This study carried out to examine the association between the haplotypes of mitochondrial control region (CR) and growth traits of two F1 progeny populations of the Red-spotted grouper. We found polymorphic patterns of the 133-bp repeat units in the CR. In the BBR01 population (70-days after hatching), a total of 1,091 F1 progeny were divided into three haplotypes (H01, H03 and H04). The significant differences were found in the levels of BL, BW and LWI (p<0.05). The F1 animals with the haplotype H01 had greater level of BL (50.815±4.586 mm) than those of H03 (47.270±6.486 mm) and H04 (47.179±6.278 mm). The H01 F1 fishes were heavier level of BW (2.270±0.559 g) than those of H03 (1.789±0.711 g) and H04 (1.797±0.706 g). In the BBR02 population (11-months after fertilization), three haplotypes H03, H04 and H05 were detected. The significant difference was found only in the BL values among three haplotypes (p<0.05). The F1 animals with the haplotype H03 had greater level of BL (19.22±2.000 cm) than those of H04 (18.64±1.964 cm) and H05 (18.86±1.512 cm). There were no significant differences in BW and LWI among haplotypes in the BBR02 population (p>0.05). These results suggested that the mitochondrial haplotypes may affect the growth traits during early developmental stage of the Red-spotted grouper. The marker-assisted selection system for broodstock animals may be helpful in improving performance traits for aquaculture of the Red-spotted grouper.

To characterize the female or male transcriptome of the Pacific abalone and further increase genomic resources, we sequenced the mRNA of full-length complementary DNA (cDNA) libraries derived from pooled tissues of female and male Haliotis discus hannai by employing the Iso-Seq protocol of the PacBio RSII platform. We successfully assembled whole full-length cDNA sequences and constructed a transcriptome database that included isoform information. After clustering, a total of 15,110 and 12,145 genes that coded for proteins were identified in female and male abalones, respectively. A total of 13,057 putative orthologs were retained from each transcriptome in abalones. Overall Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analyzed in each database showed a similar composition between sexes. In addition, a total of 519 and 391 isoforms were genome-widely identified with at least two isoforms from female and male transcriptome databases. We found that the number of isoforms and their alternatively spliced patterns are variable and sex-dependent. This information represents the first significant contribution to sex-preferential genomic resources of the Pacific abalone. The availability of whole female and male transcriptome database and their isoform information will be useful to improve our understanding of molecular responses and also for the analysis of population dynamics in the Pacific abalone.

In order to examine the effects of four different light spectra (i.e., white, red, green, blue) on the oocyte maturation in grass puffer, the change of reproductive parameters via brain-pituitary-gonad (BPG) axis were investigated in this study. After exposure to four different light spectra for 7 weeks, the abundance of gonadotropin-releasing hormone (GnRH) mRNA which is a type of seabream (sbGnRH) and two different of subunit of gonadotropin hormones mRNAs, follicle-stimulating hormone (fshβ) mRNA and luteinizing hormone (lhβ) mRNA, were analyzed in the brain and pituitary. Gonadosomatic index (GSI) and oocyte developmental stage were also investigated in the gonad based on anatomical- and histological observations. GSI value was significantly increased in the green spectrum-exposed group when compared to that of the other light-exposed groups (i.e., white, red, blue), with the presence of yolk stage oocytes (p˂0.05). The abundances of sbGnRHmRNA and fshβ mRNA in the green spectrum-exposed group were also significant higher than those of the other light spectra-exposed groups(p˂0.05). However, there was no significant difference between the abundances of lhβmRNA in all light spectra-exposed groups. These results indicate that the maturation of oocyte in grass puffercan be accelerated by exposure to the spectrum of green. The sbGnRHmRNA and fshβ mRNA may play an important reproductive parameters role in the initiation of maturation of oocyte. To better understand the molecular mechanism for the maturation of oocytein grass puffer, further study examining the relationship between oocyte development and its related genes (e.g., sbGnRHmRNA and fshβmRNA) is required.

Cryopreservation is an effective method for the long-term storage of fish sperm. However, because of a lack of methods for cryopreserving fish eggs and embryos, a technical challenge remains in preserving maternally inherited cytoplasmic compartments. Here, we aimed to generate functional eggs and sperm in sterile rainbow trout hatchlings by transplanting cryopreserved ovarian germ cells into the peritoneal cavity. Immature ovaries were isolated from 9-month-old transgenic rainbow trout, whose ovarian germ cells expressed green fluorescent protein (GFP), and cryopreservation was optimized by testing several different cryoprotective agents. Dominant orange-colored, pvasa-Gfp transgenic rainbow trout females served as donors, and wild-type triploid rainbow trout served as germ cell recipients. Viability and transplantation efficiencies of frozen ovarian germ cells were stable for up to 1,185 days. Ovarian germ cells frozen for 8months efficiently differentiated into eggs and sperm in the ovaries and testes of recipient fish. Inseminating the resultant eggs and sperm generated viable offspring displaying the donor phenotypes (orange body color, green fluorescence, XX ovaries, and matching karyotype). This study represents the first success in generating functional gametes from cryopreserved female germ cells in any fish species, which should facilitate the conservation of protogynous fishes, female heterogametic fish, and currently endangered fish.

멍게는 해양무척추동물로 유생 단계에 꼬리 근육 및 등쪽에 신경관을 갖는 등 기본적인 척삭동물의 형태를 나타낸다. 유생은 유영생활을 하다 암반 등에 부착하고 변태를 한다. 변태 과정에서 몸의 아래쪽에 심장이 형성되며, 개방혈관계를 갖는다. 심장은 주기적으로 혈류의 방향을 바꾸는 것으로 알려져 있으나 ,어떠한 방식으로 박동 방향이 역전되는지, 그 주기는 어떻게 되는지, 종에 따른 차이가 있는지 등 아직 알려지지 않은 것들이 산적해 있다. 본 연구에서는 멍게(우렁쉥이, Halocynthia roretzi)를 이용하여 이와 관련된 현상을 연구하였다. 변태 후 약 10일부터 심장 박동이 관찰되었으며, 초기 단계부터 박동과 정지, 재박동이 일어났다. 심장의 박동 횟수는 9도에서 평균 3.05초 간격으로 80회 이상 지속되었다. 이후 10초 이상의 심장 박동의 정지가 있은 후 다시 재박동이 일어났다. 재박 동 때의 혈류의 방향은 언제나 반대로 일어났다. 심장의 박동은 9도보다 13도에서 빨랐고, 유치자의 연령이 증가하면서 느려지는 경향이 관찰되었다. 멍게의 심장 형성 과정은 척추동물 심장 형성의 기본적인 모습을 하고 있어, 인간의 심장 형성 연구 및 기능 이상을 연구하는데 있어 중요한 실험재료라 할 수 있다. 앞으로 심장이 주기적으로 박동과 정지를 반복하며, 혈류의 방향이 역전되는 메커니즘에 대한 분자생물학적인 연구가 필요하다.

BACKGROUND Ca2+ oscillations during fertilization induce eggs activation and embryonic development in mammalian eggs.. The type 1 inositol 1,4,5-trisphosphate receptor (IP3R1) is in charge of Ca2+ oscillations for the release of stored Ca2+ from the endoplasmic reticulum. The capacity of this oscillation is obtained during egg maturation and corresponds with an increase in the sensitivity of the IP3R1 and their localization in cytoplasm. Cluster formation of IP3R1 in the egg cortex is important to initiation of Ca2+ oscillations during egg and sperm fusion. In this study, we investigated that cell cycle–coupled redistribution of IP3R1 and Ca2+- oscillatory activity in mouse zygotes. MATERIALS AND METHODS Metaphase II arrested eggs were collected from ICR female mouse after super ovulation induction. At 14 hr post hCG, MII eggs were collected, and artificially activated in Ca2+ free CZB medium with 10 mM SrCl2 for 2 hrs. Pronuclear zygotes (PN) were collected from Strontium activated eggs at 8 hr post activation, and the first mitotic eggs were collected at 16~17 hr post activation. To identify cell cycle coupled IP3R1 redistribution, MII eggs, zygotes, and first mitotic eggs were collected, and fixed for immunostaining with anti-IP3R1antibody (CT-1) and observed on CLSM. Ca2+-oscillatory activity was monitored with fluorescence microscope mounted SimplePCI program (Hamamatsu) after injection of cRNA of mouse phospholipase C zeta (mPLCZ). RESULT IP3R1 were shown clusters, 1~2 um in diameter, in cortex of ovulated MII eggs with high Ca2+ oscillatory activity by mPLCZ injection. These eggs represent more than 6 spikes per 60 min. However, IP3R1 clusters were disappeared in PN eggs and these eggs showed very low Ca2+- oscillatory activity by mPLCZ. In mitosis I stage eggs, clusters of IP3R1 were appeared and Ca2+-oscillatory activity was reactivated slightly (2 spikes per 60 min). CONCLUSIOINS This study introduced the redistribution of IP3R1 clusters were occurred in egg activation according to cell cycle dependent manner. Also, functional modification of IP3R1 including protein phosphorylation was associated with cortical clustering of IP3R1 in cell cycle coupled Ca2+ oscillatory activity.

Spermatogonial stem cells (SSCs; also known as Asingle [As] spermatogonia in mice) divide to self-renew or to produce progenitor cells known as Apaired(Apr) spermatogonia in basal compartment of seminiferous tubules of mammalian testis. These characterized cells are the finally differentiated product of a developmental process referred to as “spermatogenesis.” In the development of SSCs it is critical to maintain a balance between self-renewal and differentiation. because an excess of either process will lead to infertility. these two processes are tightly controlled by intrinsic signals of SSCs and extrinsic signals from the microenvironment, known as the SSC niche. The SSC niche is formed by Sertoli cells, the only somatic cells found inside the seminiferous tubules. The WNT/β-catenin pathway is known to regulate Sertoli cell functions critical to their capacity to support spermatogenesis in the postnatal testis, but The mechanisms and factors of the pathway are not well known. We found a factor TLE3 (Transducin Like Enhancer Of Split 3). The transcriptional co-repressor TLE family is known to function as transcription co-repressors within the context of Wnt signaling by interacting with histone deacetylase HDAC2. We examined the expression level of TLE3 in various mouse tissues. As a result of RT-PCR, TLE3 showed significantly higher expression in testis than that in other tissues. Immunofluorescent analysis revealed that TLE3 and HDAC2 expression are differentially regulated in the mouse testis during postnatal development. In adult testis, TLE3 and HDAC2 were co-expressed in Sertoli cells. TLE3 and HDAC2 protein are also located in nucleus in mouse TM4 Sertoli cells. Taken together, TLE3 may play a role in regulating WNT/β-catenin pathway via interaction with HDAC2 in Sertoli cell. Futher studies are needed to look into factors that regulated by siTLE3 in Sertoli cell and interated with TLE3 in WNT/β-catenin pathway.