Dislocated type-A spermatogonia in human seminiferous tubules

The access to sufficient numbers of spermatogonial stem cells (SSCs) is a prerequisite for the study of their regulation and further biomanipulation. A specialized medium and several growth factors were tested to study thein vitrobehavior of bovine type A spermatogonia, a cell population that includes the SSCs and can be specifically stained for the lectin Dolichos biflorus agglutinin. During short-term culture (2 weeks), colonies appeared, the morphology of which varied with the specific growth factor(s) added. Whenever the stem cell medium was used, round structures reminiscent of sectioned seminiferous tubules appeared in the core of the colonies. Remarkably, these round structures always contained type A spermatogonia. When leukemia inhibitory factor (LIF), epidermal growth factor (EGF), or fibroblast growth factor 2 (FGF2) were added, specific effects on the numbers and arrangement of somatic cells were observed. However, the number of type A spermatogonia was significantly higher in cultures to which glial cell line-derived neurotrophic factor (GDNF) was added and highest when GDNF, LIF, EGF, and FGF2 were all present. The latter suggests that a proper stimulation of the somatic cells is necessary for optimal stimulation of the germ cells in culture. Somatic cells present in the colonies included Sertoli cells, peritubular myoid cells, and a few Leydig cells. A transplantation experiment, using nude mice, showed the presence of SSCs among the cultured cells and in addition strongly suggested a more than 10 000-fold increase in the number of SSCs after 30 days of culture. These results demonstrate that bovine SSC self-renew in our specialized bovine culture system and that this system can be used for the propagation of these cells.

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p53-Dependent Apoptosis in the Inhibition of Spermatogonial Differentiation in Juvenile Spermatogonial Depletion (Utp14bjsd) Mice

Endocrinology ◽

10.1210/en.2007-1338 ◽

2008 ◽

Vol 149 (6) ◽

pp. 2773-2781 ◽

Cited By ~ 13

Author(s):

Gunapala Shetty ◽

Shan H. Shao ◽

Connie C. Y. Weng

Keyword(s):

Double Mutant ◽

Fas Ligand ◽

Type A ◽

Terminal Deoxynucleotidyl Transferase ◽

Seminiferous Tubules ◽

Caspase 9 ◽

Apoptotic Pathway ◽

Independent Process ◽

Old Mice ◽

Type A Spermatogonia

In adult male mice homozygous for the juvenile spermatogonial depletion (Utp14bjsd) mutation in the Utp14b gene, type A spermatogonia proliferate, but in the presence of testosterone and at scrotal temperatures, these spermatogonia undergo apoptosis just before differentiation. In an attempt to delineate this apoptotic pathway in jsd mice and specifically address the roles of p53- and Fas ligand (FasL) /Fas receptor-mediated apoptosis, we produced jsd mice deficient in p53, Fas, or FasL. Already at the age of 5 wk, less degeneration of spermatogenesis was observed in p53-null-jsd mice than jsd single mutants, and in 8- or 12-wk-old mice, the percentage of seminiferous tubules showing differentiated germ cells [tubule differentiation index (TDI)] was 26–29% in the p53-null-jsd mice, compared with 2–4% in jsd mutants with normal p53. The TDI in jsd mice heterozygous for p53 showed an intermediate TDI of 8–13%. The increase in the differentiated tubules in double-mutant and p53 heterozygous jsd mice was mostly attributable to intermediate and type B spermatogonia; few spermatocytes were present. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling staining showed that most of these differentiated spermatogonia still underwent apoptosis, thereby blocking further continuation of spermatogenesis. In contrast, the percentage of tubules that were differentiated was not significantly altered in either adult Fas null-jsd mice or adult FasL defective gld-jsd double mutant mice as compared with jsd single mutants. Furthermore, caspase-9, but not caspase-8 was immunochemically localized in the adult jsd mice spermatogonia undergoing apoptosis. The results show that p53, but not FasL or Fas, is involved in the apoptosis of type A spermatogonia before/during differentiation in jsd mice that involves the intrinsic pathway of apoptosis. However, apoptosis in the later stages must be a p53-independent process.

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Non-random distribution of spermatogonia in rats: evidence of niches in the seminiferous tubules

Reproduction ◽

10.1530/rep.0.1260669 ◽

2003 ◽

pp. 669-680 ◽

Cited By ~ 95

Author(s):

H Chiarini-Garcia ◽

AM Raymer ◽

LD Russell

Keyword(s):

Basal Lamina ◽

Random Distribution ◽

Type A ◽

Seminiferous Epithelium ◽

Seminiferous Tubules ◽

Interstitial Tissue ◽

Primitive Type ◽

Linear Index ◽

Type A Spermatogonia

The relationships and distribution of spermatogonia were studied as a function of the stage of the seminiferous epithelium cycle in rats. Primitive spermatogonia in the mouse are located along regions of the basal lamina that face the interstitium. Before studying the distribution of spermatogonia in rats, it was necessary to characterize the various types of spermatogonia, as recently performed for mice. The Strauss' linear index (Li) selectivity method was then used and spermatogonia of the A(single) (A(s)) to A(aligned) (A(al)) lineage were preferentially found to be located in regions opposing the interstitium at stages V, VII and IX of the spermatogenic cycle. Because relatively little tubule-to-tubule contact occurs in rats, the aim of this study was to determine whether tubule-to-tubule contact or tubule proximity (or alternatively, the amount of interstitium) was an important factor in spermatogonial position. In this regard, another method (tubule proximity) was devised to determine spermatogonial position that accounted for the presence of adjacent tubules. This method showed that the position of tubules, rather than tubule contact, was more accurate than the Li method in determining the location of spermatogonia in the rat. The results also showed a non-random distribution of spermatogonia resembling that of the mouse, and that tubule-to-tubule contact is not essential for the positioning of spermatogonia. In conclusion, the results of this study strongly indicate that the most primitive type A spermatogonia (A(s), A(paired) and A(al)) in rats are present in niches located in those areas of the seminiferous tubules that border the interstitial tissue.

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Changes in the cells of the Sex Cords and Seminiferous Tubules during the development of the Testis of the rat and mouse.

Australian Journal of Zoology ◽

10.1071/zo9620178 ◽

1962 ◽

Vol 10 (2) ◽

pp. 178 ◽

Cited By ~ 52

Author(s):

CS Sapsford

Keyword(s):

Basement Membrane ◽

Type A ◽

Staining Intensity ◽

Seminiferous Tubules ◽

Postnatal Life ◽

Foetal Development ◽

Cytoplasmic Inclusions ◽

Daughter Cells ◽

Early Postnatal Life ◽

Type A Spermatogonia

The sex cords of the testis of the foetal rat and mouse are made up of two types of cell- the gonocytes and the indifferent cells. During foetal and early postnatal life, the former undergo a process of maturation involving principally an increase in size and in number of cytoplasmic inclusions. Nuclear enlargement is accompanied by a diminution in staining intensity. Gonocytes are more centrally placed within the sex cords than indifferent cells, and in the later stages of foetal development they cease to divide. In the first week of postnatal life, however, they resume mitotic activity and migrate to the basement membrane of the sex cords. Gonocytes as such disappear and are replaced by smaller daughter cells, the immature type A spermatogonia. The appearance of the latter is followed by the onset of spermatogenesis. As the tubules gradually become filled with the layers of cells of the spermatogenic line, the daughter cells of the gonocytes become flattened against the basement membrane, and come to resemble the type A spermatogonia found in the adult. Indifferent cells, which are always cytologically distinct from gonocytes and spermatogonia, exist as mononucleate units. The pattern of change of these cells during the development of the testis is different from that of the germ cells. Unlike the gonocytes, the indifferent cells continue to divide during foetal and early postnatal life. Little change takes place in these cells until after the onset of spermatogenesis, when they gradually increase in size and become Sertoli cells. The latter are principally mononucleate in form.

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PSXIII-1 Age-related changes in the ratio of different spermatogonia types in the seminiferous tubules of quail testes

Journal of Animal Science ◽

10.1093/jas/skz258.744 ◽

2019 ◽

Vol 97 (Supplement_3) ◽

pp. 373-374

Author(s):

Evgeniya K Tomgorova ◽

Natalia A Volkova ◽

Anastasia N Vetokh ◽

Inna P Novgorodova ◽

Ludmila A Volkova ◽

...

Keyword(s):

Early Period ◽

Type A ◽

Seminiferous Tubules ◽

Maximum Value ◽

Age Related ◽

Spermatogonia Cells ◽

Hematoxylin Eosin ◽

Generative Cells ◽

Testis Tissue ◽

Type A Spermatogonia

Abstract Spermatogonia are early-undifferentiated germ cells, giving rise to mature male generative cells — the spermatozoa. There are two types of spermatogonia – A and B. Of greatest interest is the use of type A spermatogonia, which are the stem cells of the testes. To select the appropriate age for collecting spermatogonia А from quails it is necessary to know the specific features of spermatogenesis. Development dynamics of various spermatogonia types in the quail testicular tubules was studied. Histological studies of the quails testicular tubules at the age of 1, 2, 3, 4, 5 and 6 weeks (n = 30) were carried out. Samples of testis tissue were fixed in Bouin’s fixative. Histological sections were stained with hematoxylin-eosin. Identification of different spermatogonia types was carried out according to their morphology. Type A spermatogonia were additionally identified by immunohistochemistry using SSEA-1 antibodies. The proportion of spermatogonia in the total number of spermatogenic cells in the seminiferous tubules of quails changed with age. The maximum value was reached at the age of 3 weeks and it was 76±6%. On reaching maturity (6 weeks), this indicator decreased to 12 ± 1 %. In the early period of ontogenesis (1–2 weeks), spermatogonia cells were represented mainly by type A spermatogonia. The proportion of these cells from the total number of spermatogonia reached 80 ± 3 %. With increasing age, this indicator decreased, reaching minimum values for achieving maturity (6 weeks) - 16 ± 1 %. The percentage of type B spermatogonia in the seminiferous tubule of quails on the contrary increased with age — from 5 ± 1% at 1 week old to 70 ± 2% at maturity. Thus, the age no later than 2 weeks is the most optimal for the isolation type A spermatogonia of quails. Supported by RFBR (18-29-07079).

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11β-Hydroxylase loss disrupts steroidogenesis and reproductive function in zebrafish

Journal of Endocrinology ◽

10.1530/joe-20-0160 ◽

2020 ◽

Vol 247 (2) ◽

pp. 197-212

Author(s):

James A Oakes ◽

Lise Barnard ◽

Karl-Heinz Storbeck ◽

Vincent T Cunliffe ◽

Nils P Krone

Keyword(s):

Reproductive Function ◽

Type A ◽

Seminiferous Tubules ◽

Sex Characteristics ◽

Secondary Sexual Characteristics ◽

Secondary Sex Characteristics ◽

Sexual Characteristics ◽

Predominantly Female ◽

And Function ◽

Type A Spermatogonia

The roles of androgens in male reproductive development and function in zebrafish are poorly understood. To investigate this topic, we employed CRISPR/Cas9 to generate cyp11c1 (11β-hydroxylase) mutant zebrafish lines. Our study confirms recently published findings from a different cyp11c1−/− mutant zebrafish line, and also reports novel aspects of the phenotype caused by loss of Cyp11c1 function. We report that Cyp11c1-deficient zebrafish display predominantly female secondary sex characteristics, but may possess either ovaries or testes. Moreover, we observed that cyp11c1−/− mutant male zebrafish are profoundly androgen- and cortisol-deficient. These results provide further evidence that androgens are dispensable for testis formation in zebrafish, as has been demonstrated previously in androgen-deficient and androgen-resistant zebrafish. Herein, we show that the testes of cyp11c1−/− mutant zebrafish exhibit a disorganised tubular structure; and for the first time demonstrate that the spermatic ducts, which connect the testes to the urogenital orifice, are severely hypoplastic in androgen-deficient zebrafish. Furthermore, we show that spermatogenesis and characteristic breeding behaviours are impaired in cyp11c1−/− mutant zebrafish. Expression of nanos2, a type A spermatogonia marker, was significantly increased in the testes of Cyp11c1-deficient zebrafish, whereas expression of markers for later stages of spermatogenesis was significantly decreased. These observations indicate that in zebrafish, production of type A spermatogonia is androgen-independent, but differentiation of type A spermatogonia is an androgen-dependent process. Overall, our results demonstrate that whilst androgens are not required for testis formation, they play important roles in determining secondary sexual characteristics, proper organisation of seminiferous tubules, and differentiation of male germ cells.

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Regulation of proliferation and differentiation in spermatogonial stem cells: the role of c-kit and its ligand SCF

Development ◽

10.1242/dev.127.10.2125 ◽

2000 ◽

Vol 127 (10) ◽

pp. 2125-2131 ◽

Cited By ~ 1

Author(s):

H. Ohta ◽

K. Yomogida ◽

K. Dohmae ◽

Y. Nishimune

Keyword(s):

Stem Cells ◽

Germ Cells ◽

Type A ◽

Spermatogonial Stem Cells ◽

Mutant Mice ◽

Seminiferous Tubules ◽

Kit Receptor ◽

Proliferation And Differentiation ◽

Cell Niche ◽

Type A Spermatogonia

To study self-renewal and differentiation of spermatogonial stem cells, we have transplanted undifferentiated testicular germ cells of the GFP transgenic mice into seminiferous tubules of mutant mice with male sterility, such as those dysfunctioned at Steel (Sl) locus encoding the c-kit ligand or Dominant white spotting (W) locus encoding the receptor c-kit. In the seminiferous tubules of Sl/Sl(d) or Sl(17H)/Sl(17H) mice, transplanted donor germ cells proliferated and formed colonies of undifferentiated c-kit (−) spermatogonia, but were unable to differentiate further. However, these undifferentiated but proliferating spermatogonia, retransplanted into Sl (+) seminiferous tubules of W mutant, resumed differentiation, indicating that the transplanted donor germ cells contained spermatogonial stem cells and that stimulation of c-kit receptor by its ligand was necessary for maintenance of differentiated type A spermatogonia but not for proliferation of undifferentiated type A spermatogonia. Furthermore, we have demonstrated that their transplantation efficiency in the seminiferous tubules of Sl(17H)/Sl(17H) mice depended upon the stem cell niche on the basement membrane of the recipient seminiferous tubules and was increased by elimination of the endogenous spermatogonia of mutant mice from the niche by treating them with busulfan.

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Expression of Col1a1, Col1a2 and procollagen I in germ cells of immature and adult mouse testis

Reproduction ◽

10.1530/rep.1.00694 ◽

2005 ◽

Vol 130 (3) ◽

pp. 333-341 ◽

Cited By ~ 29

Author(s):

Zuping He ◽

Lixin Feng ◽

Xiaodong Zhang ◽

Yixun Geng ◽

Daniela A Parodi ◽

...

Keyword(s):

Germ Cell ◽

Germ Cells ◽

Adult Mouse ◽

Type A ◽

Seminiferous Tubules ◽

Positive Staining ◽

Cellular Expression ◽

Old Mice ◽

Type A Spermatogonia

The objective of this study was to compare the expression of Col1a1, Col1a2, and procollagen I in the seminiferous tubules of immature and adult mice and to characterize the cellular expression pattern of procollagen I in germ cells during spermatogenesis in order to provide necessary groundwork for further functional studies in the process of spermatogenesis. Microarray analysis demonstrated that Col1a1 and Col1a2 were abundantly expressed in the seminiferous tubules of 6-day-old mice compared with 60-day-old mice, and the expression levels of Col1a1 and Col1a2 mRNA were validated using a semi-quantitative RT-PCR assay. Western blot analysis further confirmed that procollagen I was expressed at a higher level in the seminiferous tubules of 6-day-old mice compared with 60-day-old mice. Immunohistochemical analysis revealed that type A spermatogonia were positive for procollagen I in the testis of 6-day-old mice, whereas Sertoli cells were negative for this protein. Thein vivoprocollagen I staining in type A spermatogonia was corroborated in spermatogonia exhibiting a high potential for proliferation and the ability to form germ cell colonies inin vitroculture. Moreover, procollagen I was also detected in type A spermatogonia, intermediate spermatogonia, type B spermatogonia, and preleptotene spermatocytes in the adult mouse testes, but positive staining disappeared in more differentiated germ cell lineages detaching from the basement membrane, including leptotene spermatocytes, pachytene spermatocytes, round spermatids and elongated spermatids. These data suggest that Col1a1, Col1a2 and procollagen I are associated with type A spermatogonia and play a potential role in mediating the detachment and migration of germ cells during spermatogenesis.

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ISOLATION, PURIFICATION AND CRYOPRESERVATION OF TYPE A SPERMATOGONIA FROM PREPUBERTAL RAT TESTIS

European Urology Supplements ◽

10.1016/s1569-9056(08)60084-6 ◽

2008 ◽

Vol 7 (3) ◽

pp. 91 ◽

Cited By ~ 1

Author(s):

M. Amirchaghmaghi ◽

M. Sedighi Gilani

Keyword(s):

Type A ◽

Rat Testis ◽

Type A Spermatogonia

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Studies of Sl/Sld in equilibrium with +/+ mouse aggregation chimaeras. II. Effect of the steel locus on spermatogenesis

Development ◽

10.1242/dev.102.1.117 ◽

1988 ◽

Vol 102 (1) ◽

pp. 117-126 ◽

Cited By ~ 2

Author(s):

H. Nakayama ◽

H. Kuroda ◽

H. Onoue ◽

J. Fujita ◽

Y. Nishimune ◽

...

Keyword(s):

Mast Cells ◽

Germ Cell ◽

Germ Cells ◽

Cross Sections ◽

Sertoli Cells ◽

Type A ◽

Precursor Cells ◽

Intrinsic Defect ◽

Serial Sections ◽

Type A Spermatogonia

Mutant mice of Sl/Sld genotype are deficient in melanocytes, erythrocytes, mast cells and germ cells. Deficiency of melanocytes, erythrocytes and mast cells is not attributable to an intrinsic defect in their precursor cells but to a defect in the tissue environment that is necessary for migration, proliferation and/or differentiation. We investigated the mechanism of germ cell deficiency in male Sl/Sld mice by producing aggregation chimaeras from Sl/Sld and +/+ embryos. Chimaeric mice with apparent white stripes were obtained. Two of four such chimaeras were fertile and the phenotypes of resulting progenies showed that some Sl/Sld germ cells had differentiated into functioning sperms in the testis of the chimaeras. In cross sections of the testes of chimaeras, both differentiated and nondifferentiated tubules were observed. However, the proportions of type A spermatogonia to Sertoli cells in both types of tubules were comparable to the values observed in differentiated tubules of normal +/+ mice. We reconstructed the whole length of four tubules from serial sections. Differentiated and nondifferentiated segments alternated in a single tubule. The shortest differentiated segment contained about 180 Sertoli cells and the shortest nondifferentiated segment about 150 Sertoli cells. These results suggest that Sertoli cells of either Sl/Sld or +/+ genotype make discrete patches and that differentiation of type A spermatogonia does not occur in patches of Sl/Sld Sertoli cells.

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