SPINDLE MEMBRANES DURING SPERMATOGENESIS IN DERMATOBIA-HOMINIS (DIPTERA, CUTEREBRIDAE)

During the mitotic and meiotic division in Dermatobia hominis spermatogenesis, the nuclear envelope is fragmented and membranes appear around the spindle. The membranes surrounding the mitotic spindle are formed by two layers of cisterns. The membranes of the meiotic spindle consist in at least 3 or 4 layers of long smooth cisterns which isolate the spindle from the remaining cytoplasm. The presence of this kind of membranes during meiosis seems to be usual in insect male germ cell.

Spermatogenesis and nucleolar behavior in males of aquatic Heteroptera

Aspects of spermatogenesis and nucleolar behavior were analyzed in Brachymetra albinerva, Cylindrostethus palmaris, Halobatopsis platensis, Limnogonus aduncus (Gerridae), Martarega sp (Notonectidae), Rhagovelia whitei, and Rhagovelia sp (Veliidae). The testicles are rounded (Veliidae), elongated (Gerridae) or spiral (Notonectidae) and have a transparent membrane covering them. The complement chromosome was 2n = 23 (22A + X0, L. aduncus and Rhagovelia sp), 25 (24A + X0, B. albinerva and H. platensis), 26 (22A + 2m + XY, Martarega sp), 29 (28A + X0, C. palmaris), or 39 (38A + X0, R. whitei) chromosomes, and the only species with a different sex chromosome system was Martarega sp, which showed an XY system and m-chromosomes. The meiotic behavior of all species was similar: holocentric chromosomes and heteropyknotic material at prophase, interstitial and/or terminal chiasmata, and first reductional division for the autosomes and the reverse for the sex chromosomes. The only difference observed was related to the very large size of Martarega sp cells in all stages of spermatogenesis. With regard to nucleolar behavior, the species did not show differences, except for Martarega sp with larger nucleoli than the other species. The only species in which it was clearly possible to identify the nucleolar organizer region was L. aduncus, in the region of a terminal autosome. It was also confirmed that the telomeric associations do not occur at random. In the other species, specific staining was very discrete, and the nucleolar organizer region location was not at all evident.

Cytoplasmic bridges, intercellular junctions, and individualization of germ cells during spermatogenesis in Dermatobia hominis (Diptera: Cuterebridae)

During mitotic and meiotic divisions in Dermatobia hominis spermatogenesis, the germ cells stay interlinked by cytoplasm, bridges as a result of incomplete cytokinesis. By the end of each division, cytoplasmic bridges flow to the center of the cyst, forming a complex, called the fusoma. During meiotic prophase I, spermatocytes I present desmosome-like junctions and meiotic cytoplasmic bridges. At the beginning of spermiogenesis, the fusoma moves to the future caudal end of the cyst, and at this time the early spermatids are linked by desmosome-like junctions. Throughout spermiogensis, new and sometimes broad cytoplasmic bridges are formed among spermatids at times making them share cytoplasm. In this case the individualization of cells is assured by the presence of smooth cisternae that outline then structures The more differentiated spermatids have in addition to narrow cytoplasmic bridges, plasmic membranes junctions. By the end of spermiogenesis the excess cytoplasmic mass is eliminated leading to spermatid individualization. Desmosome-like junctions of spermatocytes I and early spermatids appear during the fusoma readjustment and segregations; on the other hand, plasmic membrane junctions appear in differentiating spermatids and are eliminated along with the cytoplasmic excess. These circumstances suggest that belt desmosome-like and plasmic membrane junctions are involved in the maintenance of the relative positions of male germ cells in D. hominis while they are inside the cysts. © 1996 Wiley-Liss, Inc.

Patterns of nucleolar activity during spermatogenesis of two triatomines, Panstrongylus megistus and P. herreri

The present work analyzed spermatogenesis in two species of triatomines (genus Panstrongylus) using silver-ion impregnation. The sex chromosomes of P. megistus and P. herreri had nucleolar organizing activity and became strongly impregnated during the phases of meiotic prophase I. Fragmentation of the nucleolus occurred in both species during the meiotic cycle. The nucleolar region could be observed up to diakinesis in meiotic prophase after which only nucleolar bodies and fragments were seen. Postmeiotic reactivation of rRNA synthesis occurred in these two species and was probably related to cell differentiation.

The ultrastructure of the pre-meiotic and meiotic stages of spermatogenesis in Plagioscion squamosissimus (Teleostei, Perciformes, Sciaenidae)

Spermatogenesis of 'corvina' P. squamosissimus starts from a stem cell that gives rise to germ cells. These cells are enveloped by Sertoli cells, forming cysts. The germ cells in the cysts are all at the same stage of development and are interconnected by cytoplasmic bridges. Spermatogonia are the largest germ cells. In the cysts, these cells differentiate into primary spermatogonia and secondary spermatogonia. The primary spermatogonia are isolated in the cyst and give rise to the secondary spermatogonia. After several mitotic divisions, they produce spermatocytes I, which can be identified by synaptonemal complexes in the nucleus. The spermatocytes I enter the first phase of meiosis to produce the spermatocytes II. These are not very frequently seen because they rapidly undergo a second phase of meiosis to produce spermatids.

Kinetics of spermatogenesis in the Mongolian gerbil (Meriones unguiculatus)

The Mongolian gerbil (Meriones unguiculatus) is a small rodent native to the arid regions of Mongolia and Northeastern China. The present study provides descriptions of both the cellular associations of the seminiferous-epithelium cycle and relative frequencies of stages in the gerbil. Based on the development of the acrosomic system and the nuclear morphology changes using the PAS-H staining technique, the transformation of spermatids into spermatozoa was divided into 15 steps. The first 12 steps were used to identify 12 stages or cellular associations and the other three steps were spread among the first six stages of the cycle of the seminiferous epithelium. The relative frequencies found for stages I through XII were: 13.15; 8.06; 8.98; 6.48; 5.37; 6.71; 7.36; 7.45; 7.27; 5.83; 11.53 and 11.81, respectively. Stage I had the highest frequency while stage V proved the lowest frequency among the XII stages. The pattern of spermatogenesis is similar to those of rodents used as laboratory animals. The present description is the first for this rodent and provides the foundation for a variety of future studies of the testis in this animal. © 2002 Elsevier Science Ltd. All rights reserved.

Structural characterization of nuclear phenotypes during Scinax fuscovarius spermatogenesis (Anura, Hylidae)

In anuran amphibian Scinax fuscovarius, the spermatogenesis occurs in structures called seminiferous loculi, in which germ epithelium is organized in spermatocysts. Each cyst contains cells in the same stage of cytodifferentiation. Characteristics of each cellular type and their groups made the identification and differentiation of the germ lineage cells possible. In the basis of the epithelium there are the spermatogonia I, the biggest cells and always associated with the Sertoli cell. After the phase of mitotic proliferation, the cysts containing variable number of spermatogonia II are originated, quite smaller and with cellular boundaries a little distinct. After differentiation and growth in volume, the spermatocytes I appear, the nuclei of which are spherical and with different degrees of compaction of the nuclear material. Starting the meiotic process, the spermatocytes II are originated, which by means of the second meiotic division become haploid cells, the spermatids I. These two last spermatocysts are very similar. In this phase, the cells will go through a prominent process of differentiation until they form the spermatids II, which are elongated and begin to be organized in bundles supported by prominent Sertoli cells. With the process of spermiogenesis, spermatozoa appear, usually observed in compact bundles with tails turned to the lumen and their heads fitted in their support cells. In more advanced stages, the spermatozoa can be observed free in the locular lumen, ready to follow the spermatic path.

Cysts reabsorption in the testis of the bee wax pest Achroia grisella (Lepidoptera, Pyralidae) during spermatogenesis

The present paper reports the occurrence of testicular cysts degeneration during spermatogenesis of Achroia grisella, a Lepidoptera with dimorphic spermatogenesis, through ultrastrucutural studies. Signs of cysts degeneration can be detected in the last larval instar but it increases during pupation and early adulthood. The degeneration affects the eupyrene, as well apyrene cysts but it is not always possible to recognize which cysts are degenerating. Some morphological features of cysts degeneration resemble apoptosis.

Cytogenetic analysis in the spermatogenesis of Triatoma melanosoma (Reduviidae; Heteroptera)

Triatomines are of great concern in public health because they are vectors of Chagas' disease. This study presents an analysis of the species Triatoma melanosoma. The cytogenetic characteristics of triatomines include holocentric chromosomes, post-reductional meiosis in the sex chromosomes and nucleolar fragmentation in the meiotic cycle. The methodology utilized consisted of the techniques of lacto-acetic orcein staining and silver ion impregnation. The organs analyzed were adult testicles. The results enabled to classify the chromosomes by number and size, being three large, eight medium and one small heterochromosome. The three largest chromosomes and the heterochromosomes showed heteropyknotic chromatin in meiosis. The heterochromosomes in 8.05% of the cells in metaphase I behaved as pseudobivalents, contrasting with 91.95% of the cells with individualized sex chromosomes, confirming the achiasmatic nature of these chromosomes. However, the pseudobivalents occurred prominently in metaphase II (78.38%), this fact probably is related to the post-reductional nature of the sex chromosomes. The nucleolus in T. melanosoma persisted until the diplotene phase after which it began to fragment. Nucleolar corpuscles were observed in metaphases I and II and during anaphases I and II, these characteristics being related to the phenomenon of nucleolar persistence. In the initial spermatids, peripheral silver ion impregnation occurred, which could be analogous to the pre-nucleolar corpuscles observed after fragmentation. Thus, this study extends our knowledge of the characteristics of triatomines, in particular, heteropyknotic degree, kinetic activity, formation of sex chromosome achiasmatic pseudobivalency, confirmation of the fragmentation phenomenon, and post-meiotic nucleolar reactivation. ©FUNPEC-RP.

An overview of functional and stereological evaluation of spermatogenesis and germ cell transplantation in fish

Although there are almost thirty-thousand species of fish living in a great variety of habitats and utilizing vast reproductive strategies, our knowledge of morphofunctional and quantitative aspects of testis structure and spermatogenesis is still incipient for this group of vertebrates. In this review, we discuss aspects that are important to better understanding of testis structure and function, and of the development of germ cells (GC) during spermatogenesis. To achieve this, we have recently completed a number of studies presenting morphometric and functional data related to the numbers of GC and Sertoli cells (SC) per each type of spermatogenic cyst, the number of spermatogonial generations, the SC efficiency, and the magnitude of GC loss that normally occurs during spermatogenesis. We also investigated SC proliferation and the relationship of this important event to early spermatogenic cysts. The available data strongly suggest that SC proliferation in sexually mature tilapia is the primary factor responsible for the increase in testis size and for determination of the magnitude of sperm production. The influence of temperature on the duration of spermatogenesis in tilapia was also evaluated and we have used this knowledge to deplete endogenous spermatogenesis in this teleost, in order to develop an experimental system for GC transplantation. This exciting technique results in new possibilities for investigation of spermatogenesis and spermatogonial stem cell biology, creating also an entirely new and promising scenario in biotechnology - transgenic animal production and the preservation of the genetic stocks of valuable animals or endangered species. © Springer Science+Business Media B.V. 2008.

Spermatogenesis and karyotypes of three species of water striders (Gerridae, Heteroptera).

Although they are of economic importance, there have been few cytogenetic studies of the Gerridae (Heteroptera) in Brazil. We examined spermatogenesis (meiosis and spermiogenesis) and nucleolar behavior in three species of the family Gerridae. Brachymetra albinerva and Halobatopsis platensis were found to have a chromosome complement of 2n = 25 (24A + X0) and Cylindrostethus palmaris 2n = 29 (28A + X0) chromosomes. Fifteen individuals of these species were collected from the reservoir of São José do Rio Preto, SP, using screens and were transported in pots containing water to the laboratory, where cytogenetic preparations were made. The polyploidy nuclei are formed by several heteropyknotic regions; cells in meiotic prophase have a heteropyknotic region that is probably the sex chromosome, and the chromosomes from chiasmata. The spermatids are rounded and have a heteropyknotic region at the periphery of the nucleus; the sperm head is small, with a long tail. Silver impregnation of meiotic cells showed one or more disorganized bodies around the perichromosomal sheath. The round spermatids had two bodies next to each other, but these were elongated; one of the bodies remained in the head and the other migrated to the initial part of the tail at the end of spermagenesis, when the staining was no longer evident. The meiotic cells appear during spermatogenesis and have very similar silver-impregnation patterns in different species of Heteroptera.

Comparative study of spermatogenesis and nucleolar behavior in testicular lobes of Euschistus heros (Heteroptera: Pentatomidae)

In some testicular lobes of the Pentatomidae there may be occurrence of atypical spermatogenesis or polymegaly, leading to the production of nonfertile sperm. The comparative analysis of spermatogenesis and nucleolar behavior in testicular lobes of Euschistus heros showed cells with polymegaly in lobes 4 and 6. Generally, when these lobes are present in the same individual, there is also the formation of atypical cells in the flanking lobe. Such characteristic was not seen in E. heros. However, differences regarding the concentration of heteropyknotic chromatin and silver-positive bodies in this lobe deserve attention. This study explored the literature and demonstrated the prevalence of some lobes in the formation of differentiated cells. It was also found in the literature that there is an association of the chromocenter with the nucleolus in several species of Pentatomidae, but in E. heros this association does not appear to occur. © 2010 Hederson Vinicius de Souza and Mary Massumi Itoyama.

Chagas disease and triatomine biology (Heteroptera, Triatominae), with emphasis on aspects of spermatogenesis

A century after the discovery of Chagas disease, it is still one of the most important parasitic diseases affecting humans. The subfamily Triatominae is important in medical health, because these insects are vectors of Trypanosoma cruzi, the etiologic agent of Chagas disease. These insects are also of important cytological relevance because they have particular cell characteristics, such as persistence of nucleolar material in spermatogenesis. The germ cells of the animal kingdom have chromatoid bodies (CBs) in their cytoplasm that can originate from nucleolar material that is fragmented in the early stages of spermatogenesis and plays an important role in cellular communication between the spermatids during spermiogenesis. Currently, there are few studies on the function and formation of the CB in nucleologenesis, especially with emphasis on the ultrastructure of the cells involved in spermatogenesis of insects. Considering the importance of knowledge about the triatomine fauna, we conducted a study of the biogeography and reports of these insects and a survey of patients with Chagas disease in the northwestern region of São Paulo State. Data collected from 1995 to 2009 indicated 700 individuals with Chagas disease, demonstrating a range of 0 to 40 years, which shows that the disease may be active in this region. Moreover, of the 1150 patients treated for cardiomyopathy, 44% were chagasic. Regarding the triatomines noted and captured in the period from 2004 to 2009, the species were Triatoma sordida and Rhodnius neglectus, with T. sordida being the most abundant. In addition, some triatomines were infected by T. cruzi in various developmental stages. We also analyzed the nucleolar cycle and fibrillarin nucleolar protein expression in CB of spermatogenic cells of T. infestans and T. sordida, using histological, ultrastructural and immunocytochemical techniques. The results revealed fibrillarin nucleolar protein expression in the nucleus and in some cytoplasmic spots of germ cells during spermatogenesis in triatomines. These data suggest that fibrillarin could be a constituent of CB, which was most likely derived from nucleolar fragmentation. This is the first time that fibrillarin protein expression has been shown in CB during spermatogenesis progression in triatomines. Knowledge about the biology of triatomines was deepened in this study and, in particular, the structural and ultrastructural aspects of spermatogenesis in triatomines. This study showed that the disease may be active in the northwestern region of São Paulo and expanded our knowledge of the biology of triatomines, the main vectors of Chagas disease. © FUNPEC-RP.