Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country (UPV/EHU), Leioa, Spain.
Department of Cell Biology and Histology, School of Medicine, Regional Campus of International Excellence 'Campus Mare Nostrum', University of Murcia, IMIB-Arrixaca, Murcia, Spain.
Andrology. 2019 May;7(3):319-328. doi: 10.1111/andr.12594. Epub 2019 Feb 20.
Golgi-associated PDZ and coiled-coil motif-containing protein (GOPC) is a Golgi protein that plays a role in vesicular transport and intracellular protein trafficking and degradation. Mice deficient in GOPC protein have globozoospermia and are infertile. The germ cell nuclear factor (GCNF) is a member of the nuclear receptor superfamily which is expressed in male germ cells, from spermatocytes and spermatids, both in the nucleus and the acrosomal region. It is not known if its expression could be altered in Gopc mice with defective acrosomes.
The aim of the present work was to analyze the distribution of GCNF protein in spermatids of Gopc knockout mice.
We have analyzed the expression and distribution during spermatogenesis of GCNF and its deregulation in Gopc mutant mice by RT-qPCR, Western blot, immunohistochemistry and immunogold.
Germ cell nuclear factor was localized in the nucleus of all the cell types in the seminiferous tubules. Despite being a nuclear protein, it was also located in the acrosome and in the manchette of elongating spermatids. We have found that in the absence of GOPC, the expression of GCNF was increased in the nucleus of spermatocytes, mainly in leptotene, and in the nucleus and the manchette during the spermatid elongation.
Gopc mice have defective acrosome and manchette. The manchette is involved in the transport of proteins through the cytoplasm and the nucleus. Considering that the GCNF protein is normally transported to the acrosome and the nucleus, it can be thought that transport deficiencies in Gopc mice are responsible for the increased expression of this protein.
高尔基相关 PDZ 和卷曲螺旋结构域蛋白（GOPC）是一种高尔基体蛋白，在囊泡运输和细胞内蛋白质运输和降解中发挥作用。GOPC 蛋白缺失的小鼠表现为圆头精子症和不育。生殖细胞核因子（GCNF）是核受体超家族的成员，在精母细胞和精子细胞中表达，在细胞核和顶体区均有表达。目前尚不清楚在顶体缺陷的 Gopc 小鼠中其表达是否会发生改变。
本研究旨在分析 GOPC 基因敲除小鼠精子细胞中 GCNF 蛋白的分布。
我们通过 RT-qPCR、Western blot、免疫组织化学和免疫金技术分析了 GCNF 蛋白在 Gopc 基因敲除小鼠精子发生过程中的表达和分布，以及其在 Gopc 突变小鼠中的失调情况。
生殖细胞核因子定位于生精小管中所有细胞类型的细胞核中。尽管它是一种核蛋白，但它也位于顶体和伸长精子细胞的顶体帽中。我们发现，在 GOPC 缺失的情况下，GCNF 的表达在精母细胞的细胞核中增加，主要在细线期，在精子细胞伸长过程中在细胞核和顶体帽中增加。
Gopc 小鼠的顶体和顶体帽缺陷。顶体帽参与蛋白质通过细胞质和细胞核的运输。考虑到 GCNF 蛋白通常被运输到顶体和细胞核，因此可以认为 Gopc 小鼠的运输缺陷是导致该蛋白表达增加的原因。
