Method for constructing animal model of spermatogenesis disorder

Abstract

The present invention relates to a method for constructing an animal model of spermatogenesis disorder. The method constructs an animal model of spermatogenesis disorder by inactivating the Iqcd gene function of a male animal. The testicles and epididymis of the animal model of spermatogenesis disorder constructed by the method have no difference from those of the wild type in appearance, but spermatogenesis disorder is mainly blocked at the spermatocyte stage, so it can be used for the study of spermatogenesis disorder that blocks spermatogenesis at the spermatocyte stage.

Description

Technical Field

[0001] The invention relates to the technical field of genetic engineering, and in particular to a method for constructing an animal model of spermatogenesis disorder. Background Art

[0002] According to incomplete statistics, 8% to 12% of couples worldwide face infertility problems, of which about 50% are caused by male factors. Male infertility is a pathological disease caused by multiple factors, accounting for about 7-10% of the male population. The causes of male infertility include physical factors, chemical factors, endocrine factors, environmental factors, genetic factors, etc., and about 60-70% of male infertility is unknown. In mammals, spermatogenesis mainly includes three consecutive processes: (1) mitosis and differentiation of spermatogonial stem cells; (2) two consecutive meiotic divisions of spermatocytes; (3) deformation of spermatocytes, including acrosome formation, chromatin remodeling, histone-protamine conversion, flagella development, etc. The spermatogenesis process involves the coordinated expression of more than 2,000 genes. Male infertility caused by genetic factors is extremely complex and heterogeneous.

[0003] In recent years, more and more studies on male infertility have been conducted at home and abroad, and related animal models have developed rapidly. For example, in order to study the influence of physical factors, infertility animal models have been established through thermal effects and radiation effects; in order to study the influence of chemical factors, infertility models of mice have been induced by cottonpol, adenine, formaldehyde, etc.; in order to study the influence of endocrine factors, infertility models of mice have been induced by the estrogen diethylstilbestrol; in order to study the influence of genetic factors, infertility models of mice have been obtained by knocking out the phospholipid hydrolase glutathione peroxidase gene or androgen receptor. Among them, the infertility model of mice after knocking out the phospholipid hydrolase glutathione peroxidase gene showed a decrease in sperm activity and a significant increase in the deformity rate, while the infertility model of mice after knocking out the androgen receptor showed that the mouse testicles were atrophic, the germ cells were reduced, and the contraction force of the cells around the testicular tubules was weakened, which affected the occurrence and output of sperm. Summary of the invention

[0004] The present invention has found that after knocking out the Iqcd gene in germ cells of mouse testicular tissue, male mice were found to be infertile. The growth of mice after knocking out the Iqcd gene in germ cells, the appearance of the testicles and epididymis were no different from those of the wild type, but spermatogenesis was impaired, mainly blocked at the spermatocyte stage. Therefore, the inactivation of the Iqcd gene function can be used to construct an animal model of spermatogenesis impaired in which the blockage occurs at the spermatocyte stage.

[0005] Based on this, a method for constructing an animal model of spermatogenesis disorder that blocks spermatogenesis at the spermatocyte stage is provided.

[0006] In addition, a method for screening or identifying drugs for treating spermatogenesis disorders using a spermatogenesis disorder animal model prepared by the above-mentioned method for constructing a spermatogenesis disorder animal model is also provided.

[0007] In one embodiment, the Iqcd gene function is inactivated in the germ cells of a male animal.

[0008] In one embodiment, the animal used to prepare the spermatogenesis disorder animal model includes mice or rats.

[0009] In one embodiment, the animal is a C57BL / 6N mouse or a C57BL / 6J mouse.

[0010] In one embodiment, the animal used to prepare the spermatogenesis disorder animal model is a mouse, and the Iqcd gene function is inactivated by knocking out exon 2 of the Iqcd gene in germ cells of the mouse testicular tissue.

[0011] In one embodiment, the Cre / LoxP recombinase system is used to knock out exon 2 of the Iqcd gene in germ cells of mouse testicular tissue.

[0012] In one embodiment, the construction method comprises the following steps:

[0013] LoxP sites were inserted at both ends of exon 2 of the mouse Iqcd gene to construct LoxP homozygous female mice;

[0014] The LoxP homozygous female mice are mated with male Cre tool mice to obtain LoxP heterozygous Cre positive male mice, wherein the male Cre tool mice are male mice in which Cre recombinase is expressed in germ cells; and

[0015] The LoxP heterozygous Cre-positive male mice were mated with LoxP homozygous female mice to obtain a spermatogenesis disorder animal model.

[0016] In one embodiment, the step of constructing a LoxP homozygous female mouse comprises:

[0017] Inserting both ends of exon 2 of the mouse Iqcd gene into the LoxP site to obtain F1 LoxP heterozygous male mice and F1 LoxP heterozygous female mice;

[0018] The F1 generation LoxP heterozygous male mice and the F1 generation LoxP heterozygous female mice are mated together to obtain F2 generation LoxP homozygous male mice and F2 generation LoxP homozygous female mice; and

[0019] The F2 LoxP homozygous male mice and the F2 LoxP homozygous female mice were mated together to obtain LoxP homozygous female mice.

[0020] In one embodiment, the male Cre tool mouse is a stra-8Cre positive male mouse.

[0021] A method for screening or identifying drugs for treating spermatogenesis disorders, using the spermatogenesis disorder animal model prepared by the spermatogenesis disorder animal model construction method mentioned above to screen or identify drugs for treating spermatogenesis disorders. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is the construction principle of LoxP heterozygous mice in Example 1;

[0023] Figure 2 The tail genotype identification result of the Iqcd gene conditional knockout mice in Example 1;

[0024] Figure 3 The fertility statistics of the Iqcd gene conditional knockout mice in Example 1;

[0025] Figure 4 The results of HE staining of testicular tissue and epididymal tail of Iqcd gene conditionally knocked out male mice in Example 1. DETAILED DESCRIPTION

[0026] In order to facilitate the understanding of the present invention, the present invention will be described more fully below, and the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the disclosure of the present invention more thorough and comprehensive.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art of the technical field of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention. In this article, "optionally" means giving an example.

[0028] Gene knockout refers to a technique that integrates exogenous genes into a certain site on the target cell genome through homologous recombination to achieve site-specific modification of a gene on a chromosome. It can target a sequence with a known sequence but unknown function, change the genetic genes of an organism, disable the function of a specific gene, thereby shielding some functions, and further affect the organism, thereby inferring the biological function of the gene. Gene knockout is divided into two types: complete gene knockout and conditional gene knockout (CKD, also known as incomplete gene knockout). Complete gene knockout refers to the complete elimination of target gene activity in cells or animal individuals through homologous recombination, and conditional gene knockout refers to the realization of gene knockout at a specific time and space through a localized recombination system. The Cre / LoxP recombinase system, Gin / Gix system, FLP / FRT system and R / RS system are the four commonly used localized recombination systems, especially the Cre / LoxP recombinase system is the most widely used.

[0029] IQCD is a member of the IQ protein family. This application study found that the human Iqcd gene (Gene ID in the NCBI database: 115811) was downregulated in the sperm proteomics of patients with in vitro fertilization (IVF) failure, indicating that the Iqcd gene may be involved in spermatogenesis and affect male fertility. In addition, the present invention found that male mice were infertile after conditional knockout of the mouse Iqcd gene. The appearance of their testicles and epididymis was no different from that of the wild type, but the histomorphological results showed that spermatogenesis was impaired and mainly blocked at the sperm cell stage. Therefore, the inactivation of Iqcd gene function can be used to construct an animal model of spermatogenesis impaired in which the spermatogenesis is blocked at the sperm cell stage.

[0030] Based on this, one embodiment of the present application provides a method for constructing an animal model of spermatogenesis disorder, which inactivates the Iqcd gene function of male animals to construct an animal model of spermatogenesis disorder.

[0031] The physiological functions of the testis include spermatogenesis. The seminiferous epithelium of the seminiferous tubule is where sperm is produced, and it is composed of spermatogenic cells and supporting cells. Spermatogenic cells are not a type of cell, but a general term for multiple cells, including spermatogonia, primary spermatocytes, secondary spermatocytes, spermatids, and sperm. The former develops and differentiates into the latter in sequence, and is arranged in sequence from the base of the seminiferous epithelium to the lumen surface, and finally forms spermatozoa that enter the lumen and move to the next level of lumen with the help of the contraction of the muscle-like cells on the outside of the seminiferous epithelium. Spermatogonia replicate and proliferate continuously through mitosis, some of which become stem cells to maintain the vitality of proliferation, and other spermatogonia become primary spermatocytes, and become secondary spermatocytes through one meiosis, and secondary spermatocytes undergo another meiosis to become spermatids, and then undergo a series of morphological changes to become mature sperm. The protrusions of supporting cells surround spermatogenic cells at all levels, play a role in support, protection, and nutrition, and are conducive to the transport of substances and signal transmission between cells. The human Iqcd gene is mainly expressed in the testis. Therefore, when preparing an animal model of spermatogenesis disorder, the Iqcd gene function inactivation of germ cells in reproductive organs is achieved by conditional gene knockout, which is beneficial to the study of human spermatogenesis disorder and male infertility. Therefore, in some embodiments, an animal model of spermatogenesis disorder is constructed by inactivating the Iqcd gene function in the germ cells of male animals. Optionally, the germ cells include one or more of spermatogonia and spermatocytes.

[0032] In some embodiments, the animal used to prepare the spermatogenesis disorder animal model is a mammal. Further, the animal used to prepare the spermatogenesis disorder animal model is a rodent. Further, the animal used to prepare the spermatogenesis disorder animal model includes mice (Mus musculus) or rats (Rattus norvegicus). Optionally, the animal used to prepare the spermatogenesis disorder animal model is a C57BL / 6N mouse or a C57BL / 6J mouse.

[0033] In some embodiments, the Iqcd gene function is inactivated by knocking out the start codon of the Iqcd gene. The Iqcd gene of mice (Gene ID in the NCBI database: 75732) Iqcd is located on mouse chromosome 5, has 5 exons, the start codon is on exon 2, the stop codon is on exon 5, and exon 2 occupies the main coding region of the gene. Therefore, by knocking out exon 2 of the mouse, the Iqcd gene loses the start codon and cannot be translated, thereby inactivating the Iqcd gene function. It is understandable that when the animal used to prepare the spermatogenesis disorder animal model is not a mouse, the knockout position of the Iqcd gene is also adaptively modified to a position that allows the start codon to be inactivated.

[0034] Alternatively, the second exon of the Iqcd gene of the mouse is knocked out using the Cre / LoxP recombinase system to inactivate the function of the Iqcd gene. The Cre recombinase (Cyclization Recombination Enzyme) is encoded by the Cre gene of the Escherichia coli phage P1 and is a 38kD protein consisting of 343 amino acids. It not only has catalytic activity, but also, similar to a restriction enzyme, can specifically recognize the loxP site. The LoxP (locus of X-overP1) site is 34bp long, including two 13bp inverted repeat sequences and an 8bp spacer region. Among them, the inverted repeat sequence is the specific recognition site of the Cre recombinase, and the spacer region determines the direction of the loxP site. When there is a loxP site in the genome, once there is a Cre recombinase, it will bind to the inverted repeat sequence region at both ends of the loxP site to form a dimer. This dimer binds to the dimers of other loxP sites to form a tetramer. Subsequently, the DNA between the loxP sites is cut by the Cre recombinase, and the incision is reconnected under the action of DNA ligase. The result of the recombination depends on the position and direction of the loxP site. When the two loxP sites are located on the same DNA chain and in the same direction, the Cre recombinase knocks out the sequence between the loxP. In this embodiment, the gene knockout performed using the Cre / LoxP recombinase system is a conditional gene knockout, and the knockout of the second exon of the Iqcd gene occurs in a specific tissue or organ (such as testis) and other tissues or organs that may express the Iqcd gene are not affected, thereby reducing the impact of other tissues or organs on infertility, and then being able to more accurately establish an animal model suitable for infertility. It is understandable that in other embodiments, the method for knocking out the second exon of the Iqcd gene of the mouse is not limited to the above-mentioned Cre / LoxP recombinase system, and other systems can also be used for gene knockout. For example, the Gin / Gix system, the FLP / FRT system or the R / RS system.

[0035] Optionally, the method for constructing a spermatogenesis disorder animal model using the Cre / LoxP recombinase system includes step S110, step S120 and step S130. Specifically:

[0036] Step S110: inserting LoxP sites at both ends of exon 2 of the mouse Iqcd gene to construct LoxP homozygous female mice.

[0037] Specifically, LoxP homozygous mice, also called Flox mice, refer to mice containing paired LoxP sites on the target gene. After mating with Cre tool mice, the fragment between the LoxP sites can be knocked out in specific tissues or cells.

[0038] In some embodiments, CRISPR / Cas9 technology is used to insert LoxP sites at both ends of exon 2 of the Iqcd gene. CRISPR / Cas9 is the third generation of genome site-directed editing technology after "zinc finger endonuclease (ZFN)" and "transcription activator-like effector nuclease (TALEN)". The principle of CRISPR / Cas9 system for gene knockout is that tracrRNA-crRNA is used to guide the action of Cas9 when it is fused into a single-stranded guide RNA (i.e., sgRNA). For the target gene, the target gene sequence is identified by artificially designed sgRNA (small guide RNA, sgRNA), and the Cas9 protease is guided to effectively cut the DNA double strand to form a double-strand break of DNA. Under normal circumstances, cells will use efficient non-homologous end joining (Non-homologous End Joining, NHEJ) to repair the broken DNA, but base insertion or deletion mismatches usually occur during the repair process, causing frameshift mutations, making the target gene lose its function, thereby achieving gene knockout.

[0039] It is understandable that, in other embodiments, the method of inserting LoxP sites at both ends of exon 2 of the mouse Iqcd gene is not limited to CRISPR / Cas9 technology, but can also be other technologies.

[0040] Furthermore, if the number of LoxP homozygous female mice is more than one, it involves breeding of LoxP homozygous female mice. In this case, the step of constructing LoxP homozygous female mice includes step S111, step S112 and step S113.

[0041] Step S111: inserting both ends of exon 2 of the mouse Iqcd gene into the LoxP site to obtain F1 generation LoxP heterozygous male mice and F1 generation LoxP heterozygous female mice.

[0042] Step S112: F1 generation LoxP heterozygous male mice and F1 generation LoxP heterozygous female mice are mated together to obtain F2 generation LoxP homozygous male mice and F2 generation LoxP homozygous female mice.

[0043] Step S113: F2 generation LoxP homozygous male mice and F2 generation LoxP homozygous female mice are mated together to obtain LoxP homozygous female mice (F3 generation).

[0044] It is understandable that, when the number of LoxP homozygous female mice obtained in step S112 is sufficient to carry out steps S120 and S130, step S113 may not be performed; if the number of LoxP homozygous female mice obtained in step S113 is not sufficient to carry out steps S120 and S130, the LoxP homozygous male mice and the LoxP homozygous female mice obtained in step S113 may be mated together.

[0045] It should be noted that, in this article, "LoxP heterozygous female mice" refer to female mice in which LoxP sites are inserted at both ends of the second exon of one of the two alleles Iqcd on homologous chromosomes, while the other does not have a LoxP site inserted. "LoxP heterozygous male mice" refer to male mice in which LoxP sites are inserted at both ends of the second exon of one of the two alleles Iqcd on homologous chromosomes, while the other does not have a LoxP site inserted. "LoxP homozygous female mice" are female mice in which LoxP sites are inserted at both ends of the second exon of the two alleles Iqcd on homologous chromosomes. "LoxP homozygous male mice" are male mice in which LoxP sites are inserted at both ends of the second exon of the two alleles Iqcd on homologous chromosomes.

[0046] Step S120: The LoxP homozygous female mice are mated with male Cre tool mice to obtain LoxP heterozygous Cre positive male mice.

[0047] Specifically, the male Cre tool mouse is a male mouse in which Cre recombinase is expressed in germ cells. Alternatively, the male Cre tool mouse is a stra-8Cre positive male mouse. The stra-8Cre positive male mouse is a mouse in which Cre enzyme sequence is inserted after the stra-8 gene promoter and Cre recombinase is specifically expressed in testicular tissue.

[0048] It should be noted that the “LoxP heterozygous Cre-positive male mouse” refers to a LoxP heterozygous male mouse that can express Cre recombinase.

[0049] Step S130: The LoxP heterozygous Cre tool mice are mated with LoxP homozygous female animals in the same cage to obtain a spermatogenesis disorder animal model.

[0050] Of course, in other embodiments, the method of inactivating the function of the Iqcd gene is not limited to the above-mentioned conditional gene knockout using the Cre / LoxP recombinase system, but can also be a complete gene knockout using the CRISPR / Cas9 system. It is understandable that the method of inactivating the function of the Iqcd gene can be not only knockout, but also other techniques. For example, gene knock-in, gene silencing, etc. For example, knock-in one or more bases on the Iqcd gene promoter or knock-in one or more bases on the base corresponding to the start codon, so that at least one of the promoter and the start codon loses function, so that it cannot be transcribed or translated, thereby inactivating the function of the Iqcd gene. For example, gene silencing. In addition, the method of inactivating the Iqcd gene function of the mouse is not limited to knocking out its second exon, but can also be at other positions of the Iqcd gene.

[0051] The method for constructing the spermatogenesis disorder animal model has at least the following advantages:

[0052] (1) An animal model of spermatogenesis disorder was constructed by inactivating the Iqcd gene function in male animals, in which spermatogenesis arrest occurred at the spermatid stage.

[0053] (2) This construction method can achieve Iqcd gene knockout at a specific time and space through conditional gene knockout.

[0054] In addition, one embodiment of the present application also provides a method for screening or identifying drugs for treating spermatogenesis disorders, which method uses a spermatogenesis disorder animal model prepared by the method for constructing a spermatogenesis disorder animal model of any of the above embodiments to screen or identify drugs for treating spermatogenesis disorders.

[0055] The above-mentioned method for screening or identifying drugs for treating spermatogenesis disorders utilizes the above-mentioned spermatogenesis disorder animal model to screen or identify drugs for treating spermatogenesis disorders. Since the above-mentioned spermatogenesis disorder animal model is an animal model established for spermatogenesis disorders blocked at the spermatocyte stage, it is more accurate for screening or identifying drugs involving spermatogenesis disorders acting on the spermatocyte stage.

[0056] The following is a detailed description in conjunction with specific examples. The following examples do not include other components except for inevitable impurities unless otherwise specified. The reagents and instruments used in the examples are conventionally selected in the art unless otherwise specified. The experimental methods for which specific conditions are not specified in the examples are implemented according to conventional conditions, such as the conditions described in the literature, books, or methods recommended by the manufacturer.

[0057] Example 1

[0058] 1. Construction of Iqcd gene conditional knockout mouse model

[0059] (1) Construction of Loxp heterozygous mice

[0060] Exon 2 was selected as the knockout region, and LoxP sites were inserted at both ends of exon 2 of the Iqcd gene in C57BL / 6N mice. After knocking out exon 2, a frameshift mutation occurred, and the Iqcd gene function was inactivated, achieving the purpose of gene knockout. The principle is shown in Figure 1 The construction of Loxp heterozygous mice was commissioned to Cyyt Biotechnology (Suzhou) Co., Ltd.

[0061] (2) Preparation of Iqcd gene conditional knockout mice

[0062] A: Expansion of Loxp homozygous female mice

[0063] After obtaining LoxP heterozygous (F1 generation) female mice and LoxP heterozygous (F1 generation) male mice from Saiye (Suzhou) Biotechnology Co., Ltd., they were mated together to obtain LoxP homozygous (F2 generation) female mice and LoxP homozygous (F2 generation) male mice, and then the LoxP homozygous (F2 generation) female mice and LoxP homozygous (F2 generation) male mice were mated together to obtain a large number of LoxP homozygous (F3 generation) female mice.

[0064] B: Preparation of Loxp heterozygous stra-8Cre positive male mice

[0065] LoxP homozygous (F3 generation) female mice were mated with stra-8Cre-positive male mice (donated by Professor Tong Minghan) and LoxP heterozygous stra-8Cre-positive male mice were obtained through genotyping.

[0066] C: Preparation of Iqcd gene conditional knockout mice

[0067] The LoxP heterozygous stra-8Cre positive male mice were mated with LoxP homozygous (F3 generation) female mice, and Iqcd conditional knockout mice were obtained by genotyping, wherein the genotyping step includes:

[0068] 1) Cut about 0.5 cm of the mouse tail and extract the mouse tail DNA;

[0069] 2) Design primers and perform PCR reaction to obtain amplified products. The primers for PCR are:

[0070] Forward primer: 5'-GCAGATACCACATCTCCTGAAACT-3' (SEQ ID NO: 1),

[0071] Reverse primer: 5'-TTACAAACTCTTGCCTACGTCCT-3' (SEQ ID NO: 2);

[0072] 3) The amplified product of step 2) is subjected to agarose gel electrophoresis to identify the genotype.

[0073] Genotyping results Figure 2 shown.

[0074] exist Figure 2 In the experiment, the wild type has only the 218bp band; the genotype with two bands of 218bp and 280bp is the heterozygous type; the genotype with only the 280bp band is the homozygous type, that is, the Iqcd gene conditional knockout mouse.

[0075] 2. Identification of fertility in male mice with conditional knockout of Iqcd gene

[0076] Three 8-week-old Iqcd gene conditional knockout homozygous, heterozygous, and wild male mice were housed with 8-week-old wild female mice at a ratio of 1:3 for 5 months. The number and gender of the offspring were recorded. The statistical results are shown in the figure. Figure 3 shown.

[0077] Depend on Figure 3 It can be seen that through 5 months of fertility assessment, the Iqcd gene conditional knockout mice did not produce offspring, which shows that Iqcd gene conditional knockout male mice are infertile.

[0078] 3. Changes in testicular histology in male mice with conditional knockout of Iqcd gene

[0079] The testicular tissue and epididymal tail of 12-week-old Iqcd gene conditional knockout male mice were subjected to hematoxylin-eosin staining (HE staining). The HE staining steps include:

[0080] 1) The testicular tissue and epididymal tail of 12-week-old adult male mice were fixed with fixative and embedded into wax blocks, and sliced ​​with a slicer to a thickness of 5 μm;

[0081] 2) After the slices are removed, bake them at 60°C for 1 hour, and then dewax them in xylene and gradient ethanol;

[0082] 3) After dewaxing, wash the slides with PBS;

[0083] 4) Add eosin for staining for 10 min;

[0084] 5) Hydrochloric acid alcohol for differentiation;

[0085] 6) Add hematoxylin, stain for 10 min in dark, and rinse with running water for 1 h;

[0086] 7) After staining, dehydrate with anhydrous ethanol, make transparent with xylene, seal with neutral gum, and examine under a microscope after drying.

[0087] HE staining results Figure 4 As shown. Figure 4 In the figure, A is the HE staining of the testis morphology of heterozygous mice; B is the HE staining of the testis morphology of homozygous mice; C is the HE staining of the epididymis tail of heterozygous mice; D is the HE staining of the epididymis tail of homozygous mice.

[0088] Depend on Figure 4 It can be seen that spermatogenesis in the testes of heterozygous mice is normal, while spermatogenesis in the testes of homozygous mice is impaired; and sperm is enriched in the tail of the epididymis of heterozygous mice, while sperm is almost invisible in the tail of the epididymis of homozygous mice. This shows that spermatogenesis in male mice with conditional knockout of the Iqcd gene is blocked, and it is mainly blocked at the spermatocyte stage.

[0089] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0090] The above-described embodiments only express several implementation methods of the present invention, which is convenient for understanding the technical solutions of the present invention in detail, but cannot be understood as limiting the scope of protection of the invention patent. It should be pointed out that, for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. It should be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided by the present invention are all within the protection scope of the claims attached to the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the contents of the attached claims, and the description and drawings can be used to interpret the contents of the claims.

Claims

1. A method for constructing an animal model of spermatogenesis disorder that blocks spermatogenesis at the spermatocyte stage, characterized in that: The construction method constructs a spermatogenesis disorder animal model by inactivating the Iqcd gene function of a male animal. The animal used to prepare the spermatogenesis disorder animal model is a mouse, and the Iqcd gene function is inactivated by knocking out the second exon of the Iqcd gene in germ cells in the testicular tissue of the mouse.

2. The construction method according to claim 1, characterized in that: The animals are C57BL / 6N mice or C57BL / 6J mice.

3. The construction method according to claim 1, characterized in that: The Cre / LoxP recombinase system was used to knock out exon 2 of the Iqcd gene in germ cells of mouse testicular tissue.

4. The construction method according to claim 3, characterized in that: The construction method comprises the following steps: LoxP sites were inserted at both ends of exon 2 of the mouse Iqcd gene to construct LoxP homozygous female mice; The LoxP homozygous female mice are mated with male Cre tool mice to obtain LoxP heterozygous Cre positive male mice, wherein the male Cre tool mice are male mice in which Cre recombinase is expressed in germ cells; and The LoxP heterozygous Cre-positive male mice were mated with LoxP homozygous female mice to obtain a spermatogenesis disorder animal model.

5. The construction method according to claim 4, characterized in that: The steps for constructing LoxP homozygous female mice include: Inserting both ends of exon 2 of the mouse Iqcd gene into the LoxP site to obtain F1 LoxP heterozygous male mice and F1 LoxP heterozygous female mice; The F1 generation LoxP heterozygous male mice and the F1 generation LoxP heterozygous female mice are mated together to obtain F2 generation LoxP homozygous male mice and F2 generation LoxP homozygous female mice; and The F2 LoxP homozygous male mice and the F2 LoxP homozygous female mice were mated together to obtain LoxP homozygous female mice.

6. The construction method according to claim 4 or 5, characterized in that: The male Cre tool mouse is a stra-8Cre positive male mouse.

7. A method for screening or identifying a drug for treating spermatogenesis disorders, characterized in that: The spermatogenesis disorder animal model prepared by the method for constructing the spermatogenesis disorder animal model according to any one of claims 1 to 6 is used to screen or identify drugs for treating spermatogenesis disorder.

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