Fat body, hemolymph and ovary routes for delivery of substances to ovary in Melipona quadrifasciata anthidioides: differences among castes through the use of electron-opaque tracers.

The yolk protein precursor, vitellogenin (Vg), in bees is synthesized in the fat body trophocytes, delivered to the hemolymph and ultimately absorbed from there during the vitellogenic phase of oocytes in the active ovary. The routes tracing the material exchange that occurs between the trophocytes and the hemolymph, in addition to the transportation from the hemolymph to the ovarian follicles, were marked by alkaline phosphatase and lanthanum nitrate (LN). Active ovaries from nurse workers and physogastric queens, as well as inactive ovaries of virgin queens, were examined by transmission electron microscopy. The LN permitted better visualization of the routes of exchanges between the organs and the hemolymph. Both methods demonstrate the apparent differences between the phases of the ovary and the bee caste. In inactive ovaries of the virgin queens, the routes from the follicular epithelium to the oocyte remain closed; conversely, they are open in active ovaries of the nurse workers and physogastric queens. The differences between the methods and classes of bees are discussed.

Effects of treatment of the fat body trophocytes of Melipona quadrifasciata anthidioides nurse workers and virgin queens in culture by juvenile hormone III and ecdysterone (20-HE).

The fat body (FB) consists of two types of cells: throphocytes and oenocytes. Throphocytes are related to intermediary metabolism storing lipids, carbohydrates, and proteins while oenocytes play role in the lipids and lipoproteins production. The vitellogenin is the precursor of egg yolk (vitelline) and is synthesized on FB. The aim of this work was to analyze the effects of hormones acting in bee reproduction, as juvenile hormone (JH) and ecdisteroids (20 HE) on FB cells, where vitellogenin is synthesized. For the study were chose nurse workers that in Melipona quadrifasciata anthidioides present activated ovaries and produce eggs, and virgin queens whose ovaries are not yet activated, presenting only previtellogenic follicles. FB trophocytes from these classes of bees were cultivated in media containing different amounts of JH and 20-HE. The effects on trophocytes cytoplasm reserves of lipids, proteins, and activity of acid phosphatase were compared by observing preparations from cultured FB, treated and control, by transmission electron microscopy (TEM). The results showed that the hormones effects are related to the bee's caste and functional ovary stage. The role of acid phosphatase on mobilization of the trophocyte reserves was also determined.

Cytochemistry of fat body trophocytes and ovaries of workers and queens of Melipona quadrifasciata anthidioides (Hymenoptera: Apidae: Meliponini) during vitellogenesis.

The fat body (FB) of insects is where yolk proteins are synthesized. Therefore, relationships between the FB and oogenesis were studied in nurse workers, virgins, and physogastric queens of Melipona quadrifasciata anthidioides, a stingless bee in which the workers produce and lay eggs while provisioning brood cells. The relationships between FB and oogenesis, as well as the routes of materials from hemolymph to the oocytes, were studied through the cytochemical detection of lipids by osmium imidazole (OI), carbohydrates by ruthenium red (RR) and basic proteins by ammoniacal silver (AS). The results show differences in the presence of the studied materials in FB trophocytes and ovary of the classes of females studied and oogenesis phases. Material that tested positive for the treatments was detected among the classes of individuals studied in both, trophocytes and oocytes, and in the route of those materials from hemolymph to the oocytes. The differences found among the individual classes indicate relationships with the nutrition and adaptation to the parsimonious use of nutrients in the metabolism of reproduction.

Ovarian ultrastructure in virgin queens of Apis mellifera L. narcotized by CO2.

Ovaries of newly emerged virgin queens, aged 1, 3, 5, and 10 days, were submitted to an atmosphere of CO2 for 1 min, and studied by transmission electron microscopy. Newly emerged control and experimental queens were kept caged during 15 days with six nurse workers (changed every 2 days) and fed with sugar candy and water ad libitum. For the ultrastructural studies, the ovaries were collected when the queens were 3, 5, 10, and 15 days old. The effect of the treatment on the ovary was mainly evaluated by cell death incidence and acid phosphatase detection. Neither the confinement nor CO2 treatment interfered with the pattern of queen ovarian development for up to 5 days; that is, no differences were observed in ovarian ultrastructure or acid phosphatase activity between control and treated queens. Beyond the age of 10 days, treated queens showed a lower rate of cell death than controls, but the positivity to acid phosphatase reaction remained similar in both. The protective effect of CO2 on cells, however, did not persist, and 15-day-old treated queens showed no differences compared to the controls in the rates of cell death or enzyme reactivity. In conclusion, treatment with CO2 did not significantly change the ultrastructure of ovarian cells, or acid phosphatase activity in them, during the caging time of the queens. From an applied perspective, the short-term imprisonment of newly emerged queens outside of the colony, as is done by beekeepers, does not seem to affect their performance in terms of future fertility, although the narcosis, as applied in this study, may negatively affect their long-term performance.

Association of the time course of pulmonary arterial hypertension with changes in oxidative stress in the left ventricle.

1. This study investigates the time course of pulmonary arterial hypertension (PAH) due to monocrotaline (MCT) and its association with cardiac function and oxidative stress markers in the left ventricle (LV). 2. Male Wistar rats were divided into six groups: 7 days, 21 days, and 31 days for both control and MCT groups. Following echocardiographic analysis, the heart was removed. The LV was separated and homogenized to analyze oxidized-to-total glutathione ratio and thioredoxin reductase (TrxR) activity as well as hydrogen peroxide (H(2) O(2) ) and ascorbic acid levels. 3. There was significant (P < 0.01) cardiac and right ventricle (RV) hypertrophy and pulmonary congestion in the MCT 21 day and 31 day groups. Echocardiography showed a change in the flow wave of the pulmonary artery at 21 days after MCT treatment. There was an increase in the LV ejection time (P < 0.05) at 31 days after MCT. The LV H(2)O(2) concentration was increased (P < 0.05) in the MCT 21 day and MCT 31 day groups compared with controls. There was a reduction (P < 0.05) in the LV ascorbic acid concentration and an increase (P < 0.05) in TrxR activity in the MCT 31 day rats. 4. Our findings showed RV changes due to pulmonary hypertension at 21 days after MCT injection. There was a correlation between the degree of dysfunction and the morphometry of the heart chambers, along with impairment of the antioxidant/pro-oxidant balance in the LV 31 days after the beginning of the protocol. This study suggests that LV changes follow RV dysfunction subsequent to pulmonary hypertension.

Programmed cell death during ovarian differentiation in queens of Apis mellifera Linné, 1758 (Hymenoptera, Apini)

In this study, we examined various aspects of ovarian development in adult honey bees queens (Apis mellifera). Caged honey bee queens showed an initial programmed germ cell differentiation that was independent of any external or environmental stimulus. In young queens, division of the stem germ cells resulted in cysts of clone cells (cystocytes) that were connected through intercellular bridges and appeared as rosettes. The cystocytes started differentiate shortly before the queens reached sexual maturity (about 5 days old). The oocyte subsequently appeared as a large, stained cell connected to parallel, double rows of smaller cells, or nurse cells. If the queens did not mate, germ cell differentiation was “switched off”, and development of the ovary was interrupted in an intermediate developmental stage, without follicular cell organization. Therefore, in such queens, there were no pevitellogenic follicles. This finding could explain why wvirgin queens rarely lay eggs and why after fecundation they require several days to start laying. The absence of previtellogenic follicles may also indicate that some stimulus is required for continuation of the vitellogenesis and ovary development.