Organotypic Modeling Platform for Adverse Outcome Pathways of Male Reproductive and Developmental Processes

Thesis (Master's)--University of Washington, 2016-06

Development of the male reproductive system in Callinectes ornatus Ordway, 1863 (Brachyura: Portunidae)

This study describes the histology and histochemistry of the male reproductive system in Callinectes ornatus, comparing juvenile and adult developmental stages. We also analyzed changes in the gonadosomatic (GSI) and hepatosomatic (HSI) indices, and the weights of the testis and vas deferens during the development. The results showed that all stages, beginning with the juvenile (JUV), through developing (DEV) and mature (MAT) adult males of C. ornatus produce sperm and spermatophores. During development, testicular lobes showed the same characteristics of production and release of sperm into the seminiferous duct. The vas deferens showed little histological and histochemical change in the epithelium in juvenile and adult males. The differences consisted of the larger amount of secretion in MAT males compared to JUV and DEV ones. The chemical composition of the seminal fluid was similar, but MAT males produced a more homogeneous secretion. Morphological and physiological maturation are not synchronized in C. ornatus, since JUV males produced spermatophores similar to those in DEV and MAT males. However, these JUV are not yet able to reproduce, since they still have the abdomen attached to the cephalothoracic sternum. The increase of the GSI during development was significant for MAT males, and is related to the production of sufficient volume of seminal fluid to form the sperm plug in the female seminal receptacle. The HSI decreased from DEV to MAT adult stages, indicating that reserves from the hepatopancreas are used to develop the reproductive system after the pubertal molt.

Delayed reproductive development in pubertal male rats exposed to the hypolipemiant agent rosuvastatin since prepuberty

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Glyphosate impairs male offspring reproductive development by disrupting gonadotropin expression

Sexual differentiation in the brain takes place from late gestation to the early postnatal days. This is dependent on the conversion of circulating testosterone into estradiol by the enzyme aromatase. The glyphosate was shown to alter aromatase activity and decrease serum testosterone concentrations. Thus, the aim of this study was to investigate the effect of gestational maternal glyphosate exposure (50 mg/kg, NOAEL for reproductive toxicity) on the reproductive development of male offspring. Sixty-day-old male rat offspring were evaluated for sexual behavior and partner preference; serum testosterone concentrations, estradiol, FSH and LH; the mRNA and protein content of LH and FSH; sperm production and the morphology of the seminiferous epithelium; and the weight of the testes, epididymis and seminal vesicles. The growth, the weight and age at puberty of the animals were also recorded to evaluate the effect of the treatment. The most important findings were increases in sexual partner preference scores and the latency time to the first mount; testosterone and estradiol serum concentrations; the mRNA expression and protein content in the pituitary gland and the serum concentration of LH; sperm production and reserves; and the height of the germinal epithelium of seminiferous tubules. We also observed an early onset of puberty but no effect on the body growth in these animals. These results suggest that maternal exposure to glyphosate disturbed the masculinization process and promoted behavioral changes and histological and endocrine problems in reproductive parameters. These changes associated with the hypersecretion of androgens increased gonadal activity and sperm production.

Sex Determination and Female Reproductive Development in the Genus Schistosoma: A Review

Parasites of the genus Schistosoma were among the first metazoans to develop separate sexes, which is chromosomally determined in the fertilized egg. Despite the occurrence of specific sex chromosomes, the females of most Schistosomatidae species do not complete their somatic development and reach no sexual maturity without the presence of males. Indeed, the most controversial and at the same time most fascinating aspect about the sexual development of Schistosoma females lies on discover the nature of the stimulus produced by males that triggers and controls this process. Although the nature of the stimulus (physical or chemical) is a source of controversy, there is agreement that mating is a necessary requirement for maturation to occur and for migration of the female to a definitive final site of residence in the vascular system of the vertebrate host. It has also been proposed that the stimulus is not species-specific and, in some cases, not even genus-specific. Despite a vast literature on the subject, the process or processes underlying the meeting of males and females in the circulatory system have not been determined and as yet no consensus exists about the nature of the stimulus that triggers and maintains female development. In the studies about their role, Schistosoma males have been considered, at times pejoratively, the brother, the muscles or even the liver of females. Indeed, it still remains to be determined whether the stimulus responsible for female maturation involves the transfer of hormones, nutrients, neuromediators, mere tactile stimulation or a combination of chemotactic and thigmotactic factors

Morphological changes of Sertoli cells during the male reproductive cycle of the teleost Piaractus mesopotamicus (Holmberg, 1887)

Realizou-se uma investigação das mudanças histológicas e ultra-estruturais das células de Sertoli durante o ciclo reprodutivo de machos de Piaractus mesopotamicus. Os resultados mostraram que o desenvolvimento das células de Sertoli está estritamente relacionado à maturação das células gaméticas. Portanto, as células de Sertoli têm algum papel na maturação das células germinativas durante o ciclo reprodutivo dessa espécie, talvez formando um tecido de sustentação para os cistos espermatogênicos em desenvolvimento, ajudando a reorganização testicular para um novo ciclo reprodutivo, além de outras possíveis funções discutidas no texto.

Sex Determination and Female Reproductive Development in the GenuSchistosoma: A Review

Os parasitas do gênero Schistosoma situam-se entre os primeiros metazoários que desenvolveram sexos separados, determinado cromossomicamente no ovo fertilizado. Apesar da ocorrência de cromossomos sexuais específicos, as fêmeas de Schistosoma não atingem a maturidade somática e sexual sem a presença dos machos. Na verdade, um dos aspectos mais controversos e, ao mesmo tempo, mais fascinantes, envolvendo o desenvolvimento sexual das fêmeas está em se desvendar a natureza do estímulo que controla e mantém tal processo. Muito embora a natureza do estímulo (físico ou químico) seja motivo de controvérsia, concordam os mais diferentes autores que o acasalamento é um requisito indispensável para que ocorra a maturação e migração das fêmeas para o sítio definitivo de permanência no sistema vascular do hospedeiro vertebrado. Admite-se, ainda, que o estímulo não é espécie-específico e, em alguns casos, nem mesmo gênero-específico. Não obstante a existência de um número considerável de artigos dedicados ao tema, não há um consenso sobre o processo (ou processos) que controla(m) o encontro de machos e fêmeas no sistema circulatório do hospedeiro vertebrado, bem como está por ser determinada a natureza do estímulo, oriundo dos machos, que controla e mantém o desenvolvimento somático e sexual das fêmeas. Ao longo dos anos os machos de Schistosoma têm sido considerados, por vezes pejorativamente, os irmãos, os músculos ou o fígado das fêmeas. em síntese, resta saber se a natureza do estímulo responsável pelo desenvolvimento das fêmas envolve a transferência de hormônios, nutrientes, a mera estimulação tátil ou a combinação de dois ou mais desses fatores

Does the male reproductive performance depend on the early lactation milk in rats?

This study investigated the role of the early lactation milk (ELM) on the male reproductive performance and the participation of GnRH on this effect in Wistar rats. Newborn males were divided into three experimental groups: 1) ELM-deprived pups, 2) ELM-deprived pups treated with exogenous GnRH 1, 7, 25, and 31 h after birth, and 3) non-ELM-deprived and without exogenous GnRH. In relation to the other two groups, the ELM-deprived male rats exhibited as adults: reduced fertility, decreased weight of both vas deferens and seminal vesicle, and reduced levels of fructose in the seminal vesicle and prostate gland. We suggest that the intake of ELM during the neonatal period is important to the later sexual development of rats, and that GnRH is somehow involved in such an effect.

Structure, histochemistry and ultrastructure of the male reproductive accessory glands in the neotropical flat-faced fruit-eating bat Artibeus planirostris (Chiroptera: Phyllostomidae)

Chiroptera, the second largest mammalian order, presents different reproductive strategies and unique reproductive features. However, there are few reports regarding male reproductive accessory glands (RAGs) in Chiroptera. Thus, the aim of the present study was to characterise the RAGs of the exclusively neotropical bat Artibeus planirostris (Chiroptera: Phyllostomidae) macroscopically, microscopically and ultrastructurally. The RAGs were composed of a prostatic complex with two regions (ventral and dorsal) and paraurethral and bulbourethral glands, but no seminal vesicles. The ventral region had an undefined epithelium, with secretory and basal cells, and its secretions were periodic acid-Schiff (PAS) positive. The dorsal region received both deferens ducts, had a columnar pseudostratified epithelium with secretory and basal cells. There were two types of secretions from the dorsal region: one that was basophilic and another that was mixed PAS positive and PAS negative. The paraurethral glands were dispersed in the connective tissue of the urethra, whereas the bulbourethral glands were located in the penile root. Histological and ultrastructural data confirmed the prostatic nature of the ventral and dorsal regions and the holocrine nature of the ventral region, with the latter finding never having been described previously for the prostate gland. Our findings demonstrate the wide discrepancy of RAGs between A. planirostris and other mammals in terms of their composition, structure and morphology. © CSIRO 2013.

Dynamics of cell and tissue genesis in the male reproductive system of ticks (Acari: Ixodidae) Amblyomma cajennese (Fabricius, 1787) and Amblyomma aureolatum (Pallas, 1772): a comparative analysis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Extracellular matrix remodeling of the testes through the male reproductive cycle in Teleostei fish

During the fish reproductive cycle, testes undergo morphological changes related to germinal epithelium and remodeling of extracellular matrix components (ECM). ECM is degraded mainly by action of matrix metalloproteinases (MMPs). Due to the natural renewal of ECM in fish testes, we choose Pimelodus maculatus to study remodeling of ECM throughout reproductive cycle, using picrosirius (to identify type I, II, III collagen) and reticulin (type III collagen), and to immunolocalize MT1-MMP (membrane type 1-matrix metalloproteinase) and MMP-2 in testis cells. Testes were classified in four reproductive phases: regenerating, development, spawning capable and regressing. Picrosirius and reticulin demonstrated a differential distribution of total collagen fibers during the reproductive cycle. Immunohistochemistry showed MT1-MMP only in acidophilic granulocyte cells mainly inside blood vessels, in connective tissue of capsule close to the germinal compartment, and also infiltrated in interstitial connective tissue. MMP-2 was detected in fibroblast and endothelial cells of interstitial and capsule blood vessels, in epithelial cells of capsule, and in acidophilic granulocyte cells at same description for MT1-MMP. The fish testes ECM were remodeled throughout reproductive cycle in according to morphophysiological alterations. During reproductive season (spawning capable), the interstitium increased in total collagen fibers (type I, II, III). After spermiation period (regression and regenerating), the amount of collagen fibers decreased in response to action of MMPs on collagen degradation and other interstitial components (not assessed in this study). MMPs seem to be indispensable components for natural cyclic events of ECM remodeling of fish testes and for guarantee tissue homeostasis throughout reproductive cycle.

Structure, histochemistry and seasonal variations of the male reproductive accessory glands in the Pallas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Structure, histochemistry and seasonal variations of the male reproductive accessory glands in the Pallas’s mastiff bat, Molossus molossus (Chiroptera: Molossidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Safety assessment of after maternal exposure on male reproductive parameters in rats

Context: Hibiscus sabdariffa L. (Malvaceae) is a species widely used in folk medicine for the treatment of some disorders. Objective: This study evaluated the effects of H. sabdariffa (HS) on the development of the male reproductive tract in rats following in utero exposure. Materials and methods: Pregnant rats received 250 or 500 mg/kg of HS extract or vehicle from gestational day 12 until day 21 of lactation. Results and discussion: Both doses of HS increased the body weight of male offspring at weaning, without compromising the puberty onset parameters. At puberty, there was a significant increase in the vas deferens absolute weight and a significant reduction in the relative weight of kidney at higher dose. These animals also presented a significant reduction in the sperm number in the caput/corpus of epididymis after exposure to both doses and a reduction in the sperm number in the cauda epididymis for the lower dose. At adulthood, the highest dose significantly reduced the sperm production in relation to controls and both doses provoked a reduction in the relative sperm number in the epididymis without affecting the sperm morphology. Conclusion: These findings demonstrated that maternal exposure to H. sabdariffa can adversely influence the male reproductive system in rats.

Dynamics of cell and tissue genesis in the male reproductive system of ticks (Acari: Ixodidae) Amblyomma cajennense (Fabricius, 1787) and Amblyomma aureolatum (Pallas, 1772): a comparative analysis

Ticks are classified into three families: Argasidae, Ixodidae, and Nutalliellidae. The taxonomy and phylogeny within Ixodidae are still discussed by the specialists, thus requiring further studies. Amblyomma cajennese and Amblyomma aureolatum (Brazil) belong to two species complexes known as “cajennese” and “ovale”, respectively, and are directly related to the transmission of the Brazilian spotted fever. This confirms the medical and veterinary significance of these species, as well as the need for further morphological studies that will bring a better understanding of their taxonomy, phylogeny, and control. In this context, the present study aimed to characterize the morphology of the male reproductive system of A. cajennese and A. aureolatum when unfed and after 4 days of feeding, thereby seeking to: (a) distinguish the two species or “complexes”, and (b) study an internal system which has the potential to be targeted by acaricides. Therefore, males from both species (unfed and after 4 days of feeding) were cold-anesthetized, dissected, and had their reproductive systems removed for histological analysis. The results showed that the morphology of the male reproductive system is generally similar between both species, like in other Ixodidae ticks, exhibiting a multilobed accessory gland complex related to seminal fluid secretion, a pair of vasa deferentia and a pair of testes housing germ cells (spermatocytes) in different stages. The main differences were found in the development of the accessory gland complex cells and germ cells, showing that the maturation of the male reproductive system starts later in A. aureolatum, when compared to A. cajennese. However, during the blood meal, A. aureolatum development is increased, thus making germ cell maturation and gland complex activity higher than in A. cajennese. This study shows the differences in the development of the male reproductive systems of both species, while providing information that can assist in the establishment of new control methods.