Immunohistochemical study of seminiferous epithelium in adult bovine testis using monoclonal antibodies against Ki-67 protein and proliferating cell nuclear antigen (PCNA)
Immunohistochemical study of seminiferous epithelium in adult bovine testis using monoclonal antibodies against Ki-67 protein and proliferating cell nuclear antigen (PCNA)
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DOI：
10.1007/s004410050529
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发表时间：
1996-01
影响因子：
3.6
通讯作者：
K. Wrobel;D. Bickel;R. Kujat
中科院分区：
文献类型：
--
作者：
K. Wrobel;D. Bickel;R. Kujat
应用单克隆抗体免疫组织化学方法研究了增殖细胞核抗原（PCNA）和Ki-67蛋白在成年牛生精上皮中的分布。为两种增殖标记物中的每一种定制方法学方案是在管状切片和整体封片中获得最佳结果的必要先决条件。A-，I-和B-精原细胞显示PCNA阳性细胞核，除了在Meta，有丝分裂的ana和末期。PCNA阴性的细胞核在基底管室，不包括那些从非循环支持细胞，属于精原细胞前体细胞系。然而，在暴露于针对Ki-67蛋白的MIB-1抗体后，分别只有约30%、45%和47%的A-、I-、B-精原细胞具有阳性细胞核。具有MIB-1阴性核的精原细胞代表处于G1期的细胞。两种抗体均与细线期前初级精母细胞的细胞核发生强烈反应。PCNA反应也存在于细线期至粗线期。Ki-67蛋白表达是不存在的细线期和偶线期，但再次遇到粗线期和减数分裂I和II。抗PCNA/抗蛋白基因产物9.5双标记表明，从精原细胞前体细胞到A1-精原细胞的过渡在给定的管段中并不严格同步，这可能是牛生精小管中A-精原细胞繁殖灵活性的一个原因。
The distribution pattern of proliferating cell nuclear antigen (PCNA) and Ki-67 protein was studied in adult bovine seminiferous epithelium by means of immunohistochemistry using monoclonal antibodies. Tailoring the methodological protocol for each of the two proliferation markers was a necessary prerequisite for obtaining optimal results in tubular sections and whole-mounts. A-, I- and B-spermatogonia displayed PCNA-positive nuclei, except during meta-, ana- and telophases of mitosis. PCNA-negative nuclei in the basal tubular compartment, excluding those from non-cycling Sertoli cells, belonged to the spermatogonia precursor cell line. However, only about 30%, 45% and 47% of the respective A-, I-, B-spermatogonia had positive nuclei after exposure to the MIB-1 antibody directed against the Ki-67 protein. Spermatogonia with MIB-1-negative nuclei represented cells in the G1-phase. Both antibodies reacted intensely with the nuclei of preleptotene primary spermatocytes. PCNA reactivity was also present during leptotene through pachytene. Ki-67 protein expression was absent during leptotene and zygotene but was again encountered during pachytene and meiosis I and II. Anti-PCNA/anti-protein gene product 9.5 double-labelling indicated that the transition from spermatogonia precursor cells into A1-spermatogonia is not strictly synchronized in a given tubular segment, a possible reason for the flexibility in A-spermatogonial propagation seen in bovine seminiferous tubules.