Congenital Zika virus infection impacts on male mouse offspring's reproductive biology.

In brief: Congenital ZIKV infection promotes alarming effects on male offspring's reproductive biology. This study showed the presence of the ZIKV antigen in the testis parenchyma, decreased testosterone levels, and sperm abnormalities in male offspring born to infected mothers. Abstract: Infection with ZIKV during pregnancy is associated with fetal developmental problems. Although neurological issues are being explored more in experimental studies, limited research has focused on the reproductive health consequences for offspring born to infected mothers. In this context, this study aimed to assess the impact of ZIKV infection during pregnancy on the testes and sperm of adult male offspring. Female mice were intraperitoneally inoculated with a Brazil strain of ZIKV during the 5.5th day of embryonic gestation. The offspring were evaluated 12 weeks after birth to analyze cellular and molecular changes in the testes and sperm. A novel approach combining variable-angle spectroscopic ellipsometry and machine learning modeling was also introduced for sperm sample analysis. The study revealed the presence of ZIKV protein in the testis parenchyma of adult male offspring born to infected mothers. It was shown that the testes exhibited altered steroidogenesis and inflammatory mediators, in addition to significant issues with spermiogenesis that resulted in sperm with DNA fragmentation, head defects, and protamination failure. Additionally, sperm dielectric properties and artificial intelligence showed potential for rapid identification and classification of sperm samples from infected mice. These findings provide crucial insights into the reproductive risks for men born from ZIKV-infected pregnant women.

Testis and brown adipose tissue xenografts from yellowish myotis (Myotis levis).

Yellowish myotis present a seasonal reproduction, influenced by rainfall distribution, in which the testis mass, germ cell composition, and brown adipose tissue mass change along the reproductive stages. In the present study, tissue xenografts were performed in immunodeficient mice to investigate spermatogenesis development in a stable endocrine milieu and the possible androgenic role of brown adipose tissue. Forty-one adult male bats were captured in the Santuário do Caraça, Minas Gerais, Brazil. The gonads and brown adipose tissue were collected, weighed, and grafted under the mice's back skin. Mice biometric and hormonal data were evaluated after grafting, and the testis grafts and mice gonads were fixed for histological and immunohistochemical analyses. As a result, testis grafts from adult bats presented a continuous germ cell development in all reproductive phases, showing round spermatids in all testis tissues. Furthermore, testis fragments in the Rest stage presented elongating spermatids as the most advanced germ cell type in the seminiferous epithelium after seven months of grafting. These data indicated that yellowish myotis spermatogenesis could be continued (presenting a constant spermatogonial differentiation) in a stable endocrine milieu, as found in mice. In addition, the best spermatogenic development was achieved when testis fragments were transplanted at their lowest activity (Rest stage). Regarding the brown adipose tissue grafts, the adipose tissue consumption by mice increased seminal vesicle mass and testosterone serum levels. This data proved that the brown adipose tissue is related to testosterone synthesis, which may be critical in stimulating the differentiation of spermatogonia in yellowish myotis.

A Biosafety Level 2 Mouse Model for Studying Betacoronavirus-Induced Acute Lung Damage and Systemic Manifestations.

The emergence of life-threatening zoonotic diseases caused by betacoronaviruses, including the ongoing coronavirus disease 19 (COVID-19) pandemic, has highlighted the need for developing preclinical models mirroring respiratory and systemic pathophysiological manifestations seen in infected humans. Here, we showed that C57BL/6J wild-type mice intranasally inoculated with the murine betacoronavirus murine hepatitis coronavirus 3 (MHV-3) develop a robust inflammatory response leading to acute lung injuries, including alveolar edema, hemorrhage, and fibrin thrombi. Although such histopathological changes seemed to resolve as the infection advanced, they efficiently impaired respiratory function, as the infected mice displayed restricted lung distention and increased respiratory frequency and ventilation. Following respiratory manifestation, the MHV-3 infection became systemic, and a high virus burden could be detected in multiple organs along with morphological changes. The systemic manifestation of MHV-3 infection was also marked by a sharp drop in the number of circulating platelets and lymphocytes, besides the augmented concentration of the proinflammatory cytokines interleukin 1 beta (IL-1ß), IL-6, IL-12, gamma interferon (IFN-γ), and tumor necrosis factor (TNF), thereby mirroring some clinical features observed in moderate and severe cases of COVID-19. Importantly, both respiratory and systemic changes triggered by MHV-3 infection were greatly prevented by blocking TNF signaling, either via genetic or pharmacologic approaches. In line with this, TNF blockage also diminished the infection-mediated release of proinflammatory cytokines and virus replication of human epithelial lung cells infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Collectively, results show that MHV-3 respiratory infection leads to a large range of clinical manifestations in mice and may constitute an attractive, lower-cost, biosafety level 2 (BSL2) in vivo platform for evaluating the respiratory and multiorgan involvement of betacoronavirus infections. IMPORTANCE Mouse models have long been used as valuable in vivo platforms to investigate the pathogenesis of viral infections and effective countermeasures. The natural resistance of mice to the novel betacoronavirus SARS-CoV-2, the causative agent of COVID-19, has launched a race toward the characterization of SARS-CoV-2 infection in other animals (e.g., hamsters, cats, ferrets, bats, and monkeys), as well as adaptation of the mouse model, by modifying either the host or the virus. In the present study, we utilized a natural pathogen of mice, MHV, as a prototype to model betacoronavirus-induced acute lung injure and multiorgan involvement under biosafety level 2 conditions. We showed that C57BL/6J mice intranasally inoculated with MHV-3 develops severe disease, which includes acute lung damage and respiratory distress that precede systemic inflammation and death. Accordingly, the proposed animal model may provide a useful tool for studies regarding betacoronavirus respiratory infection and related diseases.

Sperm production and seminal analyses in a Neotropical sperm-storing vespertilionid bat yellowish myotis (Myotis levis).

Yellowish myotis is a Neotropical vespertilionid bat that presents a seasonal reproduction. The sperm is produced in the Mature stage, stored in the Regressed stage and released in the Rest stage (mating period). Aiming to understand, for the first time, the relationship between testis and epididymis physiology in yellowish myotis reproduction, the spermatogenesis length, sperm production, and seminal parameters were herein evaluated. Fifty-one adult male bats were captured in Santuário do Caraça, Minas Gerais, Brazil. The gonads were collected in the Maturing and Mature stages for histomorphometric and immunohistochemical analyses, whereas the epididymis was evaluated in all reproductive stages for seminal studies. Our results demonstrated that the yellowish myotis spermatogenic process is fast, lasting 31.70 ± 0.15 days. Despite the low Sertoli cell efficiency (6.60 ± 1.23), the high numbers of Sertoli cells per testis enable an elevated sperm production in the Mature stage. The sperm concentration, vitality, and motility presented the highest values in the Regressed stage; however, in this period, an increased incidence of sperm morphological defects was detected. In the following period (Rest stage), a drastic reduction of defective sperm was observed, suggesting quality control of sperm before the mating period. Furthermore, the epididymis ability to maintain a long-term sperm-storage was observed in 26.7% of the bats in the Maturing stage. In summary, yellowish myotis presented a fast and high sperm production during the Mature stage. These sperms are stored and selected before mating period.

Male reproductive morphofunctional evaluation of a Neotropical sperm-storing vespertilionid bat (Myotis levis) in an environmental context.

Myotis levis (yellowish myotis) is a small Neotropical insectivorous vespertilionid bat that provides valuable ecosystem services, such as control of disease vectors and agricultural pests. Aiming to describe the fluctuations of the reproductive organs throughout the year, the gonads and epididymis from 124 adult bats were histologically evaluated. These animals were captured in Santuário do Caraça, Minas Gerais, Brazil. After the initial screening, six bats per reproductive stage (in a representative month) had specific organs harvested for further investigation. The gonads, epididymis, accessory sex gland and brown adipose tissue were collected for biometric analyses. Furthermore, yellowish myotis testis was evaluated through histomorphometric and molecular assays, whereas blood samples were collected for hormonal analyses. The data were compared among the reproductive stages and correlated with rainfall distribution. As a result, we demonstrated that yellowish myotis presented a seasonal reproduction showing testis regression and rest, resembling the pattern exhibited by temperate-zone vespertilionid bats. During the Mature stage, after the peak of rainfall distribution, yellowish myotis testicles were fully developed for gamete production and maximum testosterone synthesis. These findings indicate a significant influence of this environmental factor on yellowish myotis reproduction. Following that, the accessory sex gland, brown adipose tissue and epididymis weights increased in the Regressed stage. The epididymis sperm storage occurred for at least 8 months and was observed in the Regressed, Rest and beginning of the Maturing stage. This reproductive fluctuation is interesting because the reactivation of the gonads coincided with the least amount of sperm in the epididymis.

Prepubertal PTU treatment in rat increases Sertoli cell number and sperm production.

The number of Sertoli cells (SCs) ultimately determines the upper limit of sperm production in the testis. Previous studies have shown that thyroid hormones (TH) receptors are abundantly expressed in developing SCs; therefore, it was highly significant to discover that transient neonatal hypothyroidism induced by the goitrogen 6-n-propyl-2-thiouracil (PTU) can extend SCs proliferation beyond the first 2 weeks postnatal and increase testis weight and sperm production. Further studies concluded that treatment must begin before day 8 post birth in rats. Recent studies, however, showed that SCs present in the transition region at the rete testis exhibit a more immature phenotype and have prolonged mitotic activity, which led to the hypothesis that SCs in this region will retain the capacity to respond to PTU treatment over a longer period of time. In the present study, male Wistar rats were treated with PTU from days 21 to 40 and were evaluated at 40 and 160 days of age. Similar to neonatal rat SCs, it was demonstrated that prepubertal SCs in the transition region have a high mitotic activity and are highly sensitive to TH levels. This delayed, transient hypothyroidism resulted in significantly increased testis weight, SCs number and daily sperm production. The results demonstrate for the first time that Sertoli cells showing plasticity in the transition region can be stimulated to increase proliferation and contribute to a late stage surge in testis weight and sperm output.