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Naked-backed bats of the genus Pteronotus (family Mormoopidae) occur in the Neotropics from Mexico through northern South America. These are relatively small-sized insectivorous species that frequently roost in caves. Eight specimens of naked-backed bats (Pteronotus parnellii) were live-trapped in Suriname and one in Cuba (P. quadridens). Their parotid glands were fixed in an aldehyde mixture designed for field work and postfixed in the laboratory with osmium tetroxide. Tissues were further prepared for electron microscopy by conventional means. The parotid glands of the two species of Pteronotus closely resemble each other except for the substructure of their serous secretory granules. Serous granules in P. parnellii are bizonal, with a moderately dense inner matrix and an outer, denser corona or crescent. The matrix is occupied by laminae, flakes, and filaments in random array. In contrast, serous granules in P. quadridens consist of a uniform matrix that contains dense, usually stacked toroids or tubules either in random array or packed in bundles. A parotid gland from one specimen of P. parnellii contained an endpiece that consisted of cells that contained giant (up to 9 pm in diameter) serous granules. Serous cells in both species contain aggregates of small, uniformly dense, rod-like, membrane-delimited organelles as well as occasional bundles of cytofilaments. The endpieces are separated from intercalated ducts by a ring of granulated cells that contain secretory granules that often have a bull's-eye configuration. Intercalated and striated ducts are typical in appearance, except that many of the cells in the latter contain small, dense secretory granules in their apical cytoplasm. The parotid glands in the two species of naked-baked bats differ slightly in terms of acinar secretory granule ultrastructure, but otherwise are fairly conservative. It is thought that the glands in these particular bats might represent the "basal" condition of the salivary glands of insectivorous bats and thus can serve as a reference point for making comparisons to the highly diversified (in terms of diet) phyllostomid bats.

The tent-building bat, Uroderma bilobatum, is a small, frugivorous phyllostomid bat with a broad neotropical distribution. Generally found in humid forest, this bat lives in small groups that create daytime "roosts" from large leaves of a variety of tropical plants. Fruit eating engenders a variety of ecological and physiological challenges for bats, some of which could require adaptive features in their salivary glands. The parotid salivary glands of Uroderma bilobatum were prepared for transmission electron microscopy by using methods that have become standard for field work. The parotid gland is extremely unusual in structure. Although the secretory endpieces still produce serous granules with a complex substructure, they are modified into quasi striated ducts. Their basal folds, which are extensive, occasionally harbor some vertically oriented mitochondria, imparting a resemblance to striated ducts. Other evidence for the endpiece origin of these parenchymal components is a well-developed system of intercellular canaliculi, structures that never occur in bona fide striated ducts. The long but sparse intercalated ducts consist of two types of cells, each of which elaborates a modest number of secretory granules of differing substructure. Striated ducts are of conventional morphology, except that a few dark cells shaped like wine glasses are present in their walls. The striated duct cells produce no secretory granules, but their apical cytoplasm may contain some small, empty vesicles. Capillaries lie in longitudinal grooves in the base of the duct cells, an arrangement that might enhance electrolyte exchange. Excretory ducts consist of simple cuboidal epithelium composed of cytologically unspecialized cells that sometimes includes a dark cell. It was concluded that salivary glands could have a major role in adapting species to acquire nutrients from marginal sources, such as tropical fruits, which have a low protein and sodium content. The unusual parotid acinar cells in Uroderma bilobatum are discussed in the context of salivary pH and buffering capacity. Comparisons are made with four other bat species, including an insectivorous species with a salivary pH > 8.0 and a very high buffering capacity, an intermediate species, and a fruit bat with acidic-stimulated saliva and very low buffering capability. Such interspecific comparisons provide a foundation for hypothesizing that ultrastructural features of the acinar cell basolateral membranes and intercellular canaliculi correlate with differences involving Na/H+ exchangers and release of HCO3- and, thus, are associated with the species differences that are important to diet and nutrient acquisition.

Background: Many species of bats have two sets of submandibular glands, principal and accessory. The accessory gland may resemble the principal one but more often shows wide morphological divergence. The free-tailed bat, Tadarida thersites, is very unusual in that it has two sets of parotid glands rather than binary submandibular glands. We studied the ultrastructure of the principal parotid gland to establish a baseline for comparison with the accessory parotid., Methods: Two specimens of adult free-tailed bats, one male and one female, were live-trapped in western Kenya. Parotid glands were fixed for electron microscopy using a protocol expressly designed for field fixation and then embedded by conventional means., Results: Histologically, the principal parotid is a typical serous gland. The secretory granules of the endpiece cells have an unusual substructure in that they contain variable numbers of lucent halos and one or several spherules. Intercalated duct cells contain a significant number of dense, serous-like granules. Striated ducts have the usual basal configuration of mitochondria and folded plasma membranes, but the supranuclear cytoplasm contains many small, dense granules, so that these ducts resemble the granular convoluted tubules found in the submandibular glands of many families of rodents. The apices of the duct cells have a peculiar contour--the luminal surfaces obliquely invaginate into the apical cytoplasm, so that in thin section the luminal membranes appear to be underlaid by a layer of vacuoles., Conclusion: Although the principal parotid gland of the free-tailed bat shows some distinctive, species-specific ultrastructural features, it basically is similar to the parotid gland in two other molossid bats, Tadarida brasiliensis and Molossus molossus. The distinctive features in the principal parotid gland of T. thersites might relate to its feeding on hard-bodied insects and perhaps to the production of lysozyme.

Background: Many bat species have an extra set of major salivary glands. In some species, the accessory glands are quite similar to the principal one, but in others they may be radically different. Accessory glands usually are associated with the submandibular gland, but the free-tailed bat, Tadarida thersites, also has an accessory parotid gland. In the present study, we compared the accessory parotid gland with its principal counterpart., Methods: Salivary glands were removed from two specimens of free-tailed bats, one of each sex, that had been live-trapped in western Kenya and immersion-fixed in a specially formulated mixture designed for field fixation. Once back in the laboratory, the tissue was further prepared for electron microscopy by conventional means., Results: The secretory endpieces consist of serous tubules composed of typical serous cells that contain numerous dense granules. In contrast, the intralobular duct system shows a radical departure from normal. These ducts are enormous in caliber, their lumina measuring greater than 100 microm in diameter. They appear to arise by amalgamation of the homologues of intercalated and striated ducts into macroducts. The walls of the macroducts consist of intermingled patches of simple cuboidal and simple columnar epithelia that occasionally include a tuft cell and are underlaid by an almost continuous layer of myoepithelium. A few cells have some modified basal striations, but most cells display a cytological organization that differs radically from either of their two putative ancestral cell types. Both tall and short epithelial macroduct cells have a paranuclear collection of ovate mitochondria and aggregates of what presumably are peroxisomes. Macroduct cells in both the female and male are pervaded by a system of tubular smooth endoplasmic reticulum (SER). In the female, the SER gives rise to membranous whorls that consist of numerous plies. As the macroducts approach their termini, a single row of small dense secretory granules appears just beneath their luminal surface. At the lobular periphery, the ducts taper down to become excretory ducts of normal dimensions., Conclusions: An accessory parotid gland occurs in T. thersites, but apparently is absent in the related species, T. brasiliensis. The ultrastructural data are consistent with a possible steroidogenic function, although other features of the gland might relate to the elaboration of a secretory product associated with feeding on chitinous beetles. The macroducts conceivably function as reservoirs of preformed saliva.

Background: Basal folds are slender plications at the basal surface of acinar cells in the salivary glands of many mammalian species. These largely organelle-free folds increase the surface area of the basal plasmalemma manyfold and are unquestionably involved in the translocation of organic and inorganic molecules and water into the acinar cells., Methods: Specimens of salivary glands were obtained from over 230 species of live-trapped bats from major areas of the globe. Tissues for electron microscopy were fixed and processed by conventional means., Results: A number of the bat species examined had dense material in the intercellular spaces between basal and lateral folds of serous cells in the parotid gland. This intercellular material was particularly prominent in three species of New World bats, viz., Pteronotus parnellii, P quadridens, and Phyllostomus latifolius, and in one species of Old World bats, Chalinolobus argentatus. This dense material, which has a farinaceous texture, appears not to pass through tight junctions, so it is excluded from the lumina of intercellular canaliculi and acini. The dense material originates in the acinar cells--it is carried to the membranes of the folds via coated vesicles, which empty their dense content by exocytosis into the intercellular space. Similar dense material is present in the intercellular spaces of the basal labyrinth of striated ducts in the two species of Pteronotus. The manner in which this material accumulates in the striated duct is unclear., Conclusions: Although the function of the intracellular dense material is undetermined, it appears to be placed strategically to influence molecular traffic into acinar cells or to modulate the paracellular pathway. From a comparative evolutionary perspective, we hypothesize that, in bats, the combination of basal folds and extracellular densities is associated with insectivory. Similar morphologies appear to be lacking in frugivorous or nectarivorous species.

Background: Hart's little fruit bat, Enchisthenes hartii, is uncommon and, although it has been the subject of recent molecular genetic studies, is little known biologically. Because chiropteran salivary glands vary interspecifically in ways that reflect evolutionary history and ecology, we examined the parotid gland in E. hartii to ascertain the extent to which it resembles homologous glands in species to which this bat presumably is related., Methods: The parotid glands were prepared for electron microscopic examination by conventional means., Results: The parotid gland of E. hartii is structurally unique among all previously studied species of bats (> 230 species examined) and other mammals. In contrast to the same gland in other mammals, the parotid gland in E. hartii lacks secretory endpieces. In their place, there is a type of striated duct. Thus, in this species single secretory elements consist of (proceeding in the direction of salivary flow): striated duct--intercalated duct--and a conventionally located striated duct. The proximal ducts possess microvillus-lined intercellular canaliculi, whereas the walls of the distal ducts include occasional dark cells. Some small serous-like granules may be present in the intercalated duct cells., Conclusions: The function(s) and biological role of the unique parotid gland in E. hartii are unknown. Nevertheless, the presence of two sets of striated ducts provides two separate glandular components seemingly capable of electrolyte transport. This might be of adaptive significance in enabling this species to make use of tropical nutrient resources that otherwise would be unavailable. The uniqueness of its parotid glands lends support to the current hypothesis that E. hartii should be classified as a monotypic genus rather than as a species of Artibeus, whose members it resembles morphometrically.

Background: All examined mammals have at least two sets of lingual salivary glands: von Ebner's glands and Weber's glands. A third set, the glands of Blandin and Nuhn, is present in the tongues of some but not all mammals. Vampire bats, Desmodus rotundus, are unusual in that they possess another set of lingual glands, these being in the midtongue region., Methods: The anterior half of the tongue was extirpated from several adult vampire bats, dissected, and tissue blocks derived from the midregions of the body of the tongue prepared for transmission electron microscopy by conventional means., Results: The midlingual glands are in the form of long, tubular secretory endpieces that are succeeded by ducts of simple morphology. In general, the secretory portions consist of two cell types, which may be intermingled in the same tubule or may form tubules that consist wholly of one cell type or the other. Seromucous cells usually have one or several rough endoplasmic reticulum cisternae that are hugely distended by a homogeneously dense material. Their granules have a bizonal substructure: one or several dense bands are embedded in a lighter matrix. Mucous cells are rather typical in structure, but their secretory product is different from run-of-the-mill mucous droplets. These droplets vary in density from cell to cell. In some cells, these droplets have a relatively light matrix; in other cells, the droplet is unusually dense, consisting mainly of a dark, structureless matrix with marginal lenticular lacunae of low density in which some short, irregular filaments are scattered. A rare finding is the presence of ciliated cells intermingled with secretory endpiece cells. The cilia are of conventional morphology. Secretory tubules are succeeded by ducts that resemble intercalated ducts; the epithelium of these ducts gradually increases in height to form a kind of excretory duct, without the intervention of striated ducts. As the ducts approach the lingual surface, the epithelium changes to stratified squamous., Conclusions: Saliva produced by the midlingual glands may be an aid in the reciprocal grooming behavior of vampire bats. Based on their morphology, the excurrent ducts may not modify the initial saliva elaborated by these glands and might act simply as pipelines by which the saliva reaches the mouth.

Background: In previous studies, we determined that the submandibular glands of five species of Neotropical fruit bats in the genus Artibeus had seromucous granules in their demilune cells with substructures that varied interspecifically in accordance with systematic relationships. Moreover, the striated ducts in these frugivores exhibited structural modifications that apparently are related to the consumption of a diet rich in potassium, but deficient in sodium. We now turn our attention to the parotid gland in a large number of species in this genus to determine if it follows the same structural pattern as does the submandibular gland., Methods: Members of seven different species of Artibeus were live-trapped in various Neotropical locations. The parotid glands were extirpated from euthanized bats, fixed in the field, and prepared for electron microscopic examination by conventional means. The parotid glands in all seven species were virtually identical in morphology. The acinar cells (determined to be seromucous on the basis of ultrastructural criteria) contain large numbers of what appear to be vacuoles, but which are a type of secretory granule. These granules have an electron-lucent matrix and may contain one or several circular membranous profiles arranged either concentrically or in a random array. These granules appear to form by progressive dilatation of the termini of Golgi saccules, with the nascent granules finally severing their connection with the Golgi apparatus. Many of the internal membranous profiles are formed simply by invaginations of the limiting membrane of the granule; others may result from indentation of the limiting membrane by protrusions from adjacent granules; the source of multiple internal membranes in certain granules is unclear. The exocytosis of these granules results in the acinar and intercalated duct lumina being filled with an abundance of membranous material. Such extruded membranes are present in some striated ducts, but not in others, suggesting that they are degraded during passage through the duct system. The striated ducts are of conventional appearance, lacking the frondose processes that are prominent in the submandibular glands of Artibeus., Conclusions: The parotid gland in Artibeus shows none of the interspecific ultrastructural variability that characterizes the submandibular gland in bats of this genus. The seromucous acinar cells secrete granules that release phospholipids as well as glycoconjugates into the saliva. Based on the lack of frondose processes with their sodium-transporting portasomes, the striated ducts of the parotid gland are less concerned with electrolyte homeostasis than are those in the submandibular gland.

Background: The phyllostomid fringe-lipped bat, Trachops cirrhosus, is sui generis (in a family of ca. 138 species) in that it subsists in part on tropical frogs. These amphibians frequently possess highly toxic integument. We examined the salivary glands of this bat to determine if these glands could be the source of protective factors that permit consumption of seemingly unsavory prey. The parotid and principal salivary glands of this bat are similar to homologous glands in other phyllostomids, but the accessory submandibular gland is unique., Methods: The accessory submandibular glands of live-trapped T. cirrhosus were fixed and processed for transmission electron microscopy by conventional means., Results: The accessory submandibular gland consists of follicles and ducts. The principal cells of the follicular walls have an abundance of rough endoplasmic reticulum (RER), free ribosomes, and extensive Golgi apparatuses. Typically, these cells have relatively few serous secretory granules. The ells contain collections of peculiar lipid droplets, and some of their mitochondria have dense crystalloids within expanded cristae. A layer of irregular, moderately dense bodies lies immediately subjacent to the luminal plasmalemma; it is not clear if these structures are endocytotic or exocytotic. Clusters of mucous cells, some of which have a single, hugely distended RER cisterna, are ensconced in the follicular walls; mucus from these cells reaches the lumen via intercellular canaliculi. Ducts progress from simple cuboidal to simple columnar epithelium. They lack basal striations, and their constituent cells contain relatively few mitochondria. Follicles and ducts have numerous myoepithelial cells at their periphery, and both are heavily innervated by hypolemmal nerve terminals., Conclusions: The unusual accessory submandibular gland in T. cirrhosus documents the extreme modifications in gland histology and in cell ultrastructure that have occurred in mammalian families. The cells composing the follicle walls and ducts bear little similarity to typical acinar or duct cells. Duplication of the submandibular gland in some bat lineages might be the key innovation underlying such plasticity. The heavy innervation of both follicles and ducts also implies that these structures are sensitive to and capable of responding to various inputs, perhaps including dietary factors.

Background: As part of a continuing investigation of the comparative ultrastructure of chiropteran salivary glands, we examined the submandibular glands of eight species of neotropical fruit bats in the genus Artibeus. We previously described secretory granules of unusual substructure in the seromucous demilunar cells of this organ in some species in this genus. In the present study, we turned our attention to the serous acinar cells in the same glands., Methods: Specimens of eight species of Artibeus were collected in neotropical localities. Salivary glands were extirpated in the field and thin slices were fixed by immersion in triple aldehyde-DMSO or in modified half-strength Karnovsky's fixative. Tissues were further processed for electron microscopy by conventional means., Results: In contrast to seromucous cells, which exhibit species-specific diversification in bats of this genus, the secretory apparatus and secretory granules in the serous acinar cells are highly conserved across all seven species. The single exception involves the mitochondria in one species. In this instance, some of the serous cell mitochondria in Artibeus obscurus are modified into megamitochondria. Such organelles usually have short, peripheral cristae; a laminar inclusion is present in the matrix compartment of every outsized organelle. Inclusions of this nature never are present in normal-size mitochondria in the serous cells. None of the megamitochondria were observed in the process of degeneration., Conclusions: The giant mitochondria in A. obscurus have a matrical structure that is radically different from that of the only other megamitochondria reported to occur in bat salivary glands. The factors that lead to variation in megamitochondrial substructure in different species, as well as the functional capacities of such giant organelles, are unknown.

Background: Investigations of reproductive endocrinology of flying foxes (genus Pteropus) have been hampered by inadequate information on the normal morphology of the megachiropteran pituitary., Methods: The novel technique of graphical three-dimensional (3-D) reconstruction, supported by more traditional anatomical techniques, have now been used to examine the shapes of, the interrelations between, the lobes of the pituitary of the little red flying fox, Pteropus scapulatus. Statistical analysis of data from three species tested whether there were changes in pituitary size with annual cycles in function, particularly with key stages of reproduction., Results: In the three species of Australian flying foxes examined, the hypophyseal cleft is closed; the pars intermedia extends over the rostral, ventral, and lateral surfaces of the neural lobe. The pars distalis is broad rostrally and extends over two-thirds of the lateral and ventral pars intermedia. The hypophyseal recess is broad at the median eminence, then narrows and extends through the infundibulum to, but not into, the neural lobe. In adult animals the pituitary weight was 10.0 +/- 0.3 mg (mean +/- s.e.) in P. scapulatus, 14.7 +/- 0.5 mg in Pteropus poliocephalus (greyheaded flying foxes), and 18.7 +/- 1.5 mg in Pteropus alecto (black flying foxes). Pituitary weight was not significantly affected by reproductive stage., Conclusions: Thus histologically, the adenohypophysis and neurohypophysis are similar to those of other mammals. Comparative differences in pituitary size reflected differences in species body size rather than cyclical reproduction.

Background: Hypolemmal nerve terminals, which are abundant in many types of salivary glands, are naked axons that have penetrated the basement membrane to take up a position between adjacent parenchymal cells. Although they may form vesicle-filled varicosities, there usually are no obvious morphological indications as to which cell actually is being innervated., Methods: Specimens of over 200 species of bats were live-trapped and their major salivary glands extirpated and prepared for electron microscopic examination., Results: In 13 species of bats out of the more than 200, mitochondria-hypolemmal nerve terminal complexes were observed in different sites in different salivary glands. In these 13 species, mitochondria in epithelial cells that abut nerve varicosities are closely applied to the plasma membranes of their cells to follow the contours of these nerve elements. The complexes so formed often consist of a varicosity flanked by two mitochondria in separate cells. In intensely innervated glandular segments, mitochondria may be opposed not only to varicosities, but to the axonal portions of these nervous elements as well., Conclusions: The physiological significance of such complexes is unknown; it is conceivable that the mitochondria facilitate signal transmission or play a role in local calcium homeostasis related to nerve function.

Background: Having observed the apparent absence of a bony patella in a Madagascar flying fox (Pteropus sp.), other species from the two suborders of bats (Megachiroptera and Microchiroptera) were examined to determine the presence or absence of a bony patella and the distribution of this feature among bats., Methods: Gross, radiographic, and histologic examination of seven megachiropteran species representing four genera, as well as six microchiropteran species representing six genera, was performed., Results: A bony patella was observed in all six microchiropteran and in three megachiropteran species. The tendon of the quadriceps femoris muscle in Microchiropteran species was composed mainly of dense regular connective tissue. The quadriceps tendon in Megachiropteran species with a patella contained an abundance of fibrocartilage and hyaline cartilage, unlike the quadriceps femoris tendon of the Microchiroptera or a laboratory mouse examined for comparison., Conclusions: Four species of the megachiropteran genus Pteropus lacking a bony patella displayed a similar occurrence and distribution of fibrocartilage and hyaline cartilage within the quadriceps tendon as seen in the other bats. In reference to this singular feature, Pteropus is unique among the representatives of megachiropteran and microchiropteran genera examined here.

Background: Certain species of bats possess two sets of submandibular glands, namely, principal and accessory. The ultrastructure and histochemistry of the accessory submandibular gland was examined in three species of long-winged bats., Methods: Specimens of Miniopterus schreibersi and M. magnator were live-trapped in Thailand, and of M. inflatus were live-trapped in Kenya. For electron microscopy, accessory submandibular lands were initially fixed in triple aldehyde-DMSO, postfixed in osmium tetroxide, and embedded in Epon-Maraglas. A portion of the glands collected in Thailand (M. schreibersi and M. magnator) was fixed in buffered formalin and embedded in paraffin. Sections of the latter material were subjected to a battery of histochemical tests for glycoconjugates., Results: Although in all three species the accessory submandibular glands have normal histological structure, the glands in two, M. schreibersi and M. magnator, were distinguished by possessing mucous droplets of unusual morphology. These droplets, whose identity as mucous was confirmed by histochemical tests for glygoconjugates, are delimited by manifold membranes: up to 10 in M. schreibersi and fewer, but still multiple, in M. magnator. In both species, the entire array of surface membranes may fold inward in the fashion of mitochondrial cristae, forming packets of membranes, many of which have the spurious appearance of floating free in the droplet matrix. These multipartite limiting membranes appear to originate simply by Golgi saccules and moderately large, flattened Golgi vesicles repeatedly wrapping themselves around the surface of nascent mucous droplets. During exocytosis, the outermost membrane of each mucous droplet contacts the luminal membrane, this barrier ruptures, then the remainder of the droplet--multiple membranes and matrix--either flow into the lumen or are cast out in toto. In either case, a great deal of membrane phospholipid is added to the saliva. This salivary lipid may permit these bats to consume insects that normally are able to repel predators with chemical defenses that make them unpalatable. The third species that we studied, M. inflatus, has mucous droplets of normal appearance, i.e., they have only one limiting membrane., Conclusions: The varying structure of mucous secretory products among the species of Miniopterus provides important clues as to the evolution of this genus as well as to the evolution of secretory cells in general.

Measurements of extremely high osmolalities in cauda epididymidal fluids of hibernating bat species led to an investigation of the junctional complex morphology of the epithelium of this sperm storage site. Freeze fracture replicas revealed the presence, at certain times of the year, of a tight junction architecture that resembled that traditionally thought to be exclusive to the blood-testis barrier, the strongest permeability barrier in the body. It is hypothesized that seasonal establishment of these specialized Sertoli cell-like tight junctions is necessary to the maintenance of the high osmotic state of the luminal environment, allowing for the prevention of dilution of its contents by paracellular routes and its protection from bursting under the osmotic pressure contained within.

Giant mitochondria, measuring up to 6.4 microns in diameter, are present in the seromucous secretory cells of the accessory submandibular gland of the long-haired fruit bat, Stenonycteris lanosus. These mitochondria, as well as all of the smaller ones in the same cells, contain in their matrix compartment an abundance of 33 nm threads that probably consist of protein. Some mitochondria, regardless of size, contain 5 nm helical filaments within an expanded crista. Despite their altered morphology, the enlarged mitochondria in the accessory submandibular gland of S. lanosus must be able to function normally in energy metabolism, since the secretory cells in which they are found elaborate numerous secretory granules.

The purpose of this study was to examine the morphological characteristics of the pituitary gland in the little brown bat that might influence mechanisms of hypothalamic releasing hormone transport. Paraffin sections were prepared from whole crania to examine in situ the orientations of the three parts of the adenohypophysis (pars distalis, pars intermedia, and pars tuberalis) relative to the components of the neurohypophysis (pars nervosa and infundibular stem) and the basal hypothalamus. Of particular interest was the observation that the axis of the infundibular stem is directed posteriorly from the median eminence and occupies a depression in the dorsal surface of the pars distalis as it approaches the pars nervosa. Previous studies have revealed that neuronal projections containing releasing hormones extend into the infundibular stem in this species. Therefore, we conducted a fine structural study to determine whether the zone of contact between the infundibular stem and the pars distalis could represent a site of specialized interaction between hypophysiotropic hormones and their target cells. The results show that the sparse connective tissue along this boundary contains abundant fenestrated capillaries that are exposed on one side to neurosecretory axons and on the other to cells of the pars distalis. Furthermore, secretory cells nearest these capillaries exhibit ultrastructural evidence of heightened secretory activity. We conclude that the fine structural characteristics of this zone are consistent with localized mechanisms of releasing hormone transport.

The ultrastructure of the parotid gland was examined in the little brown bat. The seromucous acinar cells contained abundant granules of variable morphology. These granules were characterized by a submembranous dense layer consisting of fine parallel slats. In some bats, the matrix of the granules was structureless, whereas in others it consisted of closely packed but randomly arranged bundles of tubules. The intercalated ducts had a highly developed rough endoplasmic reticulum, often containing large numbers of intracisternal granules. In contrast, only a few secretory granules were present in the supranuclear cytoplasm. The striated ducts, which exhibited the characteristic basal striations consisting of vertically oriented mitochondria and highly folded plasmalemmas, contained numerous small dense granules in a subluminal band. These granules had a paracrystalline substructure with a periodicity of 8 nm. Excretory ducts strongly resembled striated ducts. They showed the same kind of basal striations and about half their constituent cells contained small paracrystalline granules.

The ultrastructure of the two types of cells, ciliated and microvillous, lining the wall of the bat pituitary cleft was examined at different periods during the annual life cycle. Ciliated lining cells remained unchanged in structure throughout the yearly activity cycle. In contrast, microvillous lining cells exhibited striking cyclic morphological changes. Throughout the active phase of the life cycle and during most of the hibernating period, the structure of microvillous cells is unchanged. However, toward the end of hibernation, the cells produce and store large numbers of round to ovoid dense secretory granules. Just prior to arousal, the microvillous cells are packed with secretory granules. At arousal, the cells undergo marked degranulation; the granules appear to be discharged into the lumen of the cleft. Degranulation is followed by a period of proliferation of rough endoplasmic reticulum, the cavity of which is often filled with large paracrystalline inclusions. Within 1 month, microvillous cells return to a resting state and are devoid of secretory granules.

Pregnant Myotis lucifugus were captured in mist nets set outside a large maternity colony and, in most cases, were examined 12-15 hours later. Anterior and posterior halves of uteri were pinned to dental wax and either incubated in glyoxylic acid to produce adrenergic nerve fluorescence, or fixed in buffered glutaraldehyde for electron microscopy. Blood was collected for radioimmunoassay of plasma progesterone. We found no evidence of decreased nerve fluorescence even in late pregnancy when plasma progesterone levels were 10-20 times preovulatory values. Ultrastructural examination also showed no evidence of damage to, or destruction of, nerves in the myometrium. However, we did find gap junctions between myometrial muscle cells during the periparturient period in both normal and aborting bats. Gap junctions began to form several days before term, increased in number until parturition, then decreased dramatically within a day or two thereafter. The percentage of muscle cell plasma membrane involved in gap junction formation was closely correlated with plasma progesterone levels, although whether this is causal or coincidental is not certain. These data do not agree with the conclusions drawn from observations in other mammals that a disappearance of adrenergic nerves from the myometrium is associated with the initiation of parturition, or that gap junction formation is associated with changes in nerve function. They do, however, lend further support to the hypothesis that there is neurogenic control of myometrial contractility in M. lucifugus uteri at term.

Uteri from hibernating bats, Myotis lucifugus, collected periodically from Renfrew County, Ontario, were fixed in 2% glutaraldehyde and processed for electron microscopy or incubated in glyoxylic acid to show adrenergic nerves by fluorescence. The bat uterus is structurally typical of mammalian species; although the right uterine horn is permanently enlarged in parous bats due to hypertrophy of both myometrium and endometrium. Nerves were abundant between both longitudinal and circular layers of muscle cells. Unmyelinated, and some myelinated, axons, ranging from few to many, coursed generally parallel to the uterine long axis. Numerous axonal varicosities containing small dense-cored (adrenergic) vesicles or, less often, small agranular (cholinergic) vesicles, were found forming close nerve-muscle contacts between myometrial cells and blood vessels. Fluorescent microscopy showed a dense network of adrenergic nerves in parous uteri, but a sparse network in nulliparous uteri. A specific adrenergic nerve marker, 5-hydroxydopamine, greatly increased the density and in some instances, the size of granular vesicles, while 6-hydroxydopamine, which depletes adrenergic neutrotransmitter, reduced the number of dense-cored vesicles. Nulliparous uteri appeared unchanged by six daily injections of 0.1 microgram estradiol-17 beta; 0.25 mg progesterone, or both; but parous uteri were greatly enlarged by all regimes. Nerve ultrastructure, however, appeared unaffected by steroid treatment; nor, despite the absolute dextral bias in implantation, were left-right differences observed. Gap junctions were not found between muscle cells in myometria of any bat uteri. Based on this study, we suggest that M. lucifugus may provide a most useful model for examination of neurogenic regulation of the uterus.

Skull measurements from ten anatomically and behaviorally diverse genera of bats show marked variation in positioning of the face upon the cranium but a relative stability of the site of the mandibular fossa. Factors associated with maintaining occlusion in bats which exhibit dorsally-inclined maxillary toothrows include dorsally angulated mandibular bodies and elevated condyles. Detailed comparisons are made between the generalized morphology of Myotis lucifugus and anatomical extremes represented by Rhinolophus lepidus, Mormoops megalophylla, and Pteropus giganteus. In these four bats, masticatory movements of the teeth and temporomandibular joints, despite marked interspecific variation, appear to relate to a common pattern. The beginning of jaw opening is important for maximal occlusal shear, particularly in Pteropus. Observed differences in the histology of the temporomandibular joints reflect postulated differeces of pressure patterns within them. Differences in skeletal and dental morphology, together with variations in size and orientation of masticatory muscles, could account for known and postulated differences in the four respective chewing patterns, with no major variation from the known muscle firing sequences of Myotis lucifugus. Basic patterns of interaction between central nervous system and masticatory musculature would therefore appear to have undergone minimal modification. This accords with the concept that neural control of mastication is a relatively conservative mechanism; as such, it would appear to have imposed significant limitation upon adaptive change in bats.

A light and electron microscope study of the small intestine of the little brown bat, Myotis lucifugus, was carried out at several stages in the animal's annual life cycle. An unusual morphological observation was the presence of cells in the lamina propria of the small intestine which were packed with a conspicuous basophilic granular material that appeared crystalline. Moreover, such cells were present only during the hibernation period and were therefore called "hibernation crystalloid" (HC) cells. By light microscopy, the crystal-like material was not sudanophilic, did not stain for nucleic acids, and did not contain acid phosphatase; it did show reactivity when stained by the periodic acid-Schiff procedure. By electron microscopy, the crystal-like material was found to be present in smooth, membrane-enclosed vacuoles along with an amorphous, dense granular substance. The crystalline material occasionally formed rigid-appearing rods that reached lengths of 10 microns. The crystal-containing cells were contacted by axonal varicosities. It is suggested that these innervated HC cells represent a unique cell type with a gastrointestinal function, yet to be determined, that may be related to hibernation.

In previous studies of the adult bat, Myotis lucifugus, bone loss that occurred during the hibernating period was attributed to osteocytic osteolysis, rather than osteoclastic activity. We have used histochemistry and light and electron microscopy to determine the functional state of the skeleton during the hibernating period. We find that during hibernation the marrow cavity of the long bones is filled with lipid deposits interspersed with vascular sinusoids containing mononuclear cells and red blood cells. The lipid deposits are found within fat cells and at extracellular sites. Hematopoietic cells are absent, osteoclasts are absent, and the bone surfaces are covered with quiescent bone-lining cells. Osteocytes retain their structural integrity and maintain canalicular systems with the bone lining cells and with other osteocytes. During the arousal period, osteoclasts reappear on the bone surface, followed in time by increased numbers of osteoblast-like cells. Perivascular cells undergo structural hypertrophy. Many mononuclear cells are now found in extravascular sites. The lipid content in the marrow is gradually reduced and replaced by hematopoietic cells. Each of these events occurs in a repeatable sequence that is related to the state of hibernation of Myotis lucifugus.

The distribution and size of Boettcher cells were determined from serial sections of the temporal bones of four little brown bats and six rabbits. In addition, one harp seal and one two-toed sloth were examined. In the little brown bat, the heights of the Boettcher cells measured 4-8 micron and they were found throughout much of the length of the cochlea. There were four rows in most of the lower basal coil, five rows in the upper basal coil, and four rows in the middle coil. In the rabbit there were nine rows in the basal coil and 12 rows in the middle coil. The heights of the Boettcher cells were approximately 14 micron in the rabbit. In the two-toed sloth, Boettcher cells were sparsely distributed along the basal coil; similarly, in the harp seal, Boettcher cells were confined solely to the basal coil, where there were only three rows, which measured approximately 18 micron in height. The distribution and size of Boettcher cells in the rabbit and the little brown bat were compared to those in other mammalian species.

The reproductive biology of the bat Macrotus waterhousii was investigated anatomically and physiologically several times monthly throughout the year. The male spermatogenic cycle is initiated in June; sperm are available in August. Testicular hypertrophy is accompanied by increased levels of testicular ascorbic acid and circulating plasma testosterone. Sperm are present in the seminiferous tubules and later the epididymides from August to early December. Regression in testicular size begins in late September and testes are involuted by early December. Levels of testicular ascorbic acid and plasma testosterone decline at this time, although detectable amounts of both are present year-round. Mature sperm are absent from the male tract after late November or early December. The accessory sex gland complex and Cowper's glands undergo an annual cycle in synchrony with the testicular cycle. The accessory complex contains fructose, the concentration of which varies seasonally in direct proportion to glandular hypertrophy. Only trace levels of fructose are elaborated by Cowper's glands undergo an annual cycle in synchrony with the testicular cycle. The accessory complex contains fructose, the concentration of which varies seasonally in direct proportion to glandular hypertrophy. Only trace levels of fructose are elaborated by Cowper's glands. In contrast to vespertilionid bats, where the accessory glands remain enlarged and active when testes involute, those in Macrotus regress. The penis differs structurally from that of other temperate North American bats, primarily by lacking a bony baculum and specialized erectile accessory corpus cavernosus bodies. Males are not reproductively active in their first autumn.