Pectoral Fin Breeding Tubercle Clusters: A Method to Determine Zebrafish Sex

McMillan, Stephanie C.; Géraudie, Jacqueline; Akimenko, Marie‐Andrée

doi:10.1089/zeb.2014.1060

Cited by 34 publications

(29 citation statements)

References 8 publications

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“…Automation of analysis and classification of sex using machine learning methods directly from images is a great possibility to increase the efficiency in the prediction process (Singh & Goel, ). The current guidelines for phenotypic sex classification in zebrafish using conventional methods are subjective and rely on different phenotypic observations by humans, which are highly labor‐intensive and potentially prone to error in the prediction process (McMillan et al, ). For example, zebrafish quickly become pale after removing from the water (Hutter et al, ), which may refer to the presence or absence of iridophore pigment cell types.…”

Section: Discussionmentioning

confidence: 99%

“…However, a precise sex determination of adult fish can be performed using microscopic examination by dissecting gonad tissue, which requires killing animals (Abozaid et al, ), is labor‐intensive and requires biological expertise. Hence, the current approaches for sex determination in zebrafish are subjective and not sufficiently reliable, and highly time‐consuming (McMillan et al, ).…”

Section: Introductionmentioning

confidence: 99%

“…However, for human observers the differences in coloration between male and female zebrafish are often difficult to discriminate due to minor sexual dimorphism in body color in this species (Hutter et al, 2012). The conventional guidelines for sex determination in zebrafish by human perception include sex-related differences in various features such as color, shape, behavior, and genital papilla (McMillan, Géraudie, & Akimenko, 2015). For example, females often have a round shape and protruding abdomen compared to males, but not all females possess an obviously distended abdomen (Hutter et al, 2010;McMillan et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Efficient phenotypic sex classification of zebrafish using machine learning methods

Hosseini

Simianer

Tetens

et al. 2019

Ecology and Evolution

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Sex determination in zebrafish by manual approaches according to current guidelines relies on human observation. These guidelines for sex recognition have proven to be subjective and highly labor‐intensive. To address this problem, we present a methodology to automatically classify the phenotypic sex using two machine learning methods: Deep Convolutional Neural Networks (DCNNs) based on the whole fish appearance and Support Vector Machine (SVM) based on caudal fin coloration. Machine learning techniques in sex classification provide potential efficiency with the advantage of automatization and robustness in the prediction process. Furthermore, since developmental plasticity can be influenced by environmental conditions, we have investigated the impact of elevated water temperature during embryogenesis on sex and sex‐related differences in color intensity of adult zebrafish. The estimated color intensity based on SVM was then applied to detect the association between coloration and body weight and length. Phenotypic sex classifications using machine learning methods resulted in a high degree of association with the real sex in nontreated animals. In temperature‐induced animals, DCNNs reached a performance of 100%, whereas 20% of males were misclassified using SVM due to a lower color intensity. Furthermore, a positive association between color intensity and body weight and length was observed in males. Our study demonstrates that high ambient temperature leads to a lower color intensity in male animals and a positive association of male caudal fin coloration with body weight and length, which appears to play a significant role in sexual attraction. The software developed for sex classification in this study is readily applicable to other species with sex‐linked visible phenotypic differences.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Efficient phenotypic sex classification of zebrafish using machine learning methods

Hosseini

Simianer

Tetens

et al. 2019

Ecology and Evolution

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“…Fish were sexed based on morphologically apparent dimorphic features, such as color pattern and presence or absence of male-specific breeding tubercles, the latter of which gives pectoral fins of males a refractile appearance when viewed from above (McMillan et al, 2015). In some cases, fish were also sexed by dissection and visual examination of gonads.…”

Section: Methodsmentioning

confidence: 99%

Dmrt1 is necessary for male sexual development in zebrafish

Webster

Schach

Ordaz

et al. 2017

Developmental Biology

242

178

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The dmrt1 (doublesex and mab-3 related transcription factor 1) gene is a key regulator of sex determination and/or gonadal sex differentiation across metazoan animals. This is unusual given that sex determination genes are typically not well conserved. The mechanisms by which zebrafish sex is determined have remained elusive due to the lack of sex chromosomes and the complex polygenic nature of sex determination in domesticated strains. To investigate the role of dmrt1 in zebrafish sex determination and gonad development, we isolated mutations disrupting this gene. We found that the majority of dmrt1 mutant fish develop as fertile females suggesting a complete male-to-female sex reversal in mutant animals that would have otherwise developed as males. A small percentage of mutant animals became males, but were sterile and displayed testicular dysgenesis. Therefore zebrafish dmrt1 functions in male sex determination and testis development. Mutant males had aberrant gonadal development at the onset of gonadal sex-differentiation, displaying reduced oocyte apoptosis followed by development of intersex gonads and failed testis morphogenesis and spermatogenesis. By contrast, female ovaries developed normally. We found that Dmrt1 is necessary for normal transcriptional regulation of the amh (anti-Müllerian hormone) and foxl2 (forkhead box L2) genes, which are thought to be important for male or female sexual development respectively. Interestingly, we identified one dmrt1 mutant allele that cooperates with a linked segregation distorter locus to generate an apparent XY sex determination mechanism. We conclude that dmrt1 is dispensable for ovary development but necessary for testis development in zebrafish, and that dmrt1 promotes male development by transcriptionally regulating male and female genes as has been described in other animals. Furthermore, the strong sex-ratio bias caused by dmrt1 reduction-of-function points to potential mechanisms through which sex chromosomes may evolve.

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“…Adult zebrafish may be reliably sexed on the basis of differences in morphology and pigmentation between males and females. 2,15 Breeding trials…”

Section: Determination Of Sex Ratiosmentioning

confidence: 99%

Successful Replacement of Artemia salina nauplii with Marine Rotifers (Brachionus plicatilis) in the Diet of Preadult Zebrafish (Danio rerio)

2015

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Previously established rearing protocols for zebrafish begin feeding with marine rotifers (Brachionus plicatilis), followed by Artemia nauplii until the fish reach subadult stage, the developmental time point at which they can be most easily transitioned onto a processed diet. However, the inclusion of Artemia is less than ideal, given its fluctuating availability and high costs. We tested whether or not we could replace Artemia with rotifers during our normal rearing sequence and still meet published performance standards for (i) weaning fish onto a processed diet by 25 days postfertilization (dpf) and (ii) successful breeding by 60 dpf. Here, we present the results of trials where wild-type and casper zebrafish were fed exclusively with rotifers (R) or rotifers followed by Artemia (RA) for the first 25 dpf after which point all fish were transitioned to a processed diet (Gemma Micro 300). We measured growth and survival at days 25 and 60, and tested for reproductive capability at 60 dpf. While growth performance was significantly better in the RA groups, we were still able to meet goals for both weaning and generation time in the R groups without compromising survival or sex ratios.

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Pectoral Fin Breeding Tubercle Clusters: A Method to Determine Zebrafish Sex

Cited by 34 publications

References 8 publications

Efficient phenotypic sex classification of zebrafish using machine learning methods

Efficient phenotypic sex classification of zebrafish using machine learning methods

Dmrt1 is necessary for male sexual development in zebrafish

Successful Replacement of Artemia salina nauplii with Marine Rotifers (Brachionus plicatilis) in the Diet of Preadult Zebrafish (Danio rerio)

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