Barrière hématotesticulaire (blood–testis barrier) et variations de perméabilité du compartiment gonial dans le follicule testiculaire de Locusta migratoria implanté en milieu dépourvu d'ecdystéroïdes

1986 ◽  
Vol 64 (9) ◽  
pp. 2053-2061 ◽  
Author(s):  
C. Marcaillou ◽  
S. Lauverjat

In Locusta migratoria, the testicular follicle is made on an apical compartment that contains the youngest germ cells and a basal one in which the spermatids differentiate. As soon as it is formed during the fourth larval stage, the latter compartment becomes impervious to exogenous macromolecules. This permeability barrier, which corresponds to the blood–testis barrier defined in mammals, depends on extensive septate junctions. In this paper, testicular follicles were cultured in vivo in adult males known to be free of hemolymphatic ecdysteroids. The permeability of the apical compartment to horseradish peroxidase, used as a tracer, varied as a function of implantation time. This apical compartment became impermeable as soon as the basal compartment cells were postmeiotic. Once implanted into an adult host, a follicle that contained only gonia completed normal spermatogenesis and established a blood–testis barrier, providing a tight compartment in which meiotic processes occurred. Septate junctions developed normally. These results show that ecdysterone controls neither the differentiation of septate junctions nor the establishment of the blood–testis barrier, which appear both to be conditioned by the initiation of meiosis.

2011 ◽  
Vol 209 (3) ◽  
pp. 337-351 ◽  
Author(s):  
Linlin Su ◽  
Dolores D Mruk ◽  
Will M Lee ◽  
C Yan Cheng

The blood–testis barrier (BTB) creates an immunological barrier that segregates the seminiferous epithelium into the basal and apical compartment. Thus, meiosis I/II and post-meiotic germ cell development take place in a specialized microenvironment in the apical compartment behind the BTB and these events are being shielded from the host immune system. If unwanted drugs and/or chemicals enter the apical compartment from the microvessels in the interstitium via the basal compartment, efflux pumps (e.g. P-glycoprotein) located in Sertoli cells and/or spermatids can actively transport these molecules out of the apical compartment. However, the mechanism(s) by which influx pumps regulate the entry of drugs/chemicals into the apical compartment is not known. In this study, a solute carrier (SLC) transporter organic anion transporting polypeptide 3 (Oatp3, Slco1a5) was shown to be an integrated component of the N-cadherin-based adhesion complex at the BTB. However, a knockdown of Oatp3 alone or in combination with three other major Sertoli cell drug influx pumps, namely Slc22a5, Slco6b1, and Slco6c1, by RNAi using corresponding specific siRNA duplexes failed to perturb the Sertoli cell tight junction (TJ) permeability barrier function. Yet, the transport of [3H]adjudin, a potential male contraceptive that is considered a toxicant to spermatogenesis, across the BTB was impeded following the knockdown of either Oatp3 or all the four SLC transporters. In short, even though drug transporters (e.g. influx pumps) are integrated components of the adhesion protein complexes at the BTB, they are not involved in regulating the Sertoli cell TJ permeability barrier function, instead they are only involved in the transport of drugs, such as adjudin, across the immunological barrier at the BTB.


Endocrinology ◽  
2013 ◽  
Vol 154 (5) ◽  
pp. 1907-1920 ◽  
Author(s):  
Xiaojing Qian ◽  
Dolores D. Mruk ◽  
Elissa W. P. Wong ◽  
Pearl P. Y. Lie ◽  
C. Yan Cheng

Abstract In rat testes, the ectoplasmic specialization (ES) at the Sertoli-Sertoli and Sertoli-spermatid interface known as the basal ES at the blood-testis barrier and the apical ES in the adluminal compartment, respectively, is a testis-specific adherens junction. The remarkable ultrastructural feature of the ES is the actin filament bundles that sandwiched in between the cisternae of endoplasmic reticulum and apposing plasma membranes. Although these actin filament bundles undergo extensive reorganization to switch between their bundled and debundled state to facilitate blood-testis barrier restructuring and spermatid adhesion/transport, the regulatory molecules underlying these events remain unknown. Herein we report findings of an actin filament cross-linking/bundling protein palladin, which displayed restrictive spatiotemporal expression at the apical and the basal ES during the epithelial cycle. Palladin structurally interacted and colocalized with Eps8 (epidermal growth factor receptor pathway substrate 8, an actin barbed end capping and bundling protein) and Arp3 (actin related protein 3, which together with Arp2 form the Arp2/3 complex to induce branched actin nucleation, converting bundled actin filaments to an unbundled/branched network), illustrating its role in regulating actin filament bundle dynamics at the ES. A knockdown of palladin in Sertoli cells in vitro with an established tight junction (TJ)-permeability barrier was found to disrupt the TJ function, which was associated with a disorganization of actin filaments that affected protein distribution at the TJ. Its knockdown in vivo also perturbed F-actin organization that led to a loss of spermatid polarity and adhesion, causing defects in spermatid transport and spermiation. In summary, palladin is an actin filament regulator at the ES.


2012 ◽  
Vol 303 (8) ◽  
pp. C843-C853 ◽  
Author(s):  
Linlin Su ◽  
Dolores D. Mruk ◽  
C. Yan Cheng

The blood-testis barrier (BTB) divides the seminiferous epithelium into the basal and the adluminal compartment. It restricts paracellular diffusion of molecules between Sertoli cells, confers cell polarity, and creates a unique microenvironment in the adluminal compartment for spermatid development. However, it undergoes restructuring during the epithelial cycle so that preleptotene spermatocytes differentiated from type B spermatogonia residing in the basal compartment can traverse the BTB at stage VIII of the cycle, while the immunological barrier is maintained. Herein, coxsackievirus and adenovirus receptor (CAR), a tight junction (TJ) integral membrane protein in the testis and multiple epithelia and endothelia, was found to act as a regulatory protein at the BTB, besides serving as a structural adhesion protein. RNAi-mediated knockdown of CAR in a Sertoli cell epithelium with an established TJ-permeability barrier that mimicked the BTB in vivo resulted in a disruption of the TJ barrier and an increase in endocytosis of the TJ-protein occludin. Furthermore, such an enhancement in occludin endocytosis was accompanied by a downregulation of Thr-phosphorylation in occludin and an increase in the association of endocytosed occludin with early endosome antigen-1. These findings were confirmed by overexpressing CAR in Sertoli cells, which was found to “tighten” the Sertoli cell TJ barrier, promoting BTB function. These findings support the emerging concept that CAR is not only a structural protein, it is involved in conferring the phosphorylation status of other adhesion proteins at the BTB (e.g., occludin) possibly mediated via its structural interactions with nonreceptor protein kinases, thereby modulating endocytic vesicle-mediated protein trafficking.


Endocrinology ◽  
2009 ◽  
Vol 150 (7) ◽  
pp. 3336-3344 ◽  
Author(s):  
Erica R. Siu ◽  
Elissa W. P. Wong ◽  
Dolores D. Mruk ◽  
K. L. Sze ◽  
Catarina S. Porto ◽  
...  

Several integral membrane proteins that constitute the blood-testis barrier (BTB) in mammalian testes, in particular rodents, are known to date. These include tight junction (TJ) proteins (e.g. occludin, junctional adhesion molecule-A, claudins), basal ectoplasmic specialization proteins (e.g. N-cadherin), and gap junction proteins (e.g. connexin43). However, the regulators (e.g. protein kinases and phosphatases) that affect these proteins, such as their interaction with the cytoskeletal actin, which in turn confer cell adhesion at the TJ, remain largely unknown. We report herein that focal adhesion kinase (FAK) is a putative interacting partner of occludin, but not claudin-11 or junctional adhesion molecule-A. Immunohistochemistry and fluorescence microscopy studies illustrated that the expression of FAK in the seminiferous epithelium of adult rat testes was stage specific. FAK colocalized with occludin at the BTB in virtually all stages of the seminiferous epithelial cycle but considerably diminished in stages VIII–IX, at the time of BTB restructuring to facilitate the transit of primary leptotene spermatocytes. Using Sertoli cells cultured in vitro with established TJ-permeability barrier and ultrastructures of TJ, basal ectoplasmic specialization and desmosome-like junction that mimicked the BTB in vivo, FAK was shown to colocalize with occludin and zonula occludens-1 (ZO-1) at the Sertoli-Sertoli cell interface. When these Sertoli cell cultures were treated with CdCl2 to perturb the TJ-barrier function, occludin underwent endocytic-mediated internalization in parallel with FAK and ZO-1. Thus, these findings demonstrate that FAK is an integrated regulatory component of the occludin-ZO-1 protein complex, suggesting that functional studies can be performed to study the role of FAK in BTB dynamics.


Endocrinology ◽  
2012 ◽  
Vol 153 (10) ◽  
pp. 5023-5035 ◽  
Author(s):  
Wenhui Su ◽  
Dolores D. Mruk ◽  
Pearl P. Y. Lie ◽  
Wing-yee Lui ◽  
C. Yan Cheng

Abstract The blood-testis barrier (BTB) is an important ultrastructure in the testis. A delay in its assembly during postnatal development leads to meiotic arrest. Also, a disruption of the BTB by toxicants in adult rats leads to a failure in spermatogonial differentiation. However, the regulation of BTB assembly remains unknown. Herein, filamin A, an actin filament cross-linker that is known to maintain and regulate cytoskeleton structure and function in other epithelia, was shown to be highly expressed during the assembly of Sertoli cell BTB in vitro and postnatal development of BTB in vivo, perhaps being used to maintain the actin filament network at the BTB. A knockdown of filamin A by RNA interference was found to partially perturb the Sertoli cell tight junction (TJ) permeability barrier both in vitro and in vivo. Interestingly, this down-regulating effect on the TJ barrier function after the knockdown of filamin A was associated with a mis-localization of both TJ and basal ectoplasmic specialization proteins. Filamin A knockdown also induced a disorganization of the actin filament network in Sertoli cells in vitro and in vivo. Collectively, these findings illustrate that filamin A regulates BTB assembly by recruiting these proteins to the microenvironment in the seminiferous epithelium to serve as the building blocks. In short, filamin A participates in BTB assembly by regulating protein recruitment during postnatal development in the rat testis.


2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Minesh Kapadia ◽  
M. Firoz Mian ◽  
Donglai Ma ◽  
Craig P. Hutton ◽  
Amber Azam ◽  
...  

Abstract Background Circulating autoantibodies and sex-dependent discrepancy in prevalence are unexplained phenomena of Alzheimer’s disease (AD). Using the 3xTg-AD mouse model, we reported that adult males show early manifestations of systemic autoimmunity, increased emotional reactivity, enhanced expression of the histone variant macroH2A1 in the cerebral cortex, and loss of plaque/tangle pathology. Conversely, adult females display less severe autoimmunity and retain their AD-like phenotype. This study examines the link between immunity and other traits of the current 3xTg-AD model. Methods Young 3xTg-AD and wild-type mice drank a sucrose-laced 0.4 mg/ml solution of the immunosuppressant cyclophosphamide on weekends for 5 months. After behavioral phenotyping at 2 and 6 months of age, we assessed organ mass, serologic markers of autoimmunity, molecular markers of early AD pathology, and expression of genes associated with neurodegeneration. Results Chronic immunosuppression prevented hematocrit drop and reduced soluble Aβ in 3xTg-AD males while normalizing the expression of histone variant macroH2A1 in 3xTg-AD females. This treatment also reduced hepatosplenomegaly, lowered autoantibody levels, and increased the effector T cell population while decreasing the proportion of regulatory T cells in both sexes. Exposure to cyclophosphamide, however, neither prevented reduced brain mass and BDNF expression nor normalized increased tau and anxiety-related behaviors. Conclusion The results suggest that systemic autoimmunity increases soluble Aβ production and affects transcriptional regulation of macroH2A1 in a sex-related manner. Despite the complexity of multisystem interactions, 3xTg-AD mice can be a useful in vivo model for exploring the regulatory role of autoimmunity in the etiology of AD-like neurodegenerative disorders.


Endocrinology ◽  
2014 ◽  
Vol 155 (10) ◽  
pp. 3981-3995 ◽  
Author(s):  
N. Ece Gungor-Ordueri ◽  
Elizabeth I. Tang ◽  
Ciler Celik-Ozenci ◽  
C. Yan Cheng

Abstract During spermatogenesis, the transport of spermatids and the release of sperms at spermiation and the remodeling of the blood-testis barrier (BTB) in the seminiferous epithelium of rat testes require rapid reorganization of the actin-based cytoskeleton. However, the mechanism(s) and the regulatory molecule(s) remain unexplored. Herein we report findings that unfold the functional significance of ezrin in the organization of the testis-specific adherens junction at the spermatid-Sertoli cell interface called apical ectoplasmic specialization (ES) in the adluminal compartment and the Sertoli cell-cell interface known as basal ES at the BTB. Ezrin is expressed at the basal ES/BTB in all stages, except from late VIII to IX, of the epithelial cycle. Its knockdown by RNA interference (RNAi) in vitro perturbs the Sertoli cell tight junction-permeability barrier via a disruption of the actin microfilaments in Sertoli cells, which in turn impeded basal ES protein (eg, N-cadherin) distribution, perturbing the BTB function. These findings were confirmed by a knockdown study in vivo. However, the expression of ezrin at the apical ES is restricted to stage VIII of the cycle and limited only between step 19 spermatids and Sertoli cells. A knockdown of ezrin in vivo by RNAi was found to impede spermatid transport, causing defects in spermiation in which spermatids were embedded deep inside the epithelium, and associated with a loss of spermatid polarity. Also, ezrin was associated with residual bodies and phagosomes, and its knockdown by RNAi in the testis also impeded the transport of residual bodies/phagosomes from the apical to the basal compartment. In summary, ezrin is involved in regulating actin microfilament organization at the ES in rat testes.


Zootaxa ◽  
2021 ◽  
Vol 4990 (1) ◽  
pp. 1-22
Author(s):  
OSCAR ARRIBAS ◽  
KAMİL CANDAN ◽  
YUSUF KUMLUTA ◽  
ÇETİN ILGAZ

All the Turkish populations studied, both those previously assigned to D. dryada (Subaşı and Yoldere villages, near Hopa) and those attributed to D. clarkorum (the largest sample studied so far, 177 specimens in total), are indistinguishable from each other and therefore must all be ascribed to the natural variability of a monotypic D. clarkorum. The Georgian specimens from the Type Locality of D. dryada (Charnaly river gorge, Chevachauri district) are clearly different, so that taxon cannot be considered a simple synonym for D. clarkorum, but as a valid taxon, although its proper status (more probably as a subspecies of D. clarkorum), is yet to be clarified. It is a highly threatened population, so studies should be done in vivo or with as low intrusiveness as possible.                 Darevskia dryada is clearly larger (SVL) than any D. clarkorum studied, with strongly longer heads and pilei in adult males (and hence more teeth in dentary bone), and higher dorsalia counts. There also seem to be (but need to be studied in a larger sample) more longitudinal rows of temporal scales between tympanic and parietal plates, a tendency to have more supralabial scales; comparatively smaller values for longitudinal rows of scales on the ventral surface of the thigh between the femoral pores and the outer row of enlarged scales, and higher collaria, and circumanalia scales. Other differences in femoralia and gularia are also reflected in Darevsky & Tuniyev’s (1997) tables and should also be investigated with more Georgian specimens.                 Two supposed discriminant characters, the frontonasal index and the presence of developed masseteric, are not valid. The frontonasal index does not discriminate both taxa; D dryada specimens fall inside the variation of D. clarkorum for this character. Also the presence of a developed masseteric plate is supposed to be rare if at all in D. clarkorum but always present in D. dryada; however, it appears in nearly 75% of D. clarkorum studied and in all D. dyada, so is also no longer valid for taxa discrimination.                 Although very similar, D. clarkorum and D. dryada are morphologically different, and genetic studies (as the unpublished results mentioned by Fu, 1999) do not make the provenance of the specimens clear, and hence the correct identification of the supposed specimens of D. dryada used.                 There are no geographical clines in D. clarkorum. However, as stated by Schmidtler et al. (2002), there is an inverse relationship between altitude and dorsalia values in D. clarkorum. Both the general differentiation between populations and the scalation (dorsalia) appear statistically correlated with the altitude and also with latitude (being both factors not strictly the same). The correlation seems to be stronger with morphology in general (multiple scalation characters and head biometry) than only with dorsalia. In the case of the general differentiation among samples, it is also significantly correlated with temperatures during the activity period (April-September) and with precipitation during incubation (July-August). As these climatic parameters of temperature and precipitation are not directly correlated with the dorsalia variation, the relation with altitude (and perhaps latitude) must be linked to some other climatic parameter not studied here, perhaps solar radiation or evapotranspiration.  


2018 ◽  
Vol 115 (52) ◽  
pp. E12435-E12442 ◽  
Author(s):  
Marilyn N. Vera-Chang ◽  
Antony D. St-Jacques ◽  
Rémi Gagné ◽  
Chris J. Martyniuk ◽  
Carole L. Yauk ◽  
...  

The global prevalence of depression is high during childbearing. Due to the associated risks to the mother and baby, the selective serotonin reuptake inhibitor fluoxetine (FLX) is often the first line of treatment. Given that FLX readily crosses the placenta, a fetus may be susceptible to the disruptive effects of FLX during this highly plastic stage of development. Here, we demonstrate that a 6-day FLX exposure to a fetus-relevant concentration at a critical developmental stage suppresses cortisol levels in the adult zebrafish (F0). This effect persists for three consecutive generations in the unexposed descendants (F1 to F3) without diminution and is more pronounced in males. We also show that the in vivo cortisol response of the interrenal (fish “adrenal”) to an i.p. injection of adrenocorticotropic hormone was also reduced in the males from the F0 and F3 FLX lineages. Transcriptomic profiling of the whole kidney containing the interrenal cells revealed that early FLX exposure significantly modified numerous pathways closely associated with cortisol synthesis in the male adults from the F0 and F3 generations. We also show that the low cortisol levels are linked to significantly reduced exploratory behaviors in adult males from the F0 to F2 FLX lineages. This may be a cause for concern given the high prescription rates of FLX to pregnant women and the potential long-term negative impacts on humans exposed to these therapeutic drugs.


Author(s):  
Mengrou Liu ◽  
Chunsen Zhu ◽  
Shun Bai ◽  
Xin Li ◽  
Kaiqiang Fu ◽  
...  

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