对乌龟性别分化的影响。硕士论文，1998
15 谭立军 乌龟胚胎发育的研究。硕士论文，1988
16 沈卉君等 中华鳖的解剖研究Ⅳ：消化、呼吸、泌尿生殖系统。上海师范学院学报，1983(1)：43-50
17 叶玉珍等 稚鳖的性别分化及组织学研究。中国水产科学，1998，5(2)：11-14
18 谢万奎 不同性别鳖的生长差异及孵化温度控制鳖性别分化的初步研究。水利渔业，1993，(4)：23-24
19 罗曼等 温度对鳖卵孵化中性比影响的研究．水利渔业，1998(1)：6-7
20 任少亭等 温度对中华鳖孵育率及性别决定影响的研究。水产养殖，1998，(6)：20-21
21 阳建春等 爬行动物性别机理研究。中山大学学报论丛，1997，1：43-48
22 聂刘旺 温度依赖性别决定的分子机制。生物学杂志，1998。15(5)：35-38
23 林植华，计翔 孵化温度对北草蜥孵化卵和孵出幼体的影响．动物学研究，1998，19(6)：
439-443
24 郭超文等 三种龟类动物的细胞遗传研究．水生生物学报，1998，22(1)：17-24
25 郭超文等 中国水龟类的核型与银带带型研究。水生生物学报，1998，22(4)：314-318
26 刘丽等 加温养殖下乌龟精巢发育的研究。湛江海洋大学学报，2000，20(4)：1-4
27 侯陵 鳖胚原始生殖细胞发生的研究。两栖爬行动物学报，1987，6(2)5-9
28 刘少军 革胡子鲶原始生殖细胞的起源、迁移及性腺分化。水生生物学报，1991，15(1)：1-7
29 金岭梅等 温度对鳖卵孵化的影响．家畜生态，1998，19(4)：24-26
30 汪鸣等 中华花龟的染色体组型与Ag-NORs．安徽师范大学学报(自然科学版)，1999，22(1)：44-46
31 叶玉珍等 甲基睾丸素对中华鳖外部性征诱导作用的研究．华中农业大学学报，1999，18(1)：72-74
32 聂刘旺等 两种龟类SOX基因的PCR扩增及SSCP分析的研究．应用与环境生物学学报，1999，5(4)：378-381
33 郭超文等 中国四种龟的细胞遗传研究。遗传学报，1995，22(1)：40-45
34 张海军等 乌龟SOX gene的PCR扩增及其条件。四川动物，2001，20(2)：82-85
35 林浩然等 鳗鲡繁殖生物学研究Ⅴ．性类固醇激素诱导雌鳗鲡促性腺激素(GtH)分泌和卵巢发育的作用．水生生物学报，1994，18(3)73-78
36 毕建花 乌龟的人工繁殖．水产养殖，1997，6：9-10
37 林光华，熊敬维 革胡子鲶卵巢在第一次性周期内分化与发育的研究．动物学研究，1995，16(4)：365-372
38 邱东茹，吴振斌 环境雌激素对动物和人体的影响及其作用机制．水生生物学报，1997，21(4)：365-374
39 匡颖，翟玉梅 小鼠原生殖细胞的研究进展。动物学杂志，1999，34(1)：48-51
40 薛妹朗等 中华鳖精母细胞联会复合体的研究。遗传，1999，21(1)：6-8
41. Yntema, C.L Characteristics of gonads and oviducts in hatching and young of Chelydra serpentina resulting from three incubation temperatures.J .Morph.1981,167:297-304
42. Packard, G C et al .Influence of moisture, temperature and substrature on snapping turtle
eggs and emdryos. Ecology, 1987, 68:983-993
43 Phillips, J A et al, Influence of and temperature on eggs and emdryps of gree iguanas (Iguana iguana). Herpetologica, 1990, 46:238-245
44 Yntema, C L. Effect s of various temperature on the embryonic development of chelydra serpentina. Anat. Rec,1960,36:305-306
45 Ewert, M A. Embryology of turtles in biology of reptilia, academic press, London and New York, 1985,15:75-2667
46 Limpus, C J et al. Movement induced mortality of loggerhead eggs. Heroetologica, 1979, 35:335-338
47 Parmeter, C J. Incubation of the eggs of the gree turtle Chelonia mydas. Intorres strait Australia the effect of movement on hatchability. Aust. Wild .Res. 1980, 7:487-491
48 Pieau, C Sur la proportion sexuelle chez Ies embryous de deux cheloniens (Testudo gracea L et Emys orbicularis L.) issus d'oeufs incubes artificilellement. C. R Acad. Sci. Paris, 1971, 272D:3071-3074
49 Pieau, C. Effects de la temperature sur le developpement des glandes genitals chez les embryons de deux cheloniens, Emys orbicularis L.et testudo graeca L. C A Aca Sci. Paris. 1972, 274D: 719-722
50 Bull ,J J. Sex ditermination in reptile.Quart Rev Biol,1980,55:3-21
51 Yntema, C L. Effects of incubation temperatures on sexual defferentiation in turtle, Chelydra serpentina. J. Morph, 1976,150:453-462
52 Pieau, C. Temperature variation and sex determination in reptiles. Bioessays, 1996, 18: 19-26
53 Vogt, R C. Ecology of hatching sex ratio in map turtles. Ecology, 1984, 65(2): 582-587
54 Pieau ,C. Modalities of the action of temperature on sexual defferentiation in field-developing embryos the European pond turtle Emys orbicularis (Emydidae). J .Exp. Zool, 1982, 220:353-360
55 Georges ,A, Limpus C et al. Hatching sex in the marine turtle Caretta caretta is determined by proportion of development at a temperature, not daily duration of exposure. J.
Exp .Zool, 1994, 270: 432-444
56 Pieau, C. et al. Determination of temperature sensitive stages of sexual differentiation of the gonads in embryos of the turtle,Emys orbicularis. J .Morphol, 1981,170:373-382
57 Bull, J J ,Vogt R C. Temperature-sensitive period of sex determination in Emydid Turtles. J .Exp .Zool, 1981, 218:435-440
58 Yntema. Temperature levels and period of sex determination during incubation of eggs of Chelydra serpentina. J. Morph, 1979,159:17-28
59 Yntema,.A series of stages in the Embryonic development of Chelydra serpentina. J. Morph 1968,125:219-252
60 Crews ,D et al. Temperature-dependent sex determination in reptiles: proximate mechanisms, ultimate outcomes, and practical applications. Dev. Genet, 1994, 15: 297-312
61 Forbes, T R. Intersexuality in reptile.In intersexuality in vertebrates including Man. C.N Armstrong and A.J.Marshall, eds.Academic Press,London,1964,pp:273-283
62 Pieau C. Sur la differentiation du sexe en fonction de la temperature chez les embryons d'Emys robicularis L. (chelonien); effects des hormones sexuelles. Ann. Embryol .Morphogl, 1974, 7:365-394
63 Raynaud, A. Pieau,C.Embryonic development of the genital system.In:Biology of the Reptilia, Vol 15,Development B C.Gans and F.Billett.John Wiley and Sons. New York PP.1985:149-300
64 Pieau C. Sexual differentiation of gonads as a function of temperature in the turtli Emys orbicularis :endocrine function ,intersexuality and growth.J.Exp.Zool,1998, 281:400-408
65 Charnier, M.Action de la temperature sur la sex-ration chez I'em bryon d'Agama agama (Agamidae,Lacertlien).Soc.Biol.Af,1966,160:620-622
66 Patrick, Zaborski.et al. H-Y antigen expression in temperature sex-reverseed turtles (Emys orbicularis), differentiation ,1982,22:73-78
67 Crews,d.et al. Estrogen and sex reverseal in turtles, a dose-dependent phenomenon. Gen.Comp.Endocrinol,1991,81:357-364
68 Merchant-Larios,h. et al. Correlation among thermosensitive period, estradiol response and gonadaldifferentiation in the sea turtle Lepidochelys olivadal.Gen.Comp.Endocrinol, 1993,107:373-385
69 Dorizzi, M.et al. The ovary retains male potential after the thermosensitive period for sex determination in the turtle Emys orbicularis. Differentiation,1996,60:193-201
70 Bull, J J. et al. Sex reversal by estradiol in three reptilien orders.Gen.Comp.Endocrinol, 1988,70:425-428
71 Gutzke, W H P V. et al. Sensitive periods during embryogeny for hormonally induced sex determination in turtles.Gen.Comp.Endocrinol,1988,71:265-267
72 Smith C A. et al. Uptake of H-estradiol by embryonic crocodile gonad during the period of sexual differentiation. J.Exp.Zool,1994,270:219-224
73 Wibbels, T. et al. Synergism between temperature and estradiol : a common pathway in turtle sex determination?J.Exp.Zool,1991,260:130-134
74 Swain,A.et al.Mammalian sex determination : a molecular drama.Genes and Dev,1999,13:757-767
75 Kent, J. et al. The evolution of WT1 sequence and expression pattern in the vertebrates. Oncogene,1995,11:1781-1792
76 Westernps. Et al. Temperature-dependent sex determination: upregulation of SOX9 expression after commitment to male development. Dev.Dyn,1999,214:171-177
77 Western, P S. et al. Temperature-dependent sex determination in the American alligator. expression of SF1,WT1 and DAX1 during gonadogenesis. Gene, 2000,241:223-232
78 Thane, Wibbels.et al. Temperature-dependent sex determination in the Red-Eared Slider turtle,Trachemys scripta. J.Exp.Zool,1998,281:409-416
79 Lortta, D.et al. Sequence and expression analysis of WT1 and SOX9 in the red-eared slider turtle, Trachemys scripta. J.Exp.Zool,1998,281:417-427
80 Lynch, J P. et al. Steroidgenic factor 1, an orphan nuclear receptor, regulates the expressiong of the rat aromatase gene in gonadal tissues. Mol.Endocrinol,1993,7:776-786
81 Lala ,D S. et al. Steroidogenic factor 1, a key regulator of steroidogenic enzyme expression,
is the mouse nomologue of Fushi tarazu factor 1. Mol.Endocrinol,1992,6:1249-1258
82 Clemente, N. et al. Effect of AMH upon aromatase activity and porcine immature ovaries. Endocrine,1994,2:553-558
83 Denny, P. et al. A conserved family of genesrelated to the testis determining genes SRY. Nucl.Acid.Res,1992,20:2887-2888
84 Pieau C. Temperature-dependent sex determination and gonadal differentiation in reptiles. Genes and Mechanisms in Vertebrate Sex Determination. 2001,BIRKHAUSER Verlag Base/Switzerland: 117-141
85 Moreno-Mendoza, N. et al.Differential expression of SOX9in gonads of the sea turtle Lepidochelys olivacea at male- or female-promoting temperatures. J. Exp. Zool,1999, 284:705-710
86 Severine, N. et al. Expression of AMH, SF1, and SOX9 in gonads of genetic female chickens during sex reversal induced by an aromatase inhibitor. Dev. Dyn,2001,222:228-237
87 M oreno-Mendoza, N. et al. Temperature regulates SOXexpression in cultured gonads of Lepidochelys olivacea, a species with temperature sex determination. Dev.Biol, 2001, 229:319-326
88 Shyril O'steen. Embryonic temperature influences juvenile temperature choice and growth rate in snapping turtles Chelydra serpentina. J. Exp.Zool,1998,201:439-449
89 Kenneth, J. et al. Orientation and open-sea navigation in sea turtles. J. Exp. Biol, 1996,199:73-81
90 Bull J J. Temperature-dependent sex determination in turtles. Science,1979,206:1186-1188
91 Hugh Wallace. et al. Sex reversal of the newt Triturus cristatus reared at extreme temperature. Int J.Dev.Biol, 2000,44:807-810
92 Alice Fleming, David Crews. Estradiol and incubation temperature modulate regulation of steroidogenic factor 1 in the developing gonad of the red-eared slider turtle. Endocrinology, 2001,142(4): 1403-1411
93 Lda Torres-Maldonado. Et al. Timing of SOX9 downregulation and female sex determination in gonad of the sea turtle Lepidochelys olivacea. J.Exp.Zool,
2001,290:498-503
94 Wallace H.et al. Amphibian sex determination and sex reversal CMLS,Cell.Mol.Life Sci, 1999,55:901-909
95 Mrosovsky N, Pieau C. Transitional range of temperature, pivotal temperatures and thermosensitive stages for sex determination in reptiles. Amphibia-Reptilia, 1991, 12:169-179
96 Christian Dournon. et al. Temperature sex-reversal in amphibians and reptiles. Int.Dev.Biol, 1991,34:81-92