Sex differentiation in teleost

The term sex differentiation varies across species as it has different process to achieve the
end point which is the formation of mature sex organ (citation). Similar to livestock
mammals such as ovine, bovine and porcine, the sex of teleost plays an important role for
agriculture purposes. As discussed previously, the sex genes and its regulators are not
conserved even across the same species. There are many factors that can contribute to the sex
differentiation process of teleost which can be divided into two major categories. Firstly, the
genetic sex determination (GSD) where the entire process of sex differentiation of gonad into
mature sexual organ solely on depends the genes and its availability. Secondly, the major
signals produced from the environment or known as environmental sex determination (ESD)
that includes the extrinsic factors such as temperature, pH value of the water and social
interaction (Guiguen et al. 2019). These forms of diversity are well observed in teleost as
they have evolved independently and repeatedly.

Despite the knowledge gathered in sex differentiation in teleost across last decades, it
is still rather tough to fully comprehend the process. At cellular level, sex differentiation
process involves activation and suppression of various and complicated pathways that
eventually leads to the germ cell determination and differentiation (citation). The potency and
efficacy of the process depends on the variation of allele at the genetic loci that encodes the
components that may or may not be influenced by environmental factor. In this pathway,
some components can work with other component to create a cascade of reaction while
playing the dominant role in directing the sex determination which will result to either the
development of an ovary or a testis. In certain genetic structure, it ranges from a complete
polygenic control to a sex determining dominant factors with addition of autosomal controls
to a higher evolution of heterogametic sex chromosomes (XY) males and (ZW) females
(Devlin and Nagahama 2002).

Gonadal sexual differentiation

In gonochoristic teleost, the sex differentiation depends on genetic, environment and in some
cases both. For instance, during gonadal differentiation of tilapia, the morphological sexual
dimorphism involves the change in the number of germ cells (citation). The primordial germ
cells (PGCs) are visibly in different appearance from the somatic cells. These cells are found
in the outer circle of the lateral plate mesoderm. Nine days after sex differentiation begin, the
mitosis takes place in the XX female germ cells but there was not any proliferation seen in
XY male gonads from day nine to day fourteen. Although the growth was observed in the
germ cells, the morphology of somatic tissue in the XX and XY gonads was not visible until
a few days later. The initial appearance of differentiated gonads was recorded to be twenty
and twenty-five days after post hatching for XX gonads and XY gonads respectively. As for
meiosis in the ovaries, it started between day twenty-five to day thirty and the initiation of
spermatogenesis was observed between day fifty to day seventy (Kobayashi et al. 2013).

The Sertoli and granulosa cells express the sox9b gene in the bipotential gonadal
primordia (citation). Furthermore, this gene is expressed in both XX and XY cells of teleost.
In contrast, the expression of sox9b in mammal is only expressed in the Sertoli cells
(citation). During testicular development of teleost, the expression sox9b genes induce the
expression of dmrt1 gene. Subsequently, initiate the gonadal primordia differentiation into
Sertoli cells which are found on the interior of the lobules in the testis. Since the gonadal
primordia can differentiate into either a testis or an ovary, the differentiation process has two
different pathways. For instance, oogenesis in the ovary involved the progression from
germline stem cells to early diplotene oocytes. At this stage, the expression of foxl2
substitutes the sox9b in granulosa cells. Then, the diplotene oocytes and the surrounding
somatic cells leaves the germinal cradle and recruiting the theca cells for formation of
follicles. These follicles were end up in stromal compartment to acquire two layers of somatic
cells which comprise of an inner granulosa and outer theca cells.

Sex hormones regulating sex differentiation

The endocrine system plays an important role in sex differentiation (cite). Previously, steroids
hormone has shown to induce phenotypic sex reversal in teleost (cite). The production of
oestrogen is related to ovarian differentiation. Meanwhile, 11-oxygenated androgens are
connected to testicular differentiation. However, previous findings have shown that the
balance of these two would conclude the gonadal sex differentiation rather than the absence
or presence of these steroid hormones (cite). Study in Chinook salmon has shown that
inhibition of oestrogen synthesis with specific enzyme such as aromatase inhibitor, the
genetically female fish underwent phenotypic masculinization. This postulated that the
absence of oestrogen is adequate to steer the gonad differentiation towards testis together
with the absence of androgens hormones. To conclude, oestrogen is needed for female sex
differentiation while the lack thereof results in male sex differentiation in an oestrogen-
centric model. Although androgens might or might not be needed for testicular development,
they are certainly needed to maintain the male phenotype as they are a natural suppressor of
the production of oestrogen. Therefore, the male sex of the teleost is maintained as the
aromatase gene is repressed as well. In the absence of oestrogen, it leads to impartial or
complete functioning masculinization which leads to the assumption that maintenance of the
ovary requires constant production of oestrogen. Therefore, oestrogen hormone is an essential
hormone for gonadal differentiation into ovaries as well to maintain this sexual form. Besides
sex hormones, other studies have shown that cortisol is responsible for masculinization in
teleost (citation). This event is postulated to occur with two reasons; i) a key enzyme in
cortisol synthesis also responsible for 11-oxygenated androgen synthesis and this 11-
oxygenated androgen favours testicular differentiation, ii) cortisol induce the increased
expression of the cyp19a1a gene. In addition, this particular cyp19a1a gene experiences
epigenetic inhibition when there is an increase in temperature resulting in masculinization of
female teleost (citation).

Environmental factors influencing sex determination

There are some environmental factors that could impact the sex differentiation of teleost
which includes temperature, pH and social interactions. When the temperature of the habitat
fluctuates, the biochemical pathways were affected and resulting in a bias during sex
differentiation (citation). Generally, in the thermosensitive teleost, temperature elevation
induces this fish population leans into a higher male population. However, low water
temperature setting for this fish induces the development of ovaries (citation). This
phenomenon has been observed in a few species from the genus Apistogramma and in
Dicentrarchus labrax L., a type of sea bass. The temperature shift however is only crucial
during the very initial process of sex differentiation. Temperature sensitivity is linked to
genetic constituent because the offspring of the produced females has various reaction to the
temperature. It has been inferred that when a teleost is thermosensitive, it could be a
hereditary trait. The young of Poeciliopsis lucida is formed inside the parent. When they are
exposed to a higher temperature prior to delivery, the offspring sexual ratio skew towards
males but only in delicate lines. However, another study has shown that this temperature
factor has no effect on some teleost such as Cyprinodon variegatus and Salmonid Coregonus
hoyi.

Social factors also play a role in sex differentiation in hermaphroditic teleost. Sex
reversal is mainly regulated by this factor in response to population density and the ratio of
male to female at a given period. The precise mechanism of this effect is complex and not
fully understood. The Thalassoma bifasciatum male is produced by two different pathways; i)
they can either mature as males naturally (primary males), ii) they mature as females and then
undergo a sex reversal (secondary males). Primary males are usually generated by the high
population density. As for the Cichlasoma citrinellum, the social factor that affects the sex
differentiation is the size of juvenile where the relatively bigger fishes mature as males
(Sandra and Norma 2010).

Conclusion

The sex determining gene and the sex differentiation process are influenced by not only genes
but also other extrinsic factors. It is known that the sex of the teleost is a complex process
which depends on more than one element and component. In order to completely understand
the mechanism of sex determining gene and sex differentiation, more studies must be
conducted across the species. Future studies could be executed to identify the equivalent of
the Sry gene in mammals for teleost to understand the sex determining gene better. There are
many studies have been done on zebrafish, chichilds and rainbow trout but that is not
sufficient as it varies across genus and sometimes even species. The Malaysian mahseer Tor
tambroides; an endangered species is one of the species that have very minimal information
on their sex determining gene and sex differentiation as of now. Studies have been done on
their growth and mitochondria but not in sex determination and differentiation. Since they
take a relatively longer time to mature when compared to the examples above, knowing their
sex determining gene would make it easier to repopulate them and reduce their chances into
extinction.

References:

Devlin RH, Nagahama Y (2002) Sex determination and sex differentiation in fish: an
overview of genetic, physiological, and environmental influences Aquaculture
208:191-364
Guiguen Y, Fostier A, Herpin A (2019) Sex determination and differentiation in fish:
Genetic, genomic, and endocrine aspects Sex control in aquaculture 1:35-63
Kobayashi Y, Nagahama Y, Nakamura M (2013) Diversity and plasticity of sex
determination and differentiation in fishes Sexual Development 7:115-125
Sandra G-E, Norma M-M (2010) Sexual determination and differentiation in teleost fish
Reviews in Fish Biology and Fisheries 20:101-121