scholarly journals Prenatal and lactation nicotine exposure affects Sertoli cell and gonadotropin levels in rats

Reproduction ◽  
2016 ◽  
Vol 151 (2) ◽  
pp. 117-133 ◽  
Author(s):  
C C Paccola ◽  
S M Miraglia
Keyword(s):  
Epithelial Cells ◽  
Germ Cell ◽  
Sertoli Cells ◽  
Hormone Secretion ◽  
Seminiferous Epithelium ◽  
Nicotine Exposure ◽  
Adult Rats ◽  
Pregnant Rats ◽  
Tubular Lumen ◽  
Hormonal Analysis

Nicotine is largely consumed in the world as a component of cigarettes. It can cross the placenta and reach the milk of smoking mothers. This drug induces apoptosis, affects sex hormone secretion, and leads to male infertility. To investigate the exposure to nicotine during the whole intrauterine and lactation phases in Sertoli cells, pregnant rats received nicotine (2 mg/kg per day) through osmotic minipumps. Male offsprings (30, 60, and 90 days old) had blood collected for hormonal analysis (FSH and LH) and their testes submitted for histophatological study, analysis of the frequency of the stages of seminiferous epithelium cycle, immunolabeling of apoptotic epithelial cells (TUNEL and Fas/FasL), analysis of the function and structure of Sertoli cells (respectively using transferrin and vimentin immunolabeling), and analysis of Sertoli-germ cell junctional molecule (β-catenin immunolabeling). The exposure to nicotine increased the FSH and LH plasmatic levels in adult rats. Although nicotine had not changed the number of apoptotic cells, neither in Fas nor FasL expression, it provoked an intense sloughing of epithelial cells and also altered the frequency of some stages of the seminiferous epithelium cycle. Transferrin and β-catenin expressions were not changed, but vimentin was significantly reduced in the early stages of the seminiferous cycle of the nicotine-exposed adult rats. Thus, we concluded that nicotine exposure during all gestational and lactation periods affects the structure of Sertoli cells by events causing intense germ cell sloughing observed in the tubular lumen and can compromise the fertility of the offspring.

Correlation between blood-testis barrier development and onset of the first spermatogenic wave in normal and busulfan-treated rats

1990 ◽  
Vol 48 (3) ◽  
pp. 78-79
Author(s):  
Juan C. Cavicchia ◽  
Fabio L. Sacerdote
Keyword(s):  
Germ Cell ◽  
Tight Junctions ◽  
Basal Lamina ◽  
Sertoli Cells ◽  
Freeze Fracture ◽  
Seminiferous Epithelium ◽  
Seminiferous Tubules ◽  
Pregnant Rats ◽  
Synaptonemal Complexes ◽  
New Born

Pregnant rats (day 13) received 10mg/kg of Busulfan i.p. in order to deplete of germ cell the testes of the new born rats. The seminiferous tubules of their off spring from postnatal age 1 day up to day 35 were examined with TEM after fixation plus intercellular tracers, and with freeze-fracture techniques. During this period, the inter-Sertoli tight junctions of controls increase both in numbers and in length. Between days 10 and 13 the seminiferous cords have numerous preleptotene and leptotene spermatocytes (Fig.1) surrounded by tracer. The inter-Sertoli junctions are tortuous and predominantly perpendicular to the basal lamina. Between ages 13 and 20 days the seminiferous epithelium reaches zygotene-pachytene stages (fig.2) identified by the presence of synaptonemal complexes. The tracer is stopped at the inter-Sertoli junctions at this stage, whereas it still permeates tubules displaying preleptotene and leptotene spermatocytes.Freeze-fracture shows that the orientation of inter-Sertoli junctions has changed to parallel, both to each other and to the basal lamina (fig.3). In the Busulfan treated rats, the tubules continue having, up to postnatal day 30, only Sertoli cells and scanty spermatogonia.

The murine seminiferous epithelial cycle is pre-figured in the Sertoli cells of the embryonic testis

Development ◽  
2002 ◽  
Vol 129 (3) ◽  
pp. 635-647 ◽  
Author(s):  
Paula M. Timmons ◽  
Peter W. J. Rigby ◽  
Françoise Poirier
Keyword(s):  
Germ Cell ◽  
Sertoli Cells ◽  
Cell Function ◽  
Seminiferous Epithelium ◽  
Developmental Origins ◽  
Sperm Production ◽  
Functional Heterogeneity ◽  
Adult Testis ◽  
Germ Cell Apoptosis ◽  
Cell Lineages

The seminiferous epithelial cycle and spermatogenic wave are conserved features of vertebrate spermatogenic organisation that reflect the need for the rigorous maintenance of sperm production. Although the cycle and the wave of the adult seminiferous epithelium have been well characterised, particularly in rodent species, their developmental origins are unknown. We show that the Sertoli cells of the pre-pubertal mouse, including those of the germ cell-deficient XXSxra mutant, exhibit coordinated, cyclical patterns of gene expression, presaging the situation in the adult testis, where Sertoli cell function is coupled to the spermatogenic cycle. In the case of the galectin 1 gene (Lgals1), localised differential expression in the Sertoli cells can be traced back to neonatal and embryonic stages, making this the earliest known molecular marker of functional heterogeneity in mammalian testis cords. In addition, the timing of germ cell apoptosis in normal pre-pubertal testes is linked to the temporal cycle of the Sertoli cells. These data show that the cycle and wave of the murine seminiferous epithelium originate at a much earlier stage in development than was previously known, and that their maintenance in the early postnatal cords depends exclusively on the somatic cell lineages.

scholarly journals Dynein 1 supports spermatid transport and spermiation during spermatogenesis in the rat testis

2018 ◽  
Vol 315 (5) ◽  
pp. E924-E948 ◽  
Author(s):  
Qing Wen ◽  
Elizabeth I. Tang ◽  
Wing-yee Lui ◽  
Will M. Lee ◽  
Chris K. C. Wong ◽  
...  
Keyword(s):  
Germ Cell ◽  
Heavy Chain ◽  
Sertoli Cells ◽  
Cytoplasmic Dynein ◽  
Seminiferous Epithelium ◽  
Organelle Transport ◽  
Cellular Events

In the mammalian testis, spermatogenesis is dependent on the microtubule (MT)-specific motor proteins, such as dynein 1, that serve as the engine to support germ cell and organelle transport across the seminiferous epithelium at different stages of the epithelial cycle. Yet the underlying molecular mechanism(s) that support this series of cellular events remain unknown. Herein, we used RNAi to knockdown cytoplasmic dynein 1 heavy chain (Dync1h1) and an inhibitor ciliobrevin D to inactivate dynein in Sertoli cells in vitro and the testis in vivo, thereby probing the role of dynein 1 in spermatogenesis. Both treatments were shown to extensively induce disruption of MT organization across Sertoli cells in vitro and the testis in vivo. These changes also perturbed the transport of spermatids and other organelles (such as phagosomes) across the epithelium. These changes thus led to disruption of spermatogenesis. Interestingly, the knockdown of dynein 1 or its inactivation by ciliobrevin D also perturbed gross disruption of F-actin across the Sertoli cells in vitro and the seminiferous epithelium in vivo, illustrating there are cross talks between the two cytoskeletons in the testis. In summary, these findings confirm the role of cytoplasmic dynein 1 to support the transport of spermatids and organelles across the seminiferous epithelium during the epithelial cycle of spermatogenesis.

scholarly journals Regulation of BTB Dynamics in Spermatogenesis—Insights From the Adjudin Model

2019 ◽  
Vol 172 (1) ◽  
pp. 75-88 ◽  
Author(s):  
Bai-Ping Mao ◽  
Linxi Li ◽  
Ming Yan ◽  
Renshan Ge ◽  
Qingquan Lian ◽  
...  
Keyword(s):  
Germ Cell ◽  
Low Dose ◽  
Mammalian Target Of Rapamycin ◽  
Underlying Mechanism ◽  
Seminiferous Epithelium ◽  
Regulatory Proteins ◽  
Cell Organelles ◽  
Adult Rats ◽  
Downstream Signaling ◽  
Cellular Events

Abstract During spermatogenesis, cell organelles, and germ cells, most notably haploid spermatids, are transported across the seminiferous epithelium so that fully developed spermatids line-up at the edge of the tubule lumen to undergo spermiation at stage VIII of the cycle. Studies have suggested that the microtubule (MT)-based cytoskeleton is necessary to support these cellular events. However, the regulatory molecule(s) and underlying mechanism(s) remain poorly understood. Herein, we sought to better understand this event by using an adjudin-based animal model. Adult rats were treated with adjudin at low-dose (10 mg/kg b.w.) which by itself had no notable effects on spermatogenesis. Rats were also treated with low-dose adjudin combined with overexpression of 2 endogenously produced blood-testis barrier (BTB) modifiers, namely rpS6 (ribosomal protein S6, the downstream signaling protein of mammalian target of rapamycin complex 1 [mTORC1]) and F5-peptide (a biological active peptide released from laminin-γ3 chain at the Sertoli-spermatid interface) versus the 2 BTB modifiers alone. Overexpression of these 2 BTB modifiers in the testis was shown to enhance delivery of adjudin to the testis, effectively inducing disruptive changes in MT cytoskeletons, causing truncation of MT conferred tracks that led to their collapse across the epithelium. The net result was massive germ cell exfoliation in the tubules, disrupting germ cell transport and cell adhesion across the seminiferous epithelium that led to aspermatogenesis. These changes were the result of disruptive spatial expression of several MT-based regulatory proteins. In summary, MT cytoskeleton supported by the network of MT regulatory proteins is crucial to maintain spermatogenesis.

scholarly journals Sertoli–germ cell junctions in the testis: a review of recent data

2010 ◽  
Vol 365 (1546) ◽  
pp. 1593-1605 ◽  
Author(s):  
Ilona A. Kopera ◽  
Barbara Bilinska ◽  
C. Yan Cheng ◽  
Dolores D. Mruk
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Seminiferous Epithelium ◽  
Cell Junctions ◽  
Molecular Architecture ◽  
Future Studies ◽  
Junction Type ◽  
Stage Viii ◽  
Immunological Barrier

Spermatogenesis is a process that involves an array of cellular and biochemical events, collectively culminating in the formation of haploid spermatids from diploid precursor cells known as spermatogonia. As germ cells differentiate from spermatogonia into elongated spermatids, they also progressively migrate across the entire length of the seminiferous epithelium until they reach the luminal edge in anticipation of spermiation at late stage VIII of spermatogenesis. At the same time, these germ cells must maintain stable attachment with Sertoli cells via testis-unique intermediate filament- (i.e. desmosome-like junctions) and actin- (i.e. ectoplasmic specializations, ESs) based cell junctions to prevent sloughing of immature germ cells from the seminiferous epithelium, which may result in infertility. In essence, both desmosome-like junctions and basal ESs are known to coexist between Sertoli cells at the level of the blood–testis barrier where they cofunction with the well-studied tight junction in maintaining the immunological barrier. However, the type of anchoring device that is present between Sertoli and germ cells depends on the developmental stage of the germ cell, i.e. desmosome-like junctions are present between Sertoli and germ cells up to, but not including, step 8 spermatids after which this junction type is replaced by the apical ES. While little is known about the biology of the desmosome-like junction in the testis, we have a relatively good understanding of the molecular architecture and the regulation of the ES. Here, we discuss recent findings relating to these two junction types in the testis, highlighting prospective areas that should be investigated in future studies.

Investigation of the potential role of the germ cell complement in control of the expression of transferrin mRNA in the prepubertal and adult rat testis

1997 ◽  
Vol 19 (1) ◽  
pp. 67-77 ◽  
Author(s):  
S M Maguire ◽  
M R Millar ◽  
R M Sharpe ◽  
J Gaughan ◽  
P T K Saunders
Keyword(s):  
Germ Cell ◽  
Mrna Expression ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Direct Access ◽  
Adult Rats ◽  
Rat Testis ◽  
Adult Rat

ABSTRACT Iron is required for the normal development of germ cells during spermatogenesis. Because these cells have no direct access to systemic iron, there exists a shuttle system involving production and secretion of the iron-transporting protein transferrin by the Sertoli cells. Previous reports using cultures of immature Sertoli cells exposed to adult germ cells, or in vivo studies involving germ cell-depleted adult rat testes, concluded that production of transferrin by Sertoli cells is modulated by germ cell complement. In the present study we have used in situ hybridisation with cRNA probes directed against the 5′ and 3′ ends of transferrin mRNA to examine the pattern of expression of transferrin in the immature and adult rat testis. Adult rats were treated with ethane dimethane sulphonate or methoxyacetic acid (MAA) to manipulate their testosterone levels or germ cell complement respectively. Initial findings obtained using the 3′ probe showed a decrease in transferrin mRNA associated with round spermatid depletion. However, these data were not confirmed by in situ hybridisation when the 5′ probe was used. The specificity of the probes was examined using Northern blotting and the 3′ probe was found to hybridise to the germ cell transcript for hemiferrin even under conditions of high stringency. Examination of immature and pubertal rat testes by in situ hybridisation using the 5′ transferrin-specific probe found that as early as 14 days of age the level of expression of transferrin mRNA was clearly different between tubules, and the mRNA appeared to be expressed in Leydig cells on and after day 31. In the adult rat testis, maximal expression of transferrin mRNA was found at stages VIII-XIV, calling into question the interpretation of the results of some previous studies showing expression of transferrin mRNA at all stages of the spermatogenic cycle. This stage-specific pattern of expression was not altered by acute germ cell depletion using MAA. However, Northern blot analysis showed a statistically significant increase in transferrin mRNA expression at 7 days after MAA treatment when pachytene spermatocytes were depleted from tubules at all stages of the spermatogenic cycle at which transferrin is normally expressed. In conclusion, we found that transferrin mRNA expression was not modulated by round spermatids as has been reported previously but that meiotic germ cells may influence expression of transferrin at specific stages of the spermatogenic cycle.

scholarly journals Two distinct Sertoli cell states are regulated via germ cell crosstalk†

2021 ◽  
Author(s):  
Rachel L Gewiss ◽  
Nathan C Law ◽  
Aileen R Helsel ◽  
Eric A Shelden ◽  
Michael D Griswold
Keyword(s):  
Germ Cell ◽  
Sertoli Cell ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Seminiferous Tubule ◽  
Seminiferous Epithelium ◽  
Critical Component ◽  
Cell Transcriptome ◽  
Cell Crosstalk ◽  
Blood Testis Barrier

Abstract Sertoli cells are a critical component of the testis environment for their role in maintaining seminiferous tubule structure, establishing the blood-testis barrier, and nourishing maturing germ cells in a specialized niche. This study sought to uncover how Sertoli cells are regulated in the testis environment via germ cell crosstalk in the mouse. We found two major clusters of Sertoli cells as defined by their transcriptomes in Stages VII–VIII of the seminiferous epithelium and a cluster for all other stages. Additionally, we examined transcriptomes of germ cell-deficient testes and found that these existed in a state independent of either of the germ cell-sufficient clusters. Altogether, we highlight two main transcriptional states of Sertoli cells in an unperturbed testis environment, and a germ cell-deficient environment does not allow normal Sertoli cell transcriptome cycling and results in a state unique from either of those seen in Sertoli cells from a germ cell-sufficient environment.

scholarly journals Transcriptional Profiling of the Hormone-Responsive Stages of Spermatogenesis Reveals Cell-, Stage-, and Hormone-Specific Events

Endocrinology ◽  
2009 ◽  
Vol 150 (11) ◽  
pp. 5074-5084 ◽  
Author(s):  
Liza O'Donnell ◽  
Kyriakos Pratis ◽  
Andrea Wagenfeld ◽  
Ulrich Gottwald ◽  
Jörg Müller ◽  
...  
Keyword(s):  
Sertoli Cells ◽  
Transcriptional Profiling ◽  
Seminiferous Epithelium ◽  
Cyclic Process ◽  
Down Regulation ◽  
Adult Rats ◽  
Cell Stage ◽  
Transcriptional Changes ◽  
Pachytene Spermatocytes ◽  
The Impact

Spermatogenesis occurs within the highly complex seminiferous epithelium. This cyclic process is accompanied by dynamic stage-specific transcriptional changes and is driven by androgens and FSH by mechanisms that are unclear. Here we report the impact of acute androgen and FSH suppression on the transcriptional dynamics of the seminiferous epithelium. We used transcriptional profiling to compare the most hormone-sensitive seminiferous epithelial stages (VII and VIII) from control and hormone-suppressed adult rats, together with publicly available datasets to delineate stage- and cell-specific transcriptional changes. The analyses reveal that, in these stages, there was a hormone-responsive down-regulation of spermatogonial and Sertoli cell transcripts maximally expressed in the earlier spermatogenic stages (I–VI). Transcripts expressed in Sertoli cells from stage VII and beyond were both up- and down-regulated by hormone suppression, with lysosome function, immune system-related genes, and lipid metabolism predicted to be hormone responsive. Hormone-responsive genes with putative roles in integrin-mediated cell adhesion were also identified. In pachytene spermatocytes, there was an initiation of transcription likely important for the completion of meiosis. A transcriptional switch in round spermatids was observed, from a hormone-responsive down-regulation of transcripts expressed in steps 1–7 spermatids to a hormone-independent up-regulation of transcripts expressed in steps 8–11 and likely involved in spermatid differentiation and DNA compaction. This study points to the existence of hormone-responsive global transcriptional repressors in Sertoli cells, spermatogonia, and spermatids and reveals novel and diverse cell-specific responses of the seminiferous epithelium to hormone suppression.

Purified bovine AMH induces a characteristic freemartin effect in fetal rat prospective ovaries exposed to it in vitro

Development ◽  
1987 ◽  
Vol 100 (1) ◽  
pp. 43-55 ◽  
Author(s):  
B. Vigier ◽  
F. Watrin ◽  
S. Magre ◽  
D. Tran ◽  
N. Josso
Keyword(s):  
Epithelial Cells ◽  
Germ Cell ◽  
Sertoli Cells ◽  
Basal Membrane ◽  
Cell Depletion ◽  
Morphological Abnormalities ◽  
Fetal Rat ◽  
Fetal Rats ◽  
Clear Cytoplasm

To determine whether anti-Mullerian hormone (AMH) is responsible for the gonadal lesions observed in bovine genetic females united by placental anastomoses to male twins (freemartins), prospective ovaries of fetal rats were exposed to purified bovine AMH in vitro. In cultures initiated at 14 days p.c. and maintained 3 to 10 days, AMH consistently induced a characteristic ‘freemartin effect’, namely reduction of gonadal volume, germ cell depletion and differentiation, in the gonadal blastema, of epithelial cells with large clear cytoplasm linked by interdigitations, resembling rat fetal Sertoli cells. These cells tend to become polarized and form cords, delineated by a continuous basal membrane containing laminin and fibronectin. Such structures, resembling developing seminiferous cords, were not detected in control ovarian cultures. These data strongly suggest that AMH is the testicular factor responsible for triggering the morphological abnormalities of freemartin gonads.

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