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Meiosis is the key process for producing mature gametes. A natural fertile triploid Carassius auratus population (3nDTCC) and an artificially derived sterile triploid crucian carp (3nCC) have been previously observed, providing suitable model organisms for investigating meiosis characteristics in triploid fish. In the present study, the microstructures and ultrastructures of spermatogenesis were studied in these fishes. TdT-mediated dUTP nick end labeling detection was performed to investigate the apoptosis of spermatocytes. Fluorescence in situ hybridization was employed to trace chromatin pairing. In addition, the mRNA expressions of cell cycle-related genes (i.e., cell division control 2 and cell cycle protein B) were determined by quantitative realtime polymerase chain reaction to illustrate the molecular mechanism of abnormal meiosis in the 3nCC. The results showed that the 3nCC undergoes an irregular prophase I, with the chromosomes distributed in a unipolar radial manner and exhibiting partial pairing, hindered metaphase I, and degenerated cells in the subsequent stages. Meanwhile, the 3nDTCC presented a relatively regular meiotic prophase I with complete conjugate chromosome pairs and chromosomes distributed along the karyotheca, which were presented as a ring structure by slicing. Only the spreads with 130-150 irregular chromosomes can be easily detected in the 3nDTCC, suggesting that it may undergo an abnormal metaphase I. This study provides new insights into the meiosis of fertile and sterile triploid cyprinid fish.

The spermatogenesis of fish remained unclear about the cytological process, especially on the meiotic process of spermatocytes after meiosis I. In the present study, microstructure and ultrastructure of spermatogenesis in Cyprinus carpio (abbreviated CC) were studied, the meiosis process was traced by chromosomal observation and the corresponding immunohistochemical staining of testis where spermatocytes at different stages were distinguished. Results firstly described the telomere-led bouquet configuration of prophase I-stage cells in fish, which ensure the normal chromosomal behavior just like in other eukaryotes. Compared to the canonical meiotic process, the spermatocytes at leptotene, zygotene, pachytene, diplotene and diakinesis stages in CC were normal and also proceeded metaphase I interacting with the meiotic spindle as a unit, while the subsequently meiosis II in CC demonstrated contradicted findings in most organisms, in which no obvious spindle-dependent sister chromatids aligning and segregating could be observed, but some circular condensed nucleus structures. Furthermore, protein antibody of RAD51, DMC1, RNA-Pol II CTD P–S5, H3K36me3 were used respectively as probes, marked initially between the primary spermatocytes at prophase I and metaphase I stage, and between the spermatids and spermatozoa. The systematical study in cytological characteristics of the whole meiotic process in fish spermatogenesis in this paper, provide a standard reference for fish meiosis, which can be of great significance in the practice of fish genetics and breeding.

Polyploidy occurs naturally in fish; however, the appearance of these species is an occasional and gradual process, which makes it difficult to trace the changes in phenotypes, genotypes, and regulation of gene expression. The allotetraploid hybrids (4nAT) of red crucian carp (RCC; ♀) × common carp (CC; ♂) generated from interspecies crossing are a good model to investigate the initial changes after allopolyploidization. In the present study, we focused on the changes in the active sites of the testicular transcriptome of the allotetraploid by localization of RNA Pol II CTD YSPTSPS (phospho S5) using immunofluorescence and RNA-seq data via bioinformatic analysis. The results showed that there was no significant difference in signal counts of the RNA Pol II CTD (S5) between the different types of fish at the same stages, including RCC, CC, 2nF1, and 4nAT, which means that the number of transcriptionally active sites on germ cell chromosomes was not affected by the increase in chromosome number. Similarly, RNA-seq analysis indicated that in the levels of chromosomes and 10-kb regions in the genome, there were no significant changes in the highly active sites in RCC, 2nF1, and 4nAT. These findings suggest that at the beginning of tetraploid origin, the active transcriptome site of 4nAT in the testis was conserved in the regions of the genome compared to that in RCC and 2nF1. In conclusion, 4nAT shared a similar gene expression model in the regions of the genome with RCC and 2nF1 with significantly different expression levels.