Disruption of androgen signaling is known to cause testicular malformation and defective spermatogenesis in zebrafish. However, knockout of cyp17a1, a key enzyme responsible for the androgen synthesis, in ar-/- male zebrafish paradoxically causes testicular hypertrophy and enhanced spermatogenesis. Because Cyp17a1 plays key roles in hydroxylation of pregnenolone and progesterone (P4), and converts 17α-hydroxypregnenolone to dehydroepiandrosterone and 17α-hydroxyprogesterone to androstenedione, we hypothesize that the unexpected phenotype in cyp17a1-/-;androgen receptor (ar)-/- zebrafish may be mediated through an augmentation of progestin/nuclear progestin receptor (nPgr) signaling. In support of this hypothesis, we show that knockout of cyp17a1 leads to accumulation of 17α,20β-dihydroxy-4-pregnen-3-one (DHP) and P4. Further, administration of progestin, a synthetic DHP mimetic, is sufficient to rescue testicular development and spermatogenesis in ar-/- zebrafish, whereas knockout of npgr abolishes the rescue effect of cyp17a1-/- in the cyp17a1-/-;ar-/- double mutant. Analyses of the transcriptomes among the mutants with defective testicular organization and spermatogenesis (ar-/-, ar-/-;npgr-/- and cyp17a-/-;ar-/-;npgr-/-), those with normal phenotype (control and cyp17a1-/-), and rescued phenotype (cyp17a1-/-;ar-/-) reveal a common link between a downregulated expression of insl3 and its related downstream genes in cyp17a-/-;ar-/-;npgr-/- zebrafish. Taken together, our data suggest that genetic or pharmacological augmentation of the progestin/nPgr pathway is sufficient to restore testis organization and spermatogenesis in zebrafish with the depletion of androgen signaling.

Since 2011, ecological operation trials of the Three Gorges Reservoir (TGR) have been continuously conducted to improve the spawning quantity of the four major Chinese carp species below the Gezhouba Dam. In particular, exploring the effects of short-term water velocity stimulation on ovarian development in grass carp (Ctenopharyngodon idellus) is essential to understand the response of natural reproduction to ecological flows. We performed ovary histology analysis and biochemical assays among individuals with or without stimulation by running water. Although there were no obvious effects on the ovarian development characteristics of grass carp under short-term water velocity stimulation, estradiol, progesterone, follicle-stimulating hormone (FSH), and triiodothyronine (T3) concentrations were elevated. Then, we further explored the ovarian development of grass carp under short-term water velocity stimulation by RNA sequencing of ovarian tissues. In total, 221 and 741 genes were up- or downregulated under short-term water velocity stimulation, respectively, compared to the control group. The majority of differentially expressed genes (DEGs) were enriched in pathways including ABC transporters, cytokine-cytokine receptor interaction, ECM-receptor interaction, and steroid hormone biosynthesis. Important genes including gpr4, vtg1, C-type lectin, hsd17b1, cyp19a1a, cyp17a1, and rdh12 that are involved in ovarian development were regulated. Our results provide new insights and reveal potential regulatory genes and pathways involved in the ovarian development of grass carp under short-term water velocity stimulation, which may be beneficial when devising further ecological regulation strategies.

Ecological operation of hydraulic engineering is essential for the conservation of fishery resources. Water velocity is known to affect the spawning of fishes delivering drifting eggs. This study aims to explore the effects of water velocity stimulation on the ovarian maturation and antioxidant capacity of adult grass carp (Ctenopharyngodon idellus) through laboratory experiments in order to understand the physiological mechanism underlying the response of natural reproduction to ecological flows. We examined the histology, sex hormones and vitellogenin (VTG) concentrations of ovary, and the transcripts of key genes in the hypothalamus-pituitary-gonad (HPG) axis, as well as the antioxidant activities of ovary and liver in grass carp. The results showed that although there was no discernible difference on the ovarian development characteristics of grass carp under water velocity stimulation, estradiol, testosterone, progesterone, 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DHP), and VTG concentrations were elevated, which was related to the transcriptional regulation of the HPG axis genes. The gene expression levels (gnrh2, fshβ, lhβ, cgα, hsd20b, hsd17b3, and vtg) in the HPG axis were significantly elevated under water velocity stimulation, while those of hsd3b1, cyp17a1, cyp19a1a, hsd17b1, star, and igf3 were suppressed. In addition, appropriate water velocity stimulation could enhance body health status by increasing the activities of antioxidant enzymes in the ovary and liver. The results of this study provide the fundamental knowledge and data support for ecological operation of hydropower projects and river ecological restoration.

Understanding the early life history of the endangered largemouth bronze gudgeon (C. guichenoti) is crucial for optimizing artificial rearing and conservation strategies. To provide a unified scientific basis for these efforts, this study characterized the ontogeny of C. guichenoti larvae from hatching to 30 days post-hatch (dph) by integrating analyses of allometric growth, digestive histology, and enzyme activities. Morphometric analysis revealed that total length increased exponentially with age. Positive allometric growth was observed in swimming organs (pectoral, dorsal, and caudal fins) and eye diameter, indicating a priority in developing locomotion and sensory capabilities for survival in lotic environments. Histological examination showed that the digestive tract became patent by 4 dph, and functional structures were established by 5 dph, coinciding with yolk sac absorption and the initiation of exogenous feeding. The digestive system reached near-adult maturity by 20 dph. Enzyme activity profiles showed that trypsin activity increased continuously, peaking at 30 dph, while amylase and alkaline phosphatase peaked at 20 dph. Lipase activity remained low until a slight increase at 30 dph. These results indicate that C. guichenoti larvae adopt a “locomotion-first” strategy, with the digestive system becoming functional at 5 dph and maturing by 20 dph. These findings collectively define the critical windows for artificial propagation, specifically guiding the timing of initial feeding and feed formulation.