Fine structural and enzymehistochemical observations on the notochord of Ichthyophis glutinosus and Ichthyophis kohtaoensis (Gymnophiona,Amphibia)

Summary The notochord of Ichthyophis glutinosus and I. kohtaoensis consists of peripheral flattened cells characterized by a well-developed system of rough endoplasmic reticulum, bundles of tonofilaments, and abundant glycogen particles. These cells contain furthermore fairly high activities of -naphtyl-acetate esterase and 4-chloro-5-bromoindoxyl acetate esterase as well as acid phosphatase which was found in lysosomal localization. The huge intracellular vacuoles of the centrally situated cells possibly originate from electron translucent spaces within the glycogen fields of the peripheral cells.The notochord sheath consists of variously differentiated layers of collagen fibers and of an elastica externa. The diameters of the collagen fibers increase from the inner towards the outer region of the sheath. A peculiar feature of the Ichthyophis notochord sheath is a ring of mineralized collagen. The notochord of the caecilians investigated is compared with that of anurans, urodeles, and several groups of fish.Supported by the Deutsche Forschungsgemeinschaft.     

The influence of temperature on the thyroid gland of Chthonerpeton indistinctum (Gymnophiona,Amphibia)

Summary The thyroid gland of the tropical amphibian Chthonerpeton is markedly stimulated when the animal is exposed for two days to relatively high temperatures (36° C). Control animals were kept at 20° C. The stimulated gland exhibits collapsed follicles and a very tall follicular epithelium. At the ultrastructural level the activated epithelial cells were characterized by an increased number of apical microvilli, increased amounts of rough endoplasmic reticulum, an extended Golgi apparatus and a high number of various granular and vesicular inclusions. These differences in histology and fine structure were not paralleled by quantitative results, which were approximately the same in both groups of animals. The values of these determinations were relatively high.Gratefully dedicated to Professor Dr. Drs. h.c. W. Bargmann on the occasion of his 70th birthday.Supported by the Deutsche Forschungsgemeinschaft (We 380/5, Sto 75/3).     

Retinal projections in the caecilian Ichthyophis kohtaoensis (Amphibia,Gymnophiona)

Summary The retinal projections of the caecilian Ichthyophis kohtaoensis were investigated by anterograde transport of HRP. The optic tract forms two bundles in the diencephalon, a narrow medial bundle in the optic tectum, and a basal optic tract consisting of few fibres. Terminal fields are in the thalamus, pretectum, tectum, and as a circum-scribed basal optic neuropile in the tegmentum. Thalamic, pretectal and tectal projections are contralateral as well as ipsilateral. The reduced but existing visual projection corresponds to a reduced but existing visually guided behaviour.     

Marginal plates in hepatic peroxisomes of Ichthyophis glutinosus (Amphibia: Gymnophiona)

Summary The ultrastructure of hepatic peroxisomes was investigated in Ichthyophis glutinosus (Amphibia: Gymnophiona), employing perfusion fixation and the diaminobenzidine (DAB) technique for the visualization of catalase. The majority of peroxisomes is circular or rod-shaped, although elongated particles occasionally occur. They contain a finely granular matrix, lightly stained after the DAB procedure. Their mean diameter is approximately 0.25 m. Serial sections reveal that the circular and rod-shaped peroxisomal profiles are cross and oblique sections of highly tortuous, tubular organelles exceeding 2 m in length.In addition to tubular profiles, elongated, rectangular particles, as well as straight dumbbell-shaped organelles with distinct marginal plates are observed. They range from 900 to 1650 nm in length (mean = 1200 nm). In the flattened, thin central portion of the dumbbell-shaped particle, the peroxisomal membranes form a cisterna enclosing one or two uniformly thick marginal plates, which display a definite substructure with a periodicity of 10 nm.These findings indicate that peroxisomes in the liver of Ichthyophis exhibit a complex organization. It is suggested that the organelles undergo a specific differentiation process, morphologically characterized by the formation of enlarged segments of unusual shape.This study was supported by grants from the Deutsche Forschungsgemeinschaft, Fa 146/1-2 and Sto 75/9     

Observations on the fine structure,enzyme histochemistry,and innervation of parathyroid gland and ultimobranchial body of Chthonerpeton indistinctum (Gymnophiona,Amphibia)

Summary Fine structural and enzyme histochemical observations on ultimobranchial body and parathyroid gland of the caecilian Chthonerpeton are presented. The cell clusters and follicles of the ultimobranchial body consist mainly of granulated cells which are termed C-cells and obviously belong to the APUD cell series. In the larger follicles additional possibly exhausted degranulated cells and replacement cells occur. A rich supply of nerve fibres has been found in this gland. Frequently nerve terminals were observed to come into synaptic contact with the C-cells. Two categories of nerve fibres occur: a) fibres containing large polymorphic electron dense granules (probably purinergic fibres), b) fibres containing small electron transparent vesicles and a few electron dense granules (probably cholinergic fibres). The parathyroid gland consists of elongated cells (one cell type) poor in organelles and often containing fields of glycogen and lipid droplets. The cells are further characterized by fair amounts of lysosomal enzymes; they are interconnected by maculae adhaerentes and occludentes. No nerves and blood vessels have been found in the parathyroid gland of Chthonerpeton. This study has been supported by the Deutsche Forschungsgemeinschaft We 380/5.     

Immunohistochemical study of the adenohypophysis of Typhlonectes compressicaudus (Amphibia,Gymnophiona)

Summary The indirect immunofluorescence method was used to identify and locate LTH-, STH-, LH-, TSH-, ACTH- and MSH-immunoreactive cells in the pituitary of Typhlonectes compressicaudus (Gymnophiona). The present study defines the histological and histochemical staining properties of each cell type identified.     

Temperature dependent changes in the thyroid gland of Mertensiella caucasica (Urodela,Amphibia)

Summary The influence of temperature on the thyroid gland of the Caucasian salamander, Mertensiella caucasica has been studied (1) with the light- and electron microscopes, (2) with enzyme histochemical and quantitative methods. Animals which had been kept for years at 16° C were exposed for 48 hrs to 4° C or 27° C.At 4° C the follicular epithelium consisted of cuboidal cells with well-developed rough ER, occasional distended cisternae of the rough endoplasmic reticulum (ER) filling most of the cytoplasm, a relatively small Golgi apparatus and numerous round or oval shaped lysosomes. In animals kept at 27° C the follicular wall was composed of markedly taller epithelial cells, the apices of which bulged into the follicular lumen. Compared to the animals at 4° C, there appeared to be a decrease in the rough ER. The Golgi apparatus did not exhibit clear differences. The lysosomes increased in number and showed various shapes among which elongated ones with pointed ends were particularly striking. In the apical cytoplasm colloid droplets appeared indicating hormone release. The apical microvilli increased in number.The measurements of total T₄ have shown that the hormone level in animals kept at 27° C was about twice as high as in animals kept at 4° C.Dedicated in gratitude to Professor Dr. Drs. h.c. W. Bargmann on the occasion of his 70th birthday.Supported by the Deutsche Forschungsgemeinschaft (We 380/5).     

Fine structural studies on the thyroid gland of the normal domestic fowl

Summary The ultrastructure of the thyroid gland of the domestic fowl has been investigated and found to be similar to that of mammals. The differences were found at subcellular level in the distribution of the dark bodies which were mainly apical and in the sizes of primary lysosomes. These were found to range from 100 to 500 nm in diameter. All organelles described in mammals as being concerned with the production of thyroglobulin and the two hormones thyroxine and triiodothyronine were found to be present.     

Histological and histochemical observations on the neurosecretory cells in the diencephalon of Chthonerpeton indistinctum and Ichthyophis paucisulcus (Gymnophiona,Amphibia)

Summary In the diencephalon of two species of Gymnophiona (Amphibia) two neurosecretory nuclei were examined with histological (Alcian Blue, Aldehyde Fuchsin, Brookes Trichrome stain) and enzyme histochemical techniques (acid phosphatase, -naphthyl acetate esterase, acetylcholinesterase (AChE)). In the preoptic nucleus two categories of secretory neurons were distinguished: large and medium sized neurons. The perikarya of both cell types contain very little neurosecretory material. The Alcian Blue method stained the medium sized neurons faintly but selectively. The tractus praeopticohypophyseus is marked by the presence of Herring bodies, which, however, are relatively scarce. The neurohypophysis, in contrast, contains large amounts of neurosecretory material. Both cell types of the preoptic nucleus are characterized by their very strong AChE and -naphthylacetate esterase activity. The AChE also marks the tractus praeoptico-hypophyseus. In the large neurons acid phosphatase is present around the nucleus; in the medium sized neurons this enzyme is concentrated close to the origin of the axon. In the dorso-caudal hypothalamus a small group of neurons is stained with Alcian-Blue. These neurons, which also contain AChE, are located immediately under the ependyma which seems to be specialized in this region.The financial support of the Deutsche Forschungsgemeinschaft is gratefully acknowledged (We 380/5)     

A stem-group caecilian (Lissamphibia: Gymnophiona) from the Lower Cretaceous of North Africa

Of living amphibian groups, the limbless burrowing caecilians are amongst the most highly specialised, but are the least known. Their fossil record is extremely poor, leaving unresolved questions as to their origins, relationships and early distribution. We describe here caecilian remains from a Lower Cretaceous (Berriasian) microfossil locality near Anoual, Morocco. This material represents the second oldest record for the group, after the Jurassic Eocaecilia of North America, and the earliest caecilian record for Gondwana. It forms the basis of a new genus, Rubricacaecilia , which appears slightly more derived than Eocaecilia , but lacks major features of crown-group taxa. We support the use of Apoda Oppel, 1811 for the crown-group alone, and Gymnophiona Rafinesque 1814 for the clade comprising stem-group taxa + Apoda.     

The female cloaca of an oviparous caecilian amphibian (Gymnophiona): functional and seasonal aspects

Kuehnel, S., Herzen, J., Kleinteich, T., Beckmann, F. and Kupfer, A. 2011. The female cloaca of an oviparous caecilian amphibian (Gymnophiona): functional and seasonal aspects. —Acta Zoologica (Stockholm) 00 :1–14. Reproductive morphology is receiving increased attention in animals that have variable reproductive modes combined with internal fertilization. Exceptionally among amphibians all caecilian species practice internal fertilization via an intromittent organ: an everted part of the male cloaca (phallodeum or phallus). Because research has mostly concentrated on males, knowledge of the female cloacal morphology is scarce. Here, we present the first single‐species study of the functional morphology of the female cloaca of an oviparous, phylogenetically basal caecilian (Ichthyophis cf. kohtaoensis). We have analyzed female cloacal shape during the reproductive cycle combining conventional histology with 3D‐reconstruction. All females are similar in their overall cloacal structure with some differences in size and histology associated with the reproductive cycle. The female cloaca is divided into two distinct chambers similar to the male condition. The cranial chamber contains urogenital pockets into which oviducts and Wolffian ducts open and which may have function during oviposition. The caudal cloacal chamber bears a novel feature – dorsolateral blind sacs, which are homologous to the male condition, but are considerably smaller. The study of female cloacal morphology is essential to understanding the evolution of the caecilian reproductive system and contributes to the understanding of tetrapod genital morphology in general.     

Hair Cell Polarization in the Inner Ear of a Caecilian,Ichthyophis glutinosus (Amphibia: Gymnophiona)

The inner ear of Ichthyophis glutinosus is described with emphasis on the position of the sensory organs and the polarization of the hair cells. The hair cell polarization patterns of the maculae, cristae and papilla basilaris is similar to previous observations in other tetrapods. The papilla amphibiorum shows a simpler bidirectional polarization than described in other amphibians. The papilla neglecta, a sensory organ in the utriculus shows a unidirectional posteriorly directed polarization. A neglecta has not been found in the utriculus of anurans and urodeles previously.     

Ultrastructure of the kidney of a South American caecilian,Typhlonectes compressicaudus (Amphibia,Gymnophiona)

The ultrastructure of the renal corpuscle, the neck segment, the proximal tubule and the intermediate segment of the kidney of a South American caecilian, Typhlonectes compressicaudus (Amphibia, Gymnophiona) was examined by means of transmission electron microscopy (TEM), scanning electron microscopy (SEM) and freeze-fracture technique. The glomerular filter apparatus consists of the podocyte epithelium, a distinct basement membrane, a subendothelial space and the capillary endothelium. Emanating from the podocyte cell body, several long primary processes encircle neighboring capillaries. The short slender foot processes originating from the primary processes interdigitate with those from other primary processes, thereby forming the meandering filtration slit. Thick bundles of microfilaments are found in the primary processes, but absent in the foot processes. The basement membrane consists of a lamina rara externa and a rather thin lamina densa (50 nm thickness). The wide subendothelial space contains abundant microfibrils, a few collagen fibrils and many thin processes of mesangial cells. The endothelium is flat and fenestrated (compared to mammals displaying relatively few fenestrations); some of the fenestrations are bridged by a diaphragm. The glomerular mesangium is made up of the mesangial cells and a prominent mesangial matrix containing microfibrils and collagen fibrils. The cells of the neck and intermediate segments display numerous cilia with their microtubules arranged in the typical 9 + 2 pattern. The basal bodies of the cilia are attached to thick filaments with a clear crossbanding pattern of 65 nm periodicity. The proximal tubule is composed of cells typical for this segment (PT cells) and light cells lacking a brush border (bald-headed cells). The PT cells measure 10-25 micron in height and 15-30 micron in width and do not interdigitate at their lateral borders with each other. Their basolateral cell membrane is amplified by many folds projecting into lateral intercellular spaces and into basal recesses. The brush border is scarce and composed of loosely arranged short microvilli.     

Ultrastructure of the kidney of a South American caecilian,Typhlonectes compressicaudus (Amphibia,Gymnophiona)

Summary The ultrastructure of the distal nephron, the collecting duct and the Wolffian duct was studied in a South American caecilian, Typhlonectes compressicaudus (Amphibia, Gymnophiona) by transmission and scanning electron microscopy (TEM, SEM). The distal tubule (DT) is made up of one type of cell that has a well-developed membrane labyrinth established both by interdigitating processes and by interlocking ramifications. The processes contain large mitochondria, the ramifications do not. The tight junction is shallow and elongated by a meandering course. The connecting tubule (CNT) is composed of CNT cells proper and intercalated cells, both of which are cuboidal in shape. The CNT cells are characterized by many lateral interlocking folds. The intercalated cells have a dark cytoplasm densely filled with mitochondria. Their apical cell membrane is typically amplified by microplicae beneath which a layer of globular particles (studs) is found. The collecting duct (CD) is composed of principal cells and intercalated cells, again both cuboidal in shape. The CD epithelium is characterized by dilated intercellular spaces, which are often filled with lateral microfolds projecting from adjacent principal cells. The apical membrane is covered by a prominent glycocalyx. The intercalated cells in the CD are similar to those in the CNT. The Wolffian duct (WD) has a tall pseudostratified epithelium established by WD cells proper, intercalated cells and basal cells. The WD cells contain irregular-shaped dense granules located beneath the apical cell membrane. The intercalated cells of the WD have a dark cytoplasm with many mitochondria; their nuclei display a dense chromatin pattern.Research fellow of the Alexander von Humboldt Foundation     

鲻鱼甲状腺免疫组织化学与超微结构

用组织学、免疫组织化学和电镜技术对鲻鱼甲状腺的分布与形态结构进行研究。结果表明，鲻鱼甲状腺滤泡分散地分布在第1对至第3对鲻弓的入鳃动脉之间；构成同一滤泡的单层上皮细胞呈扁平形和立方形两种形态，反映出同一滤泡中上皮细胞生理活动的不同步，且发现不同发育时期的鲻鱼这两种形态细胞的比例有显著差异。用特异性甲状腺球蛋白(TG)抗体对甲状腺滤泡的免疫染色反应显示，阳性物质分布在滤泡上皮细胞的脑膜及围绕滤泡腔的周边与中央胶质，幼年与成年鲻鱼甲状腺滤泡的免疫阳性反应部位有所不同。两种不同活动状态的细胞，扁平形细胞和立方形细胞，在核与胞质的超微结构上有显著不同。扁平形细胞的核为扁平形或椭圆形，核内常染色质十分丰富，核膜光滑，核周腔和核孔没有扩张，胞质中各种细胞器发育较差；立方形细胞的核膜高度凹陷，将核分为2叶至3叶，核周腔和核孔扩张，胞质中各种细胞器发育良好，出现长棒形和哑铃形线粒体，且数量明显多于扁平形细胞，多层粗面内质网呈板层状排列，还有发育好的高尔基复合体。另外，我们还观察到一种介于扁平形细胞和立方形细胞之间的中间型细胞，其胞质和核的发育明显好于扁平形细胞，但比立方形细胞差。上述结果可为了解鲻鱼甲状腺滤泡细胞分泌活动的机制提供基础资料[动物学报49(2)：230—237，2003]。     

Contribution of the secretory material of caecilian (amphibia: Gymnophiona) male Mullerian gland to motility of sperm: a study in Uraeotyphlus narayani

Caecilians are a unique group of limbless burrowing amphibians with discontinuous distribution. Several caecilian species are viviparous, and all practice internal fertilization. In amniotic vertebrates the sperm undergo post-testicular physiological maturation when they are initiated into motility under the influence of an epididymal secretion. Further, during ejaculation mammalian sperm are suspended in a fluid secreted by the male accessory sex glands, viz., prostate gland and seminal vesicles. Caecilians lack comparable glands, but still practice internal fertilization. Uniquely, male caecilians retain the Mullerian ducts in the adults as a pair of functional glands. It has long been hypothesized, based on indirect evidence, that the Mullerian gland would be a male accessory sex gland, secreting a fluid in which sperm are suspended during ejaculation and which would also provide nutritional support to the ejaculated sperm. In the present study, the secretory material of the Mullerian gland of Uraeotyphlus narayani was mixed with sperm obtained from the testis, and the changes in motility were recorded. Uraeotyphlus narayani sperm possess a perforatorium of the acrosome proceeding deep into the endonuclear canal of the nucleus. The midpiece is characterized by closely applied centrioles, the anterior ends of the axoneme and axial fiber, and a mitochondrial sheath. The long tail has an undulating membrane on one side, supported by the axoneme and an axial fiber. The live sperm possess a mitochondrial vesicle, also known as the cytoplasmic droplet, anywhere along the head and the midpiece, as in anuran sperm, which is shed from sperm that have ceased motility. Uraeotyphlus narayani sperm are motile the moment they are released directly from the testis, indicating that the sperm do not require post-testicular physiological maturation. On being mixed with the secretory material of the Mullerian gland, the spermatozoa are enhanced in speed as well as duration of motility. Therefore, the caecilian male Mullerian gland is considered to be the male accessory sex gland.     

Structure and innervation of the pineal gland of the rabbit,Oryctolagus cuniculus (L.)

Summary A light microscopic investigation of the rabbit pineal gland with the aid of silver-stained sections gave the following results. In the gland a medulla and a cortex can be distinguished, the medulla containing so-called light and dark pinealocytes, the cortex only light ones. Autonomic nerve fibres reach the pineal organ by two routes: (1) via the perivascular spaces of pineal blood vessels and (2) via two distinct nerve bundles, the nervi conarii. Bilateral superior cervical ganglionectomy revealed that these pinealo-petal nerve fibres are mainly orthosympathetic postganglionic. Intramural pineal neurones with synaptic-like structures on their cell bodies and dendrites point to the presence of a parasympathetic innervation next to the orthosympathetic one. Direct afferent or efferent neural connections with the brain appeared to be absent. Acknowledgements. The author wishes to thank Professor Dr. J. Ariëns Kappers for encouragement and help, Mr. H. K. Koerten for his technical assistance and Miss A. M. Feddema for typing the manuscipt.