Manchette binding protein and effects thereof during spermiogenesis

A technology that combines proteins and sperm, applied in the biological field, can solve the problem that the molecular mechanism of sperm cells is not very clear

Inactive Publication Date: 2010-06-16
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Although so many genes have been identified to play roles in various aspects of spermatogenesis, the mol

Method used

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  • Manchette binding protein and effects thereof during spermiogenesis
  • Manchette binding protein and effects thereof during spermiogenesis
  • Manchette binding protein and effects thereof during spermiogenesis

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Embodiment approach

[0107] As an embodiment of the present invention, the RIM-BP3 protein can be directly administered to mammals (such as humans), or the gene encoding the RIM-BP3 protein can be cloned into an appropriate vector (such as In conventional prokaryotic or eukaryotic expression vectors, or viral vectors such as herpes virus vectors or lentiviral vectors), the vectors are introduced into cells that can express the RIM-BP3 protein, so that the cells express RIM-BP3 protein.

[0108] Methods well known to those skilled in the art can be used to construct an expression vector containing the sequence of the RIM-BP3 protein coding gene and appropriate transcription / translation control signals. These methods include in vitro recombinant DNA technology, DNA synthesis technology, in vivo recombination technology and the like. Said DNA sequence can be operably linked to an appropriate promoter in the expression vector to direct mRNA synthesis. The expression vector also includes a ribosome b...

Embodiment 1

[0146] Example 1.RIM-BP3 is specifically expressed in the testes of mice, and has the highest abundance in sperm cells

[0147] The mouse RIM-BP3 is an intronless gene encoding 1606 amino acids. It has three SH3 (SrcHomology 3) domains and two carboxy-terminal FNIII (fibronectin type III) domains ( figure 1 A). This overall domain distribution is very similar to RIM-BP1 and RIM-BP2 (Mittelstaedt, T. and Schoch, S. (2007). Structure and evolution of RIM-BP genes: identification of a novel family member. Gene 403 , 70-79). At the N-terminal of RIM-BP3, it has a unique SbcC domain, suggesting ATPase activity (Connelly, J.C. et al. (1998). The SbcCD nuclease of Escherichia coli is a structural maintenance of chromosomes (SMC) family protein that cleaves hairpin DNA. Proc. Natl. Acad. Sci. U.S.A. 95, 7969-7974).

[0148] The inventors detected the transcription of RIM-BP3 in various tissues of adult mice by means of Northern blot hybridization ( figure 1 B), it was found that ...

Embodiment 2

[0152] Example 2. RIM-BP3 protein localization on the sperm collar

[0153] Next, the present inventors used mouse seminiferous tubule display technology (Kotaja, N. et al. (2004). Preparation, isolation and characterization of stage-specific spermatogenic cells for cellular and molecular analysis. Nat Methods 1, 249-254) , the subcellular localization of RIM-BP3 in developing spermatids was analyzed by immunofluorescence. RIM-BP3 first appeared in the sperm cells at the ninth stage, continued to express until the 13th stage, and finally disappeared at the 16th stage ( figure 2 A). By co-staining with an antibody against α-tubulin, it was observed that RIM-BP3 colocalizes with the sperm collar in a large area. At the same time, no signal was detected in samples from RIM-BP3 knockout mice, which indicated that the specificity of the antibody was very good ( figure 2 B). The expression pattern of RIM-BP3 is in good agreement with the dynamic distribution of the sperm colla...

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Abstract

The invention relates to a manchette binding protein and effects thereof during spermiogenesis, and discloses application of rab interaction molecule binding protein 3 (RIM-BP3) or an accelerator or an inhibitor thereof in preparing a composition for regulating the spermioteleosis of male mammals. The invention also discloses a compound for regulating the spermioteleosis of the male mammals, which comprises the RIM-BP3 and Hook1 protein. The invention also discloses a method for selecting the potential substance for regulating the spermioteleosis of the male mammals.

Description

technical field [0001] The invention belongs to the field of biotechnology; more specifically, it relates to a sperm collar binding protein, its action mechanism and application. Background technique [0002] Normal fertility in male mammals depends on the successful production of motile sperm containing an intact paternal genome and capable of fertilizing eggs. Round spermatids must go through a series of dramatic morphological changes to become mature, motile sperm. This process is called spermiogenesis. Currently, little is known about the mechanism of sperm morphogenesis. Spermatogenesis is a very complex and tightly regulated process, which can be divided into the following three phases: mitosis (self-renewal and proliferation of spermatogonia), meiosis (reduction of chromosome number from diploid to haploid) body) and spermatogenesis (deformation of haploid spermatozoa to mature spermatozoa). Among them, during spermatogenesis, haploid round spermatozoa must underg...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61K38/17A61K31/7088A61K48/00A61P15/00A61P15/08G01N33/68
Inventor 徐国良周静杜雅蕊黄燕女
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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