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

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.

scholarly journals Ferredoxin 1b Deficiency Leads to Testis Disorganization, Impaired Spermatogenesis, and Feminization in Zebrafish

Endocrinology ◽  
2019 ◽  
Vol 160 (10) ◽  
pp. 2401-2416 ◽  
Author(s):  
James A Oakes ◽  
Nan Li ◽  
Belinda R C Wistow ◽  
Aliesha Griffin ◽  
Lise Barnard ◽  
...  
Keyword(s):  
Sex Differentiation ◽  
Reproductive Function ◽  
Gonadal Development ◽  
Sex Characteristics ◽  
Vitro Fertilization ◽  
Sex Development ◽  
Receptor Mutant ◽  
Predominantly Female ◽  
And Function

Abstract The roles of steroids in zebrafish sex differentiation, gonadal development, and function of the adult gonad are poorly understood. Herein, we used ferredoxin 1b (fdx1b) mutant zebrafish to explore such processes. Fdx1b is an essential electron-providing cofactor to mitochondrial steroidogenic enzymes, which are crucial for glucocorticoid and androgen production in vertebrates. Fdx1b−/− zebrafish mutants develop into viable adults in which concentrations of androgens and cortisol are significantly reduced. Adult fdx1b−/− mutant zebrafish display predominantly female secondary sex characteristics but may possess either ovaries or testes, confirming that androgen signaling is dispensable for testicular differentiation in this species, as previously demonstrated in androgen receptor mutant zebrafish. Adult male fdx1b−/− mutant zebrafish exhibit reduced characteristic breeding behaviors and impaired sperm production, resulting in infertility in standard breeding scenarios. However, eggs collected from wild-type females can be fertilized by the sperm of fdx1b−/− mutant males by in vitro fertilization. The testes of fdx1b−/− mutant males are disorganized and lack defined seminiferous tubule structure. Expression of several promale and spermatogenic genes is decreased in the testes of fdx1b−/− mutant males, including promale transcription factor sox9a and spermatogenic genes igf3 and insl3. This study establishes an androgen- and cortisol-deficient fdx1b zebrafish mutant as a model for understanding the effects of steroid deficiency on sex development and reproductive function. This model will be particularly useful for further investigation of the roles of steroids in spermatogenesis, gonadal development, and regulation of reproductive behavior, thus enabling further elucidation of the physiological consequences of endocrine disruption in vertebrates.

scholarly journals Propagation of bovine spermatogonial stem cells in vitro

Reproduction ◽  
2008 ◽  
Vol 136 (5) ◽  
pp. 543-557 ◽  
Author(s):  
Pedro M Aponte ◽  
Takeshi Soda ◽  
Katja J Teerds ◽  
S Canan Mizrak ◽  
Henk J G van de Kant ◽  
...  
Keyword(s):  
Stem Cells ◽  
Growth Factor ◽  
Cultured Cells ◽  
Somatic Cells ◽  
Type A ◽  
Spermatogonial Stem Cells ◽  
Seminiferous Tubules ◽  
Type A Spermatogonia ◽  
Stimulation Of

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.

scholarly journals Types, norms, and normalisation: Hormone research and treatments in Italy, Argentina, and Brazil, c. 1900–50

2020 ◽  
pp. 095269512094119
Author(s):  
Chiara Beccalossi
Keyword(s):  
Latin American ◽  
Medical Science ◽  
Physiological Model ◽  
The Body ◽  
Sex Characteristics ◽  
Medical Doctors ◽  
Secondary Sexual Characteristics ◽  
Hormone Research ◽  
Sexual Characteristics ◽  
Hormone Therapies

Displacing the physiological model that had held sway in 19th-century medical thinking, early 20th-century hormone research promoted an understanding of the body and sexual desires in which variations in sex characteristics and non-reproductive sexual behaviours such as homosexuality were attributed to anomalies in the internal secretions produced by the testes or the ovaries. Biotypology, a new brand of medical science conceived and led by the Italian endocrinologist Nicola Pende, employed hormone research to study human types and hormone treatments to normalise individuals who did not conform to accepted medical norms. Latin American medical doctors, eugenicists, and sexologists took up biotypology with enthusiasm. This article considers the case studies of Italy, Argentina, and Brazil, and analyses the work of medical doctors who adopted a biotypological mode of reasoning and employed to various extents hormone therapies in their practice. By focusing on hormone therapies that aimed to normalise secondary sexual characteristics and the sexual instinct, the article suggests that while the existence of normality was contested to the point that a number of medical scientists argued that no such thing existed, the pursuit of normality was carried out in very practical terms through the new medical technologies hormone research had introduced.

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

Endocrinology ◽  
2008 ◽  
Vol 149 (6) ◽  
pp. 2773-2781 ◽  
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.

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

Reproduction ◽  
2003 ◽  
pp. 669-680 ◽  
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.

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

1962 ◽  
Vol 10 (2) ◽  
pp. 178 ◽  
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.

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

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).

Dislocated type-A spermatogonia in human seminiferous tubules

1984 ◽  
Vol 236 (1) ◽  
pp. 35-40 ◽  
Author(s):  
A.F. Holstein ◽  
E. Bustos-Obreg�n ◽  
M. Hartmann
Keyword(s):  
Type A ◽  
Seminiferous Tubules ◽  
Type A Spermatogonia

scholarly journals Distribution of GFRA1-expressing spermatogonia in adult mouse testis

Reproduction ◽  
2012 ◽  
Vol 143 (3) ◽  
pp. 325-332 ◽  
Author(s):  
Margherita Grasso ◽  
Andrea Fuso ◽  
Lisa Dovere ◽  
Dirk G de Rooij ◽  
Mario Stefanini ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adult Mouse ◽  
Pcr Analysis ◽  
Seminiferous Tubules ◽  
Self Renewal ◽  
Expression Of Genes ◽  
Specific Manner ◽  
A Cell ◽  
And Function ◽  
Type A Spermatogonia

In mice and other mammals, spermatogenesis is maintained by spermatogonial stem cells (SSCs), a cell population belonging to undifferentiated type A spermatogonia. In the accepted model of SSC self-renewal, Asingle (As) spermatogonia are the stem cells, whereas paired (Apaired (Apr)) and chained (Aaligned (Aal)) undifferentiated spermatogonia are committed to differentiation. This model has been recently challenged by evidence that As and chained (Apr and Aal), undifferentiated spermatogonia are heterogeneous in terms of gene expression and function. The expression profile of several markers, such as GFRA1 (the GDNF co-receptor), is heterogeneous among As, Apr and Aal spermatogonia. In this study, we have analysed and quantified the distribution of GFRA1-expressing cells within the different stages of the seminiferous epithelial cycle. We show that in all stages, GFRA1+ chained spermatogonia (Apr to Aal) are more numerous than GFRA1+ As spermatogonia. Numbers of chained GFRA1+ spermatogonia are sharply reduced in stages VII–VIII when Aal differentiate into A1 spermatogonia. GFRA1 expression is regulated by GDNF and in cultures of isolated seminiferous tubules, we found that GDNF expression and secretion by Sertoli cells is stage-dependent, being maximal in stages II–VI and decreasing thereafter. Using qRT-PCR analysis, we found that GDNF regulates the expression of genes such as Tex14, Sohlh1 and Kit (c-Kit) known to be involved in spermatogonial differentiation. Expression of Kit was upregulated by GDNF in a stage-specific manner. Our data indicate that GDNF, besides its crucial role in the self-renewal of stem cells also functions in the differentiation of chained undifferentiated spermatogonia.

scholarly journals Management of the patients with stage I germ cell ovarian tumors

2019 ◽  
Vol 21 (2) ◽  
pp. 37-39
Author(s):  
Innesa V Nechushkina ◽  
Valentina M Nechushkina ◽  
Elena I Boychenko ◽  
Natalia A Susuleva
Keyword(s):  
Germ Cell ◽  
Ovarian Function ◽  
Disease Risk ◽  
Germ Cell Tumors ◽  
Reproductive Function ◽  
Ovarian Tumors ◽  
Stage I ◽  
Secondary Sexual Characteristics ◽  
Sexual Characteristics ◽  
Single Ovary

The goal of the treatment of children and adolescents with germ cell ovarian tumors is to save both life and its quality. One of the quality of life issue in pediatric patients is fertility preservation. The removal of the ovary on the affected side allows saving the ovary on the opposite side. The single ovary is able to save further full development of female secondary sexual characteristics and will play important role as psychological value. In addition, the reproductive function will be stored. Chemotherapy may badly impact the function of a single ovary. To find out in advance how chemotherapy will affect ovarian function is impossible, so it is necessary for patients with Stage I ovarian germ cell tumors to decide the possibility of dismissing chemotherapy from the treatment. Under certain conditions unilateral salpingo-oophorectomy may be sufficient for stage I disease. Risk factors which should be considered when refusing chemotherapy are discussed.

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