Cell death in ovarioles causes permanent sterility in Frieseomelitta varia worker bees

Frieseomelitta varia worker bees do not lay eggs even when living in queenless colonies, a condition that favors ovary development and oviposition in the majority of highly social bees. The permanent sterility of these worker bees was initially attributed to a failure in ovary morphogenesis and differentiation. Using transmission electron microscopy we found that at the beginning of the pupal phase the ovaries of F. varia workers are formed by four ovarioles, each of them composed of 1) a terminal filament at the apex of the ovarioles, containing juxtaposed and irregularly shaped cells, 2) a germarium with clusters of cystocytes and prefollicular cells showing long cytoplasmic projections that envelop the cystocyte clusters, 3) fusiform interfollicular and basal stalk precursor cells, and 4) globular, irregularly contoured basal cells with large nuclei. However, during the pupal phase an accentuated and progressive process of cell death takes place in the ovarioles. The dying cells are characterized by large membrane bodies, electron-dense apoptotic bodies, vacuoles, vesiculation, secondary lysosomes, enlarged rough endoplasmic reticulum cisternae, swollen mitochondria, pycnotic nuclei, masses of chromatin adjacent to the convoluted nuclear envelope, and nucleoli showing signs of fragmentation. Cell death continues in ovarioles even after the emergence of the workers. Once they become nurse bees, the ovaries have become transformed into a cell mass in which structurally organized ovarioles can no longer be identified. In F. varia workers, ovariole cell death most certainly is part of the program of caste differentiation.

Bupivacaína, Ropivacaína e Levobupivacaína em Analgesia e Anestesia de Parto. Repercussões Materno-Fetais

Background and Objectives - Bupivacaine has been a very useful local anesthetic in Obstetrics in spite of its potential cardiotoxicity. In obstetric analgesia, ropivacaine is preferred to bupivacaine, and levobupivacaine is less cardiotoxic than the racemic mixture. The aim of this study was to compare the maternal-fetal effects of bupivacaine, ropivacaine and levobupivacaine in obstetric analgesia and anesthesia. Methods - Participated in this study 33 term pregnant women, physical status ASA I and II submitted to epidural anesthesia who received 18.75 mg (in 15 ml completed with 9% saline) of: GI - bupivacaine, GII - ropivacaine, and GIII - levobupivacaine. Pain intensity, sensory block level, onset time, quality of analgesia, motor block intensity, analgesia duration and time for labor resolution were evaluated. For vaginal delivery, 40 mg (in 8 ml of saline) of the same local anesthetic were used; for cesarean delivery, the dose has been mg in 20 ml solution. Newborns of these mothers were evaluated through the Apgar score in minutes 1, 5 and 10, and through the Amiel-Tison method (neurologic and adaptative capacity score - NACS) at 30 min, 2 h, and 24 h. Results - There were no significant statistical differences among groups as to sensory block level, onset time, quality of analgesia, labor analgesia duration, time for labor resolution, and Apgar scores at minute 1. Ast to motor block, GIII > GII and GI was intermediary. In relation to pain intensity, there was a trend for GI > GIII. For Apgar scores in minute 5, GII > (GI = GIII), and in minute 10 (GI = GII) < GIII. NACS at 2 h showed, GII > GI > GIII, and at 24 h, GII > GIII > GI. Conclusions - Ropivacaine has relieved maternal pain with less motor block. Newborns of GII mothers (ropivacaine) showed the best Apgar and NACS scores.

Regression of the lateral oviducts during the larval-adult transformation of the reproductive system of Melipona quadrifasciata and Frieseomelitta varia

Toward the end of the larval phase (pre-pupa), the reproductive systems of Melipona quadrifasciata and Frieseomelitta varia workers are anatomically similar. Scanning electron microscopy showed that during this developmental phase the right and left ovaries are fused and form a heart-shaped structure located above the midgut. Each ovary is connected to the genital chamber by a long and slender lateral oviduct. During pupal development, the lateral oviducts of workers from both species become extremely reduced due to a drastic process of cell death, as shown by transmission electron microscopy. During the lateral oviduct shortening, their simple columnar epithelial cells show some signs of apoptosis in addition to necrosis. Cell death was characterized by cytoplasmic vesiculation, peculiar accumulation of glycogen, and dilation of cytoplasmic organelles such as mitochondria and rough endoplasmic reticulum. The nuclei, at first irregularly contoured, became swollen, with chromatin flocculation and various areas of condensed chromatin next to the nuclear envelope. At the end of the pupal phase, deep recesses marked the nuclei. At emergence, worker and queen reproductive systems showed marked differences, although reduction in the lateral oviducts was an event occurring in both castes. However, in queens the ovarioles increased in length and the spermatheca was larger than that of workers. At the external anatomical level, the reproductive system of workers and queens could be distinguished in the white- and pink-eyed pupal phase. The metamorphic function of the death of lateral oviduct cells, with consequent oviduct shortening, is discussed in terms of the anatomical reorganization of the reproductive system and of the ventrolateral positioning of adult worker bee ovaries. (C) 2000 Wiley-Liss, Inc.

Morphology of testis and epididymis in an ethanol-drinking rat strain (UChA and UChB)

The objective of the present study was to analyze the prospective alterations of the testis and epididymis in a defined strain of alcoholic rats in order to contribute to our understanding of the effects of chronic alcoholism on reproduction. The testis and epididymis of the animals were submitted to morphological analysis by macroscopy, light microscopy and electron microscopy and to morphometric analysis. The UCh rats showed atrophy of the epithelium and reduction of testis and epididymis weight, liver hypertrophy and fat infiltration and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the weight and in the epithelium of the testis and epididymis and in the hypothalamus-pituitary axis of the UCh rats.

Ultrastructure of Sorubim lima (Teleostei, Siluriformes, Pimelodidae) spermiogenesis

The spermiogenesis and the spermatozoon ultrastructure of Sorubim lima were studied. Our observations showed that early spermatids are round-shaped cells, have spherical nucleus with diffuse chromatin, small quantity of mitochondria and large amount of vesicles in the cytoplasm. During the differentiation process in the nucleus, chromatin compacts in a progressive and homogeneous way, and the flagellum is formed. In the cytoplasm the vesicles, that have double membranes, aggregate and fuse on the plasma membrane. The spermatozoa of 5. lima have no acrosome and show spherical nucleus with homogeneous and highly compacted chromatin, intermediary piece with mitochondria and double wall vesicles contiguous to the plasma membrane, as well as a flagellum formed by a basic axoneme (9 + 2).

Endocytic activity in the thrombocytes of the turtle Phrynopys hilarii (freshwater South American species)

The phagocytic process in cells depends on lysosomal enzymes, high-energy metabolism and cellular recognition. In this paper, we investigated the presence of energy and recognition factors in thrombocytes of turtle Phrynopys hilarii (a freshwater South American species). Turtle thrombocytes (P. hilarii) present glycogen - possibly β particles - dispersed in their cytoplasm and glycoproteins in the cell surface, as well as a large number of enzymes involved in the endocytic process (Pellizzon, 1996). The activity of these enzymes depends on high-energy metabolism and on cellular recognition provided by specific glycoconjugates (Alberts et al., 1994). This metabolic characterization is demonstrated by the large amount of glycogen particles observed in the cytoplasm by Thiéry's method. Glycogen labeling was also observed when concanavalin A-peroxidase was used as a marker for thrombocytes and for endocyted charcoal particles. Our results show that these cells have phagocytic ability, suggesting that their function in blood circulation is not limited to aggregation but may also involve a great potential for phagocytosis.

Effects of chronic experimental alcoholism on the epithelium of the uterine horn of rats (Rattus norvegicus albinus)

Sixty adult tats (Rattus norvegicus albinus) of the same age (3 months) and with a mean body weight of 228 g were divided into two experimental groups. The control group received solid diet (Purina rat chow) and tap water ad libitum. The other (alcoholic group), received the same solid diet and was allowed to drink only sugar cane brandy dissolved in 30° Gay Lussac (v/v). At the end of periods of 90, 180 and 270 days of treatment, the animals were anaesthetized with ethyl ether during estrus, weighed and sacrificed. The final mean body weights were similar in the control and alcoholic groups. The results showed intense atrophy on the lining epithelium of the endometrium of uterine horns in the alcoholic group. Important ultrastructural epithelial alterations were also observed in the female alcoholic group, such as: intense lipid droplet accumulation, increased rough endoplasmic reticulum cisternae and mitochondrial size and presence of intraepithelial neutrophils. The secretory activity of these rats was reduced. Therefore, we concluded that alcohol acts as a toxin on the epithelial layer of the rat endometrium.

Morphology of the ventral lobe of the prostate and seminal vesicles in an ethanol-drinking strain of rats (UChA and UChB)

Chronic alcoholism alters reproduction and therefore may be responsible for alterations of prostate and seminal vesicles, which are the subject of this analysis in UCh ethanol-drinking rats. The prostate and seminal vesicles of 20 animals were submitted to macroscopic, light microscopy, electron microscopy and morphometric analysis. The UCh rats showed atrophy of the epithelium and reduction of the weight of the prostate and seminal vesicle, liver hypertrophy and fat infiltration and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the weight and in the epithelium of prostate and seminal vesicles and hypothalamus-pituitary axis of UCh rats.

Testicular ultrastructure and morphology of the seminal pathway in Prochilodus scrofa

The seminiferous tubules of Prochilodus scrofa present a coiled morphological arrangement with intertubular anastomoses and unrestricted spermatogonial distribution. The structural pattern of the seminiferous tubules is cystic, with cysts formed by cytoplasmic prolongations of Sertoli cells. Inside the cysts are observed different types of germ cells. The seminiferous tubules open individually on the ventral surface of the main testicular duct present in each testis. Each main testicular duct prolongs as a spermatic duct, fusing with the spermatic duct of the opposite side to form the common spermatic duct which opens into the urogenital papilla. The mature sperm cysts break and extravasate their content into the lumen of the seminiferous tubules from which the seminal fluid and the spermatozoa penetrate the main testicular duct, the spermatic duct and the common spermatic duct for semen ejaculation.

Morphology and histochemistry of the hyolingual apparatus in chameleons

We reexamined the morphological and functional properties of the hyoid, the tongue pad, and hyolingual musculature in chameleons. Dissections and histological sections indicated the presence of five distinctly individualized pairs of intrinsic tongue muscles. An analysis of the histochemical properties of the system revealed only two fiber types in the hyolingual muscles: fast glycolytic and fast oxidative glycolytic fibers. In accordance with this observation, motor-endplate staining showed that all endplates are of the en-plaque type. All muscles show relatively short fibers and large numbers of motor endplates, indicating a large potential for fine muscular control. The connective tissue sheet surrounding the entoglossal process contains elastin fibers at its periphery, allowing for elastic recoil of the hyolingual system after prey capture. The connective tissue sheets surrounding the m. accelerator and m. hyoglossus were examined under polarized light. The collagen fibers in the accelerator epimysium are configured in a crossed helical array that will facilitate limited muscle elongation. The microstructure of the tongue pad as revealed by SEM showed decreased adhesive properties, indicating a change in the prey prehension mechanics in chameleons compared to agamid or iguanid lizards. These findings provide the basis for further experimental analysis of the hyolingual system. © 2001 Wiley-Liss, Inc.

Exocrine glands of Schwarziana quadripunctata (Hymenoptera, Apinae, Meliponini)

This article describes the location, anatomy, histology and ontogeny of adult Schwarziana quadripunctata exocrine glands. These glands appear either as individualized organs (salivary gland system and Dufour gland) or as epidermis differentiation (tegumentary glands). Variations in the occurrence and degree of development among colony components with regard to their degree of maturity are also described.

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.

The ultrastructural aspects of vitellogenesis or oocyte secondary growth in Serrasalmus spilopleura (Teleostei, Characiformes, Serrasalminae)

Oocyte secondary growth in S. spiloleura corresponds to the period in which different vesicular structures are formed, including the cortical alveoli and the yolk granules. The oocytes with cortical alveolus formation show vesicular structures with filamentous content in the cortical cytoplasmic region, which are the cortical alveolus precursors. In these oocytes, electron-dense vesicles of heterogenous content are dispersed in the inner cytoplasmic region and their nuclei are irregular, showing many nucleoli of different sizes. The oocytes in vitellogenesis are filled with many vesicles. The cortical alveolus precursors are in the peripheral region, and electron-dense granules are seen near to the nucleus. These fuse and form yolk granules. The oocytes in vitellogenesis show a very irregular nucleus that has nucleoli of different sizes. In the oocytes in final vitellogenesis, the yolk granules are scattered throughout the cytoplasm, displacing the cortical alveoli toward cell periphery. The nucleus is similar to the other stages.

Morphology and histoenzymology of eosinophilic granulocytes in the circulating blood of the turtle (Chrysemys dorbignih)

The studies on the characterization of eosinophils and neutrophils/ heterophils of turtles are contradictory. Some authors have pointed out the existence of two distinct cell types: eosinophils and heterophils. Other authors have proposed that eosinophils and heterophils may be the same cells in different stages of maturation. These interpretations are based only on a morphological analysis. In the blood of the turtle (Chrysemys dorbignih), a South American freshwater species, there are two types of granulocytes with eosinophilic staining pattern: the first with round cytoplasmic granules and the second with ellipsoidal cytoplasmic granules. In the present study by using histoenzymological methods for the analyses of enzymological cellular content, we found that the cells with round cytoplasmic granules were positive for nonspecific esterase and the cells with ellipsoidal granules were positives for acid phosphatase, alkaline phosphatase, nonspecific esterase and peroxidase. The results show that these cells are distinct cells and that the cells with ellipsoidal cytoplasmic granules have the same histoenzymological characteristics as the neutrophils/heterophils of mammalians and other vertebrates.

Mouse extensor digitorum longus and soleus show distinctive ultrastructural changes induced by veratrine

We investigated whether veratrine (5 μl, 10 ng/kg) injected into the mouse extensor digitorum longus (EDL) (fast-twitch) and soleus (SOL) (slow-twitch) muscles provokes distinctive ultrastructural disturbances 15, 30 and 60 min later. The mitochondria in SOL were affected earlier (within 15 min) than in EDL. Swelling of the sarcoplasmic reticulum terminal cisternae was more marked in EDL than in SOL and caused distortion of sarcomeres so that fragmentation of myofilaments was more pronounced in EDL. Hypercontracted sarcomeres were seen mainly in SOL and veratrine caused infoldings of the sarcolemma only in this muscle. In both muscles, the T-tubules remained unaffected and by 60 min after veratrine most of the above alterations had reverted to normal. Pretreatment with tetrodotoxin prevented the alterations induced by veratrine. This suggests that most of the alterations resulted from the enhanced influx of Na+ into muscle fibers. These results emphasize the importance of considering the type of muscle when studying the action of myotoxic agents.