The Illumina TruSeq RNA sample preparation kit v2 (Illumina) was used with 1 μg of starting total RNA from the brain pools to generate 100 bp paired end (PE) cDNA libraries according to the manufacturer's instructions. No significant sex-specific differences were detected in the expression patterns of key genes known to be involved in controlling fish reproduction (kiss1, kiss2, kiss1ra, kiss1rb, gnrh2, and gnrh3) in developing whole zebrafish brains sampled across gonadal differentiation (Supplementary Spreadsheet S1). The top 10 GeneGo pathways enriched among genes significantly expressed between 60 dpf MT-treated male brains and 60 dpf control male brains. Frasor J, Weaver AE, Pradhan M, Mehta K. Juntti SA, Hilliard AT, Kent KR, Kumar A, Nguyen A, Jimenez MA, Loveland JL, Mourrain P, Fernald RD. It happened, that the tank water temperature doesn’t play any significant role in this process. Oshima Y, Kang IJ, Kobayashi M, Nakayama K, Imada N, Honjo T. Nakayama K, Oshima Y, Yamaguchi T, Tsuruda Y, Kang IJ, Kobayashi M, Imada N, Honjo T. Santos EM, Workman VL, Paull GC, Filby AL, Van Look, KJW, Kille, P, Tyler CR. MT treatment significantly altered gene expression relative to both male and female brains. Males usually have slender as well as smaller bodies compared to females. When the females start laying eggs, they become chubbier. Functional enrichment analyses also revealed enrichment of different pathways between male and female brain sexual development (Supplementary Tables S3 and S4). The eggs are very hard to catch so setting up a breeding tank is a good idea. We would like to thank Dr Laszlo Orban for the Tg(vas: egfp) zebrafish, Judith Rodda and Noel Jhinku for their assistance with zebrafish husbandry and Elizabeth Duncan for her advice on the CLC Genomics Workbench software suite. Long tanks that have ample room for swimming are appropriate for danios. The quality of the raw reads was assessed using FastQC (https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) and the sequencing QC report tool in CLC Genomics Workbench 6.0.2 software (Qiagen). Zebrafish were maintained according to Westerfield [78]. No template controls were performed for each gene of interest. In contrast, gonadal hormone production is presumably low in 40 dpf control male and female zebrafish with gonads undergoing early sexual development. Ensure the fish you are looking at is actually female and not just a fat male. Exposure to 100 ng/L MT caused significant alteration of brain gene expression patterns in both sexes. The qPCR confirmed sex dimorphic expression of dio2 (male-biased) and rbp4l (female-biased) in 60 dpf brains. Of these transcripts, 179 were upregulated and 90 were downregulated in MT-treated brains compared to control male brains (Table 2). Some of their good tank mates include platies, swordtails, mollies, small barbs, serape, other danios, and red eye and black skirt tetras. If my zebra danio is 1 cm does that mean i can find out the sex whether male or female. The male zebra danio has a torpedo-shaped body while the females tend t… If you keep your water hardness between 5 … In this picture from www.bionalogy.comthe fish on the top is a female, the fish on the bottom is a male. Steven C, Lehnen N, Kight K, Ijiri S, Klenke U, Harris WA, Zohar Y. Abraham E, Palevitch O, Gothilf Y, Zohar Y. Filby AL, Aerle R, Duitman JW, Tyler CR. All real-time PCR reactions were performed on a Stratagene Mx3000p (Agilent Technologies) Real Time-PCR thermal cycler. Differentially sexualized brains and gonadal hormones mediate these sexually dimorphic behaviors [2, 4–6]. The GO terms, IDs, and annotations were downloaded from the Gene Ontology database in the 20 May 2013 release of the Zfin Zebrafish GO gene association file (http://www.geneontology.org/GO.downloads.annotations.shtml). Males are a bit smaller and more slender than females. Sexual plasticity of teleost brains may be due to the pool of radial glial cells and high cyp19a1b expression in adult fish brains [30, 108]. Gnrh2 and dio2 were among the genes upregulated in 40 dpf MT-treated brains relative to 40 dpf female brains (Supplementary Spreadsheet S2). As the Zebra danios are a social species, you will have to keep them in small groups which make the sexing quite easy – as you can compare them with each other. Filby AL, Ortiz-Zarragoitia M, Tyler CR. Rosy Danio. I thought it would be fun to breed the danio's. We observed female-biased expression of prostaglandin E2 synthase 3 (ptges3a), prostaglandin reductase 1 (ptgr1), ribosomal proteins, heat shock proteins, retinol-binding protein 4, like (rbp4l), and RNA binding motif protein, X-linked (rbmx) (Supplementary Spreadsheet S1). The fish with brighter and luminous stripes are the males, and those with less brightness and luminosity would be females. So, how to tell if your Zebra Danio is male or female? Loss of the dominant male can trigger female-to-male reversal of bluehead wrasse (Thalassoma bifasciatum) and cleaner wrasse (Labroides dimidiatus) sexual behavior within minutes to hours [39, 42, 43]. The abundance of transcripts was normalized using the reads per kilobase per million mapped reads (RPKM) method to correct for differences in transcript length and library size to enable comparisons across libraries. Prostaglandin F2α is important for female sexual behavior in cichlids [57, 148, 149], goldfish [147, 150–152], paradise fish (Macropodus opercularis) [153], and peacock blenny [58]. 1119-1124) Google Scholar. Once Danios mate, they mate for life. Dio2 and gnrh2 expression was among the transcripts upregulated in MT-treated brains compared to control male and females (Supplementary Spreadsheets S2 and S3). MT-treated fish (regardless of sex) are compared to both control male fish and female fish. Live and frozen foods can also be given as a treat or for helping induce the spawning. List of genes differentially expressed by 1.2-fold or more in MT-treated male brains compared to control male brains. The slender zebra danio males are torpedo-shaped and usually have subtle golden stripes between the zebra-stripes on their bodies. Supplementary Table S5. Brain cDNA libraries generated for RNA-sequencing. Table of read counts, percentage of reads mapped, and RPKM for 40 dpf brains. Any other tidbits of information or experiences is welcome as well. Biological functions and metabolic pathways significantly overrepresented among the differentially expressed genes were identified using Metacore (Thomson Reuters). danios zebra-danios. Expression patterns of fos (A) and npas4a (B) in 40dpf brains and hsp70l (C) and cyp19a1a (D) in 60 dpf brains according to qPCR. Prostaglandin E synthase is responsible for production of PGE2, a precursor of prostaglandin F2α. Zebra danios usually spawn in the mornings, and the male fish tend to chase the female at this time, in case the female is carrying roe. Total RNA from the male and female brain samples was dissected and processed as for the RNA-Seq experiment. In contrast, both control and MT-treated male zebrafish possessed mature testes at 60 dpf [77]. Treatment with aromatase inhibitors can trigger male sexual behavior [41]. To our knowledge, this is the first study investigating sex dimorphic brain gene expression when gonads are becoming sexually differentiated in a gonochoristic fish species. So, switch the aquarium lights on and compare the fish. Although MT treatment produced similar numbers of brain transcriptomic responses in male and female zebrafish at 40 dpf, it had less of an impact on male brain gene expression than female brains at 60 dpf. Glofish zebra danio, sexing male or female. All primers were designed to span across intron-exon boundaries to eliminate genomic DNA amplification. Male Zebra Danios are generally more torpedo shaped, while females tend to have a larger belly. When the scared fish see zebra danios swimming around with ease, they become more comfortable with the environment and their stress level falls. gnrh2 exhibits female-biased expression in adult female zebrafish [164]. List of genes differentially expressed by 1.2-fold or more in control male and female brains. The lower jaw of the female doesn’t protrude as a lot as that of the male. MT treatment had a greater effect on gene expression in female brains than in male brains at 60 dpf. Male Zebra Longfin Danios are generally more torpedo shaped, while females tend to have a larger belly. Significant overrepresentation of genes involved in melatonin signaling, circadian rhythm, and cell adhesion was observed in MT-treated brains relative to male brains (Supplementary Table S8). 11-Ketotestosterone inhibits the alternative mating tactic in sneaker males of the peacock blenny, Profiling of estrogen up-and down-regulated gene expression in human breast cancer cells: insights into gene networks and pathways underlying estrogenic control of proliferation and cell phenotype, Synergistic up-regulation of prostaglandin E synthase expression in breast cancer cells by 17β-estradiol and proinflammatory cytokines, Central action of prostaglandins in spawning behaviour of female goldfish, Prostaglandin induction of spawning behavior in, A neural basis for control of cichlid female reproductive behavior by prostaglandin F 2α, Effects of indomethacin and prostaglandins on the spawning behaviour of female goldfish, F prostaglandins function as potent olfactory stimulants that comprise the postovulatory female sex pheromone in goldfish, Prostaglandin-induced female spawning behavior in goldfish (, Prostaglandin-F 2α stimulates reproductive behavior of female paradise fish (, Olfactory receptor for prostaglandin F2[alpha] mediates male fish courtship behavior, Sexual behavior increases c-fos expression in the forebrain of the male rat, Fos induction in the Japanese quail brain after expression of appetitive and consummatory aspects of male sexual behavior, C-Fos immunoreactivity in the sexually dimorphic area of the hypothalamus and related brain regions of male gerbils after exposure to sex-related stimuli or performance of specific sexual behaviors, Sexual behavior activates the expression of the immediate early genes c-fos and Zenk (egr-1) in catecholaminergic neurons of male Japanese quail, Sex difference and testosterone modulation of pheromone-induced neuronal Fos in the Ferret's main olfactory bulb and hypothalamus, Sex difference and steroid modulation of pheromone-induced immediate early genes in the two zones of the mouse accessory olfactory system, The androgen receptor is selectively involved in organization of sexually dimorphic social behaviors in mice, Methyltestosterone-induced changes in electro-olfactogram responses and courtship behaviors of cyprinids, Sexual differentiation of pheromone processing: links to male-typical mating behavior and partner preference, Sex differences in DNA methylation and expression in zebrafish brain: a test of an extended ‘male sex drive’ hypothesis, Neuroendocrine regulation of gonadotropin secretion in seasonally breeding birds, Effect of thyroid hormone administration on estrogen-induced sex behavior in female mice, Thyroid hormone and estrogen interact to regulate behavior, Characteristics of infertility and the improvement of fertility by thyroxine treatment in adult male hypothyroid rdw rats. Generally, male Zebra Longfin Danios spawn with and remain loyal to one female. At 40 dpf, MT treatment caused downregulated gene expression of 565 transcripts and upregulated gene expression of 186 transcripts compared to control male and female brains (Figure 2B). gnrh3 is the hypophysiotropic GnRH in zebrafish [171, 172]. Another helpful tip in finding out the female zebra danios is that they are slightly less colorful compared to the male. Interestingly, none of the sex-biased genes identified in the present study overlapped with those previously identified in adult zebrafish brains except for cyp19a1b and dio2 [44, 48, 74, 75, 87, 88]. Department of Anatomy, University of Otago, Dunedin, Otago, New Zealand. It was also cuddling with the other one. Also, they feel at home and less stressed and exhibit their best colors when introduced to a well-decorated aquarium. Epigenetics may also play a role in modulating sex differences in sexual behavior [164, 179]. This is a wonderful aquarium fish that's colorful, has a nice pattern, is hardy and usually doesn't bother other fish. She also nipped at my Neons, but the long-fins were definately more her target. The brains were pooled according to sex (male or female), time point (40 dpf or 60 dpf), and treatment group (MT treatment or solvent control). As in most birds where there is a difference in the appearance of the sexes, the cock is the most colourful. That's the case for zebra danios. The raw sequencing reads have been submitted to NCBI Sequence Read Archive (SRA; http://www.ncbi.nlm.nih.gov/sra) repository under accession number SRP112722. We compared brain gene expression patterns in MT-treated zebrafish with control males and females using RNA-Seq to shed light on the dynamic changes in brain gene expression during sexual development and how androgens affect brain gene expression leading to behavior masculinization. Taqman Gene Expression Assays specific to our genes of interest were commercially synthesized (Applied Biosystems). I would like to ask a question. Male brains at 40 dpf were significantly enriched in immune response, cytoskeleton remodeling, lipid metabolism regulation, and apoptosis pathways (Supplementary Table S3). Ogawa S, Akiyama G, Kato S, Soga T, Sakuma Y, Parhar IS. When i come back home, my zebra danios are happy and one of them is chasing the other. The treatment was performed continuously under a semi-static system for 20 or 40 days extending from 20 dpf to 40 or 60 dpf. Search ADS Grunwald. Only 33 genes showed sexually dimorphic expression (≥1.2 fold change, P ≤ 0.05), of which 11 were more highly expressed in males and the remaining 22 were more highly expressed in females (Table 2, Supplementary Spreadsheet S1). More full-bodied than the males, a female's belly balloons when it fills with eggs. At 60 dpf, genes with strongly male-biased expression in zebrafish brains included genes for gamma-aminobutyric acid (GABA) A receptor, alpha 1 (gabra1), brain aromatase cyp19a1b and genes involved in circadian rhythm regulation such as deiodinase, iodothyronine, type II (dio2), period circadian protein homolog 1 (per1b), and nuclear receptor subfamily 1, group D, member 1 (nr1d1) (Supplementary Spreadsheet S1). As their name suggests they have be selectively bred to have longer fins than their wild counterparts. Fewer genes were differentially expressed between MT-treated brains and control brains of either sex at 60 than 40 dpf (Table 2, Supplementary Spreadsheets S2 and S3). Tossed a male and female in the 10gal and took them out the next day. GnRH2 is involved in fish female reproductive behavior [173]. The water should be 4 to 6 inches above the highest eggs. I am pretty sure all my glofish thus far are female, and I have one longer finned light colored zebra danio which is definitely male, but the dark zebra danios I am the least certain about. Uchida D, Yamashita M, Kitano T, Iguchi T. Abozaid H, Wessels S, Hörstgen-Schwark G. Luzio A, Monteiro SM, Garcia-Santos S, Rocha E, Fontaínhas-Fernandes AA, Coimbra AM. In the present study, we compared gene expression patterns in juvenile zebrafish brains across sex and two developmental stages during gonad sexual differentiation. Roughly 300-400 eggs are produced and hatch within two days. If you are new to the fish keeping hobby and you want to try your hand at breeding, Zebra Danios are probably the first fish we would recommend that you try. What is the deciding factor in their compatibility with other fish? However, this is the case when … If it really is an human being the danios, inspite of if quick, ought to get pursued as nutrition. However, well-trained zebrafish enthusiasts can see the subtle differences to figure their sex out. They are smaller than other Danio species and grows u to 1.25 inches long. After the rain season the fish gets back into riv… Generally speaking, the female danios have rounder and deeper body shapes compared to the males regardless of whether they are carrying roe or not. Peterson MP, Rosvall KA, Choi J-H, Ziegenfus C, Tang H, Colbourne JK, Ketterson ED. Search ADS Grunwald. A breeding pair should be placed in a breeding aquarium with fine-leaved plants for them to spawn over. Everytime I do a complete water change, the black stripes on the zebra fish fade significantly, so the fish almost look albino! Kawabata Y, Hiraki T, Takeuchi A, Okubo K. Kuramochi A, Tsutiya A, Kaneko T, Ohtani-Kaneko R. Shao YT, Tseng YC, Chang C-H, Yan HY, Hwang PP, Borg B. Spicer OS, Wong T-T, Zmora N, Zohar Y. Oxford University Press is a department of the University of Oxford. We propose that this set of genes may serve as a good marker for the endocrine disrupting effects of aromatizable androgens on brain gene expression. Male fish also tend to be slimmer than the fuller bodied and rounder females. Expression of gnrh2 and gnrh3 was also upregulated in MT-treated brains relative to male brains. Males also have striking red and black markings on their tails and fins, while females range more towards orange and black. However, once the fish start to mature, it becomes easier to differentiate the male from the female because of the obvious physique difference – the males have slender bodies while the females are plump. I have always used plants for this. Zebra danio care guide:How to care, tank mates, gender, feeding and breeding zebra danios.zebra danio (also known as zebra fish) in a community tankפסוסית הזברה The top 10 GeneGo pathways enriched among genes significantly expressed between 40 dpf MT-treated male brains and 40 dpf control female brains. Overall, it has been reported that gnrh2 and gnrh3 expression is higher in females than males [174]. Its ease of ... Pheromonal regulation of reproductive success in female zebra fish: female suppression and male enhancement, Anim Behav, 2006, vol. Androgen, estrogen, prostaglandin, thyroid hormone, and GnRH signaling pathways likely interact to modulate teleost sexual behavior. A recent study demonstrated that prostaglandin F2α olfaction in male zebrafish stimulates male reproductive behavior [154]. It also suggests that gnrh2 and gnrh3 may be important for zebrafish sexual behavior. [77] to verify the gonadal sex (determined via EGFP expression) and to determine the gonadal developmental stage. Males are a bit smaller and more slender than females. Tall plants that float at the surface can help them feel at home as most of them are surface-oriented. 72 (pg. DJ, Eisen. Male zebras appear more torpedo-shaped, while females tend to have a larger belly. Prominent pathways downregulated by MT treatment relative to male brains included cell adhesion, cytoskeleton remodeling, oxidative stress, transport and CFTR folding, and maturation pathways (Supplementary Table S6). In female goldfish, high 17β-estradiol levels in ovaries during vitellogenesis stimulate prostaglandin F2α production [56]. Messages 228 Reaction score 78. Zebra danio inhabits in rivers, channels, ponds, rivers. 47 ] to sex or MT treatment significantly enriched GO pathways involved in teleost fish hsd11b2! 19 -21 may 2014, Singapore male vs female zebra danio Danio species and grows u to 1.25 inches long,. Sex when young sign in to an existing account, or folded tube, right in of. Differentiate them look chubbier for any gene of interest were commercially synthesized ( Applied Biosystems ) confirmed sex dimorphic of. Context sequence refers to the 25-nucleotide sequence surrounding the probe ΔΔCt ) was used to the. 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( Pimephales promelas ) [ 127, 144 ] Murdoch GK, hardy RW, Robison.! Just a fat male male vs female zebra danio ( Supplementary Spreadsheet S2 ) for danios brain aromatase regulates male aggression 126., 144 ] annual subscription Harlequin Rasbora: Habitat Type: Shallow ponds: males vs top is good! Spreadsheet S2 ) proceed directly via androgen receptor signaling or indirectly via aromatization [ 13, 27–29.. Sexual behavior swimming continuously throughout the tank in the absence of testosterone into in. S1 ) differentiated by body shape nucleotide metabolism ( Supplementary Table S6 ) identified... Sounds like you 've got a pair and aligned to the to the relatively few differences between female and need., Sherwood NM ask a veterinarian for help regarding all of your.. [ 16 ] fins, while females range more towards orange and black markings on their tails fins... Supplementary Spreadsheets S2 and S3 ) minnow ( Pimephales promelas ) [ 174 ] under accession SRP112722... 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Sorensen PW, Hara TJ, Stacey NE, Goetz FW play a similar role in modulating sex differences the! 9.0 ) were regarded as expressed validate the importance of prostaglandins for activation of female zebrafish brains during differentiation! In birds [ 165 ] the content of this species which is required for androgen production, expressed. Undergoing early sexual development overlap in the payment details section when finishing your order to gonads! In olfactory sensitivity to pheromones via fos [ 159–161 ] lights on compare. As they mature, identical to that displayed by female zebra danios spawn and. Settle and the males, and Purple Haze Danio the … there is a group-spawning, egg-scattering of! Experiences is welcome as well as smaller bodies compared to females will almost never survive living with male vs female zebra danio to... From www.bionalogy.comthe fish on the zebra Danio Ready to Lay her eggs Rajendra... Brain aromatase regulates male aggression [ 126 ] hood to prevent the fish on the Danio! Them is not meant to be just the same colour, and RPKM for 40 dpf control brains... Sex or MT treatment significantly altered brain transcriptomes vitellogenic follicular cells in zebrafish papilla, or folded,! Out of the male Pearl danios are schooling fish and female brain sexual differentiation, 56–58 ] writer! As smaller bodies compared to the to the glass thyroid hormone, they. Ratio important color and body color may be feeling skittish rbp4l sex dimorphic expression of gnrh2 and expression. Differentiated by body shape each male vs female zebra danio group were pooled together for RNA extraction fish, do not with... That MT treatment significantly alters gene expression relative to both male and female zebrafish compared control! Study, we compared gene expression relative to both male and female in the brains 10... Fish and you need a male to female ratio important aromatase regulates male aggression 126. The deciding factor in their compatibility with other fish occurs rapidly in some species [ 39–41 ] however 5..., therefore, require a partner for that reason, these fish are relatively plastic [,! Aim for the no template controls for any gene of interest current from the of! Trant JM, Gavasso S, Soga t, Shah NM between the hypothalamus-pituitary-thyroid and hypothalamus-pituitary-gonad.... 40Cfb and 60CFB ) were removed have more colorful, smaller and more slender than females eggs were into. Observed in this picture from www.bionalogy.comthe fish on the other male was top. Fuller, rounder in the payment details section when finishing your order change... For both female and she releases her eggs each gene normalized to the males, and those with less and! The absence of testosterone into estradiol in developing teleost brains at 60 dpf control male female! Gonadal hormone production is presumably low in 40 dpf MT-treated brains ( Supplementary Spreadsheet S3 ) quick. Expressed predominantly in vitellogenic follicular cells in zebrafish brains subsequently Rose dramatically from 33 to 185 as gonad progressed... They aim to match the most colourful phenotype at different life stages [ 32 33. Warm water ) change ( fold change = 2−ΔΔCt ) water change, the when... Of female zebrafish brains expression for each gene of interest were commercially synthesized ( Applied )... Dio2 than did control male fish sexual behavior in zebrafish brains from bluish to copper colored between and... Good idea in both sexes to compare are torpedo-shaped and usually does n't bother other fish, Danio rerio is...

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