This report describes the sex differentiation of the Korean rose bitterling, Rhodeus uyekii, from hatching to 170 days post-hatch (DPH) in relation to total length (TL), body weight (BW), and integral water temperature (IWT). The growth curve of TL from just hatching to 83 DPH was 5.144e0.045t (R² = 0.961; t, time), and that of BW was 2.398e0.086t (R² = 0.725). Primordial germ cells (PGCs) were observed at 17 DPH (7.9 mm TL, 3.74 mg BW, 374°C IWT), and thereafter began to protrude into the peritoneal cavity. At 21 DPH (9.2±0.14 mm TL, 4.8±0.07 mg BW, 462°C IWT), some PGCs contained condensed chromatin and oocyte were observed in meiotic prophase. In contrast to the ovaries, which grew gradually after sexual differentiation, testes began multiplying at 25 DPH (10.1 mm TL, 5.42 mg BW, 550°C IWT), when testicular differentiation was first identified, and multiplied continuously thereafter. At 33 DPH (11.2 mm TL, 10.5 mg BW, 726°C IWT), the developing testes contained spermatogonia that exhibited mitotic activity. No spermatocyte or sperm cell was observed until 83 DPH (18.9 TL, 48.2 mg BW, 1,826°C IWT). At 170 DPH (32.5 mm TL, 270.1 mg BW, 3,740°C IWT), which was the end point of this study, the mature ovaries showed germinal vesicle breakdown, while the mature testes contained observable spermatocytes and sperm cells. These results allow us to identify the sex differentiation type of the Korean rose bitterling as differentiated gonochoristic.

Heat shock protein (HSP) 70, the highly conserved stress protein families, plays important roles in protecting cells against heat and other stresses in most animal species. In the present study, we identified and characterized four Hsp70 (RuHSP4, RuHSC70, RuHSP12A, RuGRP78) family proteins based on the expressed sequence tag (EST) analysis of the Korean rose bitterling R. uyekii cDNA library. The deduced RuHSP70 family has high amino acid identities of 72-99% with those of other species. Phylogenetic analysis revealed that RuHsp70 family clustered with fish groups (HSP4, HSC70, HSP12A, GRP78) proteins. Quantitative RT-PCR analysis showed the specific expression patterns of RuHsp70 family members in the early developmental stages and several tissues in Korean rose bitterling. The expression of 4 groups of Hsp70 family was detected in all tested tissue. Particularly, Hsp70 family of Korean rose bitterling is highly expressed in hepatopancreas and sexual gonad (testis and ovary). The expression of Hsp70 family was differentially regulated in accordance with early development stage of Rhodeus uyekii.

Serine-arginine-rich nuclear protein LUC7L plays an important role in the regulation of myogenesis in mice. In the present study, we isolated and characterized the Korean rose bitterling Rhodeus uyekii Luc7l cDNA, designated RuLuc7l. The RuLuc7l cDNA is 1,688 bp long and encodes a 364-amino-acid polypeptide containing serine/arginine-rich region at the C-terminus. The deduced RuLuc7l protein has high amino acid identity (71-97%) with those of other species including human. Phylogenetic analysis revealed that RuLUC7L clustered with fish LUC7L proteins. The expression of RuLuc7l mRNA was high in the brain, kidney, and stomach of Korean rose bitterling. Expression of the RuLuc7l mRNA was detected from 1 day post-fertilization (dpf) and moderately increased until 21 dpf during the early development. Further investigations are required to elucidate the functional role of RuLUC7L in myogenesis in R. uyekii.

We investigated the process of yolk absorption in Korean rose bitterling, Rhodeus uyekii, and determined the changes in its morphometric characteristics. The R. uyekii from 1 days post hatching (DPH) to 21 DPH, the eye diameter (ED) was 5.4 at 5 DPH. Thereafter, the ED/total length (TL) ratio increased to 10.7 at 21 DPH (p<0.05). The yolk length (YL) decreased from 95.4 to 1.1 by 21 DPH, and this rate of decrease was greater than that for any other dimension (p<0.05). 12 morphometric dimensions/TL for the R. uyekii were measured at each sampling day from 21 DPH to 170 DPH. At just hatching, the average TL and BW were 6.1±0.09 mm and 4.9±0.07 mg, respectively. At 53 DPH, the average TL was 12.9±0.28 mm and the average BW was 14.7±0.72 mg; the total length growth equation was TL=5.507e0.038t (R²=0.916). Further, the body weight growth equation was BW=3.3647e0.0296t (R²=0.9354). The dimensions with regard to body depth showed the greatest growth rates in the external characteristics of the fish (p<0.05). The patterns of the morphometric characteristics measured in this study can be classified in three ways. The patterns were shown to be increased (y=0.0992x+ 12.07, R²=0.8333), decreased (y=–0.0569x+42.029, R²=0.8395) or maintained (y=0.005x+18.263, R²=0.3678) from 21 DPH to 170. These results will provide useful indices for the successful rearing of the R. uyekii.