A 3-month-old American Cocker spaniel was presented at the Veterinary Teaching Hospital, Chungbuk National University, for examination of urinary tract after dissection of vaginal mass at local clinic before 10 days. Clinical examination of the affected bitch revealed a normal sized vulva in a normal anatomical position with a grossly enlarged clitoris, which contained an os clitoris. On examinations of the genital gland, there were testis, epididymis, ductus deferens and uterus. The histology of both gonads was primarily testis. Seminiferous tubules were divided into many parts by fibrous connective tissue. A small number of spermatogonia was present, but large numbers of Leydig's cells were existed. A normal female karyotype (78, XX) was detected in metaphase spreads obtained from cultured peripheral lymphocytes. Y chromosome specific sequences were not detected in genomic DNA by PCR. After 27 months, the os clitoris was larger than 3-month-old dog and os bone was more calcified than young age. Combining the results of cytogenetic, molecular genetic and histological examinations, the dog was diagnosed as a female hermaphrodite with Sry-negative XX sex reversal.

We previously reported that DNA hypermethylation of SRY promoter is associated with emergence of male-to-female sex reversal. The normality of offspring is achieved by relatively complete and correct nuclear reprogramming during somatic cell nuclear transfer and cloning process. The purpose of this study is to determine whether DNA demethylation of SRY promoter induced by 5-aza-2'-deoxycytidine (AzC) DNA methylation inhibitor may get back phenotypic XY sex reversal female to normal male in SCNT cloning. Canine femoral skin fibroblast cells were established from SCNT-cloned XY sex reversed female (GSF335). Using bisulfite genomic sequencing analysis, DNA methylation levels of SRY promoter in non-treated (normal) and 1uM AzC-treated cells were 88.4% and 55.3% in treatment for 4 days respectively. Seven SCNT-cloned puppies were cloned using the AzC-treated cells as donor cell. Six of those clones showed phenotypically normal male, through one puppy (GSF451) was only observed into male-to-female sex reversal with female genitalia. In umbilical cord tissue, DNA methylation levels on SRY promoter of GSF451 clone and the other clones were 79.2% and 5.7% to 62.2% respectively, which was approximately similar to those of non-treated (normal) and AzC-treated cells. Also, cloned puppies originated from AzC-treated cells implied significantly multiple body weight and height compared to age-matched SCNT-cloned control, which may be underlying in size-effect of AzC-treatment. Our findings suggest that DNA demethylated status of SRY promoter induced by AzC is likely to facilitate normal development including sex differentiation through epigenetic alteration of donor cells.

This study attempts to propose the possibility of the sex reversal in Sinonovacula constricta and Gomphina veneriformis by confirming the changes in the sex ratio with the shell length in the same population level. For analysis of sex ratio, 100 individuals of S. constricta (SL 26.5-95.0 mm) and 2385 individuals of G. veneriformis (SL 15.1-60.1 mm) were used. Sex was analyzed histologically. Both species displayed the tendency of increase in the female proportion with increase in shell length. In this study, changes in the sex ratio in accordance with the growth of S. constricta and G. veneriformis are determined to be indirect evidence that signifies their sex reversal.

To understand the sex reversal characteristics in the longtooth grouper (Epinephelus bruneus), this study examined the sex reversal and artificial masculinization of wild caught E. bruneus reared in indoor rearing tank after a 17α-methyltestosterone injection. To domesticate a broodstock, 64 wild caught E. bruneus, between 47.0 to 110.0 cm in total length and from 1.5 to 21.4 kg in body weight, were reared in indoor rearing tank (4.0 to 5.0 m wide, and 2.5 to 3.0 m depth) for four years. Seven specimens showed sex reversal from female to male during indoor rearing condition, whose total length and body weights were from 63.0 to 99.0 cm and from 4.4 to 13.2 kg, respectively. After inducing artificial masculinization in 14 female E. bruneus with a 17α-methyltestosterone (2.0 mg/kg BW) implants for 3 years, spermiation occurred in 9 specimens (total length: 54.0 to 68.0 cm, body weight: 2.3 to 4.3 kg). Among the female to male sex reversals, two specimens returned back to being female, whose body weights were 2.8 kg (initially 2.6 kg) and 2.7 kg (initially 2.3 kg). Therefore, this study suggested that E. bruneus (> 3.0 kg) was more effective in masculinizing by 17α-methyltestosterone implants.

The sex ratio (F:M) in the same population of oyster, Crassostrea gigas at the commencement of the study (2007) was 1:1.0, but changed to 1:2.8 by the end of the study (2008). The sex reversal rate in two-year-old oysters was 40.2%. Specifically, female to male sex reversal rate was 66.1%, which is higher than the male to female sex reversal rate of 21.1%. The sex reversal pattern of C. gigas appears to go from male female male, and as such is determined to be rhythmical hermaphroditism.

This study attempts to propose the possibility of the sex reversal in Tegillarca granosa and Ruditapes philippinarum by confirming the changes in the sex ratio with the shell length (SL) in the same population level. For analysis of sex ratio with SL, 1500 individuals of T. granosa (SL 10.1-45.0 mm) and 712 individuals of R. philippinarum (SL 15.1-70.0 mm) were used. Sex was analyzed histologically. The average sex ratios (F:M) of T. granosa and R. philippinarum were 1:1.22 and 1:0.96, respectively. However, sex ratio was found to differ when the clams were divided into groups according to SL in 5.0 mm intervals. Both species displayed the tendency of increase in the proportion of female with increase in SL. In this study, changes in the sex ratio in accordance with the growth of T. granosa and R. philippinarum are determined to be indirect evidence that signifies their sex reversal.

대마의 육종 및 재종상의 큰 장애인 "자웅이주성"을 극복할 수 있는 "자성종자" 생산기술을 개발하기 위해 수행한 일련의 연구결과를 요약하면 다음과 같다. 1. 전 생육기간 자연 단일조건에 재배한 개체들은 질산은을 처리하더라도 성전환 식물체 비율이 5%이고 웅화의 발생정도가 극히 적었으며 자화보다 24일 늦게 개화하여 수정이 되지 않았다. 2. 재배환경을 3엽기, 6엽기에 장일에서 단일조건으로 급격하게 변화시킨 자주는 웅화의 개화량은 적었지만 성전환 개체 비율이 각각 75%, 100%로 높고 자화와 개화기 차이가 각각 10일, 3일로 비슷한 시기에 개화하여 인공수정이 가능하였다. 3. 영양생장기로 판단되는 7월 2일과 7월 12일에 질산은과 단일을 동시에 처리한 개체들은 모두 성이 전환되어 성전환 액아 비율이 각각 63.2%, 61.9%로 높았고 무처리 자주와 동일한 시기에 개화하여 자연적인 풍매로 수정되었다. 또한 상위 제 1절에서 성전환 액아의 비율이 각각 80.5%, 77.0%로 가장 높아 풍매에 유리하도록 바람이 잘 통하는 식물체 상부에서 성전환 액아의 비율이 높았다. 4. 생식생장기로 판단되는 7월 22일에 처리한 개체들은 성전환 액아의 비율이 4.4%로 낮고 자화보다 16일 늦게 개화하여 수정되지 않았다. 5. 일반종자는 자주 비율이 49%로 성비가 1:1에 근접하기 때문에 종실수량이 96 kg/10a였으나 자성종자는 자주 비율이 100%로서 종실수량이 141 kg/10a로 일반 종자에 비해 47% 증수되었다.

We investigated the androgenic effects of 11-ketotestosterone (11-KT) on gonadal sex reversal and spermatogenesis in honeycomb grouper Epinephelus merra by method of gonadal biopsy. 11-KT was injected intramuscularly at a concentration of 1 and body weight. The proportion of cross sectional area of the gonad occupied by each germ cell type was measured and compared pre- and post-injection group. During the sex change phase, the distribution ratio of oocytes was decreased in all fish of 11-KT treatment group while the distribution ratio of spermatocytes was increased than pre-injection group. In male phase, all fish of 11-KT treatment group shown the increased distribution ratio of spermatocytes, but the distribution ratio of spermatozoa was decreased than pre-injection group. The present results suggest that 11-KT can stimulate degeneration of oocytes, proliferation of spermatocytes and spermiation in honeycomb grouper.

Eleven wrasse species inhabit the coastal waters of Jeju Island, Korea. They are the target of leisure fishing and are considered good eating. We investigated the distribution of standard length (SL) by sex of wrasse in Jeju coastal waters for Halichores poecilopterus, H. tenuispinis, Pseudolabrus japonicus, and Pteragogus flagellifera. A cross-section of the ovary showed the ovarian cavity and ovarian lamellae containing oocytes. A cross-section of the testis showed many lobules containing spermatogonia and spermatocytes. A cross-section of a gonad undergoing sex reversal showed the regression or reduction of oocytes and some spermatocytes located in the ovarian lamellae. A cross-section of a sex-reversed testis showed the primary structure of the ovary, with spermatocytes distributed in the epithelium of the lamellae, and reformed seminiferous ducts in the basement lamellae. (중략)