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Afferent connections to the dorsal nucleus of the lateral lemniscus of the mustache bat: evidence for two functional subdivisions.

The dorsal nucleus of the lateral lemniscus (DNLL) of the mustache bat, Pteronotus parnellii, was found to consist of two divisions. The neurons in each division were distinguished by their temporal discharge patterns evoked both by tone bursts and sinusoidal amplitude-modulated (SAM) signals. Neurons in the anterior one-third of the DNLL responded to tone bursts with an onset discharge pattern and only phase-locked to SAM signals with low modulation frequencies (< 300 Hz). Neurons in the posterior two-thirds of the DNLL responded to tone bursts with a sustained discharge pattern and phase-locked to SAM signals with much higher modulation frequencies (400-800 Hz). In addition, there was a different frequency representation in the two divisions. The frequency representation in the posterior division was from about 15 to 120 kHz, whereas in the anterior division it was only up to 62 kHz. The physiological differences were further supported by data from experiments that revealed the sources of afferent projections to the two DNLL divisions. Retrograde labeling showed that the afferent projections to the two divisions were from different neuronal populations. Input differences were of two types. Some nuclei projected to one or the other DNLL division, but not to both. For instance, the ventral nucleus of the lateral lemniscus projected predominately to the anterior DNLL and provided little or no inputs to the posterior DNLL, whereas the medial superior olive innervated the posterior but not the anterior DNLL. Other lower nuclei projected to both DNLL divisions. These include the contralateral cochlear nucleus, the ipsi- and contralateral lateral superior olives, the intermediate nucleus of the lateral lemniscus, and the contralateral DNLL. However, the projections to each division of the DNLL originate from different neuronal subpopulations in each lower nucleus. The functional implications of these findings are discussed in the context of the possible impacts that the two DNLL divisions exert on their postsynaptic targets in the inferior colliculus.

Acoustic Stimulation↗

Crystalloids in the excretory ducts of the accessory submandibular gland of the long-winged bat, Miniopterus magnator.

Cytoplasmic crystalloids are abundant in the excretory ducts of the accessory submandibular gland of the long-winged bat, Miniopterus magnator. The crystalloids, which always lack a membranous enclosure, may have an intricate silhouette. They consist of parallel linear densities with a 12.5 nm periodicity. These densities actually may be thin-walled tubules. In some crystalloids, intersecting subcrystalloids produce a complex pattern of decussate densities. In a few rare instances, continuities were detected between a crystalloid and a smooth-surfaced cisternal element. In other mammalian species, similar crystalloids connected to smooth endoplasmic reticulum play a role in steroid metabolism. We postulate that the ductular crystalloids in M. magnator might be involved in production of a factor that influences mating behavior.

Animals↗

Formation of reticulated endoderm, Reichert's membrane, and amniogenesis in blastocysts of captive-bred, short-tailed fruit bats, Carollia perspicillata.

BACKGROUND AND METHODS: As part of an effort to develop the short-tailed fruit bat (Carollia perspicillata) as a new animal model for the study of interstitial implantation and trophoblast-uterine interactions, early embryogenesis was examined histologically and ultrastructurally in captive-bred females at different intervals after the first appearance of spermatozoa in daily vaginal smears (day 1 postcoitum [p.c.]). RESULTS: In most of the early uterine embryos examined on days 16-18 p.c., much of the endoderm appeared as a reticulated meshwork; however, a unilocular yolk sac was formed prior to the development of any mesoderm. Early blastocysts of Carollia were also unusual in that endoderm surrounded much of the inner cell mass (ICM), Reichert's membrane continued over the dorsal side of the ICM, and basal laminalike material was observed around many of the endoderm and epiblast cells. A primordial amniotic cavity was formed between days 19 and 26 p.c. by cavitation. The first mesoderm appeared between days 23 and 26 p.c., concommitant with the development of an embryonic shield. CONCLUSION: The unusual reticulated appearance of early endoderm in Carollia, which is reminiscent of that seen in early human blastocysts, may be attributable to constraints imposed on growth of the blastocyst by the site and mode of implantation, temporary retardation of trophoblastic invasion by the basal laminae of endometrial epithelial elements, and endodermal proliferation in anticipation of rapid yolk sac expansion. Reichert's membrane appears to play an important role in this species in tethering the ICM and embryonic shield to the developing placenta prior to the formation of significant amounts of mesoderm.

Amnion↗

Giant, accordioned sperm acrosomes of the greater bulldog bat, Noctilio leporinus.

Sperm of the greater bulldog bat Noctilio leporinus display an architecture that is totally unique among mammalian spermatozoa. The sperm head of Noctilio is extraordinarily large and flat and lies eccentrically with respect to the sperm tail. The major portion of the atypically large acrosome lies anterior to the nucleus and is shaped into a dozen accordionlike folds that run parallel to the long axis of the sperm. The ridge of each fold is shaped into approximately 60 minute, evenly spaced rises that extend along the entire length of the fold. We speculate that acrosome ridges may serve to strengthen the sperm head during transport.

Acrosome↗

Secretory activity of gonadotropin and the responsiveness of gonadotrophs to gonadotropin-releasing hormone during the annual reproductive cycle of male bats, Rhinolophus ferrumequinum: analysis by cell immunoblot assay.

The purpose of this study was to examine secretory activity of gonadotropin (Gn) and the responsiveness of Gn secretion to Gn-releasing hormone (GnRH) in male horseshoe bats, Rhinolophus ferrumequinum, during the annual reproductive cycle. Anterior pituitary cells were monodispersed and subjected to cell immunoblot assay for Gn. Cell blots specific for follicle stimulating hormone (FSH) or luteinizing hormone (LH) were quantified using a microscopic image analyzer. The percentages of LH- or FSH-secreting cells detected as immunoreactive cell blots were markedly increased in the spermatogenic period (summer) and decreased in the hibernation period (winter). The mean Gn secretion from individual cells and total Gn secretion per unit area of the transfer membrane also showed similar changes. The responsiveness of Gn secretion to GnRH was greater in the spermatogenic period than in other seasons. On the other hand, although the secretory activity of Gn was markedly decreased during hibernation, a stimulatory effect of GnRH on Gn secretion was observed. These findings suggest that seasonal changes in the release of Gn required for gametogenesis and gonadal steroidogenesis varied depending on the reproductive activity and seasonal changes in Gn sensitivity to stimulatory effects of GnRH due to alterations in GnRH receptor numbers and/or in postreceptor events of gonadotrophs.

Animals↗

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.

Animals↗

Preimplantation development, fate of the zona pellucida, and observations on the glycogen-rich oviduct of the little bulldog bat, Noctilio albiventris.

The reproductive biology of Noctilio albiventris was investigated histologically in 112 females collected at the start of their synchronized breeding season during two different sampling years in the Cauca Valley of Colombia. Both ovaries were functional, but the animals were generally observed to be monovular. Embryonic development in the oviduct was found to proceed to blastocyst formation and loss of the zona pellucida. In 22 animals the discarded zona had been left behind in the oviduct upon passage of the embryo into the uterus and was still undissolved at the time of amniogenesis, the latest stage examined. The zonae which had passed into the uterus with the embryos often exhibited signs of dissolution. Most of the early blastocyts were morphologically distinctive, lacking a typical inner cell mass and being instead largely bilaminar. Degenerating ova from previous ovulations were found in the oviducts of two pregnant bats, suggesting that Noctilio may be another species in which the embryo stimulates its own escape from the oviduct. During tubal passage of the embryo the secretory cells of the oviductal ampulla and isthmus exhibited a transient engorgement with glycogen, particularly on the side ipsilateral to the new corpus luteum.

Animals↗

Changes in the fine structure of tanycytes during the annual reproductive cycle of the male little brown bat Myotis lucifugus lucifugus.

The hypothalamic arcuate nucleus in the male little brown bat Myotis lucifugus lucifugus was studied with the electron microscope. Animals were killed by intracardial perfusion at each season throughout the year so that the arcuate nucleus could be examined for seasonal variations in morphology. Striking seasonal changes in the fine structure of ependymal tanycytes lining the arcuate nucleus were observed. Tanycytes in animals collected in the fall and early winter exhibited pale processes characterized by a scant internal framework of microtubules and fine filaments. These processes, which were found throughout the arcuate nucleus, exhibited simple irregular shapes. In animals collected between January and June, tanycyte processes contained dense accumulations of fine filaments intermingled with microtubules, and projected long attenuated extensions that often formed multilamellar sheets around axodendritic terminals or other neuronal elements. Tanycyte processes of animals collected in July and August were densely packed with microtubules and fine filaments. The processes radiated elaborate multilamellar extensions that encapsulated axons, dendrites and even entire neuronal perikarya. Multilamellar sheets consisted of as many as 10 or 12 closely spaced gyres. The seasonal variations in tanycyte structure are suggestive of astrocyte-like behaviour. These changes are discussed with respect to seasonal changes in hypothalamic neuroendocrine activity.

Animals↗

Identification of prolactin and growth hormone cells in the pars distalis of the California leaf-nosed bat, Macrotus californicus.

Prolactin (PRL) and growth hormone (GH) cells were immunostained in the anterior pituitary lobe of the California leaf-nosed bat with antisera against rat PRL and GH by the unlabeled immunoperoxidase technique. The PRL antiserum reacts with a cell that is carmoisinophilic and pleomorphic and occurs in small groups within the ventro-, dorsocentral, and posteriolateral regions of the pars distalis. The cells that stain with the anti-GH serum are orangeophilic, smaller than the PRL cells, and randomly scattered throughout the pars distalis, singly or in palisades near capillaries. Morphometric analysis of the immunoreactive PRL and GH cells revealed an inverse relationship between these two cell types.

Animals↗

Appearance and disappearance of tubular paracrystalline structures in somatotrophs and lactotrophs during the annual life cycle of the bat (Myotis lucifugus).

The fine structure of the endocrine (granulated) cells of the adenohypophysis of the bat was examined at different periods of the annual life cycle. During the active homeothermic period (early spring until late summer) the endocrine cells were heavily granulated, and evidence of granule release was commonly seen. At this time, but not at other times (hibernation and arousal), prominent paracrystalline structures were numerous and transiently appeared in the cytoplasm of acidophils (apparently both lactotrophs and somatotrophs) but not in the cytoplasm of basophils. These paracrystalline bodies were composed of densely packed, parallel arrays of tubules. The paracrystalline structures in lactotrophs differed in size, shape, and the pattern of arrangement of component tubules from those of somatotrophs. During hibernation no evidence of exocytosis was seen, and secretory granules accumulated in the cytoplasm. Prior to arousal, secretory granules were removed by autophagy.

Animals↗

Studies on prolonged spermatozoa survival in Chiroptera: a morphological examination of storage and clearance of intrauterine and cauda epididymal spermatozoa in the bats Myotis lucifugus and M. velifer.

The cauda epididymidis, uterine corpus, and cornua and uterotubal junction of Myotis function to retain and preserve normal spermatozoa throughout hibernation. In none of the sites do spermatozoa show features that might account for their extended viability. Spermatozoa stored in the uterus and epididymis show no special orientation toward the epithelium lining these sites, whereas an intimate relationship is established between some sperm and the epithelial cells of the uterotubal junction which might either account for extended postcoital sperm survival or forecast their removal from further participation. Transmission and scanning electron microscopic observations do not disclose any morphological changes in stored luminal spermatozoa. A low rate of phagocytosis of sperm is evident in the female tract during hibernation. However, spermatozoa are evidently not vulnerable to being removed from the storage sites until spring arousal when ovulation occurs. Both uterotubal epithelial cells and phagocytes appear to be involved in the disposal of spermatozoa in the female, whereas epididymal spermatozoa apparently are primarily voided during urination. A mechanism that delays capacitation must underlie the ability of spermatozoa to survive in the female reproductive tract of the hibernating bat.

Animals↗

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.

Animals↗

Reproductive biology of the male little mastiff bat, Mormopterus planiceps (Chiroptera:Molossidae), in southeast Australia.

The anatomy, biology, and chronology of reproduction in the male of the long penile form of Mormopterus planiceps was studied in southeast South Australia and Victoria. In the morphology of its primary and accessory reproductive organs, M. planiceps was generally reminiscent of other Molossidae; however, in the specialized (sebaceous) nature of the Cowper's gland ducts, in the presence of para-anal glands, and in the unusual, horizontally bifid glans penis and the greatly elongated os penis, it was distinct from other Molossidae studied to date. Young of the year were not reproductively active. Adults displayed a single annual spermatogenic cycle that commenced in spring (September/October) and culminated in spermiogenesis in autumn (February-May), during which period plasma levels of testosterone overtook androstenedione. Thereafter, spermatogenesis appeared to cease (though scattered sperm were seen in the seminiferous tubules until August), but abundant epididymal sperm reserves persisted until September/(October). The accessory glands were hypertrophied during this period, becoming involuted by October. Although the numbers of animals available for study were small, these observations, together with the appearance of spermatozoa in the ductus deferens in August/September suggested that mating could occur during the interval from autumn to spring. Late winter/spring insemination is normal for molossids from temperate environments. However, protracted spermatogenesis commencing in spring that is not accompanied by the availability of spermatozoa until autumn, and a subsequent apparent extension of fertility (epididymal sperm storage, accessory gland hypertrophy) beyond the testicular gametogenic phase, are aspects of the male reproductive cycle in M. planiceps that have not heretofore been described in another molossid bat.

Animals↗

Reproductive biology of the female little mastiff bat, Mormopterus planiceps (Chiroptera: Molossidae) in southeast Australia.

The reproductive biology of the female little mastiff bat (Mormopterus planiceps) was studied from specimens obtained throughout the year in southeast Australia, within the region occupied only by the long penile form of this species. Mormopterus planiceps appeared to undergo a single pregnancy each year and was monotocous. Conception occurred during late winter/early spring after a protracted proestrus, during which the uterine/vaginal epithelia attained an extraordinary thickness; spermatozoa were present in the uterine corpus, vagina, and intramural oviduct for at least 2 months prior to ovulation, although only those present in the oviducts were entire and thus appeared to be viable. Following ovulation, a massive postovulatory infiltration of phagocytes occurred; and the thickness of the uterine corpus epithelium was dramatically reduced. As in other molossids, the tract was bicornuate and dextrally functional. The length of gestation was difficult to determine because early embryonic stages, up to implantation, appeared to span several months (late July/August/September) as did parturition (December/January). Growth of the young was slow; nevertheless, females attained sexual maturity in their first year. Several unusual features included the presence of a long os clitoridis, and tubuloalveolar sudoriferous and associated lobulated, sebaceous, paravaginal glands, which surrounded and emptied into the lower vagina. A deep fornix anterior and lateral to the cervix probably serves to receive the secondary glans penis. The epithelium of the uterine corpus was stratified and indistinguishable, in its cytology and cyclicity, from that of the vagina; furthermore, it lacked a glandular endometrium. This portion of the female tract likely receives the elongated primary glans. These findings are discussed in relation to other Molossidae and to the reproductive biology of male M. planiceps. Although the number of animals sampled was relatively small, the data suggest that this species does not exhibit the usual temperate molossid pattern of late winter/spring coincidence of spermatogenesis and ovulation. It would seem that pregnancy may begin, at least in some individuals, during the inhospitable winter months (when epididymal and uterine spermatozoa are abundant but spermatogenesis has largely terminated) and that additional conceptions continue into the early spring. The occurrence of sperm storage in both sexes of this species is unique among Molossidae studied to date.

Animals↗

Distribution of somatostatinlike immunoreactivity in the brain of the little brown bat (Myotis lucifugus).

The distribution of somatostatinlike immunoreactive (SLI) perikarya, axons, and terminals was mapped in subcortical areas of the brain of the little brown bat, Myotis lucifugus, using light microscopic immunocytochemistry. A preponderance of immunoreactivity was localized in reticular, limbic, and hypothalamic areas including: 1) in the forebrain: the bed nucleus of the stria terminalis; lateral preoptic, dorsal, anterior, lateral and posterior hypothalamic areas; amygdaloid, periventricular, arcuate, supraoptic, suprachiasmatic, ventromedial, dorsomedial, paraventricular, lateral and medial mammillary, and lateral septal nuclei; the nucleus of the diagonal band of Broca and nucleus accumbens septi; 2) in the midbrain: the periaqueductal gray, interpeduncular, dorsal and ventral tegmental, pretectal, and Edinger-Westphal nuclei; and 3) in the hindbrain: the superior central and parabrachial nuclei, nucleus incertus, locus coeruleus, and nucleus reticularis gigantocellularis. Other areas containing SLI included the striatum (caudate nucleus and putamen), zona incerta, infundibulum, supramammillary and premammillary nuclei, medial and dorsal lateral geniculate nuclei, entopeduncular nucleus, lateral habenular nucleus, central medial thalamic nucleus, central tegmental field, linear and dorsal raphe nuclei, nucleus of Darkschewitsch, superior and inferior colliculi, nucleus ruber, substantia nigra, mesencephalic nucleus of V, inferior olivary nucleus, inferior central nucleus, nucleus prepositus, and deep cerebellar nuclei. While these results were similar in some respects to those previously reported in rodents, they also provided interesting contrasts.

Animals↗

Implantation, development of the fetal membranes, and placentation in the captive black mastiff bat, Molossus ater.

Uterine events during pregnancy were examined histologically in laboratory-bred black mastiff bats (Molossus ater) as part of an effort to develop this species as a model for studies of the factors controlling trophoblastic growth. Embryos entered the uterus at the morula stage and in most cases shed their zonae pellucidae reasonably intact, apparently as blastocyst expansion occurred. Implantation was superficial and observed to occur only in the right uterine horn. During implantation to the endometrium by both blastocyst expansion and closure of the uterine lumen. A decidual reaction was evident at an early stage of uterine epithelial displacement and spread rapidly through the endometrium. Initial trophoblastic proliferation occurred along the uterine lumen and into the glands, while its invasion of the endometrial stroma was delayed. Although one or several primordial cavities have been observed to develop within the epiblast during implantation, these subsequently opened to a trophoepiblastic cavity, and the definitive amnion was formed by folding. A choriovitelline placenta was present briefly at thesomite stage, but disappeared as the exocoelom enlarged and the yolk sac collapsed. The latter persisted through pregnancy, however, as a glandular-appearing body. As the yolk sac retracted from the chorion, it was replaced by allantoic mesoderm, creating a diffuse labyrinthine endotheliodichorial placenta. This was prominent during mid-gestation, but was supplanted by the discoidal hemochorial placenta as the major site of feto-maternal exchange during late pregnancy.

Animals↗

Development of the discoidal hemochorial placenta in the black mastiff bat, Molossus ater: evidence for a role of maternal endothelial cells in the control of trophoblastic growth.

In an effort to define further the factors that can influence trophoblastic growth, development of the discoidal chorioallantoic placenta was examined histologically in laboratory-bred black mastiff bats, Molossus ater. This placenta normally forms only at the cranial end of the right uterine horn. Its positioning was found to be related to the development of an unusual vascular tuft in the endometrium at this site following ovulation. When a decidual reaction occurred, the endothelial cells of the tuft vessels hypertrophied and their basal laminae became more prominent in sections stained for glycoproteins. Cytotrophoblast then proliferated preferentially around the tuft in the right horn and its vessels became surrounded by trophoblastic cuffs. A functional placenta was formed when trophoblastic tubules grew out from these cuffs, became interconnected, developed lumina, and began to carry maternal blood. Maternal endothelial cells generally persisted only in portions of the larger vascular tubules. These observations suggest that the endothelial cells of the tuft vessels may be secreting factors that influence early trophoblastic growth and are at least partially incorporated into the basal laminae of the endothelial cells. Evidence is reviewed which raises the possibility that endometrial endothelial cells might have a similar morphogenetic role in many other mammals with invasive trophoblast. Mitotic activity in the trophoblastic tubules of M. ater ceased when the tubules became patent and began to carry blood. Further growth was then accomplished by hypertrophy of the existing cytotrophoblast cells. Electron microscopic examination of near-term placentas confirmed that the interhemal barrier was hemomonochorial and lacked a continuous layer of syncytiotrophoblast.

Animals↗

Lipid droplet accumulation in cardiac muscle cells of the bat: potential auto-toxicity of the cardiac sympathetic innervation.

The previously described ability of reserpine and parachlorophenylalanine to induce the accumulation of lipid droplets in ventricular cardiac muscle cells of the bat was investigated. Lipid droplet accumulation was assessed qualitatively by light microscopy and quantitatively by morphometric analysis of electron micrographs. An hypothesis that the action of the drugs was an indirect one, mediated by the cardiac adrenergic innervation, was framed and tested. Lipid droplet accumulation occurred during a time of intense sympathetic activity, that of arousal from hibernation. The ability of the two drugs to produce the effect was antagonized by prior sympathetectomy with 6-hydroxy-dopamine. The effect was mimicked by administration of exogenous norepinephrine together with inhibitors of its catabolic enzymes, monoamine oxidase and catechol-o-methyl transferase. These observations are all consistent with the initial hypothesis and raise the possibility that endogenous norepinephrine in the cardiac sympathetic innervation might be, at least potentially, auto-toxic.

Animals↗