Skin glands of an aquatic salamander vary in size and distribution and release antimicrobial secretions effective against chytrid fungal pathogens.

Amphibian skin is unique among vertebrate classes, containing a large number of multicellular exocrine glands that vary among species and have diverse functions. The secretions of skin glands contain a rich array of bioactive compounds including antimicrobial peptides (AMPs). Such compounds are important for amphibian innate immune responses and may protect some species from chytridiomycosis, a lethal skin disease caused by the fungal pathogens Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal). While the bioactivity of skin secretions against Bd has been assessed for many amphibian taxa, similar studies are lacking for Bsal, a chytrid fungus that is especially pathogenic for salamanders. We studied the skin glands and their potential functions in an aquatic salamander, the three-toed amphiuma (Amphiuma tridactylum). Skin secretions of captive adult salamanders were analyzed by RP-HPLC and tested against the growth of Bd and Bsal using in vitro assays. We found that compounds within collected skin secretions were similar between male and female salamanders and inhibited the growth of Bd and Bsal. Thus, skin secretions that protect against Bd may also provide protection against Bsal. Histological examination of the skin glands of preserved salamanders revealed the presence of enlarged granular glands concentrated within caudal body regions. A site of potential gland specialization was identified at the tail base and may indicate specialized granular glands related to courtship and communication.

The Lack of Nasolacrimal Ducts in Plethodontid Salamanders?

Nasolacrimal ducts are a terrestrial vertebrate adaptation and appear to have co-evolved with orbital glands. Although plethodontid salamanders possess orbital glands, a recent study concluded that plethodontid salamanders lack nasolacrimal ducts. Functionally, the absence of nasolacrimal ducts closes the route for orbital gland secretion passage into the nasal and vomeronasal organ cavities. Orbital glands have been implicated in enhancement of vomeronasal function so loss could have important implications for communication. Multiple older studies depict or discuss nasolacrimal ducts in plethodontid salamanders. Interestingly, the only consensus between recent and older literature is that Desmognathus lacks nasolacrimal ducts. To determine if plethodontid salamanders truly lack nasolacrimal ducts, we sectioned plethodontid salamander heads for general histological examination of species from the majority of the plethodontid tribes. From our representative sample, we found only two species that completely lacked nasolacrimal ducts (Desmognathus fuscus and Eurycea tynerensis) and one species that possessed nasolacrimal ducts that ended blindly before reaching the nasal cavities (E. spelaea). Bayesian ancestral state reconstruction resulted in the presence of nasolacrimal ducts on the branch leading to Plethodontidae and both subfamilies within Plethodontidae, with two independent losses in Desmognathus and Eurycea. Anat Rec, 301:765-775, 2018. © 2017 Wiley Periodicals, Inc.

Skin of the Red Eye Tree Frog Agalychnis Callidryas (Hylidae, Phyllomedusinae) Contains Lipid Glands of the Type Described in the Genus Phyllomedusa.

Several anuran species of the genus Phyllomedusa are known to possess specialized cutaneous glands producing lipids and exhibit a peculiar wiping behavior. This behavior is a stereotyped repertory of fore and hind limb movements distributing hydrophobic molecules onto the body surface and reducing evaporative water loss. No reports are presently available on the occurrence of lipid glands in other phyllomedusine genera, and data on the structure of the secretory units specialized for the production of cutaneous lipids are still unclear. The present report is aimed to answer both questions: it describes lipid glands of the Phyllomedusa type in Agalychnis callidryas and provides light and transmission electron microscope evidence of the syncytial structure of their secretory units, a typical feature of serous glands in anuran skin. This morphological trait supports the hypothesis that lipid glands are a specialized subset of the anuran serous glands, and underlines their flexible role in the skin adaption to sub-aerial environments. Anat Rec, 300:503-506, 2017. © 2016 Wiley Periodicals, Inc.

Ultrastructure of spermatid development within the testis of the Yellow-Bellied Sea Snake, Pelamis platurus (Squamata: Elapidae).

Little is known about spermatid development during spermiogenesis in snakes, as there is only one complete study in ophidians, which details the spermatid ultrastructure within the viperid, Agkistrodon piscivorus. Thus, the following study will add to our understanding of the ontogenic steps of spermiogenesis in snakes by examining spermatid maturation in the elapid, Pelamis platurus, which were collected in Costa Rica in 2009. The spermatids of P. platurus share many similar ultrastructural characteristics to that described for other squamates during spermiogenesis. Three notable differences between the spermatids of P. platurus and those of other snakes is a round and shorter epinuclear lucent zone, enlarged caudal nuclear shoulders, and more prominent 3 and 8 peripheral fibers in the principal and endpieces. Also, the midpiece is much longer in P. platurus and is similar to that reported for all snakes studied to date. Other features of chromatin condensation and morphology of the acrosome complex are similar to what has been observed in A. piscivorus and other squamates. Though the spermatids in P. platurus appear to be quite similar to other snakes and lizards studied to date, some differences in subcellular details are still observed. Analysis of developing spermatids in P. platurus and other snakes could reveals morphologically conserved traits between different species along with subtle changes that could help determine phylogenetic relationships once a suitable number of species have been examined for ophidians and other squamates.

Ultrastructural analysis of spermiogenesis in the Eastern Fence Lizard, Sceloporus undulatus (Squamata: Phrynosomatidae).

Studies on reptilian sperm morphology have shown that variation exists at various taxonomic levels but studies on the ontogeny of variation are rare. Sperm development follows a generalized bauplan that includes acrosome development, nuclear condensation and elongation, and flagellar development. However, minute differences can be observed such as the presence/absence of manchette microtubules, structural organization during nuclear condensation, and presence/absence of a nuclear lacuna. The purpose of this investigation was to examine sperm development within the Sceloporus genus. The process begins with the development of an acrosomal complex from Golgi vesicles followed by nuclear condensation and elongation, which results in the presence of a nuclear lacuna. As the acrosomal complex differentiates, flagellar development commences with elongation of the distal centriole. Spermatid development culminates in a mature spermatid with a highly differentiated acrosomal complex, a condensed nucleus with a nuclear lacuna, and a differentiated flagellum. Although the overall developmental pattern is consistent with other squamate species, minute differences are observed, even within the same genus. For example there is variation in the presence/absence of an endoplasmic reticulum complex during acrosome development, presence/absence of a nuclear lacuna, and presence/absence of manchette microtubules within the three species of Sceloporus studied to date. Future studies concerning sperm morphology in closely related species will aid in our understanding of variation in sperm development and may prove to be useful in testing phylogenetic and evolutionary hypotheses.

Phylogeny of Mental Glands, Revisited.

Mental glands and their associated delivery behaviors during courtship are unique to the plethodontid salamanders. Because previous interpretations of the evolution of these features were conducted using older phylogenetic hypotheses, we reanalyzed these traits with newer courtship descriptions and contemporary phylogenetic methods. Using Bayesian ancestral state reconstruction methods that have been developed since the first phylogenetic analyses were conducted in the mid-1990s, we reconstructed mental gland and courtship behavior evolution on a Bayesian phylogeny of the nuclear gene Rag1. The most probable ancestral condition for plethodontids was resolved as presence of a mental gland. Loss of a mental gland occurred in each subfamily and was recovered as the most probable ancestral condition for the Spelerpinae. In contrast, parsimony reconstruction recovered the presence of a mental gland in the ancestor to Spelerpinae with multiple secondary losses. We hypothesize that that absence of a mental gland is possibly ancestral in some clades (i.e., Spelerpinae) and secondary in others (e.g., paedomorphic Eurycea). The most probable ancestral form of the mental gland is likely to be the large pad-type distributed extensively in Plethodontinae and Bolitoglossinae. Desmognathans have the most unique mental glands, occurring in an anterior protrusion or bifurcated form (in Desmognathus wrighti). Fan-shaped mental glands evolved independently in Eurycea and Oedipina. Small pads arose independently in Bolitoglossinae, Plethodontinae, and Spelerpinae. Head-rubbing behavior for mental gland delivery mode was recovered as the most probable and parsimonious ancestral state for the Plethodontidae, with independent losses of this behavior in Plethodontinae and Spelerpinae. Because head-rubbing was observed in outgroups, we hypothesize that head-rubbing behavior predated mental gland evolution. Pulling, snapping, slapping, and biting behaviors evolved independently in the Plethodontinae and Spelerpinae and are not homologous with head-rubbing. All hypotheses of mental gland and courtship evolution invoke homoplasy.

Histology and ultrastructure of the caudal courtship glands of the red-backed salamander, Plethodon cinereus (Amphibia: Plethodontidae).

Caudal courtship glands (CCGs) are sexually dimorphic glands described in the skin of the dorsal tail base of some male salamanders in the genera Desmognathus, Eurycea, and Plethodon in the family Plethodontidae. These glands are believed to deliver pheromones to females during courtship, when the female rests her chin on the dorsal tail base during the stereotypic tail straddling walk unique to plethodontids. Although CCGs have been studied histologically, no investigations of their ultrastructure have been made. This article presents the first study on the fine structure and seasonal variation of CCGs, using the plethodontid Plethodon cinereus. The CCGs vary seasonally in height and secretory activity. The mature secretory granules observed in males collected in October and April consist of oval, biphasic granules that are eosinophilic and give positive reactions to periodic acid-Schiff for neutral carbohydrates but do not stain for acidic mucosusbtances or proteins with alcian blue and bromphenol blue, respectively. Granular glands, some of which contain mucous demilunes, are twice as large as CCGs, are syncytial (unlike CCGs), and stain for proteins. Mucous glands are similar in size to CCGs, but are basophilic, show no seasonal variation in secretory activity, and stain positive for acidic mucosubstances. CCGs do not resemble cytologically the sexually dimorphic mental glands of some plethodontids, which contain round or oval granules filled with an electron-dense amorphous substance. The CCGs are similar histologically to sexually dimorphic skin glands described in some anurans, but more comparative work is needed.

Spermatogenic cycle of a plethodontid salamander, Eurycea longicauda (Amphibia, Urodela).

Previous investigators have described the spermatogenic cycles of numerous species of plethodontid salamanders. Most studies describe a fairly stereotypical cycle with meiotic divisions of spermatogenesis commencing in the spring/summer. However, many studies lack details obtainable from histological examination and/or testicular squashes and, instead, provide only mensural data from the testes. Studies that lacked microscopic evaluation often revealed spermatogenic cycles that varied greatly from that of the stereotypical cycle with meiotic divisions commencing in the fall/winter. Those studies hamper comparisons between the spermatogenic cycles of different species and their environments, as they do not provide a correlation between testicular size and any aspect of the spermatogenic cycle. In the following manuscript, we elucidate the spermatogenic cycle of Eurycea longicauda longicauda in an effort to outline an appropriate protocol for analyzing spermatogenesis in salamanders that will facilitate future comparative studies. Like many Nearctic plethodontids, E. l. longicauda exhibits a meiotic wave that travels through the testes during the summer; this process is followed by spermiogenesis, spermiation, and recrudescence in the fall, winter, and spring.

Agkistrodon piscivorus spermatogenesis addendum: The effect of Hurricane Katrina on spermatogenesis of the western cottonmouth snake.

Recent studies detailed the spermatogenic cycle of the Western Cottonmouth Snake, Agkistrodon piscivorus and noted that spermatogenesis is bimodal, with active periods during March-June and August-October in southeastern Louisiana. However, only spermatogonia were present in September in the only specimen that was captured and the authors state that the individual "should have a high testis volume and also show spermiogenic activity." The specimen in their study was caught immediately following Hurricane Katrina outside of its normal habitat. Therefore, in order to verify their assumption, individuals were captured during September of 2008 and the testes were spermatogenically active with spermatogonia, spermatocytes, and mature spermatozoa being present in the seminiferous epithelium of the testes. These data indicate that Hurricane Katrina could have had an impact on the spermatogenic cycle in Cottonmouths, resulting in stress-induced testicular regression.

The testicular sperm ducts and genital kidney of male Ambystoma maculatum (Amphibia, Urodela, Ambystomatidae).

The ducts associated with sperm transport from the testicular lobules to the Wolffian ducts in Ambystoma maculatum were examined with transmission electron microscopy. Based on the ultrastructure and historical precedence, new terminology for this network of ducts is proposed that better represents primary hypotheses of homology. Furthermore, the terminology proposed better characterizes the distinct regions of the sperm transport ducts in salamanders based on anatomy and should, therefore, lead to more accurate comparisons in the future. While developing the above ontology, we also tested the hypothesis that nephrons from the genital kidney are modified from those of the pelvic kidney due to the fact that the former nephrons function in sperm transport. Our ultrastructural analysis of the genital kidney supports this hypothesis, as the basal plasma membrane of distinct functional regions of the nephron (proximal convoluted tubule, distal convoluted tubule, and collecting tubule) appear less folded (indicating decreased surface area and reduced reabsorption efficiency) and the proximal convoluted tubule possesses ciliated epithelial cells along its entire length. Furthermore, visible luminal filtrate is absent from the nephrons of the genital kidney throughout their entire length. Thus, it appears that the nephrons of the genital kidney have reduced reabsorptive capacity and ciliated cells of the proximal convoluted tubule may increase the movement of immature sperm through the sperm transport ducts or aid in the mixing of seminal fluids within the ducts.

Observations on variation in the ultrastructure of the proximal testicular ducts of the Ground Skink, Scincella lateralis (Reptilia: Squamata).

The North American Ground Skink, Scincella lateralis, is a member of the most speciose family of lizards, the Scincidae. The only descriptions of the testicular ducts of skinks concern the light microscopy of 13 species in eight other genera. We combine histological observations with results from transmission electron microscopy on a sample of skinks collected throughout the active season. The single rete testis has squamous epithelium with a large, indented nucleus and no junctional complexes between cells or conspicuous organelles. Nuclei of sperm in the rete testis area are associated with cytoplasmic bodies that are lost in the ductuli efferentes. The ductuli efferentes have both ciliated and nonciliated cells and show little seasonal variation except for the narrowing of intercellular canaliculi when sperm are absent. When the ductus epididymis contains sperm, the anterior one-third lacks copious secretory material around luminal sperm, whereas in the posterior two-thirds sperm are embedded in a dense matrix of secretory material. Light and dark principal cells exist and both contain saccular, often distended rough endoplasmic reticula, and widened intercellular canaliculi that bridge intracellular spaces. Junctional complexes are lacking between principal cells except for apical tight junctions. Electron-dense secretory granules coalesce at the luminal border for apocrine release. The cranial end of the ductus deferens is similar in cytology to the posterior ductus epididymis. Each of the nine squamates in which the proximal testicular ducts have been studied with electron microscopy has some unique characters, but no synapomorphies for squamates as a group are recognized.

Observations on the sexual segment of the kidney of snakes with emphasis on ultrastructure in the yellow-bellied sea snake, Pelamis platurus.

The sexual segment of the kidney (SSK) is an accessory sex structure in male lizards and snakes (Squamata). We describe histology of the SSK in 12 species of snakes, including one from the basal Scolecophidia, Leptotyphlops dulcis, and from the more advanced Alethinophidia, species from the Acrochordidae (Acrochordus granulatus), Homalopsidae (Cerberus rynchops), Uropeltidae (Teretrurus sanguineus), and eight species from the Elapidae, including six species of sea snakes. We also describe the ultrastructure of the SSK of the sea snake, Pelamis platurus. The SSK of L. dulcis does not include the ureter but does include distal convoluted tubules (DCTs) and collecting ducts. In all other snakes examined, the SSK is limited to the DCTs and does not differ in histology by any consistent character. We found apparently mature individuals of several species with inactive SSKs. Hypertrophied SSKs give positive reactions for protein secretions but variable reactions for carbohydrates. Ultrastructure of the SSK of P. platurus reveals nuclei situated medially in the epithelium and mature electron dense secretory vacuoles in other areas of the cytoplasm. Product release is apocrine. Junctional complexes only occur at the luminal border, and intercellular canaliculi become widened and are open basally. No cytologically unique characters occur in the SSK of P. platurus. The ancestral condition of the SSK in squamates is the presence of simple columnar epithelium specialized for secretion of a protein + carbohydrate product that matures and is released seasonally.

Observations on the anterior testicular ducts in snakes with emphasis on sea snakes and ultrastructure in the yellow-bellied sea snake, Pelamis platurus.

The anterior testicular ducts of squamates transport sperm from the seminiferous tubules to the ductus deferens. These ducts consist of the rete testis, ductuli efferentes, and ductus epididymis. Many histological and a few ultrastructural studies of the squamate reproductive tract exist, but none concern the Hydrophiidae, the sea snakes and sea kraits. In this study, we describe the anterior testicular ducts of six species of hydrophiid snakes as well as representatives from the Elapidae, Homolapsidae, Leptotyphlopidae, and Uropeltidae. In addition, we examine the ultrastructure of these ducts in the yellow-bellied Sea Snake, Pelamis platurus, only the third such study on snakes. The anterior testicular ducts are similar in histology in all species examined. The rete testis is simple squamous or cuboidal epithelium and transports sperm from the seminiferous tubules to the ductuli efferentes in the extratesticular epididymal sheath. The ductuli efferentes are branched, convoluted tubules composed of simple cuboidal, ciliated epithelium, and many species possess periodic acid-Schiff+ granules in the cytoplasm. The ductus epididymis at the light microscopy level appears composed of pseudostratified columnar epithelium. At the ultrastructural level, the rete testis and ductuli efferentes of P. platurus possess numerous small coated vesicles and lack secretory vacuoles. Apocrine blebs in the ductuli efferentes, however, indicate secretory activity, possibly by a constitutive pathway. Ultrastructure reveals three types of cells in the ductus epididymis of P. platurus: columnar principal cells, squamous basal cells, and mitochondria-rich apical cells. This is the first report of apical cells in a snake. In addition, occasional principal cells possess a single cilium, which has not been reported in reptiles previously but is known in some birds. Finally, the ductus epididymis of P. platurus differs from other snakes that have been studied in possession of apical, biphasic secretory vacuoles. All of the proximal ducts are characterized by widening of adjacent plasma membranes into wide intercellular spaces, especially between the principal cells of the ductus epididymis. Our results contribute to a larger, collaborative study of the evolution of the squamate reproductive tract and to the potential for utilizing cellular characters in future phylogenetic inferences.

Ultrastructural description of spermiogenesis within the Mediterranean Gecko, Hemidactylus turcicus (Squamata: Gekkonidae).

We studied spermiogenesis in the Mediterranean Gecko, Hemidactylus turcicus, at the electron microscope level and compared to what is known within other Lepidosaurs. In H. turcicus germ cells are connected via cytoplasmic bridges where organelle and cytoplasm sharing is observed. The acrosome develops from merging transport vesicles that arise from the Golgi and subsequently partition into an acrosomal cap containing an acrosomal cortex, acrosomal medulla, perforatorium, and subacrosomal cone. Condensation of DNA occurs in a spiral fashion and elongation is aided by microtubules of the manchette. A nuclear rostrum extends into the subacrosomal cone and is capped by an epinuclear lucent zone. Mitochondria and rough endoplasmic reticulum migrate to the posterior portion of the developing germ cell during the cytoplasmic shift and the flagellum elongates. Mitochondria surround the midpiece as the anlage of the annulus forms. The fibrous sheath begins at mitochondrial tier 3 and continues into the principal piece. Peripheral fibers associated with microtubule doublets 3 and 8 are grossly enlarged. During the final stages of germ cell development spermatids are wrapped with a series of Sertoli cell processes, which exhibit ectoplasmic specializations and differing cytoplasmic consistencies. The results observed here corroborate previous studies, which show the conservative nature of sperm morphology. However, ultrastructural character combinations specific to sperm and spermiogenesis seem to differ among taxa. Further studies into sperm morphology are needed in order to judge the relevance of the ontogenic changes recorded here and to determine their role in future studies on amniote evolution.

The sexual segment of Hemidactylus turcicus and the evolution of sexual segment location in squamata.

The kidneys of the Mediterranean Gecko, Hemidactylus turcicus (Gekkonidae), were investigated using light and electron microscopy with the primary focus placed on morphology of the sexual segment of the kidney. The nephrons of male H. turcicus are composed of five distinct regions: 1) a renal corpuscle and glomerulus, 2) a proximal convoluted tubule, 3) an intermediate segment, 4) a distal convoluted tubule, and 5) the sexual segment of the kidney/collecting duct. Female H. turcicus is similar but lack a sexual segment of the kidney. The sexual segment of the kidney is hypertrophied during the months of March through August, which corroborates previous reports of reproductive activity. During inactive months, the sexual segment of the kidney is nondiscernable from the collecting ducts. The sexual segment consists of tall columnar epithelial cells with basally positioned nuclei. Perinuclear Golgi complexes and rough endoplasmic reticulum are present. Secretory granules, which fill the apices of the epithelial cells, are electron dense and released into the lumen by a merocrine secretory process. Narrow intercellular canaliculi separate each epithelial cell and are sealed by tight junctions at the luminal aspect. Basally, leukoctyes are observed within the intercellular canaliculi and outside the basal lamina. Mast cells can be found just outside the basal lamina in close association with renal capillaries. The sexual segment of the kidney of H. turcicus is similar to that of three unrelated lizards for which ultrastructure was investigated with secretion mode being the major difference Also, H. turcicus is similar to most other lizards in that complete regression occurs during reproductive inactivity, but differs in this trait from the skink, Scincella lateralis, and most snakes which display a hypertrophied sexual segment of the kidney throughout the entire year. Although some unique similarities appear during the optimization, no direct patterns or directions are observed, and only the molecular based phylogeny resolves the ancestral condition of the Squamata as the sexual segment of the kidney being observed in the distal convoluted tubule, collecting duct, and ureter.

Proximal testicular ducts of the Mediterranean gecko (Hemidactylus turcicus).

The efferent ducts of the Mediterranean Gecko, Hemidactylus turcicus (Gekkonidae) were investigated using light and electron microscopy. The seminiferous tubules unite into a single rete testis tubule. The rete testis divides into 3-4 ductuli efferentes which all drain into the cranial portion of the ductus epididymis. All efferent ducts are most active during the months of December to August. The rete testis is composed of a simple squamous epithelium with bifurcated nuclei and a labyrinthine network of intercellular canaliculi. Ciliated and nonciliated cells are present, and more than one cilium extends from the scattered ciliated cells. The presence of small clear vesicles and widened intercellular canaliculi suggest that cells of the rete testis are responsible for intake of luminal fluids. The ductuli efferentes are composed of a simple cuboidal epithelium consisting of ciliated and nonciliated cells, and ciliated cells are the dominant cell type. During the inactive season the number of lysosomes increases and the cells become spermiophagic. The ductus epididymis is composed of a tall pseudostratified columnar epithelium with relatively scarce basal cells. No evidence for regionalization was observed. The ductus epididymis is highly secretory during the active season with numerous electron-dense secretory granules, whose glycoprotein products are released by merocrine secretion. Basally, the active epididymis has swollen intercellular canaliculi and enlarged cisternae of rough endoplasmic reticulum. During the inactive season the secretory activity decreases and membranous structures and fibrous material are observed within widened intercellular canaliculi apical to the basal cells.

The pelvic kidney of male Ambystoma maculatum (Amphibia, urodela, ambystomatidae) with special reference to the sexual collecting ducts.

This study details the gross and microscopic anatomy of the pelvic kidney in male Ambystoma maculatum. The nephron of male Ambystoma maculatum is divided into six distinct regions leading sequentially away from a renal corpuscle: (1) neck segment, which communicates with the coelomic cavity via a ventrally positioned pleuroperitoneal funnel, (2) proximal tubule, (3) intermediate segment, (4) distal tubule, (5) collecting tubule, and (6) collecting duct. The proximal tubule is divided into a vacuolated proximal region and a distal lysosomic region. The basal plasma membrane is modified into intertwining microvillus lamellae. The epithelium of the distal tubule varies little along its length and is demarcated by columns of mitochondria with their long axes oriented perpendicular to the basal lamina. The distal tubule possesses highly interdigitating microvillus lamellae from the lateral membranes and pronounced foot processes of the basal membrane that are not intertwined, but perpendicular to the basal lamina. The collecting tubule is lined by an epithelium with dark and light cells. Light cells are similar to those observed in the distal tuble except with less mitochondria and microvillus lamellae of the lateral and basal plasma membrane. Dark cells possess dark euchromatic nuclei and are filled with numerous small mitochondria. The epithelium of the neck segment, pleuroperitoneal funnel, and intermediate segment is composed entirely of ciliated cells with cilia protruding from only the central portion of the apical plasma membrane. The collecting duct is lined by a highly secretory epithelium that produces numerous membrane bound granules that stain positively for neutral carbohydrates and proteins. Apically positioned ciliated cells are intercalated between secretory cells. The collecting ducts anastomose caudally and unite with the Wolffian duct via a common collecting duct. The Wolffian duct is secretory, but not to the extent of the collecting duct, synthesizes neutral carbohydrates and proteins, and is also lined by apical ciliated cells intercalated between secretory cells. Although functional aspects associated with the morphological variation along the length of the proximal portions of the nephron have been investigated, the role of a highly secretory collecting duct has not. Historical data that implicated secretory activity concordant with mating activity, and similarity of structure and chemistry to sexual segments of the kidneys in other vertebrates, lead us to believe that the collecting duct functions as a secondary sexual organ in Ambystoma maculatum.

Ultrastructure of the reproductive system of the black swamp snake (Seminatrix pygaea). VI. Anterior testicular ducts and their nomenclature.

In this study, the anterior testicular ducts of the North American natricine snake Seminatrix pygaea are described using light and electron microscopy. From the seminiferous tubules, the rete testis passes into the epididymal sheath, a structure along the medial border of the testis heavily invested with collagen fibers. The rete testis consists of simple, nonciliated cuboidal epithelium (principal cells). The intratesticular ducts of the rete testis are narrow (50-70 microm) at their junction with the seminiferous tubules, widen (80-100 microm) as they extend extratesticularly, and divide into smaller branches as they anastomose with the next tubules, the ductuli efferentes. The ductuli efferentes are lined by simple cuboidal epithelium but possess nonciliated principal cells as well as ciliated cells. These are the only ducts in the male reproductive system with ciliated cells. The ductuli efferentes are narrow (25-45 microm), divide into numerous branches, and are highly convoluted. The ductus epididymis is the largest duct in diameter (240-330 microm), and the diameter widens and the epithelium thins posteriorly. The ductus epididymis is lined by nonciliated, columnar principal cells and basal cells. No regional differences in the ductus epididymis are apparent. Ultrastructural evidence suggests that all of the nonciliated principal cells in each of the anterior testicular ducts function in both absorption and secretion. Absorption occurs via small endocytic vesicles, some of which appear coated. Secretion is by a constitutive pathway in which small vesicles and a flocculent material are released via a merocrine process or through the formation of apocrine blebs. The secretory product is a glycoprotein. Overall, the characteristics of the anterior testicular ducts of this snake are concordant with those of other amniotes, and the traditional names used for snakes are changed to conform with those used for other sauropsids and mammals.

Ultrastructure of spermiogenesis in the Cottonmouth, Agkistrodon piscivorus (Squamata: Viperidae: Crotalinae).

To date multiple studies exist that examine the morphology of spermatozoa. However, there are limited numbers of data detailing the ontogenic characters of spermiogenesis within squamates. Testicular tissues were collected from Cottonmouths (Agkistrodon piscivorus) and tissues from spermiogenically active months were analyzed ultrastructurally to detail the cellular changes that occur during spermiogenesis. The major events of spermiogenesis (acrosome formation, nuclear elongation/DNA condensation, and flagellar development) resemble that of other squamates; however, specific ultrastructural differences can be observed between Cottonmouths and other squamates studied to date. During acrosome formation vesicles from the Golgi apparatus fuse at the apical surface of the nuclear membrane prior to making nuclear contact. At this stage, the acrosome granule can be observed in a centralized location within the vesicle. As elongation commences the acrosome complex becomes highly compartmentalized and migrates laterally along the nucleus. Parallel and circum-cylindrical microtubules (components of the manchette) are observed with parallel microtubules outnumbering the circum-cylindrical microtubules. Flagella, displaying the conserved 9 + 2 microtubule arrangement, sit in nuclear fossae that have electron lucent shoulders juxtaposed on either side of the spermatids basal plates. This study aims to provide developmental characters for squamates in the subfamily Crotalinae, family Viperidae, which may be useful for histopathological studies on spermatogenesis in semi-aquatic species exposed to pesticides. Furthermore, these data in the near future may provide morphological characters for spermiogenesis that can be added to morphological data matrices that may be used in phylogenetic analyses.

Seasonal variation of the oviduct of the American alligator, Alligator mississippiensis (Reptilia: Crocodylia).

The annual oviductal cycle of the American alligator, Alligator mississippiensis, is described using light and electron microscopy. Previous work done by Palmer and Guillette ([1992] Biol Reprod 46:39-47) shed some light on the reproductive morphology of the female alligator oviduct; however, their study was limited and did not report details relating to variation across the reproductive season. We recognize six variable regions of the oviduct: infundibulum, tube, isthmus, anterior uterus, posterior uterus, and vagina. Each area shows variation, to some degree, in the histochemistry and ultrastructure of oviductal secretions. Peak secretory activity occurs during the months of May and June, with the greatest variation occurring in the tube and anterior uterus. During the month of May, high densities of neutral carbohydrates and proteins are found within the tubal and anterior uterine glands. The epithelium of the entire oviduct secretes neutral carbohydrates throughout the year, but many regions lack protein secretions, and the posterior uterine glands show little secretory activity of any type throughout the year. After oviposition, secretory activity decreases drastically, andthe oviduct resembles that of the premating season. This study also provides evidence to support the homology between alligator and bird oviducts. Sperm were observed in glands at the tubal-isthmus and utero-vaginal junctions in preovulatory, postovulatory and postovipository females.