Fetal development, and placental and maternal plasma concentrations of progesterone in the little brown bat (Myotis lucifugus).

Progesterone concentrations measured in plasma samples from 280 bats captured during pregnancy or early lactation were related to fetal attributes indicative of stage of pregnancy. Fetal weight increased exponentially from 40 mg at crown-rump length of 6 mm to 2000 mg at 23 mm (term). Fetal weights at term accounted for up to 35% of the weight of intact pregnant animals. Progesterone concentrations increased from less than 5 ng/ml at 2 mm estimated crown-rump length to plateau values of approximately 65 ng/ml (geometric means) from 16 mm crown-rump length until the most advanced stages of pregnancy. Mean concentration in 8 post-partum bats, most of which were actively lactating, was 8.4 ng/ml; 11.6 ng/ml was measured in one animal that was carrying a wet neonate when sampled yet was still pregnant when captured 5 h earlier. Placental concentrations of progesterone ranged from 43 to 964 ng/g wet weight of tissue and mean values increased in a similar fashion though were about 4-fold greater than changes in plasma concentrations of the steroid. The concentrations in placental tissue were at least 15- to 20-fold higher than could be expected from blood contamination, indicating that placental steroidogenesis is likely to occur in this species.

Plasma membrane structure of bat spermatozoa: observations on epididymal and uterine spermatozoa in Myotis lucifugus.

Plasma membrane structure of bat spermatozoa was examined utilizing electron microscopy of thin sections and freeze-fracture replicas. Notable membrane features observed in replicas from cauda epididymal spermatozoa included specialized particle aggregates at the junction between the acrosomal and postacrosomal region of the head (a membrane structure not previously described in mammalian spermatozoa) and another row of rod-like particles just anterior to the posterior ring. Both of these specializations in fractured plasma membranes correspond with regions where the membrane is closely apposed to underlying structures when viewed in thin sections. The postacrosomal sheath appears to be composed of an array of longitudinally oriented filamentous components. Characteristic ordering of intramembranous particles was also noted in replicas from the midpiece region and the annulus. Major changes in plasma membrane structure were not seen in spermatozoa stored in the female reproductive tract; however, the appearance of linear particle aggregations in the principal piece membrane was noted. No evidence was obtained to suggest that an acrosome reaction had occurred in spermatozoa stored in females.

Annual skeletal changes in the little brown bat, Myotis lucifugus lucifugus, with particular reference to pregnancy and lactation.

Studies of bone from summer-active little brown bats, Myotis lucifugus lucifugus, have demonstrated sex differences in the renewal of skeletal mineral reserves following spring-arousal from hibernation. Patterns of bone remodeling in both sexes of bats indicate that new bone formation does not occur during hibernation: All new bone formation occurs during the summer-active season. Results show that a short period of time elapses after hibernation before the initial demands of a large fetus and rapidly growing neonate are expressed on maternal skeletal reserves. Bone loss in summer-active females was associated with pregnancy and lactation, whereas summer-active males did not show evidence of bone loss but, instead, uninterrupted bone accretion throughout the summer-active season. Osteoclasts and bone-forming osteoblasts, absent during the hibernation period, reappeared on bone surfaces following spring-arousal from hibernation. There was no apparent increase in osteoclast numbers or activity during lactation but resorption cavities were found in deep cortical lamellae distant from bone surfaces. The increase in bone resorption in lactating bats appeared to be by osteocytic osteolysis, suggesting that it might be a significant mechanism of bone/calcium regulation in this hibernating mammal throughout the year.

Morphology of thyroid C-cells and parathyroid glands in summer-active little brown bats, Myotis lucifugus lucifugus, with particular reference to pregnancy and lactation.

Morphological studies of thyroid C-cells and morphometric analyses of parathyroid glands in summer-active little brown bats indicated concomitant regulatory endocrine functions correlating with bone remodeling. C-cells apparently maintain maximal activity throughout the summer-active period in all bats. However, the hyperactivity of the parathyroid glands in summer-active female bats can be correlated with the maintenance of plasma calcium concentrations during lactation, when the female skeleton undergoes a period of bone demineralization. In summer-active male bats, which did not lose bone, the parathyroid glands did not show morphological evidence of hyperactivity; instead they were found to exhibit moderate parathyroid activity.

Distribution of iron in the gastrointestinal tract of the common vampire bat: evidence for macrophage-linked iron clearance.

Iron in the tissues of the digestive tract of the common vampire bat (Desmodus rotundus) has been studied using histochemical, electron microscopic, and autoradiographic methods. This animal is an obligate sanguivore and has a daily intake of dietary iron 800 times that of man. The amount and distribution of tissue iron is not affected by either a single blood meal or starvation but does reflect the degree of siderosis of each animal's liver and spleen. By 7 days after the injection of a trace amount of 55Fe into the peritoneal cavity, labelled siderotic macrophages are present both on the serosa and within the walls of the stomach and intestine. In the lower intestine, such cells can be derived from the germinal centers of Peyer's patches. Siderotic macrophages are often situated in the lamina propria under areas of siderotic epithelium. Label is also present in the apical cytoplasm of mucosal epithelial cells, a region of abundant siderosomes. The ultrastructure of these organelles is extremely variable. Accumulations of membranous whorls and stacks, "stippled bodies," ferritin molecules, and larger "ferruginous" complexes are bounded by one or a number of membranes. Iron is excreted when these epithelial cells are desquamated into the gut lumen. Similar Prussian blue-positive granules are present in the feces. Unlike other glandular cells, the parietal cells of the fundic caecum are siderotic. Their cytoplasm contains abundant siderosomes and ferritin with accumulations of amembranous ferritin bodies in the intracellular canalicular spaces. Prussian blue-positive granules are present in the lumens of fundic glands.

Structure and morphogenesis of the uterus, placenta, and paraplacental organs of the neotropical disc-winged bat Thyroptera tricolor spix (Microchiroptera: Thyropteridae).

The structure and development of the female reproductive tract, fetal membranes and placental have not previously been recorded for any member of the family Thyropteridae. Recently implanted embryos were obtained in late January, limb-bud stages in March, and full-term fetuses in late May, suggesting a possible gestation length of approximately five months. It is likely, however, that Thyroptera experiences at least two breeding cycles per year. The uterus was narrowly bicornuate; the corpus uteri was unusually large and lacked the glandular density observed in the cornua. The cervix was long, pleated, and relatively aglandular. The oviducts opened at the apices of the cornua; oviductal papillae were absent. A bursa ovarii surrounded the ovary, but there was a small pore opening to the peritoneal cavity adjacent to the fimbriated end of the oviduct. Never more than a single embryo or fetus was present, and only a single corpus luteum was observed; thus Thyroptera, like most bats, is monovular. Ovoimplantation was interstitial; a decidua capsularis was present early but disappeared by the late limb-bud stage. The decidual reaction involved both glandular epithelium and stromal cells, but most of the decidua was destroyed by term. Amniogenesis was initiated after implantation, by cavitation. Primitive entoderm was formed precociously above, as well as below, the presumptive embryonic disc, and a thin extension of Reichert's membrane passed over the cell mass, separating it from the cytotrophoblast of the chorionic placenta. During the amniogenic period, the yolk-sac entoderm fused to the parietal trophoblast via an intervening Reichert's membrane, forming an extensive bilaminar omphalopleure; this was rapidly converted to a trilaminar structure in early post-implantation stages. An avascular chorio-vitelline relationship involved most of the chorionic wall in early post-implantation stages and persisted to term in the abembryonic hemisphere after the partial inversion of the yolk-sac roof in late presomite embryos. The invaginated yolk-sac roof (splanchnopleure) also persisted to term as a viable paraplacental component. A small sac-like allantois was formed between late presomite and early limb-bud stages but disappeared by the late limb-bud stage. Development of the definitive chorioallantoic placenta resembled that in other bats, but the maternal endothelium disappeared relatively early, and trophoblastic differentiation was precocious. The ultrastructural organization of the interhemal membrane was hemodichorial, and otherwise generally resembled the organization previously described in vespertilionid bats. Similarities and differences in the structure of the uterus, placenta, and paraplacental organs of Thyropteridae, in comparison with other families of bats, are discussed. On the basis of fetal membrane characteristics, the Thyropteridae show closer affinities with the Phyllostomatoidea than with the Vespertilionoidea, to which they are presently assigned.

Reproductive asymmetry and unilateral pregnancy in Chiroptera.

Bats are noteworthy in that anatomical and/or functional asymmetries of the female reproductive organs are far more frequent and profound than recorded for any other mammalian Order, being observed in representatives of all 13 familis so far examined. Asymmetry is expressed as a unilateral dominance of an ovary, the uterus, or both. Dextral dominance is most frequent, but cases of sinistral dominance are recorded. In general 6 basic patterns of asymmetry are recognizable. These are briefly described. Physiological factors which may condition expressions of asymmetry in specific cases are discussed, and some avenues for future research are suggested.

The annular hematoma of the shrew yolk-sac placenta.

The annular hematoma of the shrew, Blarina brevicauda, is a specialized portion of the yolk-sac wall. In this study, we have examined the fine structure of the different cellular components of the anular hematoma. Small pieces of the gestation sacs from seven pregnant shrews were fixed in glutaraldehyde and osmium tetroxide and processed for transmission electron microscopy. In the area of the trophoblastic curtain, the maternal capillary endothelial cells were hypertrophied and syncytial trophoblast surrounded the capillaries. Cellular trophoblast covered part of the luminal surface of the curtain region, whereas masses of apparently degenerating syncytium were present on other areas of the surface. Maternal erythrocytes, released into the uterine lumen from the curtain region, were phagocytized and degraded by the columnar cells of the trophoblastic annulus. No evidence of iron or pigment accumulation was evident in the parietal endodermal cells underlying the annular trophoblast. Parietal endodermal cells were characterized by cuboidal shape, widely dilated intercellular spaces, and cytoplasm containing granular endoplasmic reticulum. Endodermal cells of the visceral yolk-sac accumulated large numbers of electron-dense granules as well as glycogen in their cytoplasm. Hemopoietic areas and vitelline capillaries were found subjacent to the visceral endoderm. The various portions of the yolk-sac wall of Blarina appear to perform complementary functions which are probably important in maternal-fetal iron transfer.

Calcitonin and hibernation bone loss in the bat (Myotis lucifugus).

Inhibition of osteocytic osteolysis in bats given calcitonin injections during late hibernation supported previous morphologic evidence that the bone loss during hibernation is associated with decreased calcitonin and increased parathormone activity.

Cytological development of yolk sac endoderm and protein-absorptive mesothelium in the little brown bat, Myotis lucifugus.

The yolk sac of the little brown bat is unusual in that during the course of gestation both the inner endodermal cells (bordering the yolk sac cavity) and outer mesothelium (facing the exocelom) form simple columnar epithelia which persist throughout gestation. These endodermal cells develop an extensive system of agranular endoplasmic reticulum, numerous lipid droplets and unusual "giant" mitochondria. During development the Golgi apparatus changes position from the apical to the basal side of the nucleus, reversing the polarity of the cells. In general, the endodermal cells have cytological features suggestive of synthetic or secretory activity. The mesothelial cells develop an extensive "absorptive apparatus" in their apices, while large crystalloid-containing granules become numerous in their basal cytoplasm. The mesothelial cells have large deposits of glycogen, especially during mid-gestation, but few mitochondria and little granular endoplasmic reticulum. Endodermal cells do not absorb exogenous protein (peroxidase) even if it is injected directly into the yolk sac cavity. However, placement of peroxidase either in the exocelom or in the maternal vascular system results in the appearance of this protein in the "absorptive apparatus" of mesothelial cells as well as in macrophages in the stroma of the yolk sac. While evidence of absorption was clear, no direct evidence of transport of tracer to fetal blood vascular system was obtained. It is postulated that a major function of the hypertrophied mesothelial cells during gestation is the absorption of proteins and possibly other substances from the exocelomic fluid. The major function of the hypertrophied endodermal cells may be synthesis and secretion of substances into the fetal circulation.