The process of morphological development of a differentiated gonad from an undifferentiated primordium is a very important step of gonadogenesis. Studies on sexually dimorphic gene expression are important to increase our understanding of this process and to investigate how environmental factors such as temperature can regulate gonadal development. The aim of this study was to identify putative genes involved in sex differentiation in pejerrey (Odontesthes bonariensis) reared at male- and female-producing temperatures (MPT and FPT, respectively) using a microarray heterologous from the medaka (Oryzias latipes), a closely phylogenetic species. Genes related to numerous processes presented higher expression at MPT, including those involved in muscular contraction, metabolic pathways, developmental processes, and reproduction. Genes induced by FPT were classified under the gene ontology terms of response to stimulus, transport and proteolysis. From genes selected for validation, at MPT ndrg3 expression was observed in the somatic cells, whereas pen-2 was detected in germ cells in the caudal portion of the gonads, where no apoptotic signals were observed. Finally, hsp90 was highly expressed in somatic cells of the gonads at the FPT. The results suggest that the interplay of pro-apoptotic and anti-apoptotic genes is important during the masculinization process and for the prevention of sterility following exposure to warm temperatures.

Keywords:
Apoptosis, Gonadogenesis, Microarray, Pejerrey
1.
Bar-Or C, Czosnek H, Koltai H: Cross-species microarray hybridizations: a developing tool for studying species diversity. Trends Genet 23:200–207 (2007).
2.
Baron D, Houlgatte R, Postier A, Guiguen Y: Large-scale temporal gene expression profiling during gonadal differentiation and early gametogenesis in rainbow trout. Biol Reprod 73:959–966 (2005).
3.
Barske LA, Capel B: Blurring the edges in vertebrate sex determination. Curr Opin Genet Dev 18:499–505 (2008).
4.
Beato M, Klug J: Steroid hormone receptors: an update. Hum Reprod Update 6:225–236 (2000).
5.
Blasco M, Fernandino JI, Guilgur LG, Strussmann CA, Somoza GM, Vizziano-Cantonnet D: Molecular characterization of cyp11a1 and cyp11b1 and their gene expression profile in pejerrey (Odontesthes bonariensis) during early gonadal development. Comp Biochem Physiol A Mol Integr Physiol 156:110–118 (2010).
6.
Buchner J: Hsp90 & co. – a holding for folding. Trends Biochem Sci 24:136–141 (1999).
7.
Campbell WA, Yang H, Zetterberg H, Baulac S, Sears JA, et al: Zebrafish lacking Alzheimer presenilin enhancer 2 (pen-2) demonstrate excessive p53-dependent apoptosis and neuronal loss. J Neurochem 96:1423–1440 (2006).
8.
Chen WJ, Bonillo C, Lecointre G: Repeatability as a criterion of reliability of new clades in the Acanthomorph (Teleostei) radiation. Syst Biol 26:262–288 (2003).
9.
Choi CY, An KW, Choi YK, Jo PG, Min BH: Expression of warm temperature acclimation-related protein 65-kda (Wap65) mRNA, and physiological changes with increasing water temperature in black porgy, Acanthopagrus schlegeli. J Exp Zool A Ecol Genet Physiol 309:206–214 (2008).
10.
Clark MS, Burns G: Characterisation of the warm acclimated protein gene (wap65) in the Antarctic plunderfish (Harpagifer antarcticus). DNA Seq 19:50–55 (2008).
11.
Colell A, Ricci JE, Tait S, Milasta S, Maurer U, et al: GAPDH and autophagy preserve survival after apoptotic cytochrome c release in the absence of caspase activation. Cell 129:983–997 (2007).
12.
Conesa A, Gótz S, García-Gómez JM, Terol J, Talón M, Robles M: Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics 21:3674–3676 (2005).
13.
Cool J, Capel B: Mixed signals: development of the testis. Semin Reprod Med 27:5–13 (2009).
14.
Douglas SE: Microarray studies of gene expression in fish. OMICS 10:474–489 (2006).
15.
Ferguson-Smith M: The evolution of sex chromosomes and sex determination in vertebrates and the key role of DMRT1. Sex Dev 1:2–11 (2007).
16.
Fernandino JI, Hattori RS, Kimura H, Strüssmann CA, Somoza GM, et al: Dimorphic expression of dmrt1 and cyp19a1 (ovarian aromatase) during early gonadal development in pejerrey, Odontesthes bonariensis. Sex Dev 2:316–324 (2008a).
17.
Fernandino JI, Hattori RS, Kimura H, Strüssmann CA, Somoza GM: Expression profile and estrogenic regulation of anti-müllerian hormone during gonadal development in pejerrey Odontesthes bonariensis, a teleost fish with strong temperature-dependent sex determination. Dev Dynam 237:3192–3199 (2008b).
18.
Garcia-Reyero N, Griffitt RJ, Liu L, Kroll KJ, Farmerie WG, et al: Construction of a robust microarray from a non-model species (Largemouth bass) using pyrosequencing technology. J Fish Biol 72:2354–2376 (2008).
19.
Guo YL, Wang S, Cao DJ, Colman RW: Apoptotic effect of cleaved high molecular weight kininogen is regulated by extracellular matrix proteins. J Cell Biochem 89:622–632 (2003).
20.
Hattori RS, Fernandino JI, Kishii A, Kimura H, Kinno T, et al: Cortisol-induced masculinization: does thermal stress affect gonadal fate in pejerrey, a teleost fish with temperature-dependent sex determination? PLoS One 4:e6548 (2009).
21.
Hiramatsu R, Matoba S, Kanai-Azuma M, Tsunekawa N, Katoh-Fukui Y, et al: A critical time window of sry action in gonadal sex determination in mice. Development 136:129–138 (2009).
22.
Huxley AF: Muscular contraction. J Physiol 243:1–43 (1974).
23.
Inesi G, Prasad AM, Pilankatta R: The Ca2+ ATPase of cardiac sarcoplasmic reticulum: physiological role and relevance to diseases. Biochem Bioph Res Co 369:182–187 (2008).
24.
Ito LS, Yamashita M, Takashima F, Strüssmann CA: Dynamics and histological characteristics of gonadal sex differentiation in pejerrey (Odontesthes bonariensis) at feminizing and masculinizing temperatures. J Exp Zool A Comp Exp Biol 303:504–514 (2005).
25.
Ito LS, Takahashi C, Yamashita M, Strüssmann CA: Warm water induces apoptosis, gonadal degeneration, and germ cell loss in subadult pejerrey Odontesthes bonariensis (Pisces, Atheriniformes). Physiol Biochem Zool 81:762–774 (2008).
26.
Karube M, Fernandino JI, Strobl-Mazzulla P, Strüssmann CA, Yoshizaki G, et al: Characterization and expression profile of the ovarian cytochrome p-450 aromatase (cyp19a1) gene during thermolabile sex determination in pejerrey, Odontesthes bonariensis. J Exp Zool A Ecol Genet Physiol 307:625–636 (2007).
27.
Kikuchi K, Yamashita M, Watabe S, Aida K: The warm temperature acclimation-related 65-kDa protein, Wap65, in goldfish and its gene expression. J Biol Chem 270:17087–17092 (1995).
28.
Kinoshita M, Ozato K: Cytoplasmic microinjection of DNA into fertilized medaka (Oryzias latipes) eggs. Fish Biol J Medaka 7:59–64 (1995).
29.
Kurokawa H, Saito D, Nakamura S, Katoh-Fukui Y, Ohta K, et al: Germ cells are essential for sexual dimorphism in the medaka gonad. Proc Natl Acad Sci USA 104:16958–16963 (2007).
30.
Lee MO, Kim EO, Kwon HJ, Kim YM, Kang HJ, et al: Radicicol represses the transcriptional function of the estrogen receptor by suppressing the stabilization of the receptor by heat shock protein 90. Mol Cell Endocrinol 188:47–54 (2002).
31.
Li C, Lu G, Orti G: Optimal data partitioning and a test case for ray-finned fishes (Actinopterygii) based on ten nuclear loci. Syst Biol 57:519–539 (2008).
32.
Martyniuk CJ, Xiong H, Crump K, Chiu S, Sardana R, et al: Gene expression profiling in the neuroendocrine brain of male goldfish (Carassius auratus) exposed to 17alpha-ethinylestradiol. Physiol Genomics 27:328–336 (2006).
33.
Miura T, Miura C, Konda Y, Yamauchi K: Spermatogenesis-preventing substance in Japanese eel. Development 129:2689–2697 (2002).
34.
Nelson JS: Fish of the World, ed 4 (John Wiley & Sons Inc., New York, 2006).
35.
Newsholme EA, Beis I, Leech AR, Zammit VA: The role of creatine kinase and arginine kinase in muscle. Biochem J 172:533–537 (1978).
36.
North RA: Molecular physiology of P2X receptors. Physiol Rev 82:1013–1067 (2002).
37.
Ohta H, Aizawa S, Nishimune Y: Functional analysis of the p53 gene in apoptosis induced by heat stress or loss of stem cell factor signaling in mouse male germ cells. Biol Reprod 68:2249–2254 (2003).
38.
Ospina-Álvarez N, Piferrer F: Temperature-dependent sex determination in fish revisited: prevalence, a single sex ratio response pattern, and possible effects of climate change. PLoS One 3:1–11 (2008).
39.
Pearl LH, Prodromou C, Workman P: The HSP90 molecular chaperone: an open and shut case for treatment. Biochem J 410:439–453 (2008).
40.
Piferrer F, Guiguen Y: Fish gonadogenesis. Part II: molecular biology and genomics of sex differentiation. Rev Fish Sci 16:33–53 (2008).
41.
Popesku JT, Martyniuk CJ, Menningen J, Xiong H, Zhang D, et al: The goldfish (Carassius auratus) as a model for neuroendocrine. Mol Cell Endocr 293: 43–56 (2008).
42.
Rodríguez-Marí A, Yan YL, Bremiller RA, Wilson C, Cañestro C, Postlethwait JH: Characterization and expression pattern of zebrafish anti-Müllerian hormone (Amh) relative to sox9a, sox9b, and cyp19a1a, during gonad development. Gene Exp Patterns 5:655–667 (2005).
43.
Sauer U, Preininger C, Hany-Schmatzberger R: Quick and simple: quality control of microarray data. Bioinformatics 21:1572–1578 (2005).
44.
Setiamarga DH, Miya M, Yamanoue Y, Mabuchi K, Satoh TP, et al: Interrelationships of atherinomorpha (medakas, flyingfishes, killifishes, silversides, and their relatives): the first evidence based on whole mitogenome sequences. Mol Phylogenet Evol 49:598–605 (2008).
45.
Shoemaker CM, Queen J, Crews D: Response of candidate sex-determining genes to changes in temperature reveals their involvement in the molecular network underlying temperature-dependent sex determination. Mol Endocrinol 21:2750–2763 (2007).
46.
Silva MT, do Vale A, dos Santos NM: Fish and apoptosis: studies in disease and pharmaceutical design. Curr Pharm Design 14:170–183 (2008).
47.
Smith DF, Toft DO: Minireview: the intersection of steroid receptors with molecular chaperones: observations and questions. Mol Endocrinol 22:2229–2240 (2008).
48.
Strüssmann CA, Ito LS: Where does gonadal sex differentiation begin? Gradient of histological sex differentiation in the gonads of pejerrey, Odontesthes bonariensis (Pisces, Atherinidae). J Morphol 265:190–196 (2005).
49.
Strüssmann CA, Calsina Cota JC, Phonlor G, Higuchi H, Takashima F: Temperature effects on sex differentiation of two South American atherinids, Odontesthes argentinensis and Patagonina hatcheri. Environ Biol Fish 47:143–154 (1996).
50.
Strüssmann CA, Saito T, Usui M, Yamada H, Takashima F: Thermal thresholds and critical period of thermolabile sex determination in two atherinid fishes, Odontesthes bonariensis and Patagonina hatcheri. J Exp Zool 278:167–177 (1997).
51.
Strüssmann CA, Kitahara A, Yamashita M: Role of apoptosis in temperature-dependent sex determination of pejerrey Odontesthes bonariensis. Cybium 32:77–79 (2008).
52.
Tacchini L, Radice L, Bernelli-Zazzera A: Differential activation of some transcription factors during rat liver ischemia, reperfusion, and heat shock. J Cell Physiol 180:255–262 (1999).
53.
Tusher VG, Tibshirani R, Chu G: Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci USA 98:5116–5121 (2001).
54.
Uchida D, Yamashita M, Kitano T, Iguchi T: Oocyte apoptosis during the transition from ovary-like tissue to testes during sex differentiation of juvenile zebrafish. J Exp Biol 205:711–718 (2002).
55.
Uchida D, Yamashita M, Kitano T, Iguchi T: An aromatase inhibitor or high water temperature induce oocyte apoptosis and depletion of P450 aromatase activity in the gonads of genetic female zebrafish during sex-reversal. Comp Biochem Physiol A Mol Integr Physiol 137:11–20 (2004).
56.
von Schalburg K, Cooper G, Leong J, Robb A, Lieph R, et al: Expansion of genomics research of Atlantic salmon Salmo salarL. proyect (grasp) microarray tools. J Fish Biol 72:2051–2070 (2008).
57.
Walter I, Tschulenk W, Schabuss M, Miller I, Grillitsch B: Structure of the seminal pathway in the European chub, Leuciscus cephalus (Cyprinidae); Teleostei. J Morphol 263:375–391 (2005).
58.
Wang W, Li Y, Li Y, Hong A, Wang J, et al: NDRG3 is an androgen regulated and prostate enriched gene that promotes in vitro and in vivo prostate cancer cell growth. Int J Cancer 124:521–530 (2009).
59.
Wallis MC, Waters PD, Graves JA: Sex determination in mammals-before and after the evolution of SRY. Cell Mol Life Sci 65:3182–3195 (2008).
60.
Wilhelm D, Palmer S, Koopman P: Sex determination and gonadal development in mammals. Physiol Rev 87:1–28 (2007).
61.
Williams D, Li W, Hughes M, Gonzalez S, Vernon C, et al: Genomic resources and microarrays for the common carp (Cyprinus carpio L.). J Fish Biol 72:1–32 (2008).
62.
Yang YH, Dudoit S, Luu P, Lin DM, Peng V, et al: Normalization for cDNA microarray data: a robust composite method addressing single and multiple slide systematic variation. Nucleic Acids Res 30:e15 (2002).
63.
Yao HHC, Capel B: Temperature, genes, and sex: a comparative view of sex determination in Trachemys scripta and Mus musculus. J Biochem 138:5–12 (2005).
64.
Yoshinaga N, Shiraishi E, Yamamoto T, Iguchi T, Abe SI, Kitano T: Sexually dimorphic expression of a teleost homologue of Müllerian inhibiting substance during gonadal sex differentiation in Japanese flounder, Paralichthys olivaceus. Biochem Bioph Res Co 322:508–513 (2004).
65.
Zhang D, Popesku JT, Martyniuk CJ, Xiong H, Duarte-Guterman P, et al: Profiling neuroendocrine gene expression changes following fadrozole-induced estrogen decline in the female goldfish. Physiol Genomics 38:351–361 (2009).
66.
Zhao W, Tang R, Huang Y, Wang W, Zhou Z, et al: Cloning and expression pattern of the human NDRG3 gene. Biochim Biophys Acta 1519:134–138 (2001).
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