Vitellogenin Synthesis Research Articles

FOXL2 down-regulates vitellogenin expression at mature stage in Eriocheir sinensis.

Ovarian development in crustaceans is characterized by rapid production of egg yolk protein in a process called vitellogenesis. In the present study, we investigated the involvement of a DEAD (Asp-Glu-Ala-Asp) box RNA helicase 20 (DDX20), forkhead transcription factor (FOXL)2 and fushi tarazu factor (FTZ-F)1in the regulation of vitellogenesis. Based on ESTs from the testis and accessory gland of Eriocheir sinensis, we cloned the full-length cDNAs of foxl2 and fushitarazu factor 1 (ftz-f1), which include the conserved structural features of the forkhead family and nuclear receptor 5A (NR5A) family respectively. The expression of foxl2 mRNA surged at the mature stage of the ovary, when vtg mRNA swooped, suggesting that foxl2 negatively affects the vitellogenin (VTG) synthesis at this developmental stage. Etoposide (inducing germ cell apoptosis) treatment up-regulated FOXL2 and DDX20 at both the mRNA and the protein levels, primarily in the follicular cells as shown by immunofluorescence analysis. Furthermore, foxl2, ddx20 and ftz-f1 mRNA levels increased significantly with right-eyestalk ablation. Interactions between FOXL2 and DDX20 or FTZ-F1 were confirmed by co-immunoprecipitation and the forkhead domain of FOXL2 was identified as the specific structure interacting with FTZ-F1. In conclusion, FOXL2 down-regulates VTG expression by binding with DDX20in regulation of follicular cell apoptosis and with FTZ-F1 to repress the synthesis of VTG at the mature stage. This report is the first to describe the molecular mechanism of VTG synthesis in E. sinensis and may shed new light on the regulation of cytochrome P450 enzyme by FOXL2 and FTZ-F1in vitellogenesis.

Termite queens have disproportionately more DNA in their fat body cells: reproductive division of labor and endoreduplication

AbstractIncreases in DNA content caused by endoreduplication are widely observed in the metabolically active tissues of plants and animals. During egg production, insect females synthesize very large amounts of vitellogenin in their fat bodies, and female fat bodies of some insects become polyploid to accelerate vitellogenin production. Social insects have developed reproductive division of labor, wherein queens lay most of the eggs while other individuals have reduced fertility and undertake tasks required for maintaining the colony. Therefore, only queens are engaged in vitellogenin synthesis for egg production in social insects. Here, we show that termite queens have disproportionately more DNA in their fat body cells. Our DNA content analysis using flow cytometry demonstrated that more cells contained 4C‐DNA than 2C‐DNA in the fat bodies of Reticulitermes speratus queens. This high level of endoreduplication was not found in the fat body cells of other castes. This caste‐dependent doubling of DNA content in fat body cells suggests that termites exploit endoreduplication to boost egg production, in conjunction with the development of reproductive division of labor. This study highlights nuclear polyploidization as an adaptive strategy in social insects.

Epigenetic and endocrine determinants of lifespan differences between the castes of social insects

Social insects are promising model organisms in the study of longevity mechanisms. They have a caste system, in which the same genomes may produce phenotypes significantly distinguished by longevity. The ontogenetic trajectories of bees depend on the duration of supply by a specific nutrient mixture (royal jelly) at the larval stage. Longer feeding by the royal jelly leads to the formation of the queen epigenome, which is different from the epigenome of a working bee. Such epigenetic differences, in turn, induce endocrine changes manifested in increased synthesis of juvenile hormone and activation of TOR (target of rapamycin) signaling pathway, as well as in the modulation of insulin/IGF-1 pathway in queen-destined larvae. In adults, these processes influence the synthesis of vitellogenin (egg yolk precursor determining many aspects of insect ontogenesis). Epigenetic and andocrine mechanisms that underlie differences in longevity among the castes of social insects are discussed.

Reproductive health indicators of immature common carp exposed to municipal wastewater of Behbahan, Iran

Exogenous estrogens or pollutants with estrogen-like activity can induce vitellogenin (VTG) synthesis in male and juvenile fish, making this protein a useful indicator of chemicals that mimic estrogenic activity. The purpose of this study was to investigate the impact of municipal wastewater on blood biochemical parameters of common carp (Cyprinus carpio). Under experimental conditions, biomarkers such as sex steroid levels, alkali-labile phosphate levels, cholesterol and triglycerides, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were assessed in immature fish exposed to municipal wastewaters collected from a sewage canal in Behbahan, Khuzestan Province, Iran. No significant changes were found in testosterone levels on day 21; however, estradiol, alkali-labile phosphate, triglycerides, cholesterol, and LDL-cholesterol significantly increased in the fish exposed to municipal wastewater compared with the control group. A significant decrease in HDL-cholesterol levels was observed in the fish exposed to municipal wastewater at the end of the experiment. In conclusion, the results of the present study indicated that sewage effluent of Behbahan may contain endocrine disrupters and exposure to sublethal concentrations of municipal wastewater may cause dysfunction in reproductive health indicators of common carp

In vitro and in vivo studies of the endocrine disrupting potency of cadmium in roach (Rutilus rutilus) liver

Cadmium has been reported to exert estrogenic, antiestrogenic or both effects in vertebrate species. To elucidate the endocrine disrupting action of CdCl2, ex vivo and in vivo experiments were performed in roach (Rutilus rutilus). Roach liver explants were exposed to a range of CdCl2 concentrations alone (0.1–50μM) or with an effective concentration (100nM) of 17β-estradiol (E2). In addition, juvenile roach were intraperitoneally injected with CdCl2 (0.1–2.5mg/kg) with or without 1mg E2/kg. Subsequent analysis evaluated the effect of CdCl2 on vitellogenin (VTG) synthesis both at the mRNA and protein level, on estrogen receptors (erα and erβ1) and on androgen receptor (ar) mRNA expression. Ex vivo and in vivo experiments indicated that CdCl2 is strongly anti-estrogenic as, when co-exposed to E2, CdCl2 significantly inhibited VTG production as well as vtg and erα mRNA expressions. Moreover, CdCl2 compromised the E2-mediated induction of the ar mRNA expression in vivo.

Expression of RXR, EcR, E75 and VtG mRNA levels in the hepatopancreas and ovary of the freshwater edible crab, Oziothelphusa senex senex (Fabricius, 1798) during different vitellogenic stages.

The objective of the present study was to investigate the expression profile of retinoid X receptor (RXR), ecdysone receptor (EcR) and ecdysone inducible gene (E75) in the hepatopancreas and ovary of Oziothelphusa senex senex during different vitellogenic stages. RXR, EcR and E75 complementary DNAs (cDNAs) were isolated from the ovaries, while vitellogenin (VtG) cDNA was isolated from the hepatopancreas of vitellogenic female crab. Deduced amino acid sequence of the messenger RNAs (mRNAs) of RXR, EcR and E75 showed more than 80% identity with their respective mRNAs of other brachyurans. VtG mRNA was not detected in the ovary throughout vitellogenic stages. RXR and EcR were significantly increased in the ovaries during vitellogenic stage I. The levels of EcR, E75 and VtG in the hepatopancreas elevated significantly during vitellogenic stages I and II, whereas the levels of RXR elevated only in vitellogenic stage I. During vitellogenic stage III, the levels of RXR, EcR and VtG in the hepatopancreas were significantly decreased. Immunoprecipitation analysis revealed the presence of VtG in the haemolymph, hepatopancreas and ovary extracts from the females but absent in haemolymph and hepatopancreas extract of males. It can be inferred that RXR, EcR and E75 are involved in the regulation of synthesis of VtG in hepatopancreas, whereas in ovary, it is hypothesized that they play an important role in the uptake of VtG from the haemolymph, probably by regulating the levels of vitellogenin receptor. These are the first data showing an association between the expression levels of RXR, EcR and E75 and vitellogenesis and provide an alternative molecular intervention mechanism to the traditional eyestalk ablation to induce vitellogenesis and ovarian maturation in crustaceans.

Hexestrol- and nonylphenol-induced differential vitellogenin synthesis in female and male barfin plaice Liopsetta pinnifasciata

The effects of hexestrol (HXS) and nonylphenol (NP) on plasma vitellogenin (Vtg) concentration in barfin plaice Liopsetta pinnifasciata was studied during spring and autumn experiment. In L. pinnifasciata two “complete” forms of Vtgs, namely VtgAa and VtgAb, were previously described which may be separated due to molecular mass of their largest polypeptide in SDS-PAGE. In spring, the injection of HXS led to an increase in Vtg concentrations in both females and males. SDS-PAGE analysis of plasma from HXS-exposed fish produced only one prominent band at a molecular mass of 180kDa that corresponds to an increase in VtgAb levels. NP injected in fish in spring induced statistically significant increasing of Vtg concentration in males, and only one type of Vtg, as in case of HXS, accumulated in plasma. In autumn, the injection of HXS results to the increase of Vtg concentration in the plasma of females and males, electrophoretic analysis of plasma proteins showed that only a 98kDa polypeptide, corresponding to the VtgAa-type showed a significant increase. The blood plasma ratios of VtgAa and VtgAb in experimental fish are discussed in relation to the season and stage of reproductive cycle.

Long-term effects of binary mixtures of 17α-ethinyl estradiol and dibutyl phthalate in a partial life-cycle test with zebrafish (Danio rerio).

Using 17α-ethinyl estradiol (EE2) and dibutyl phthalate (DBP) as a typical estrogen and phthalate ester, respectively, their combined in vivo effects on zebrafish (Danio rerio) were investigated from the juvenile state to the adult stage. The authors spiked EE2 (5 ng/L and 20 ng/L) and DBP (0.1 mg/L and 0.5 mg/L) either individually or in mixture. At 45 d postfertilization (dpf), the survival rate of zebrafish was comparable in all treatments. Dibutyl phthalate did not induce vitellogenin (VTG) synthesis, and no interaction was found between EE2 and DBP on VTG induction. At 90 dpf, both liver and gill were subject to more severe damage (lipid vacuoles of hepatocytes, amalgamation of gill lamellae, and clubbing at the tips of the secondary lamellae) when coexposed to these 2 chemicals, compared with single exposure. At 115 dpf, generally none of the binary mixture groups showed significantly different growth and sex ratios compared with the corresponding EE2 alone groups. In conclusion, no obvious interactions were detected between EE2 and DBP on the growth, VTG induction, or sex ratio of zebrafish, and they may act independently. However, the influence on morphology of gonad, liver, and gill induced by exposure to the mixture of EE2 and DBP was generally more potent than that by single exposure to EE2 or DBP, indicating the combined long-term harmful effects of EE2 and DBP on the development of zebrafish. Environ Toxicol Chem 2015;34:518-526. © 2014 SETAC.

Juvenile hormone downregulates vitellogenin production in Ectatomma tuberculatum (Hymenoptera: Formicidae) sterile workers.

In the ant Ectatomma tuberculatum (Olivier 1792), workers have active ovaries and lay trophic eggs that are eaten by the queen and larvae. Vitellogenins are the main proteins found in the eggs of insects and are the source of nutrients for the embryo in the fertilized eggs and for adults in the trophic eggs. In social insects, vitellogenin titres vary between castes and affect reproductive social status, nursing, foraging, longevity, somatic maintenance, and immunity. In most insects, vitellogenin synthesis is mainly regulated by juvenile hormone. However, in non-reproductive worker ants, this relationship is poorly characterized. This study determined the effects of juvenile hormone on vitellogenin synthesis in non-reproductive E. tuberculatum workers. Juvenile hormone was topically applied onto workers, and the effect on vitellogenin synthesis in the fat body and vitellogenin titres in the haemolymph were analysed by ELISA and qPCR. Juvenile hormone downregulated protein synthesis and reduced vitellogenin titres in the haemolymph, suggesting that in workers of E. tuberculatum, juvenile hormone loses its gonadotrophic function.

Analysis of proteomic profile changes of zebrafish embryos during exposure to doxorubicin, built-in the phospholipid transport nanosystem

The proteome profile of Danio rerio embryos grown in the medium containing doxorubicin, included in the phospholipid transport nanosystem (doxolip) has been investigated using combination of 1D-electrophoresis with subsequent MALDI-TOF-PMF mass spectrometry. Cultivation of growing of D. rerio embryos in the medium with doxolip caused a substantial increase in expression of the cytoskeletal proteins, a decrease in the number of nuclear proteins involved in DNA and RNA synthesis and disappearance of vitellogenin 2 in comparison with control (the cultivation medium containing the phospholipid transport nanosystem). Analysis of the proteomic profiles of doxolip-treated embryos suggests lower toxicity of doxorubicin incorporated in the phospholipid nanosystem.

In vivo and in vitro inhibitory action of 17β-estradiol and environmental estrogen 4-nonylphenol on gonad-inhibiting hormone (GIH) expression in the eyestalks of Litopenaeus vannamei.

The gonad-inhibiting hormone (GIH) belongs to a neuropeptide family synthesized and released in an X-organ sinus gland complex of crustacean eyestalks. GIH inhibits crustacean ovarian maturation by suppressing vitellogenin (Vtg) synthesis, whereas estrogen is responsible for the stimulation of vitellogenesis (not established). In this study, the effects of 17β-estradiol (E2, 10(-6) M), estrogen receptor antagonist tamoxifen (TAM, 10(-6), 10(-7), and 10(-8) M), and the environmental estrogen nonylphenol (NP, 1 μg/L and 100 μg/L) on LvGIH expression in the eyestalks of shrimp were determined by quantitative real-time PCR. Results showed that LvGIH expression decreased significantly during the L. vannamei ovarian maturation cycle. E2 and NP significantly reduced LvGIH transcripts in vivo, but TAM neutralized the inhibitory action of E2 in a dose-dependent manner (P < 0.05). In addition, the LvGIH expression levels decreased significantly in a time-dependent manner (P < 0.05) when ovary fragments were cultured in vitro with E2. The results of this study suggested that estrogen regulates GIH expression in L. vannamei eyestalks. E2 promoted ovarian development not only by directly upregulating vitellogenesis in the hepatopancreas, but it was also capable of downregulating LvGIH expression, which indirectly resulted in the stimulation of L. vannamei vitellogenesis.

In Memoriam

In Memoriam

Effects of selected xenobiotics on hepatic and plasmatic biomarkers in juveniles of Solea senegalensis

In recent years, Solea senegalensis has increasingly been used in pollution monitoring studies. In order to assess its response to some particular widespread pollutants, juveniles of S. senegalensis were administered an intraperitoneal injection of the model aryl hydrocarbon receptor agonist β-naphtoflavone (βNF) and chemicals of environmental concern, such as the fungicide ketoconazole (KETO), the lipid regulator gemfibrozil (GEM), the surfactant nonylphenol (NP) and the synthetic hormone ethinylestradiol (EE2). Two days after injection, the effect of these chemicals was followed up as alterations of hepatic microsomal activities of the cytochrome P450 (CYPs) and associated reductases, carboxylesterases (CbEs) and the conjugation enzyme uridine diphosphate glucuronyltransferase (UDPGT). In the cytosolic fraction of the liver, the effect on CbEs, glutathione S-transferase (GST) and antioxidant activities was also considered. Alterations on the endocrine reproductive system were evaluated by plasma levels of vitellogenin (VTG) and the sex steroids estradiol (E2), testosterone (T), 11-ketotestosterone (11KT) and the progestin 17α,20β-dihydroxy-4-pregnen-3-one (17,20β-P). Injection with the model compound βNF induced the hydrolysis rate of the seven CYP substrates assayed. The xenobiotic GEM induced three CYP-related activities (e.g. ECOD) and UDPGT, but depressed antioxidant defenses. EE2 induced four CYPs, more significantly ECOD and BFCOD activities. The xenoestrogens NP and EE2 altered the activities of CbE in microsomes and catalase, and were the only treatments that induced de novo VTG synthesis. In addition, the progestin 17,20β-P, was induced in NP-injected fish. None of the treatments caused statistically significant effects on steroid plasma levels. In conclusion, the CYP substrates assayed responded specifically to treatments and juveniles of S. senegalensis appear good candidates for assessing xenobiotics exposure.

Female qualities in males: Vitellogenin synthesis induced by ovary transplants into the male silkworm, Bombyx mori

Female qualities in males are common in vertebrates but have not been extensively reported in insects. Vitellogenin (Vg) is highly expressed in the female fat body and is generally required for the formation of yolk proteins in the insect egg. Vg upregulation is generally regarded as a female quality in female oviparous animals. In this study, we found that Bombyx mori Vg (BmVg) is especially highly expressed in the female pupa. Downregulation of the BmVg gene in the female pupa by RNA interference (RNAi) interfered with egg formation and embryonic development, showing the importance of BmVg in these processes. So, we used BmVg as a biomarker for female qualities in the silkworm. Hematoxylin-eosin staining and immunofluorescence histochemistry showed that ovary transplants induced BmVg synthesis in the male pupa fat body. Ovaries transplanted into male silkworms produced only a few eggs with deformed yolk granules. These results suggested that the amount of BmVg in the male silkworm was insufficient for eggs to undergo complete embryonic development. After 17-beta-estradiol was used to treat male pupae and male pupal fat bodies, BmVg was upregulated in vivo and in vitro. These findings indicated that the male silkworm has innate female qualities that were induced by a transplanted ovary and 17β-estradiol. However, in silkworms, female qualities in males are not as complete as in females.

Molecular analysis of Aedes aegypti classical protein tyrosine phosphatases uncovers an ortholog of mammalian PTP-1B implicated in the control of egg production in mosquitoes.

BackgroundProtein Tyrosine Phosphatases (PTPs) are enzymes that catalyze phosphotyrosine dephosphorylation and modulate cell differentiation, growth and metabolism. In mammals, PTPs play a key role in the modulation of canonical pathways involved in metabolism and immunity. PTP1B is the prototype member of classical PTPs and a major target for treating human diseases, such as cancer, obesity and diabetes. These signaling enzymes are, hence, targets of a wide array of inhibitors. Anautogenous mosquitoes rely on blood meals to lay eggs and are vectors of the most prevalent human diseases. Identifying the mosquito ortholog of PTP1B and determining its involvement in egg production is, therefore, important in the search for a novel and crucial target for vector control.Methodology/Principal FindingsWe conducted an analysis to identify the ortholog of mammalian PTP1B in the Aedes aegypti genome. We identified eight genes coding for classical PTPs. In silico structural and functional analyses of proteins coded by such genes revealed that four of these code for catalytically active enzymes. Among the four genes coding for active PTPs, AAEL001919 exhibits the greatest degree of homology with the mammalian PTP1B. Next, we evaluated the role of this enzyme in egg formation. Blood feeding largely affects AAEL001919 expression, especially in the fat body and ovaries. These tissues are critically involved in the synthesis and storage of vitellogenin, the major yolk protein. Including the classical PTP inhibitor sodium orthovanadate or the PTP substrate DiFMUP in the blood meal decreased vitellogenin synthesis and egg production. Similarly, silencing AAEL001919 using RNA interference (RNAi) assays resulted in 30% suppression of egg production.Conclusions/SignificanceThe data reported herein implicate, for the first time, a gene that codes for a classical PTP in mosquito egg formation. These findings raise the possibility that this class of enzymes may be used as novel targets to block egg formation in mosquitoes.

An integrated approach combining chemical analysis and an in vivo bioassay to assess the estrogenic potency of a municipal solid waste landfill leachate in Qingdao.

Various adverse effects related to landfill leachate have made leachates an important issue in past decades, and it has been demonstrated that landfill leachate is an important source of environmental estrogens. In this study, we employed chemical analysis of some already evaluated estrogenic substances, in combination with a bioassay using several specific biomarkers (e.g., plasma vitellogenin and sex steroids, enzyme activity of gonad gamma-glutamyl transpeptidase, and gonadosomatic index) to evaluate the estrogenic activities in outlets from different stages of the leachate treatment process. The results indicated that 5 environmental estrogens (4-t-octylphenol, bisphenol A, di-ethyl phthalate, di-n-butyl phthalate, and diethylhexyl phthalate) were detected by a gas chromatography-mass spectrometry, and the concentrations in leachate samples were 6153 ng/L, 3642 ng/L, 2139 ng/L, 5900 ng/L, and 9422 ng/L, respectively. Leachate (1∶200 diluted) induced the synthesis of plasma vitellogenin and led to decreased enzyme activity of gonad gamma-glutamyl transpeptidase and gonadosomatic index in male goldfish (Carassius auratus) after a 28-day exposure, while increased circulating 17β-estradiol level was also observed in males exposed to treated effluent. Although the target EEs were partially removed with removal rates varying from 87.2% to 99.77% by the “membrane bioreactor+reverse osmosis+aeration zeolite biofilter” treatment process, the treated effluent is still estrogenic to fish. The method combined chemical techniques with the responses of test organisms allowing us to identify the group of estrogen-like chemicals so that we were able to evaluate the overall estrogenic effects of a complex mixture, avoiding false negative assessments.

Hepatopancreas but not ovary is the site of vitellogenin synthesis in female fresh water crab, Oziothelphusa senex senex

The objective of the present study was to explore the site of synthesis of vitellogenin (Vtg) in fresh water edible crab, Oziothelphusa senex senex. Vtg cDNA fragments were isolated from the hepatopancreas of female crabs using RT-PCR method, and the deduced amino acid sequence of O. senex senex showed more than 60% identity with other brachyuran Vtg sequences. RT-PCR analysis showed that Vtg mRNA can be detected only in hepatopancreas of female Oziothelphusa but not in other tissues including eyestalks, Y-organs, mandibular organs, thoracic ganglion, hypodermis and ovary. Antibodies were raised against vitellin purified from the ovary of O. senex senex. Immunoprecipitation analysis revealed the presence of Vtg in the hepatopancreas of vitellogenic stage I females and in the hemolymph, hepatopancreas and ovary extracts from vitellogenic stage II females but absent in hemolymph and hepatopancreas extract of males. These results suggest that Vtg is synthesized only in hepatopancreas but not in the ovaries of O. senex senex. In addition, Vtg synthesized in hepatopancreas is transported to ovary through hemolymph.

Analysis of proteomic profile changes of Danio rerio embryos during exposure to doxorubicin, incorporated in the phospholipid transport nanosystem

The proteome profile of Danio rerio embryos grown in the medium containing doxorubicin, included in the phospholipid transport nanosystem (doxolip) has been investigated using combination of 1D-electrophoresis with subsequent MALDI-TOF-PMF mass spectrometry. Cultivation of growing of D. rerio embryos in the medium with doxolip caused a substantial increase in expression of the cytoskeletal proteins, a decrease in the number of nuclear proteins involved in DNA and RNA synthesis and disappearance of vitellogenin 2 in comparison with control (the cultivation medium containing the phospholipid transport nanosystem). Analysis of the proteomic profiles of doxolip-treated embryos suggests lower toxicity of doxorubicin incorporated in the phospholipid nanosystem.

AMPK and PTP‐1B stand at the gates of immunity, metabolism and egg formation in mosquitoes. (LB224)

Mosquitoes are major vectors of human diseases. This is mainly due to their requirement for a blood meal to lay eggs which allows the transmission of pathogens to the human being. Our aim is to identify proper molecular targets in order to block blood feeding and impair egg formation.In their early days of adult life mosquitoes feed on vegetal fluids and their midgut is continuously loaded with several bioactive compounds including polyphenols. We observed that polyphenol‐fed mosquitos dramatically increase their average lifespan (Control T 1/2 ± 15‐18 days, Polyphenol‐fed T1/2± 20‐32 days). Polyphenols also reduced the body triglyceride content from 20‐75 %. They trigger midgut autophagy which leads to a 40 % decrease on bacterial loads. Such mechanisms are mimicked by the activation of the mosquito AMPK and abolished by RNAi‐mediated silencing of this gene. Thus the presence of polyphenol on early mosquito meals optimizes mosquito metabolism and immunity and its chances of finding its first blood meal. After a blood meal digestion will take place in mosquito midgut. Blood‐derived nutrients allow yolk synthesis and egg formation. Blood feeding affects the expression of the mosquito PTP‐1B (AaPTP1B) in tissues critically involved in the synthesis and storage of vitellogenin (Vg), the major yolk protein. The protein tyrosine phosphatase inhibitor sodium orthovanadate or the PTP substrate DiFMUP decreased Vg synthesis and egg production. Also, silencing of AaPTP1B using by RNAi assays suppressed by 30 % egg production.In conclusion, the above results show that mosquito signaling pathways regulated by AMPK and PTP1B are proper targets to block blood feeding and egg formation. Future research strategies should be designed to generate novel cell signaling‐based insecticides that simultaneously decrease mosquito lifespan and reproduction.Grant Funding Source: Supported by CNPq, INCT‐EM, FAPERJ

The Reproductive Biology of Shirbot (Barbus grypus Heckel, 1843) in the Maroon River, Iran

Shirbot ( Barbus grypus ) is one of the species in south and southwest of Iran which is greatly favorable to residents of the region. Unfavorable ecological conditions in habitat of this species and overfishing have led to the reduction of the population of shirbot. Therefore, to restore the natural stock of this species, identifying its reproductive cycle associated with its habitat is of a great importance. In this study, the reproductive status of shirbot in the Maroon River in Khuzestan Province was studied in six sampling steps during four seasons. Also, morphological indicators, sex ratio, age of fish, gonadosomatic and hepatosomatic indices, histological changes in the testis and ovary of the fish were studied. The ratio of male fish to female was 2.35 to 1. The maximum value of gonadosomatic index (GSI) is among the specimens aged 3 to 5 years and in March and April. An increase of hepatosomatic index (HIS) during March may indicate the increased activity of liver during vitellogenesis and vitellogenin synthesis which is well verified by histological results of ovarian tissue. Based on our findings we recommend that the maximum reproductive activity of shirbot in the Maroon River starts around the end of March and continues to middle of July.