Localization of apoptotic and proliferating cells and mRNA expression of caspases and Bcl-2 in gonads of chicken embryos

The aim of the present study was to analyze participation of apoptosis and proliferation in gonadal development in the chicken embryo by: (1) localization of apoptotic (TUNEL) and proliferating (PCNA immunoassay) cells in male and female gonads and (2) examination of mRNA expression (RT-PCR) of caspase-3, caspase-6 and Bcl-2 in the ovary and testis during the second half of embryogenesis and in newly hatched chickens. Apoptotic cells were found in gonads of both sexes. At E18 the percentage of apoptotic cells (the apoptotic index, AI) in the ovarian medulla and the testis was lower (p < 0.05) than in the ovarian cortex. In the ovarian medulla, the AI at E18 was lower (p < 0.05) than on E12. In the testis, the AI was significantly lower (p < 0.05) at E18 than at E15 and 1D. The percentage of proliferating cells (the proliferation index: PI) within the ovary significantly increased from E15 to 1D in the cortex, while proliferating cells in the medulla were detected only at E15. In the testis, the PI gradually increased from E12 to 1D. The mRNA expression of caspase-3 and -6 as well as Bcl-2 was detected in male and female gonads at days 12 (E12), 15 (E15) and 18 (E18) of embryogenesis and the day after hatching (1D). The expression of all analyzed genes on E12 was significantly higher (p < 0.05) in female than in male gonads. This difference was also observed at E15 and E18, but only for the caspase-6. The results obtained showed tissue- and sex-dependent differences in the number of apoptotic and proliferating cells as well as mRNA expression of caspase-3, -6 and Bcl-2 genes in the gonads of chicken embryos. Significant increase in the number of proliferating cells in the ovarian cortex and lack of these cells in the ovarian medulla (stages E12, E18, 1D) simultaneous with decrease in the intensity of apoptosis only in the medulla indicates that proliferation is the dominant process involved in the cortical development, which constitutes the majority of the functional structure of the fully developed ovary. No pronounced changes in the expression of apoptosis-related genes found during embryogenesis suggest that they cannot be considered as important indicators of gonad development. The molecular mechanisms of the regulation of balance between apoptosis and proliferation in developing avian gonads need to be further investigated.     

Fertility preservation through gonadal cryopreservation

Fertility preservation is an area of immense interest in today's society. The most effective and established means of fertility preservation is cryopreservation of gametes (sperm and oocytes) and embryos. Gonadal cryopreservation is yet another means for fertility preservation, especially if the gonadal function is threatened by premature menopause, gonadotoxic cancer treatment, surgical castration, or diseases. It can also aid in the preservation of germplasm of animals that die before attaining sexual maturity. This is especially of significance for valuable, rare, and endangered animals whose population is affected by high neonatal/juvenile mortality because of diseases, poor management practices, or inbreeding depression. Establishing genome resource banks to conserve the genetic status of wild animals will provide a critical interface between ex-situ and in-situ conservation strategies. Cryopreservation of gonads effectively lengthens the genetic lifespan of individuals in a breeding program even after their death and contributes towards germplasm conservation of prized animals. Although the studies on domestic animals are quite promising, there are limitations for developing cryopreservation strategies in wild animals. In this review, we discuss different options for gonadal tissue cryopreservation with respect to humans and to laboratory, domestic, and wild animals. This review also covers recent developments in gonadal tissue cryopreservation and transplantation, providing a systematic view and the advances in the field with the possibility for its application in fertility preservation and for the conservation of germplasm in domestic and wild species.     

Cryopreservation of testis tissues and in vitro spermatogenesis

Cancer treatments, either chemo‐ or radiotherapy, may cause severe damage to gonads which could lead to the infertility of patients. In post‐pubertal male patients, semen cryopreservation is recommended to preserve the potential to have their own biological children in the future; however, it is not applicable to prepubertals. The preservation of testis tissue which contains spermatogonial stem cells (SSCs) but not sperm would be an alternative measure. The tissues or SSCs have to be transplanted back into patients to obtain sperm; however, this procedure remains experimental, invasive, and is accompanied with the potential risk of re‐implantation of cancer cells. Recently, we developed an organ culture system which supports the spermatogenesis of mice up to sperm formation from SSCs. It was also shown that the tissues could be frozen for later sperm production, which resulted in the generation of offspring. Thus, it could be useful as a clinical application for preserving the reproductive potential of male pediatric cancer patients. The establishment of an optimized cryopreservation method and the development of a culture system for human testis tissue are expected in the future.     

Validation of Surveillance,Epidemiology, and End Results TNM staging for testicular germ cell tumor

ObjectivesTo evaluate the accuracy of testicular germ cell tumor category in the Surveillance, Epidemiology, and End Results (SEER) database following the 2010 American Joint Committee of Cancer revision of the TNM staging criteria.MethodsWe performed a retrospective review of our testicular cancer database from January 2010 to July 2011. Registrar extracted data on 76 patients were entered into the Cancer Surveillance Program database from 2 hospitals. We reviewed the SEER coding for each patient, including T, N, M, and S and overall stage group, as well as the range and S value given for tumor markers (lactate dehydrogenase, beta-human chorionic gonadotropin, and α-fetoprotein) both preorchiectomy and postorchiectomy. We then compared these values with the actual staging and tumor markers determined by patient medical record review by a single urologist.ResultsA high proportion of registry records were found to have inaccurate values of category: 71% of S category entries and 34% of N category entries, leading to an overall group stage inaccuracy of 77% in SEER data. Accuracy of overall combined stage group was significantly different between hospitals, with a higher percentage of errors at Hospital A (P< 0.05).ConclusionDespite improvements made to the SEER criteria for extracting data used to code testicular germ cell tumor TNM stage, considerable errors were identified, most notably in tumor marker and nodal status, resulting in an overwhelming number of errors in overall stage. Our findings suggest caution when utilizing SEER data for review of patients with testicular cancer and their staging.     

绿麦隆和阿特拉津联合染毒对小鼠睾丸的影响

目的 研究绿麦隆和阿特拉津单体系及联合作用对小鼠睾丸形态及结构的影响.方法 将昆明种小鼠按灌胃农药和剂量随机分为20组[1个对照组,4个绿麦隆染毒组(321.5、1250、2500、5000mg/kg),3个阿特拉津染毒组(218.75、875、1750mg/kg),12个联合染毒组],每组10只.连续25 d经口灌胃,每天1次,每次1ml.并用光学显微镜、电子显微镜对小鼠睾丸病理学变化进行观察.结果 农药单体系及联合染毒体系各剂量组对小鼠睾丸均有不同程度的损伤.与对照组比较,光学显微镜观察所见为各剂量组出现不同程度生精上皮细胞排列疏松、紊乱,生精细胞脱落,层次减少,病变严重;电子显微镜观察为各剂量组出现不同程度生精细胞线粒体呈空泡样改变,核膜肿胀、弯曲,支持细胞功能低下随染毒剂量增加,上述病理学变化有加重趋势,联合染毒组病理变化比单体系染毒组更明显.结论 绿麦隆和阿特拉津对小鼠睾丸的毒性与染毒剂量相关,联合作用体系加重了对小鼠睾丸的毒性效应.     

促性腺激素释放激素拮抗剂对大鼠睾丸支持细胞雄激素受体和Fas配体基因表达的影响

本实验利用注射促性腺激素释放激素拮抗剂（ＧｎＲＨ Ａ）引起睾丸生精细胞程序化死亡的大鼠模型，用地高辛标记的大鼠雄激素受体和Ｆａｓ配体的ＲＮＡ探针对大鼠睾丸石蜡切片进行原位杂交，观察雄激素受体和Ｆａｓ配体基因在生精细胞凋亡中的变化。结果显示，ＧｎＲＨ Ａ处理后曲细精管有退行性变，原位杂交表明此时Ｓｅｒｔｏｌｉ细胞的雄激素受体表达明显减低，而Ｆａｓ配体表达显著升高。提示Ｆａｓ配体的表达可能与生精上皮细胞程序化死亡关系密切。     

A Case Report of Testicular Sparganosis Misdiagnosed as Testicular Tumor

Sparganosis is a parasitic infestation of human by plerocercoid larvae. Sparganum is usually reported to be found in the subcutaneous tissues as well as other organs, including scrotum. However, testicular sparganosis is extremely rare, because of strong capsule of tunica albuginea. An urban-living 54-yr-old Korean man presented with left scrotal pain for 6 yr. Both testes look normal physically. Ultrasonography revealed poorly defined, heterogeneous mass with increased echogenicity in the left testis. This case was misdiagnosed as testicular tumor and underwent orchiectomy, but was diagnosed as testicular sparganosis by histopathology. Sparganosis should be included for differential diagnosis of testis tumor in countries where sparganosis is prevalent.

Graphical Abstract

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Division of the genitofemoral nerve in normal scrotal testicular rats

The role of the genitofemoral nerve (GFN) on testicular descent has been clearly shown. It has also been suggested that in unilateral cryptorchid rats, after division of the ipsilateral GFN fertility rates are higher, i.e., transection of the GFN prevents contralateral testicular damage, but the mechanism is unclear. The purpose of this study was to investigate the effect of dividing the GFN on the normal scrotal testes. Thirty male Wistar albino rats were divided into three groups: group A, transection of right GFN; group B, bilateral transection of the GFN; and group C, sham operations, all at the age of 30 days. The animals were killed at 90 days of age and the testes were removed. Each excised testis was weighed and fixed for histological studies. Mean seminiferous tubular diameter was measured and germinal epithelium maturity was determined using the modified Johnsen testicular-biopsy score. In all groups, all three parameters were similar, suggesting that division of the GFN had no effect on normal testes. Accepted: 12 January 2000