Institute for Veterinary Anatomy, Histology und Embryology, Justus Liebig University, Giessen, Germany.
Institute for Veterinary Anatomy, Histology und Embryology, Justus Liebig University, Giessen, Germany.
Fertil Steril. 2017 Jul;108(1):47-54. doi: 10.1016/j.fertnstert.2017.05.019. Epub 2017 Jun 7.
To define the precise cellular localization of ciliated bronchial epithelium 1 (CBE1) in the human testis and test its relationship to impaired spermatogenesis.
Gene expression analysis, and histologic and immunohistochemical evaluation.
University research laboratories and andrologic outpatient clinic.
PATIENT(S): Forty-three human testicular biopsies: 12 biopsies showing normal spermatogenesis (NSP), 8 with maturation arrest at level of spermatocytes (STA), 8 with maturation arrest at level of spermatids (SDA), 4 with scattered elongating spermatids, and 12 with Sertoli cell-only syndrome, with an additional 5 semen samples from healthy donors.
INTERVENTION(S): None.
MAIN OUTCOME MEASURE(S): Evaluation of CBE1 expression in normal as well as impaired spermatogenesis on mRNA (quantitative reverse-transcription polymerase chain reaction and in situ hybridization) and protein level (immunohistochemistry, Western blot analysis).
RESULT(S): In normal spermatogenesis, CBE1 mRNA was expressed in late pachytene spermatocytes, and the protein was localized within the flagellum of elongating spermatids from stage V up to the spermiation in stage II. Immunoelectron microscopy showed CBE1 clearly associated with microtubules at the manchette, the head-tail coupling apparatus, and the flagellum, but the protein was absent in spermatozoa. Compared with normal spermatogenesis, CBE1 mRNA was statistically significantly reduced in samples with a maturation arrest at the level of round spermatids and primary spermatocytes, and was absent in samples showing Sertoli cell-only syndrome. CBE1 protein was completely missing in SDA samples showing few elongating spermatids.
CONCLUSION(S): Our data strongly suggest an influence of CBE1 in ciliogenesis in spermatids due to the localization at the microtubules of the elongating spermatids, indicating a role in the intramanchette and/or intraflagellar transport mechanism. The absence of CBE1 in spermatozoa suggests that CBE1 is important for the spermatid development but not for the maintenance of mature spermatozoa as a component of the flagellum.
确定人类睾丸中纤毛支气管上皮 1（CBE1）的精确细胞定位，并研究其与精子发生障碍的关系。
基因表达分析以及组织学和免疫组织化学评估。
大学研究实验室和男科门诊。
43 个人类睾丸活检样本：12 个活检样本显示正常精子发生（NSP），8 个样本在精母细胞水平出现成熟阻滞（STA），8 个样本在精细胞水平出现成熟阻滞（SDA），4 个样本出现散在的伸长精子，12 个样本为唯支持细胞综合征，另有 5 个来自健康供体的精液样本。
无。
在正常和精子发生障碍的情况下，通过 mRNA（定量逆转录聚合酶链反应和原位杂交）和蛋白质水平（免疫组织化学、Western blot 分析）评估 CBE1 的表达。
在正常精子发生中，CBE1 mRNA 表达于晚期粗线期精母细胞，蛋白质定位于 V 期至 II 期精子发生的伸长精子的鞭毛内。免疫电子显微镜显示 CBE1 与中心粒周围物质、头部-尾部连接装置和鞭毛中的微管明显相关，但在精子中不存在。与正常精子发生相比，在圆形精子和初级精母细胞水平出现成熟阻滞的样本中，CBE1 mRNA 统计学上显著降低，在仅表现为支持细胞综合征的样本中则不存在。在 SDA 样本中几乎没有伸长精子，CBE1 蛋白完全缺失。
我们的数据强烈表明 CBE1 影响精子细胞中的纤毛发生，因为它定位于伸长精子的微管上，表明其在中心粒周围物质和/或鞭毛内运输机制中发挥作用。精子中不存在 CBE1 表明 CBE1 对精子细胞的发育很重要，但对成熟精子的维持不重要，因为它是鞭毛的组成部分。
