Cell Interactions in the Reproductive System

Gametogenesis in both sexes involves a nurse-cell relationship between an epithelium and the developing gametes. In the female, a lifetime supply of oocytes is formed in the embryo and no new germ cells are formed postnatally. The granulosa cells of the ovarian follicles are believed to be responsible for maintaining the arrest of the oocytes in the fetal ovary in the dictyate stage of meiosis, in which stage they remain until shortly before ovulation many months or years later depending upon the species. Responding to gonadotropic hormones, the granulosa cells then terminate their inhibition, permit maturation of the oocyte, and participate in the reorganization of the follicle that is necessary for ovulation. In the male, the Sertoli cells which are also responsive to hormonal stimulation, maintain in the seminiferous epithelium a special microenvironment which is essential for germ cell differentiation. The cooperative motor activity and associated shape changes of these supporting cells is responsible for the adluminal translocation of the developing germ cells and for the separation of individual spermatozoa from long chains of interconnected cell bodies during sperm release. This paper will review the types and locations of the communicating and occluding junctions that are essential for the complex and highly integrated activities of the epithelia in the mammalian gonads.

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A close correlation in the expression patterns of Af-6 and Usp9x in Sertoli and granulosa cells of mouse testis and ovary

Reproduction ◽

10.1530/rep.1.00060 ◽

2004 ◽

Vol 128 (5) ◽

pp. 583-594 ◽

Cited By ~ 5

Author(s):

Takeshi Sato ◽

Yoshiakira Kanai ◽

Takashi Noma ◽

Masami Kanai-Azuma ◽

Shinichiro Taya ◽

...

Keyword(s):

Germ Cells ◽

Granulosa Cells ◽

Sertoli Cells ◽

Translational Regulation ◽

Expression Patterns ◽

Intracellular Localization ◽

Close Correlation ◽

Cell Junction ◽

Supporting Cells ◽

Junction Protein

Usp9x, an X-linked deubiquitylating enzyme, is stage dependently expressed in the supporting cells (i.e. Sertoli cells and granulosa cells) and germ cells during mouse gametogenesis. Af-6, a cell junction protein, has been identified as a substrate of Usp9x, suggesting a possible association between Usp9x and Af-6 in spermatogenesis and oogenesis. In this study, we examined the expression pattern of Af-6 and Usp9x and their intracellular localization in testes and ovaries of mice treated with or without pregnant mare serum gonadotropin (PMSG), an FSH-like hormone. In both testes and ovaries, Af-6 expression was predominantly observed in supporting cells, as well as in steroidogenic cells, but not in any germ cells. In Sertoli cells, Af-6 was continuously expressed throughout postnatal and adult stages, where both Af-6 and Usp9x were enriched at the sites of Sertoli–Sertoli and Sertoli–spermatid junctions especially at stages XI–VI. In the granulosa cells, Af-6, as well as Usp9x, was highly expressed in primordial and primary follicles, but its expression rapidly decreased after the late-secondary follicle stage. Interestingly, in PMSG-treated mice, the expression levels of Af-6 and Usp9x were synchronously enhanced, slightly in Sertoli cells and strongly in granulosa cells of the late-secondary and Graafian follicles. Such closely correlated expression patterns between Af-6 and Usp9x clearly suggest that Af-6 may be deubiquitylated by Usp9x in both Sertoli and granulosa cells. It further suggests that the post-translational regulation of Af-6 by Usp9x may be one potential pathway to control the cell adhesion dynamics in mammalian gametogenesis.

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Cell-type-specific regulation of genes involved in testicular lipid metabolism: fatty acid-binding proteins, diacylglycerol acyltransferases, and perilipin 2

Reproduction ◽

10.1530/rep-13-0199 ◽

2013 ◽

Vol 146 (5) ◽

pp. 471-480 ◽

Cited By ~ 16

Author(s):

Gerardo M Oresti ◽

Jesús García-López ◽

Marta I Aveldaño ◽

Jesús del Mazo

Keyword(s):

Fatty Acid ◽

Cell Differentiation ◽

Germ Cell ◽

Germ Cells ◽

Sertoli Cells ◽

Fatty Acid Binding Proteins ◽

Cell Type ◽

Fatty Acid Binding ◽

Germ Cell Differentiation ◽

Perilipin 2

Male germ cell differentiation entails the synthesis and remodeling of membrane polar lipids and the formation of triacylglycerols (TAGs). This requires fatty acid-binding proteins (FABPs) for intracellular fatty acid traffic, a diacylglycerol acyltransferase (DGAT) to catalyze the final step of TAG biosynthesis, and a TAG storage mode. We examined the expression of genes encoding five members of the FABP family and two DGAT proteins, as well as the lipid droplet protein perilipin 2 (PLIN2), during mouse testis development and in specific cells from seminiferous epithelium.Fabp5expression was distinctive of Sertoli cells and consequently was higher in prepubertal than in adult testis. The expression ofFabp3increased in testis during postnatal development, associated with the functional differentiation of interstitial cells, but was low in germ cells.Fabp9, together withFabp12, was prominently expressed in the latter. Their transcripts increased from spermatocytes to spermatids and, interestingly, were highest in spermatid-derived residual bodies (RB). Both Sertoli and germ cells, which produce neutral lipids and store them in lipid droplets, expressedPlin2. Yet, whileDgat1was detected in Sertoli cells,Dgat2accumulated in germ cells with a similar pattern of expression asFabp9. These results correlated with polyunsaturated fatty acid-rich TAG levels also increasing with mouse germ cell differentiation highest in RB, connecting DGAT2 with the biosynthesis of such TAGs. The age- and germ cell type-associated increases inFabp9,Dgat2, andPlin2levels are thus functionally related in the last stages of germ cell differentiation.

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RUNX1 maintains the identity of the fetal ovary through an interplay with FOXL2

Nature Communications ◽

10.1038/s41467-019-13060-1 ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 8

Author(s):

Barbara Nicol ◽

Sara A. Grimm ◽

Frédéric Chalmel ◽

Estelle Lecluze ◽

Maëlle Pannetier ◽

...

Keyword(s):

Transcription Factor ◽

Granulosa Cell ◽

Sertoli Cells ◽

Transcriptional Control ◽

Cell Lineage ◽

Supporting Cell ◽

Supporting Cells ◽

Fate Specification ◽

Fetal Ovary

Abstract Sex determination of the gonads begins with fate specification of gonadal supporting cells into either ovarian pre-granulosa cells or testicular Sertoli cells. This fate specification hinges on a balance of transcriptional control. Here we report that expression of the transcription factor RUNX1 is enriched in the fetal ovary in rainbow trout, turtle, mouse, goat, and human. In the mouse, RUNX1 marks the supporting cell lineage and becomes pre-granulosa cell-specific as the gonads differentiate. RUNX1 plays complementary/redundant roles with FOXL2 to maintain fetal granulosa cell identity and combined loss of RUNX1 and FOXL2 results in masculinization of fetal ovaries. At the chromatin level, RUNX1 occupancy overlaps partially with FOXL2 occupancy in the fetal ovary, suggesting that RUNX1 and FOXL2 target common sets of genes. These findings identify RUNX1, with an ovary-biased expression pattern conserved across species, as a regulator in securing the identity of ovarian-supporting cells and the ovary.

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A novel Amh-Treck transgenic mouse line allows toxin-dependent loss of supporting cells in gonads

Reproduction ◽

10.1530/rep-14-0171 ◽

2014 ◽

Vol 148 (6) ◽

pp. H1-H9 ◽

Cited By ~ 8

Author(s):

Mai Shinomura ◽

Kasane Kishi ◽

Ayako Tomita ◽

Miyuri Kawasumi ◽

Hiromi Kanezashi ◽

...

Keyword(s):

Germ Cells ◽

Granulosa Cells ◽

Sertoli Cells ◽

Seminiferous Tubules ◽

Specific Cell ◽

Partial Recovery ◽

Dependent Manner ◽

Mouse Line ◽

Cell Degeneration

Cell ablation technology is useful for studying specific cell lineages in a developing organ in vivo. Herein, we established a novel anti-Müllerian hormone (AMH)-toxin receptor-mediated cell knockout (Treck) mouse line, in which the diphtheria toxin (DT) receptor was specifically activated in Sertoli and granulosa cells in postnatal testes and ovaries respectively. In the postnatal testes of Amh-Treck transgenic (Tg) male mice, DT injection induced a specific loss of the Sertoli cells in a dose-dependent manner, as well as the specific degeneration of granulosa cells in the primary and secondary follicles caused by DT injection in Tg females. In the testes with depletion of Sertoli cell, germ cells appeared to survive for only several days after DT treatment and rapidly underwent cell degeneration, which led to the accumulation of a large amount of cell debris within the seminiferous tubules by day 10 after DT treatment. Transplantation of exogenous healthy Sertoli cells following DT treatment rescued the germ cell loss in the transplantation sites of the seminiferous epithelia, leading to a partial recovery of the spermatogenesis. These results provide not only in vivo evidence of the crucial role of Sertoli cells in the maintenance of germ cells, but also show that the Amh-Treck Tg line is a useful in vivo model of the function of the supporting cell lineage in developing mammalian gonads.

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1 XENOGRAFTING OF ADULT MAMMALIAN TESTIS TISSUE

Reproduction Fertility and Development ◽

10.1071/rdv19n1ab1 ◽

2007 ◽

Vol 19 (1) ◽

pp. 119

Author(s):

L. Arregui ◽

R. Rathi ◽

W. Zeng ◽

A. Honaramooz ◽

M. Gomendio ◽

...

Keyword(s):

Cell Differentiation ◽

Germ Cell ◽

Germ Cells ◽

Sertoli Cells ◽

Human Testis ◽

Seminiferous Tubules ◽

Donor Tissue ◽

Germ Cell Differentiation ◽

Sexually Mature ◽

Testis Tissue

Testis tissue grafting presents an option for preservation of genetic material when sperm recovery is not possible. Grafting of testis tissue from sexually immature males to immunodeficient mice results in germ cell differentiation and production of fertilization-competent sperm from different mammalian species (Honaramooz et al. 2002 Nature 418, 778–781). However, the efficiency of testis tissue xenografting from adult donors has not been critically evaluated. Spermatogenesis was arrested at meiosis in grafts from mature horses (Rathi et al. 2006 Reproduction 131, 1091–1098) and hamsters (Schlatt et al. 2002 Reproduction 124, 339–346), and no germ cell differentiation occurred in xenografts of adult human testis tissue (Schlatt et al. 2006 Hum. Reprod. 21, 384–389). The objective of this study was to investigate survival and germ cell differentiation of testis xenografts from sexually mature donors of different species. Small fragments of testis tissue from 10 donor animals of 5 species were grafted under the back skin of immunodeficient, castrated male mice (n = 37, 2–6/donor). Donors were pig (8 months old), goat (18 months old and 4 years old) (n = 2), bull (3 years old), donkey (13 months old), and rhesus monkey (3, 6, 11, and 12 years old). At the time of grafting, donor tissue contained elongated spermatids, albeit to different degrees (>75% of seminiferous tubules in testis tissue from pig, goat, bull, and 6–12-year-old monkeys, and 33 or 66% of tubules in tissue from donkey or 3-year-old monkey, respectively). Grafts were recovered <12 weeks (n = 14 mice), 12–24 weeks (n = 16 mice), and >24 weeks (n = 7 mice) after grafting and classified histologically as completely degenerated (no tubules found), degenerated tubules (only hyalinized seminiferous tubules observed), or according to the most advanced type of germ cell present. Grafts from pig, goat, bull, and 6–12-year-old monkeys contained >60% degenerated tubules or were completely degenerated at all time points analyzed. In contrast, in grafts from the 3-year-old monkey, only 18% of tubules were degenerated, 14% contained Sertoli cells only, 64% contained meiotic, and 4% haploid germ cells at 24 weeks after grafting. Similarly, donkey testis grafts recovered 12–24 weeks after grafting contained <2% degenerated tubules, 46% of tubules had Sertoli cells only, 45% contained meiotic, and 7% haploid germ cells. These results show that survival and differentiation of germ cells in testis grafts from sexually mature mammalian donors is poor. However, better graft survival and maintenance of spermatogenesis occurred in donor tissue from donkey and 3-year-old monkey that were less mature at the time of grafting. Therefore, species and age-related differences appear to exist with regard to germ cell survival and differentiation in xenografts from adult donors. This work was supported by USDA/CSREES 03-35203-13486, NIH/NCRR 5-R01-RR17359-05, the Spanish Ministry of Education, and Science (BES-2004-4112).

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Structure and Function of Sertoli Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100090063 ◽

1976 ◽

Vol 34 ◽

pp. 16-17

Author(s):

M. Dym

Keyword(s):

Germ Cells ◽

Sertoli Cells ◽

Structure And Function ◽

Seminiferous Epithelium ◽

Cell Type ◽

Germ Cell Differentiation ◽

Full Control ◽

Tubule Lumen ◽

Strategic Position ◽

And Function

The Sertoli cells perform an impressive array of functions in the testis. It is possible that the full control of germ cell differentiation is mediated by this elaborate cell type (Fig. 1). On the basis of its shape and strategic position within the seminiferous epithelium the functions of (1) support and nutrition have been assigned. Fawcett and Phillips (J. Reprod. Fert. 6: 405, 1969) demonstrated that the Sertoli cells engineer the (2) release of late spermatids into the tubule lumen; other data suggest that they are instrumental in the migration of the germ cells from the basal lamina to the lumen. Tight junctions between adjacent Sertoli cells subdivide the seminiferous epithelium into two compartments, basal and adluminal. These junctions form the (3) morphological basis of the blood-testis barrier . The Sertoli cells are capable of (4) phagocytizing vast numbers of degenerating germ cells and sperm residual bodies.

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Neurotropins and Their Receptors Are Expressed in the Human Fetal Ovary

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jcem.87.2.8221 ◽

2002 ◽

Vol 87 (2) ◽

pp. 890-897 ◽

Cited By ~ 38

Author(s):

Richard A. Anderson ◽

Lynne L. L. Robinson ◽

Julie Brooks ◽

Norah Spears

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Germ Cells ◽

Granulosa Cells ◽

Ovarian Development ◽

Growth Factor Receptor ◽

Primordial Follicle ◽

Receptor Protein ◽

Nerve Growth ◽

Fetal Ovary

Mammalian ovarian development is characterized by a sequential pattern of mitotic proliferation of oogonia, initiation then arrest of meiosis, and primordial follicle formation. The factors regulating these processes are poorly understood. The neurotropins are survival and differentiation factors in the nervous system, acting via high affinity receptors of the trk protooncogene family and the low affinity p75 nerve growth factor receptor, and have also been described in the rodent ovary, where changes in NT4/TrkB gene expression have been detected at the time of primordial follicle formation. There are no data on neurotropin expression in the normal human ovary. We have investigated the expression and localization of neurotropins and their receptors in the midtrimester human fetal ovary (13–21 wk gestation). Expression of mRNA for neurotropins and their receptors was detected by RT-PCR. Clusters of oogonia were found to be the predominant site of NT4 mRNA expression using in situ hybridization. However, at later gestations granulosa cells of primordial follicles showed increased expression, with lesser expression in the enclosed oocytes. NT4 protein was also localized to the granulosa cells by immunohistochemistry and at earlier developmental stages to epithelioid cells, which were mingled with clusters of oogonia not expressing NT4. TrkB receptor protein was localized by immunohistochemistry to germ cells at all gestations examined. The p75 nerve growth factor receptor protein was exclusively expressed in the ovarian stroma. These data demonstrate the expression of neurotropins and their receptors within the human fetal ovary. Developmental changes in the pattern of expression of NT4 around the time of primordial follicle formation suggest that neurotropins may be involved in signaling between somatic cells and germ cells at this crucial stage of ovarian development.

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Sertoli-secreted FGF-2 induces PFKFB4 isozyme expression in mouse spermatogenic cells by activation of the MEK/ERK/CREB pathway

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00381.2011 ◽

2012 ◽

Vol 303 (6) ◽

pp. E695-E707 ◽

Cited By ~ 10

Author(s):

Marta Gómez ◽

Anna Manzano ◽

Agnes Figueras ◽

Francesc Viñals ◽

Francesc Ventura ◽

...

Keyword(s):

Gene Expression ◽

Conditioned Medium ◽

Cell Lines ◽

Germ Cells ◽

Sertoli Cells ◽

Hormonal Regulation ◽

Neutralizing Antibodies ◽

Spermatogenic Cell ◽

Hormonal Stimulation ◽

Paracrine Regulation

Sertoli cells play a central role in the control and maintenance of spermatogenesis by secreting growth factors, in response to hormonal stimulation, that participate in the paracrine regulation of this process. In this study, we investigated how the hormonal regulation of spermatogenesis modulates 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB) isozyme expression in two mouse spermatogenic cell lines, GC-1 spg and GC-2 spd (ts). For this purpose, TM4 Sertoli cells were used to obtain conditioned medium that was treated or not with dihydrotestosterone for 2 days [dihydrotestosterone conditioned medium (TCM) and basal conditioned medium (BCM), respectively]. We observed an increase in the expression of PFKFB4 along with a decrease in PFKFB3 in spermatogenic cell lines treated with TCM. These effects were inhibited by the antiandrogen drug flutamide and by heat-inactivated TCM, indicating the protein nature of the TCM mediator and its dependence on Sertoli cell stimulation by dihydrotestosterone. In addition, adult rat testes treated with the GnRH antagonist Degarelix exhibited a reduction in the expression of PFKFB4 in germ cells. Addition of exogenous FGF-2 mimicked the changes in the Pfkfb gene expression, whereas neutralizing antibodies against FGF-2 abolished them. Interestingly, similar effects on Pfkfb gene expression were observed using different MAPK inhibitors (U-0126, PD-98059, and H-89). Luciferase analysis of Pfkfb4 promoter constructs demonstrated that a putative CRE-binding sequence located at −1,463 relative to the transcription start site is required to control Pfkfb4 gene expression after TCM treatment. Pulldown assays showed the binding of the CREB transcription factor to this site. Altogether, these results show how the paracrine regulation orchestrated by Sertoli cells in response to testosterone controls glycolysis in germ cells.

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The rete testis harbors Sertoli-like cells capable of expressing DMRT1

Reproduction ◽

10.1530/rep-19-0183 ◽

2019 ◽

Vol 158 (5) ◽

pp. 399-413 ◽

Cited By ~ 1

Author(s):

Ekaterina A Malolina ◽

Andrey Yu Kulibin

Keyword(s):

Germ Cells ◽

Sertoli Cells ◽

Adult Mouse ◽

Rete Testis ◽

Seminiferous Tubules ◽

Rna Seq ◽

Supporting Cells ◽

Testicular Cells ◽

Qpcr Analysis ◽

Different Levels

Sertoli cells (SCs) are supporting cells in the mammalian testis that proliferate throughout fetal and postnatal development but exit the cell cycle and differentiate at puberty. In our previous study, we isolated a population of highly proliferative Sertoli-like cells (SLCs) from the region of the adult mouse testis containing the rete testis and adjacent seminiferous tubules. Here RNA-seq of the adult SLC culture as well as qPCR analysis and immunofluorescence of the adult and immature (6 dpp) SLC cultures were performed that allowed us to identify SLC-specific genes, including Pax8, Cdh1, and Krt8. Using these, we found that SLCs are mostly localized in the rete testis epithelium; however, some contribution of transitional zones of seminiferous tubules could not be excluded. The main feature of SLCs indicating their relationship to SCs is DMRT1 expression. More than 40% of both adult and immature SLCs expressed DMRT1 at different levels in culture. Only rare DMRT1+ cells were detected in the adult rete testis, whereas more than 40% of cells were positively stained for DMRT1 in the immature rete testis. One more SC protein, AMH, was found in some rete cells of the immature testis. It was also demonstrated that SLCs expressed such SC genes as Nr5a1, Dhh, Gdnf, and Kitl and interacted with germ cells in 3D co-culture with immature testicular cells. All these similarities between SLCs and rete cells on one the hand and SCs on the other, suggest that rete cells could share a common origin with SCs.

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318 EFFECT OF 6-N-PROPYL-2-THIOURACIL-INDUCED HYPOTHYROIDISM IN THE HOST MOUSE ON THE DEVELOPMENT OF BOVINE TESTIS XENOGRAFTS

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab318 ◽

2010 ◽

Vol 22 (1) ◽

pp. 315

Author(s):

J. R. Rodriguez-Sosa ◽

G. M. J. Costa ◽

R. Rathi ◽

L. R. França ◽

I. Dobrinski

Keyword(s):

Cell Differentiation ◽

Germ Cell ◽

Sertoli Cell ◽

Germ Cells ◽

Sertoli Cells ◽

Cell Maturation ◽

Germ Cell Differentiation ◽

Host Mouse ◽

Thyroid Hormone Levels ◽

Collection Time

In rodents, thyroid hormones inhibit Sertoli cell proliferation, promote Sertoli cell differentiation, and accelerate lumen formation in the seminiferous tubules. Conversely, transient hypothyroidism prolongs Sertoli cell proliferation, leading to increased Sertoli cell number and testicular size. In order to evaluate whether 6-N-propyl-2-thiouracil (PTU)-induced hypothyroidism in the host mouse would affect seminiferous tubule development and germ cell differentiation, and subsequently increase spermatogenesis in bovine testis xenografts, fragments (∼1 mm3) of testes from 1-wk-old Holstein calves (n = 6) were transplanted ectopically to castrated immunodeficient male mice (n = 6/donor). Mice (n = 3/donor) were treated with 0.1% (w/v) PTU in drinking water for 4 weeks or left as control. At 5 and 7 months after grafting, grafts were analyzed by morphometry and immunohistochemistry for expression of protein gene product 9.5 (PGP 9.5) as a germ cell marker, and Mullerian-inhibiting substance (MIS) and androgen receptor (AR) to assess Sertoli cell maturation. For each variable, averages of each group were compared at each collection point by t-test PTU treatment to the drinking water for 1 month suppressed thyroid hormone levels (T4) in host mice without negative systemic effects (0.3 ± 0.2 v. 4 ± 0.3 μg dL-1 at 4 weeks in treated v. control mice, respectively, P < 0.05). Spermatogenesis in recovered grafts was arrested at meiosis regardless of treatment and collection time. Graft weight was lower in treated mice than in controls (21 ± 4 v. 42 ± 5 and 24 ± 9 v. 51 ± 5 mg, at 5 and 7 months, respectively, P < 0.05). Volume density of the tubular and intertubular compartments, and seminiferous epithelium, was not affected by treatment (P > 0.05); however, treatment reduced lumen density compared to controls (9 ± 2 v. 19 ± 3 and 12 ± 1 v. 24 ± 4%) and tubular diameter (121 ± 3 v. 140 ± 7 and 144 ± 2v. 170 ± 2 (im, at 5 and 7 months, respectively (P < 0.05). Tubule length per milligram was not different at 5 months between control and treated groups (P > 0.05) but was increased at 7 months in the treated grafts (50 ± 1 v. 30 ± 1 cm, P < 0.05). Number of Sertoli cells per milligram was not affected by treatment (P > 0.05). However, Sertoli cell volume was increased in controls (440 ± 19 v. 341 ± 14 and 504 ± 6 v. 388 ± 18 μm3, at 5 and 7 months, respectively, P < 0.05). The number of germ cells per 100 Sertoli cells was not different between groups at any collection time (P > 0.05). Sertoli cells showed variable MIS expression and lack of or weak AR expression regardless of treatment and collection time, indicating an immature phenotype. In conclusion, suppression of thyroid hormone levels in host mice affects seminiferous tubule development in bovine testis xenografts, demonstrating that endocrine manipulation of the mouse host will affect xenografts in a predictable manner. However, treatment did not affect number and differentiation of germ cells. Rather, incomplete Sertoli cell maturation appears to lead to incomplete germ cell differentiation in bovine testis xenografts. Supported by USDA (2007-35203-18213).

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