ALBERT

Notes: Abstract Tissue maps, and cell characteristics and properties were recorded in a study under the optical microscope of the development of the mouse calvarium from pre-natal to 26 days. Osteocyte populations in left and right halves of the calvarium were similar, but decreased with time for a given volume. Small isolated areas of bone matrix stained for phosphate (or carbonate) in a more readily available form from that in the surrounding matrix, which could be stained after sectioning but failed to stain in bulk. Osteocyte types were defined on the basis of histochemical methods for calcium and phosphate, which were associated inside bone cells in a complex manner, varying with time and position. The calcium and phosphate were not always present within the cell in discrete regions and were not always present in the same place in a given cell type. On the basis of a study of changes in cell types with time in selected sites a sequence of “loading” and “unloading” is proposed.

Notes: Abstract Mucopolysaccharides (MPS) and collagen have been analyzed in different types of cartilage and some other tissues at various stages of prenatal and postnatal human development. The concentration of the chondroitin sulfate-type MPS increased from the 38th embryonal day, the 6-sulfate exceeding in amount the 4-sulfate. The increase in collagen concentration was accompanied by a decrease in the proportion of neutral salt-soluble collagen. There was no accumulation of chondroitin sulfates in the calvaria. A maximal concentration of MPS of a low molecular weight occurred in the skin and in the ear pinna after the 10th prenatal week. Postnatally, the MPS-concentration in the iliac crest and in the tibial articular cartilages decreased gradually, the highest values being reached in the newborn; the collagen-concentration showed an approximately reciprocal course.

Notes: Summary When the complexity of a developmental system evolves to a certain point, appreciable variation must occur in the process. The problem the biologist faces is whether this point constitutes a limit to the evolution of complexity in developmental systems. If not, what mechanisms are employed to cope with the problem ? The problem—essentially one in “custom fitting” of parts, — and the possible solution(s) to it that have evolved are discussed. The antibody producing system appears to be one that “solves” the custom-fitting problem.

Notes: Abstract Naphthylamidase activity was studied in rat molar and incisor teeth at different stages of development. L-leucyl-4-methoxy-2-naphthylamide, L-alanyl-4-methoxy-2-naphthylamide, L-leucyl-2-naphthylamide and DL-alanyl-2-naphthylamide were used as substrates and Fast blue B and Fast Garnet GBC as diazonium salts. Naphthylamidase was not demonstrable in the teeth during enamel matrix formation. After the termination of this stage, naphthylamidase was present in the ameloblasts in their distal ends close to the enamel surface. The enzyme activity retained this localization until the dental epithelium fused with the oral epithelium at the time of tooth eruption into the oral cavity. Naphthylamidase was not found in other dental tissues, but marked activity was found in osteoclasts at the resorbing surfaces of alveolar bone surrounding the developing and erupting teeth and in certain areas of the connective tissue.

Notes: Abstract The effect of tetracycline HCl on synthesis of calcium sulphate statoliths inAurelia is reported. Tetracycline inhibits synthesis of statoliths and nematocysts when administered at an early stage of strobilation. The tetracycline, however, is not incorporated into the developing statoliths or nematocysts. As the tetracycline does not combine with the calcium of the calcium sulfate dihydrate statoliths ofAurelia, an explanation for its inhibitory effects on statoliths and nematocyst differentiation apparently does not rest with an incorporation-related factor. In vitro studies of four inorganic calcium systems and tetracycline revealed that calcium sulfate dihydrate (gypsum) did not incorporate tetracycline nor did its isostructural equivalent, calcium hydrogen phosphate dihydrate (brushite). Calcium carbonate and calcium phosphate (apatite) did incorporate tetracycline. The explanation for these different behaviors of calcium can be found in the crystal structure of the respective compounds, namely, whether or not the Ca ion is readily available to react with tetracycline.

Notes: Abstract A serial longitudinal study was undertaken to determine the activities of the alkaline and acid phosphatases in the long bones and mandibles. The optimum pH of the two enzymes was recorded at 10.2 and 5.4 for alkaline and acid phosphatase, respectively. Synchronized and randomized litters of rats were killed, 1 litter daily, starting at day 1 to day 25 post partum. Samples were analyzed for protein concentration and activity of the phosphatases. A pattern of low and high activity was observed in both bony tissues, as well as a pattern of low alkaline phosphatase activity during acid phosphatase activity peaks, and vice versa. The observed peaks suggest a correspondence between phosphatase activity and the other biochemical changes occurring in the growing bone, i.e., organic matrix and mineral formation. A separate study, considering the possibility of rhythmic features of the enzyme activity suggests that there may be a small diurnal rhythm at an early age.

Notes: Abstract Some of the morphological and physiological parameters of stipe growth or elongation inCoprinus radiatus were investigated. During the development of the fruit body the number of cells in a row in the growing portion of the stipe doubled during the development of the button, and again during the phase of rapid stipe elongation. Also during the stage of rapid elongation the cells in the upper 2/3 of the stipe increased 6–8 fold in length. The existence of a growth regulator synthesized in the cap and exerting control over the stipe was demonstrated through decapitation experiments. The cap appears to be required for normal stipe development until the stipe reaches about 1/4 of its final length. Through decapitation and cap-stipe exchanges it was found that the cap produced growth regulator up to the time of autodigestion; however, the stipe responded to the regulator only during a brief period at the onset of elongation.

Notes: Abstract Observations of sporophore development in fresh and glutaraldehydeosmium sequentially-fixed material ofProtophysarum phloiogenum show the following sequence. Small plasmodia cease streaming and round up. Food vacuoles collect in the lower center of the cytoplasmic mass. As the cytoplasm rises the food vacuolar contents are excluded from the plasmalemma and become the stalk core. A continuous, fibrous peridium and stalk tube enclose cytoplasm and stalk core respectively. Capillitial formation just precedes spore cleavage. Sporophore development is marked by autophagic activity and calcium deposition. Stalks of dried herbarium specimens of seven additional species have been examined. A mature stalk morphology very similar toProtophysarum with recognizable remnants of microorganismal food material is seen in all of them. It is thought that this marker is indicative of non-stemonitaceous stalk development.

Notes: Abstract In cultures ofacanthamoeba castellanii the stationary growth phase in which trophozoites develop to cysts, was induced in three ways: by transferring cells from a logarithmic growing culture into a nutrient-free medium, by growth in nutrient medium to high cell density and by inhibition of the mitochondrial DNA synthesis with ethidium bromide. In all cases, the intracellular concentration of adenosine-3′,5′-monophosphate (cAMP) rises by a factor of two to three from the end of the logarithmic phase to the beginning of the stationary phase. The results show that this rise may be more a consequence of an increased adenylate cyclase activity than of a diminished cAMP phosphodiesterase activity. No extracellular cAMP could be measured in cultures of the logarithmic and stationary growth phase, perhaps because of the extracellular cAMP phosphodiesterase. Because theophylline also induces the stationary phase and encystation, cAMP seems to be of importance for the development ofAcanthamoeba castellanii, especially for the degradation of glycogen.

Notes: Abstract The concentration of adenosine tri-, adenosine di-, and adenosine monophosphate in cells of Acanthamoeba castellanii was measured during the logarithmic growth phase and the stationary growth phase in which trophozoites were transformed into cysts. This developmental process was induced in three ways: by growth in nutrient medium to high cell density, by transferring cells in the logarithmic phase into a nutrient-free medium, and by mixing logarithmically growing cells with ethidium bromide. In all cases, encystment is accompanied with a reduction of total adenosine phosphate content to about 85%, mainly because of a depletion of cellular ATP. The value of the adenosine phosphate energy charge in logarithmically growing amoebae is 0.83. During development the energy charge becomes stabilized at different values (between 0.58 and 0.81), characteristic to the mode of encystation. A possible functional relationship between changes of the adenosine phosphate concentration and developmental processes of the amoeba is discussed.

Notes: Summary The epicuticle of adult Tenebrio consists of two distinct layers named outer and inner epicuticle. The outer epicuticle is the first cuticular layer to be deposited in form of small patches on top of the microvilli. These initial patches are composed of four dense laminae (A, B, C1 and C2) separated by three light spaces. The outer epicuticle grows by densification of diffuse material at the edges of the patches until the entire area is covered. The thickness of outer epicuticle remains constant (175 Å) during the development of the pharate adult, lamina B however rapidly disappears. Thus, the adult outer epicuticle is fivelayered (three dense laminae: A, C1 and C2). After being deposited, the inner epicuticle shows a complex laminar structure interpreted to represent a highly organized molecular system. The laminae are masked during the formation of the first procuticle lamellae. During the deposition of the epicuticle, lamina A is covered by a component of the moulting fluid, forming an irregular dense layer which disappears after the resorption of this fluid. Perhaps this layer protects the new epicuticle from lytic enzymes of the moulting fluid. In adult animals, there is an additional superficial layer, the signification of which is not clear. The possibility of remains of cement or wax is discussed. The development of the surface patterns of the sternal and pleural cuticle is determined before the epicuticle formation by the shape of the epidermal surface. The rate of outer epicuticle deposition appears to depend on the size of the microvilli: epicuticle deposition seems to proceed faster over high microvilli.

Notes: Summary The development of the seminal vesicle from the Wolffian duct and of the prostate from the urogenital sinus has been studied in rat fetuses from day 14 of gestation to birth with the use of the electron microscope. Prior to the onset of androgen secretion, the cells of the urogenital sinus and the caudal part of the Wolffian duct have a simple undifferentiated appearance. After the onset of androgen secretion by the fetal testes at day 15, “intracytoplasmic confronting cisternae” of the granular reticulum appear in both urogenital sinus and Wolffian duct. Portions of the granular endoplasmic reticulum of the urogenital sinus become distended with a finely granular, moderately dense material. In the urogenital sinus, many hemidesmosomes are formed at the basal surface of the epithelium. Specializations of the extracellular materials are present opposite the hemidesmosomes. The formation of the seminal vesicles and the prostate begins at day 18–19 of gestation. The cells of the seminal vesicle are taller than the Wolffian duct cells from which they arise, the granular endoplasmic reticulum increases moderately in amount, and a patent lumen is formed. The cells of the fetal prostate do not differ greatly from those of the urogenital sinus from which they arise except that the prostatic cells initially lack hemidesmosomes. The fine structural changes are discussed in relation to the onset of fetal androgen secretion, the formation of the organs, and the functions of the cells in adult life.

Notes: Summary Ultrastructural studies have been carried out on the innervation of the interstitial gland of the mouse ovary. In addition Falck's fluorescence method was applied. 1. Fluorescence microscopy: In the ovarian stroma green fluorescent nerve fibers are frequently to be found in the surroundings of large and small vessels. Seldom small fibers invade blocks of interstitial cells; however, their final ramification is not discernible. 2. Electron microscopy: Terminal fibers of the autonomic nervous system reach the cells of the interstitial gland from all sides. They may be independent from the course of the vessels. Many axons penetrate the basal membrane and come into close contact with interstitial cells, partly by forming large swellings (boutons), which may be deeply embedded into the cytoplasm of the innervated cell. The synaptic cleft is about 200 Å wide. Specialized pre- and postsynaptic membranes have not been found. The innervated cells show no peculiarities. The possible function of the synapses is discussed.

Notes: Summary The fine structure of the nerves of the ovarian stroma of the domestic fowl is described for the first time. In the fowl, the nerves are concentrated upon blood vessels, smoth muscles and mainly, the thecal gland with the steroid-producing cells. Myelinated as well as unmyelinated nerve fibers were observed. Numerous axon terminals representing adrenergic and also presumptive cholinergic nerve fibers are regularly seen in membranous contact with steroid-producing cells. In these axon terminals microvesicles are oriented towards the steroid-producing cells indicating a specialization of the surface from axon-to-cell contact. Evidence has been presented that there is a membranous neuro-humoral contact between the peripheral autonomie nervous system and the steroid-producing cells in the ovary. The present investigation has demonstrated that there is morphologic evidence for a nervous control of steroid-producing cells. The physiological importance of this neuro-humoral contact is discussed.

Notes: Summary Pituicytes of Rana pipiens could be classified into two types, pale and dense, according to their relative densities of cytoplasm and the populations of free ribosomes and cell organelles. An intermediate type of pituicyte was also recognized. Lipid droplet such as are typical in the cytoplasm of mammalian pituicytes, are not in the cytoplasm of either types of frog pituicyte. Both types have long cytoplasmic processes which run among the nerve fibers, and some of them end at the pericapillary space. Nerve endings making synapse-like contacts with the cell bodies or the processes of the pituicyte are frequent. According to the structures and sizes of granules and vesicles in the nerve endings, these endings are classified into one of three types: 1) A, which appears to be a peptidergic neuronal ending containing dense granules 1,200–2,000 Å in diameter and small clear vesicles 300–400 Å in diameter; 2) B, which appear to be monoaminergic endings containing cored vesicles 600–1,000 Å in diameter and small clear vesicles 300–500 Å in diameter; 3) C, which appear to be cholinergic endings containing only small clear vesicles. Type C endings are relatively rare. In the synaptic area the axonal membranes appose those of the pituicytes across a gap of about 200 Å and numerous “presynaptic” vesicles are clustered or accumulated near the presynaptic membranes.

Notes: Summary The differentiation of the podocytes was studied by scanning electron microscopy on kidneys of 10 days old rats. The results were compared with transmission electron microscopic pictures from the same kidneys. There are three ways of forming processes by the podocytes: 1. Slits within the cytoplasm give rise to cytoplasmic bridges which further divide themselves and finally build up a meshwork of processes within a cell. 2. Thick and sometimes very long processes originate from the cell border. Their smaller branches may interdigitate with those of distant podocytes or with other processes out of the same cell. 3. Finger-like processes of neighbouring cells interdigitate as soon as they develop. In spite of numerous desmosomal structures between neighbouring podocytes the cell processes and their interdigitations develop mostly independently from each other and only seldom after the expected rules of epithelial vicinity (type 3). These findings are interpreted as indication that flooded and growing capillaries induce the differentiation of podocytes and that they influence the orientation of their processes.

Notes: Summary The pars tuberalis of the hypophysis of Rana temporaria presents the general structural and the cytological characteristics of an endocrine gland. It is composed of elongated cells with long, branching processes ending on the external basement membrane of the pericapillary space. The pars tuberalis cells produce secretory granules which are accumulated in the pericapillary endings of the processes. Corresponding to its separate localization, the pars tuberalis of Rana temporaria has a separate vascularization of which the efferent capillaries anastomose with the capillary plexus of the median eminence. The general direction of the blood flow of the pars tuberalis is towards the capillaries of the median eminence. Also, the secretory products of the pars tuberalis pass into the blood stream of the hypophysial portal system. Several characteristics of the pars tuberalis show that its function must be different from that of the pars distalis of the hypophysis. Moreover, in contrast with the pars distalis, the activity of the pars tuberalis is not regulated by neurohumoral factors. The results show that a role of the pars tuberalis in the regulation of the activity of the pars distalis of the hypophysis is not excluded.

Notes: Summary The innervation of the adrenal cortex of the rat and the pig is investigated with the electron microscope. Nerve fibers containing synaptic and two types of dense-cored vesicles come into contact with endocrine cells. There are no specialized pre- and postsynaptic membranes. The synaptic cleft is about 200 Å wide. Generally the basement membrane between nerve and cell is absent. These observations are discussed on the base of more recent experimental findings. Small fibers having an average diameter of about 0.2 to 0.5 μ and containing only tubules and filaments are considered to represent parts of an afferent nervous system.

Notes: Summary The innervation of the mesenteric microvasculature was studied in fetal and neonatal rabbits with the aid of methods demonstrating fluorescence of catecholamines and cholinesterase activity as well as a silver impregnation procedure. The results showed that: (1) adrenergic nerve fibers were present, coursing independently in the mesentery by day twenty-one of gestation, and were found routinely in the adventitia of arterioles and venules by day 25 of gestation; (2) cholinesterase positive cells and fibers of the myenteric plexus were present by day 18 of gestation but cholinergic fibers were not present in the mesentery until day 26; the latter not being associated with blood vessels; and (3) nerve fibers in the mesentery thought to be sensory stained positively with the Holmes silver method on day 18 of gestation.

Notes: Summary Quantitative electron microscopic studies have been carried out on the human thymus. According to the equation L v =(2n)/F (Hennig, 1963) we have calculated that there is less than 0.204 mm nerve per 1 mm3 thymus tissue inside the blood-thymus-barrier (level of significance of 0.95). This result is compared to the degree of innervation in brown adipose tissue, which contains more than 160 mm nerve per 1 mm3 tissue. The biological significance of the paucity of neuronal elements in the thymus is undetermined.

Notes: Summary 1. The Harderian gland in rabbits, representing the type of a tubulo-alveolar gland, is located on the medial and posterior aspect of the eyeball and consists of two different parts, a small white lobe and a larger red one. The secretory cells in the tubular endpieces of both lobes are lipids containing cells. The lipid droplets can be stained with Sudan IV and Sudan black B. The luminal surface of both cell types is characterized by an alcianophilia at pH 2,5. 2. The tubules of both lobes have a single layer of columnar epithelium. The lipid vacuoles in the cells of the red lobe are large, these of the white lobe small. The multilocular cytoplasm of all cells contains many free ribosomes and high amounts of mitochondria lying very closely together. All cells exhibit numerous and large Golgi-zones but only few ergastoplasm membranes. 3. The lateral surfaces of the secretory cells are connected by elaborate junctional complexes (Zonulae occludentes, zonulae adhaerentes, desmosomes). These lateral surfaces are increased by intercellular canaliculi. 4. Before being released into the glandular lumen, the limiting membranes of adjacent lipid droplets fuse, thus forming a large lipid vacuole. Extrusion generally is characterized by the coalescence of the limiting membrane with the plasmalemma, the formation of an opening at the cell surface and the discharge of the secretory lipid material. In the course of another mechanism of extrusion, the fat vacuoles are transported to the apical part of the cell where consequently the plasmamembrane bulges into the lumen. Eventually the fat vacuole is pinched off surrounded by a thin cytoplasmic envelope. 5. Terminal fibers of the autonomic nervous system penetrate the basal membrane and can be found closely attached to the secretory or myoepithelial cells, partly by forming large swellings, which may be deeply embedded into the cytoplasm of the innervated cell. These terminal parts of the axons contain groups of synaptic and dense-cored vesicles, mitochondria and neurotubuli. Specific pre- and postsynaptic membranes have not been observed. The possible function of the harderian gland is discussed.

Notes: Summary The basic structure of the terminal sensilla of Locusta migratoria resembles that of Schistocerca gregaria. There are commonly six or ten neurons whose dendrites extend almost to the opening of the peg. Proximally the dendrites are clothed by a neurilemma cell which also encloses a basal cavity through which their ciliary region passes. The tormogen cell encloses the receptor-lymph cavity and actively secretes material into it. The receptor-lymph cavity and the basal cavity are quite separate. The development of new pegs at a moult is described. After apolysis the scolopale extends across the subcuticular space and protects the dendrites, which remain in a functional condition until shortly before ecdysis. As the trichogen cell grows out to form a new peg the tip is surrounded by a mass of electron-dense material, probably derived from the receptorlymph cavity. The function of this material is unknown. Regeneration of the dendrites is considered. The possible mechanism by which the tip of the peg opens and closes is considered and the general structure of the organule is discussed in relation to functioning.

Notes: Summary Electron microscopic studies have been carried out on the innervation of the mammalian anterior pituitary and parathyroids. The total area of grid squares (2.25·10−2mm2) examined was 2000 per gland and species. In the pituitary pars distalis and in the parenchyma of the parathyroid gland we did not observe a single axon profile. According to the equation $$L_V = \frac{{2n}}{F}$$ proposed by Hennig (1963) we have calculated that there might be—if any—0.133 mm of nerves per 1 mm3 tissue in those two endocrine glands (level of significance 0.95). Comparing these results to the degree of innervation in brown adipose tissue containing more than 160 mm nerve per 1 mm3 tissue we can not imagine that such a small degree of innervation is of any biological importance. In the pituitary pars tuberalis two types of axon terminals have been found both inside and outside the basement membrane surrounding the epithelial complexes. One type contains “synaptic” and two populations of smaller dense-cored vesicles, the other one contains a population of larger granules which have some properties of the classical elementary granules. Further investigations have to clarify the functional significance of those nerve endings.

Notes: Summary The penis retractor muscle of Helix pomatia is passed by thick nerve trunks, which probably are nerve nets. In the contracted muscle, they are folded meanderlike, in the extended they are pulled smooth. Four types of vesicles different in size and contents can be distinguished in the nerves. Varicose fibres accompany the muscle cells. Frequently the terminals are running in a groove of the muscle fibre. In certain axon types occur presynaptic accumulations of vesicles and thickenings of the terminal membrane. Terminal and muscle cell are separated by a cleft of 300 AE width. The muscle fibre membrane has no subsynaptic infoldings. Beside these axons thin, naked neurites are running through the connective tissue. They are characterized by a high content of neurotubuli. One part of the axons presumably possesses a monoaminergic transmitter. After the glutaraldehyde-dichromate-reaction they contain dense grana, whose diameters are mainly below 1000 AE. The nerve trunks fluoresce after exposure to paraformaldehyde vapour, excited with UV-light, green to green-yellow. The maximum of the excitation was determined at 413 nm and the maximum of emission at 496 respectively at 510 nm. It is concluded, that both, a catecholamine and 5-HT are responsible for the fluorescence. Extraction and paperchromatographic separation lead to the opinion, that the catecholamine is dopamine.

Notes: Summary The fat body sheath which surrounds the ventral nerve cord of the adult stick insect, Carausius morosus, is absent in the hatchling. Since the haemolymph sodium ion compositions of the two stages are similar, it is suggested that the fat body sheath is not a site of a blood-brain barrier dealing with extra-axonal regulation of sodium. Similarities of ultrastructure in connectives of adults and hatchlings are demonstrated, and tight junctions are shown to occur in perineural cells of both stages. The possible significance of this to the blood-brain barrier of this insect is discussed.

Notes: Summary In the ovary of Chironomus during phase 1 of the fourth larval instar, polygonally flattened “inner cells” are surrounded by smaller “outer cells” which contain bacteroids and phagosomes. Irregular cell remnants (“germ line accompanying substances”) lie among the inner cells. At the beginning of ovariole formation in phase 3, two layers of outer cells are separated by the formation of fissures. The inner layer of these cells (follicle- and egg-passage epithelium) forms regular invaginations. Cell pairs, identified as oocytes and nurse cells by “synaptic complexes” or multiple chromatin structures, wander from inside into the invaginations. Frequently between the two cells are fusomes, which later close in a characteristic manner. During phase 5, an egg passage is formed as a fissure among the egg-passage cells. During phase 7, the egg passage cells are conspicuously full of glycogen. Shortly before vitellogenesis membrane systems and annulated lamellae appear in the region of the oocyte. Accessory nuclei are formed by a “tieing-off” of projections of the the oocyte nucleus. During phase 9, microvilli and pinocytotic vesicles can be seen at the periphery of the oocyte. The distal cells of the ovariole are of oocyte or nurse cell nature, but in Ch. melanotus they are not surrounded by follicle cells and are reduced during further ovariole growth. In spite of the extremely small number of nurse cells in the follicle, the Chironomus ovary apparently does not differ functionally from other polytrophic meroistic insect ovaries.

Notes: Summary Ultrastructural diversification of muscle fibers, with regard particularly to myofibrillar changes, has been investigated in the fast-twitch extensor digitorum longus (EDL) and the slow-twitch soleus muscles of the rat during fetal and postnatal development in the presence and in the absence of motor innervation. The band pattern and the shape of the myofibrils were uniform in fetal and neonatal muscle fibers and underwent differential changes during the first weeks after birth, concomitantly with fiber type specialization. The most evident variations in myofibrillar structure arising in this period concern the thickness of the Z band and the arrangement of the myofibrils. Myofibril formation was at first not impaired by denervation of rat muscles performed in utero and, although focal disintegration of myofibrils and detachment and loss of orientation of filaments became apparent by one week, atrophic muscle fibers with well-organized myofibrils could be seen as late as 2 months after birth. However, denervated muscle fibers of EDL and soleus did not display any significant and consistent difference in myofibrillar band pattern and shape. No variation in mitochondrial content and sarcoplasmic reticulum development was likewise seen in muscle fibers of EDL and soleus after fetal denervation. The findings emphasize the importance of neuromuscular interactions in muscle differentiation.

Notes: Summary The tracheal organ of the mesothoracic tibia of Teleogryllus is located in a corresponding position to the tympanal organ of the prothoracic tibia. The mesothoracic organ contains an average of only 12 scolopidia, the location of which corresponds to that of the proximal group A and proximal main group in the prothoracid tympanal organ. There are no scolopidia corresponding to the distal group of the tympanal organ. The variability in number of scolopidia is much greater in the mesothoracic organ than in the prothoracic organ. The adult tracheal system of the mesothoracic leg resembles the early nymphal tracheal system in both pro- and meso-thoracic legs. The development of the tracheal organ is usually complete by the sixth instar. The mesothoracic tracheal organ of the adult is broadly equivalent to the prothoracic tympanal organ of a fifth instar animal.

Notes: Summary The ultrastructure and protein content of the five subfractions of the crude mitochondrial fraction from the brain stem of the 1-day old and adult rat was examined. The morphological composition of the subfractions after fixation in glutaraldehyde and osmiumtetroxide in the adult rat brain stem resembled that previously reported for the whole brain; synaptosomes sedimented in a sucrose gradient in subfractions C and D. In the 1-day old rat, mature synaptosomes were found in subfractions A, B, C and D; E contained mainly free mitochondria. 80–95% of the processes in the adult and 10–30% in the 1-day old rat contained synaptic vesicles which were of four types: (1) small agranular vesicles (2) large dense core vesicles (3) large agranular vesicles (4) coated vesicles. Pre- and postsynaptic membrane thickenings were demonstrated in many nerve-ending particles. In the subfractions of the 1-day old rat the protein content was one half and the distribution resembled that in the adult. Evidently nerve endings develop faster in the brain stem than in cortical areas; a serotoninor adrenergic origin of the early synaptosomes is suggested.

Notes: Summary Muscle spindles of limb muscles were deefferented in neonatal rats by sectioning ventral roots or by removal of the lumbosacral spinal cord. Ten to 56 days after the operation, muscle spindles were examined in the medial gastrocnemius, extensor digitorum longus and soleus muscles. The differentiation of muscle spindles was not affected by deefferentation. The number of spindles in the investigated muscles was not reduced. Intrafusal fibres increased in number from two at birth to four per spindle on the average, as in normal muscles. The characteristic ultrastructural distinctions of nuclear bag and nuclear chain fibres developed as under normal conditions. However, intrafusal fibres atrophied slowly after fusimotor denervation, their polar zones becoming reduced in diameter by about 25% in comparison with control fibre diameters. Spindle capsules, on the other hand, increased in size and attained diameters comparable with normal spindles, appearing even somewhat distended. As intrafusal fibres degenerate after complete denervation at birth (Zelená, 1957), but differentiate in the absence of fusimotor innervation, it can be concluded that sensory nerve terminals induce and support their development. It is assumed that the morphogenetic influence of sensory terminals is mediated by release and uptake of a trophic substance at the synaptic junction. The occurrence of light and dense core vesicles in the sensory terminals and of coated invaginations and vesicles at both the axonal and plasma membrane speak in favour of such a possibility.

Notes: Summary The development of the submandibular gland of the mouse was studied by means of electron microscopy, from the 14th day of gestation up to birth. In the first two days the acini are solid and their cells contain polyribosomes and a few lamellae of endoplasmic reticulum. Beginning with the 16th day secretory granules appear and rapidly fill an increasing number of cells. The different electron density of the granules makes it possible to distinguish 1. two types of granules, dense and pale, and sometimes intermediate ones, 2. “polymorphic” granules. The latter consists of electron dense and electron pale parts combined in different configurations. The possible significance of the various types of granules is discussed.

Notes: Summary The perinatal development of the Juxtaglomerular apparatus (JGA), with particular reference to the epithelioid cells, was studied. Electron microscopic studies were carried out on 18 gravid rats and 65 embryos with corresponding determinations of renin concentration and the renin contents by bio-assay. Specific secretory granules of the epithelioid cells of the JGA of juxtamedullary nephrons are first observed on the 18th day of gestation. The classification into three types of secretory granules suggests that they undergo a structural and probably also a functional transformation. All three types of secretory granule may be regarded as specific, renin-containing, cytoplasmic organelles. During the last days of gestation, the rough-surfaced endoplasmic reticulum and the Golgi apparatus become hypertrophied in epithelioid cells, indicating an increased protein synthesis as well as transport and concentration processes. After birth, there is a definite increase in the number of Type III secretory granules-which may be regarded as mature granules. The small Type I secretory granulesnot previously described-are found more frequently in the immature epithelioid cells. Determinations of the renin distribution in the plasma and in fractions of cytoplasmic constituents (total homogenate, secretory granules) prepared by differential centrifugation showed striking congruence with the morphological findings. It is generally assumed that “soluble renin” occurs in the endoplasmic reticulum and “bound renin” in secretory granules. A shift from extragranular to granular (bound) renin, occurs with increasing age, having its morphological equivalent in the relative decrease of the endoplasmic reticulum and increase in mature secretory granules.

Notes: Summary Qualitative and quantitative studies were made to determine the amount of nerve fiber supplying corpora lutea (CL) of rats during the oestrous cycle and pregnancy and sow CL during days 4–6 after ovulation. Fluorescence microscopy of freeze-dried, paraformaldehyde treated (Falck-Hillarp method) rat ovaries reveals adrenergic nerve fibers which run along with vessels and form a network among interstitial gland cells. Nerve fibers do not enter the granulosa cell layer in follicles or CL. In the CL circumference both vascular and non-vascular nerves occur the latter being related to the fibromuscular layer and probably innervating smooth muscle cells. No striking differences exist between the innervation of the ovary in non-pregnant and pregnant rats. Bodian and methylene blue staining did not contribute to a more detailed knowledge of rat ovary nerve supply. Electron microscopic quantitative analysis of rat and pig CL (rat: day 18 of pregnancy; pig: day 4–6 after ovulation) revealed no axon profiles in 2.000 grid squares (one square measuring 2.25×10-2 mm2) of randomly taken CL sections. Thus it was possible to calculate an upper limit of 133 μm of nerve fibers per 1 mm3 CL tissue, in case there were any at all.

Notes: Summary Hind limb muscles were de-efferented in 19 new-born rats by removal of the lumbosacral spinal cord, with preservation of spinal ganglia and their peripheral branches. The juxtaequatorial and polar zones of muscle spindles were studied in different leg muscles 3 to 9 weeks after the operation in order to establish whether intrafusal fibre types would become differentiated after permanent motor denervation. De-efferented intrafusal fibres developed into distinct ultrastructural fibre types similar to those found in control muscles. The nuclear bag type had confluent myofibrils with ill-defined M lines and relatively few mitochondria. The nuclear chain type had discrete myofibrils with prominent M lines, numerous large mitochondria and a more developed sarcotubular system. The fibre type characteristics were sometimes blurred by disarranged cross striation, but they were clearly discernible in 59 out of 69 de-efferented fibres of 31 spindles investigated in the electron microscope. A sample of 220 de-efferented spindles from leg muscles of 6 rats was examined in the light microscope on transverse sections stained for ATPase activity. The difference in the ATPase activity among intrafusal fibre types was marked in about 70% spindles; in contrast to this, no distinct fibre types could be discerned in the population of extrafusal fibres which were stained rather uniformly. In de—efferented spindles-as in normal control spindlesnuclear chain fibres always exhibited high ATPase activity, whereas one of the nuclear bag fibres had low ATPase and the other either low or medium to high ATPase activity. However, the ATPase activity of de-efferented muscles was generally lower than that of normal muscles. It can be concluded that intrafusal fibres do acquire their fibre type characteristics after fusimotor denervation despite complete deprivation of nerve impulse activity during the postnatal period when intrafusal fibre types differentiate in normal spindles.

Notes: Summary The innervation of the pituitary gland of Carassius auratus was studied by light and electron microscopy under various physiological and experimental conditions to investigate whether or not neurosecretory fibres play a role in regulating pars distalis function. Two types of neurosecretory fibre (Type A and Type B) were distinguished. Prolactin, ACTH and TSH1 cells were innervated by Type B fibre terminals separated from the endocrine cells by a continuous basal lamina (“indirect contacts”). Gonadotropic, STH and TSH2 cells were innervated by Type A as well as Type B neurosecretory fibres, mostly without an intervening basal lamina (“direct contacts”). The assessment of the amount of neurosecretory granules and microvesicles in nerve terminals during the pre-spawning, spawning and postspawning seasons and following the administration of Oestradiol, Thyroxine, Thiourea and Metopiron respectively revealed convincing evidence for a participation in pars distalis control for Type A and Type B fibres innervating gonadotropic cells and STH cells and Type B fibres innervating TSH2 and ACTH cells. Immediately after spawning both nerve fibre types innervating gonadotropic cells and Type A fibres innervating STH cells showed a striking decrease in the amount of dense core vesicles. During the spawning season nerve fibres innervating somatotropic cells, TSH2 cells and ACTH cells also undergo changes suggesting that prior to spawning major changes in the endocrine system of the goldfish take place.—These results point to a dual control, by peptides and amines, of teleost pars distalis function.

Notes: Summary During the development of the intermediate and late gastrula of Pomatoceros triqueter, the terminal and channel cells grow towards each other and form the protonephridium. They interdigitate through lateral microvilli. The junctions between the cells form filtration clefts, which are bridged by fibrils.

Notes: Summary An electron microscopical study has been made of the cervical spinal cord of Xenopus laevis embryos, from the time that the neural tube closes until the larvae were hatched and could swim. Sections of the whole cord were searched for intercellular junctions during this period. Two nonsynaptic types were found, the first were widely distributed puncta adherentia, the second were rare and similar to “gap” junctions. Membrane specializations with synaptic vesicles were first found when the neural folds had fused; “membrane-vesicle clusters” which looked like the presynaptic half of a synaptic junction were present, together with synaptic junctions lacking any postsynaptic membrane thickening or cytoplasm density. About four hours later, mature synaptic junctions with full thickening of the postsynaptic membrane, dense cytoplasm and striated or dense material in the synaptic cleft were present. Presynaptic mitochondria, dense-cored and flattened vesicles, fibre to fibre and fibre to cell body synapses were present from the first, as were synapses onto very fine dendrites which might be filopodia from dendritic growth cones. Synaptogenesis may start with the accumulation of vesicles in dense cytoplasm near a thickened cell membrane; the postsynaptic element becomes associated with this “membrane-vesicle cluster” and matures by increasing cleft and cytoplasmic density, and by membrane thickening.

Notes: Summary The olfactory mucosa of frog has been studied at an ultrastructural level to confirm previous light microscope observations in regard to the presence, in the sensory epithelium, of nerve fibres not belonging to the first cranial nerve proper. It has been observed that both myelinated and unmyelinated nerve fibres are present in the lamina propria and that eventually these fibres terminate inside the epithelium. Unmyelinated fibres usually contain dark core vesicles and similar content is seen in their intraepithelium terminals. Terminals containing only clear vesicles are also observed in the epithelium and they are believed to represent the terminals of the myelinated fibres. The significance of these ultrastructural findings is discussed in view of their functional meaning.

Notes: Summary In Xenopus laevis the development of hypothalamic monoaminergic cells was studied in relation to adaptation to background colour. The first melanophores appear at stage 33/34 (normal table of Nieuwkoop and Faber, 1956), gradually increasing in number. The melanine granules are dispersed throughout the cell, irrespective of the background colour. The dispersion apparently is caused by MSH released by the developing pars intermedia cells. Between stage 39 and stage 41, larvae placed on a white background changed colour from black to white due to aggregation of the melanine granules within the melanophores. With Falck's method for demonstrating monoamines, a small number of fluorescent cells was observed in the hypothalamus simultaneously with the first background-dependent colour change. These cells were arranged in a paired nucleus, bordering the third ventricle. Initially, the nucleus extends from 50 microns behind the optic chiasma to the lateral dilatations of the third ventricle; 8–10 hours later, similar cells were also found at the lateral dilatations and in the dorso-lateral part of the infundibular lobe. The cells have apical processes protruding in the ventricular lumen. Fluorescent axons, originating from the cells, were occasionally observed. Considering the above-mentioned results in combination with the electron microscopical data of Nyholm (1972), it is concluded that the MSH producing cells are under monoaminergic nervous control from the beginning of background colour adaptation.

Notes: Summary The Octopus iris is composed of five different layers: A, the external epithelium; B, the chromatophore layer; C, the iridocyte layer; D, the layer of muscles and collagen strands; E, the pigment epithelium. The nerves innervating the sphincter and the chromatophore muscles are identified and their neuromuscular junction is described. The motor endings of chromatophore nerves have an additional ending in presynaptic position which probably functions as a modifier of neuromuscular transmission. The chromatophores are naked and exhibit a tubular channel system between plasmalemma and pigment container which looks similar to the T-system of muscle cells.

Notes: Summary In the tortoise Testudo graeca, the lizards Lacerta dugesi and Lacerta pityusensis, and the snake Natrix natrix, the innervation of the testicular interstitial tissue was studied by light and electron microscopy, the acetylcholinesterase (ache) technique, the Falck-Hillarp method for the detection of catecholamines, and the application of 6-hydroxydopamine. The intertubular spaces of the reptilian testes studied contain adrenergic nerve fibers the amount and distribution of which varies considerably both in various species and in various stages of the reproduction cycle. Nerve fibers do not enter the seminiferous epithelium. Fluorescence microscopy of the lizard testis reveals catecholaminergic varicosities which are mainly arranged around blood vessels, but do not show obvious connexions to Leydig cells. Ache-positive fibers are equally distributed in lizard testes surrounding each seminiferous tubule. In Natrix natrix ache-positive fibers are irregularly spread among groups of tubules, without showing a definite relation to Leydig cells either. By electron microscopy bundles of unmyelinated axons and axon terminals can be more easily detected in the testes of immature animals than in adult. Terminals of nerve fibers containing small (400–500 Å in diameter) and large (800–1400 Å) dense-cored vesicles and sometimes small clear vesicles establish contacts with Leydig cells. Three types of contact are described. 1. “Contacts” par distance at a distance of about 2000 Å and basal lamina interposed; 2. membranous contacts having a 200 Å gap only between axolemma and Leydig cell plasmalemma; 3. invaginations of terminals into Leydig cell perikarya. The latter may exhibit surface specialisations, which strongly resemble postsynaptic membrane thickenings. Experiments using 6-hydroxydopamine underline the adrenergic character of testicular nerve fibers, which can be regarded as another example of non-cholinergic, ache-positive neurons. In the testis of the immature tortoise profiles of axons occur which probably represent purinergic, ache-positive neurons.

Notes: Summary Terminal axons emerging from the inner plexiform layer of the cat retina reach the wall of the arteria centralis retinae, as revealed by electron microscopy. Numerous unusually large dense core vesicles (about 1000 Å in diameter), of different electron densities, occur in the varicosities of these axons. These observations may be compatible with the idea of an innervation of the central artery of the retina which is non-autonomic, possibly intrinsic in nature.

Notes: Summary The ultrastructure of the lamina propria in the ovine seminiferous tubule was studied from 54 days (postcoitum) fetuses up to sexual maturity. In young animals (aged about two months) the effect of hypophysectomy and/or hormones was also analyzed. Initially, large areas of the lamina propria already show signs of morphological differentiation. The non cellular component is a single or sometimes double lamella. Outwardly, the cellular component is represented by rounded connective tissue cells or by elongated cells, exhibiting 80 Å filaments in the cytoplasm and arranged in 3 to 4 layers. By the first week after birth, the non cellular component is formed by 8 to 10 lamellae and the cellular component shows typical contractile cells, the fibroblasts being now located only in the periphery. Basically, the same aspect is found in the adult ram, thus differing from the more extensively studied prototypes of lamina propria architecture of rodents or primates. In as much as the sheep lamina propria differentiates precociously, probably induced by fetal testicular hormones, it is relatively insensitive to hypophysectomy or hormonal substitution. Hypophysectomy and cyproterone acetate administration cause intracytoplasmic lipid deposition in the contractile cells, the other components being not changed. Some questions are raised concerning the role of this peculiar lamina propria in the function of the blood-testis barrier.

Notes: Summary Neural elements within the parenchyma of the sebaceous gland have not been reported previously. Nerve endings have been observed only in the connective tissue surrounding the gland or in close association with the undifferentiated basal cells. In this study, electron microscopy revealed the possible presence of nerve endings (or terminal portions of neural elements) in the suprabasal level of functional sebaceous glands of pinnae of white rats. Morphologically, there are two distinct types of nerve endings. Type 1 is bordered by a membrane of relatively irregular contour and contains a single mitochondrion, various-sized vesicles, numerous microtubules, fine neurofilament-like fibrils, and occasional ribosome-like granules. Type II is also bordered by a membrane, but its contour was relatively smooth and rounded. Moreover, Type II contains many mitochondria, varying in size, density, and the arrangement of cristae. While ribosome-like granules are scattered throughout the structure in relative abundance, there are scarcely any fine neurofilament-like fibrils or microtubules. Whether these two structures are sensory or autonomic fibers could not be determined by electron microscopic examination.

Notes: Summary A light and electron microscope study of secondary lamellae of trout developing at 10° C is described. Collagen is secreted by mesenchyme cells in the connective tissue of the developing gill filament. This becomes enclosed in infoldings around the peripheries of mesenchyme cells. These cells become aligned in a single plane within folds of basement membrane and epithelium. The basement membrane of opposite sides of the fold becomes connected by a thick layer of collagen. Blood spaces form around the margin of the fold, connecting afferent and efferent filament blood vessels. Endothelial granules form in cells lining the outer border of the marginal channel when blood flow begins. Rows of pillar cells separate from the proximal layer of mesenchyme cells as the secondary lamellae develops further. New secondary lamellae are added at the filament tips. At 28 days, the basement membrane consists of 2 layers, a fine fibrous layer and 5–15 orthogonally arranged layers of collagen fibres. By 31 days, the collagen is arranged at random and the layer is thinner. A clear layer is also present by 67 days. A multilayered epithelium containing mucous and chloride cells is present at 28 days. By 102 days, the secondary lamellae are covered by 2 layers of epithelial cells only. Chloride cells are present in much greater numbers in developing gills than in the adult.

Notes: Summary The ultrastructure of the pineal gland of 18 human fetuses (crown-rump lengths 30–178 mm) was investigated. The pineal gland exhibits a pyramidal shape and consists of an anterior and posterior lobe. Only one parenchymal cell type, the pinealocyte, was observed. Few neuroblasts were seen between the pinealocytes and in the extended perivascular space. The pinealocytes possess all the organelles necessary for hormone synthesis. No specific secretory granule could be observed. The organ is abundantly vascularized and richly innervated. The morphology of the capillaries indicates the existence of a blood-brain barrier. The ultrastructure of the human fetal pineal gland suggests that the gland has a secretory function in early intrauterine life.

Notes: Summary The innervation of the brown, red and yellow chromatophore muscle cells in Loligo vulgaris shows quantitative and qualitative differences. The nerve bundles regularly contain axons with electronlucent vesicles of about 300 Å diameter, and occasionally axons with dense core vesicles of about 600 Å diameter. The myofibrils of the contractile cortex show a staggered arrangement and are wound in a spiral with respect to the axis of the muscle cell. In the axial sarcoplasm there is additionally a bundle of thin filaments of about 70 Å thickness. The bundle is orientated in parallel with the longitudinal axis of the muscle cell. Its function may be to maintain tonic contractions.

Notes: Summary In the blowfly Phormia terrae-novae the development of myofibrils of indirect flight muscles can be divided into periods of predisposition (anlage) and of growth. At the beginning of the growth period microtubules are disrupted by injection of colchicine. This disruption is followed by the formation of atypical ramifications of myofibrils at Z-discs leading to numerous disoriented myofibrils in late developmental stages. A possible mechanism preventing these alterations during normal development is discussed.

Notes: Summary The ultrastructure of the innervation of the anterior cerebral artery of the rat was studied in control animals and in animals after superior cervical ganglionectomy. Fluorescence histochemistry shows a periarterial network of intensely fluorescent fibers which are divided into two groups, adventitial and periadventitial. The fluorescence begins to decrease 26 hours after, and completely disappears about 32 hours after, ganglionectomy. Fine structural changes are first observed 18 hours after ganglionectomy, when the axoplasm of degenerating axons becomes electron dense. This density gradually increases up to about 32 hours. By 32 hours most axons with disintegrating axolemmas become inclusion bodies of the Schwann cells. At this stage, synaptic vesicles can still be distinguished as less dense areas, but the membrane structures of synaptic vesicles and mitochondria are difficult to recognize. The degenerating axons are gradually absorbed and by 38 hours dense, residual bodies are observed in the Schwann cells. Generally speaking, the degeneration occurs first in the adventitial fibers and then in the periadventitial fibers. The transient appearance of small, granular vesicles is noticed in axon terminals about 18 hours after denervation, although very few small, granular vesicles are seen in control tissue or at later stages of degeneration.

Notes: Summary The granular pneumocytes, one of the main cellular types of the lung epithelium, are characterized by the presence of large osmiophilic lamellar inclusions. The appearance and origin of these inclusions has been studied in the epithelium of chick embryonic lung at different developmental stages. Lamellar inclusions are first seen in the lung of 16 day old embryos. At this stage, few concentric lamellae surround a large amorphous center; the periphery of the inclusions always contains small granular structures. In the following days, the number of cells containing these lamellar inclusions increases rapidly, while their lamellae progressively become more numerous. In 19 day old embryos, the lamellar inclusions are similar to those in the lungs of adult animals. From the earliest formation of the bronchial primordia, their epithelial cells contain a number of typical “granular” inclusions. These organelles are characterized by a granular center, enclosed in a membranous system. This structure becomes more complex as the embryo develops; in the 16 day old embryo, the multilayered membranous system coils around the granular center. At the time when lamellar inclusions first appear, granular inclusions increase rapidly in number and are often found in close association with lipidic vacuoles. The relationships between lamellar inclusions, granular inclusions and lipidic vacuoles are discussed. The evidence suggests that a lamellar inclusion arises from the cooperation of several granular inclusions and a lipidic vacuole.

Notes: Summary Nerve fibers were demonstrated in the glycogen body of the avian lumbar spinal cord by using silver-impregnation and fluorescence microscopic (Falck-Hillarp) techniques. These nerve fibers originated from nuclear areas in lateral juxtaposition with the glycogen body. The fluorescence of the nuclear area was strongest near the polar regions of the glycogen body. It was suggested that the aminergic neurons of the avian lumbar spinal cord may influence the glycogen body. The avian glycogen body was compared with other storage sites of glycogen within the central nervous system.

Notes: Summary The fine structure of the pterin layer was investigated in both wild type Rana pipiens and Rana pipiens homozygous for the speckle mutant gene. No difference in morphology of the layer was noted between the wild type and mutant. The layer lines the outer surface of the stratum compactum of the dermis and separates this stratum from the stratum spongiosum. The pterin layer consists of extra-cellular material and contains membrane-bounded granules filled with fine spicules. Many of the spicules are somewhat similar in appearance to the initial calcification loci present in developing membrane bone. The layer first appears in the tadpole at approximately stage 14 (Taylor and Kollros, 1946); subsequent developmental stages are described.

Notes: Summary By means of electron microscopy “light” and “dark” pinealocytes can be distinguished in the guinea-pig pineal gland. Glial cells are rare. In the “light” pinealocyte. the most frequent cell type, some “vesicle-crowned rodlets” (VCR) show circumscribed thickenings. From these structures “vesicle-crowned balls” (VCB) have to be clearly distinguished. Furthermore “cylinders” occur, which, it is suggested, are precursors of VCB. “Dark” pinealocytes characterized by chromatin-rich nuclei and electron-dense cytoplasm are rare and contain fewer vesicles, VCR, VCB and “cylinders” than “light” pinealocytes. Numerous non-myelinated nerve fibres are situated within perivascular spaces, a few also in the parenchyma. Synapses between nerve fibres and pinealocytes were not observed. In the pineal gland of pregnant guinea-pigs the following changes can be observed in the second half of gestation. The “light” cells show many nuclear indentations and an increase of “active” zones, mitochondria, smooth ER, agranular vesicles, VCR, VCB, and “cylinders” respectively. The “dark” cells increase in number. After birth these changes reverse to normal within one week. Constant darkness leads to an activation of the “light” cells accompanied by an increase of the VCR and to an increase in number of the “dark” cells. Under constant illumination the “light” cells show a decrease of their organelles and a strong increase of the VCR. After 70 days the VCR also show a change in shape. Following reserpine treatment the VCR decrease in number and show signs of degeneration. It is discussed that the VCR function as pre- or postsynaptic structures and that they are involved either in transmitting impulses from nerve fibres to pinealocytes or from one pinealocyte to the other.

Notes: Summary The postnatal development of the rat prostate has been studied with the electron microscope. Major developmental changes begin during the second week after birth and involve organelles associated with the formation of secretions. The amount of granular endoplasmic reticulum and the size of the Golgi complex increase greatly. Large vacuoles that probably contain secretory material are formed, and the lumen of the prostatic acini appears to contain secreted material. Large lysosomes with polymorphic interiors are present as early as 10 days after birth, and they become numerous by the end of the third week. Differences in fine structure between the different lobes of the prostate are detectable in 10–14 day old rats. The subsequent differentiation of the granular endoplasmic reticulum into the forms characteristic of the different prostatic lobes is described. The initial changes in the prostate occur in advance of sexual maturity of the animal, and the adult appearance of the gland is attained by 4–5 weeks after birth.

Notes: Summary The origin of membranes required for the formation of microvilli has been investigated electronmicroscopically in enterocytes of fetal rat small intestine. It is assumed that horseshoe-like structures consisting of unit membranes covered with electron-dense material, which probably represent longitudinal sections through cap-like structures, are incorporated into the apical cell membrane and give rise to the tips of microvilli. This assumption is based chiefly on the almost identical diameters of “horseshoes” and microvilli, the localization of “horseshoes” in the terminal web, and the time of appearance and disappearance of “horseshoes” with regard to development of microvilli. There are indications that the “horseshoes” originate in the Golgi apparatus.

Notes: Summary This investigation is concerned with pineal organs of human embryos 60 to 150 days old. At every stage central nerve fibres enter the pineal organ by way of the habenular commissure, but are restricted to the pineal's proximal part. On about the 60th day of the development the sympathetic nervus conarii grows into the distal pole of the pineal organ from a dorso-caudal direction and plays the predominant part in the innervation of the pineal organ. After penetrating, it soon branches out and forms a network in the pineal tissue. Much later, not until the 5th embryonic month, sympathetic nerves appear accompanying the supplying vessels in the perivascular spaces. After a short time these nerves pierce the outer limiting basement membrane and penetrate the parenchyma. Towards the end of the 5th embryonic month the axons of the sympathetic nerves form varicosities containing clear and dense core vesicles. At this point large amounts of laminated granules appear primarily in cell processes, probably of pinealocytes. Isolated granules also occur in the varicosities of axons. The granules encountered here are most likely secretory granules.

Notes: Summary Nervous (sensory) pathways of the pineal complex were studied in Lampetra planeri. The following observations were made: (1) The pineal tract, containing more than 600 unmyelinated fibres, connects directly or indirectly to the posterior commissure. (2) Connections of the nervous region (NR) adjoining the parapinéal organ (“ganglion parapinéales” of Tretjakoff or “division of the left habenular ganglion” of Studnička) have been re-examined; our study shows that the rostral division of NR is connected to the parapineal organ by a short tract and to the left habenular ganglion (LHG) by a nervous layer (NL). The NR, NL, and LHG contain the same following elements: (1) A collection of typical neurones (which are rare in the NL) in an abundant meshwork of nerve fibres. (2) At least two main types of axons: (a) Type 1 axons, containing few granules (1,000–1,300 Å diameter). (b) Type 2 axons, containing numerous granules (900–2,500 Å diameter), and resembling neurosecretory axons. Type 2 axons are not observed in the pineal and parapinéal organs. (3) Axo-dendritic and axo-axonic(?) synapses. Our study concludes that NR and NL represent the rostral part of LHG; nevertheless, NR and NL may contain the parapinéal tract. The problem of possible incorporation of epithalamic elements (e.g., elements of the habenular ganglia) into the parapinéal and pineal organs in other animals has been considered phylogenetically.

Notes: Summary Umbilical vessels of guinea-pig fetuses were studied shortly before birth. In all umbilical cords investigated an innervation of the umbilical vessels is lacking. The intrafetal parts of the umbilical vessels on the other hand are richly innervated. A marked difference in the amount of nerve fibres and the pattern of innervation is found between artery and vein. The artery is supplied by a dense nerve plexus which spins around the media and which originates from nerve bundles within the outer adventitial layers. The comparatively scanty innervation of the vein exhibits a more coarsely meshed net pattern. The nerve bundles in the vein exhibit a close affinity to the vasa vasorum. Number and type of the close contacts between the muscle cells are different in the various sections of the umbilical vessels. Similar to the distribution of nerves they are almost absent in the vessels of the umbilical cord, numerously, however, in the intrafetal parts. Contrary to the innervation, the close contacts in the vein are developed more numerously and more broadly than in the corresponding artery.

Notes: Summary The development of adrenergic nerves to the anterior eye segment was studied in human and guinea-pig embryos. Adrenergic terminals had already appeared in the earliest human embryos available (4–6 cm). They first appeared mainly in nerve trunks in the primitive chorioid, especially in the region of the developing ciliary body. Adrenergic nerves then grow into different structures of the eye as these develop, but typical terminals in contact with effector cells appeared late during the development, about the 25–30 cm stage. No adrenergic nerves were observed in the chamber angle. Corneal adrenergic nerves (also intraepithelial terminals) appeared much more frequently in embryos than in adults. No adrenergic neurons were observed in the retina. In the guinea-pig, the first adrenergic fibres were observed at about gestation day 35. The general principle of the development was very similar to that of the humans. At gestation day 45 to 50, the supply of adrenergic fibres was essentially that of the adult animal, except that the corneal adrenergic fibres were increasing until just before birth and that the adrenergic terminals of the chamber angle appeared shortly before term.

Notes: Summary Nerve fibres containing granular vesicles first appear in the median eminence of the rat on the 16th foetal day while secretory granules in the cells of the adenohypophysis are not present till the 17th foetal day. These observations suggest that the differentiation and early activity of pars distalis cells may depend on substances elaborated at nerve terminals in the median eminence. Although the loops of the primary plexus of portal vessels do not develop until the 4th postnatal day, substances released by nerve fibres in the neurohypophysis could reach the pars distalis through vessels already present at the 15th foetal day in the mesenchyme between the diencephalon and the adenohypophysis. This view is supported by the fact that the earliest cells to exhibit ultrastructural evidence of secretory activity are in the rostral pole of the pars distalis, the first region of the gland to become vascularized. The earliest granules to appear in the cells of the pars distalis correspond to those which are considered to contain mucoprotein hormones; somatotrophin type granules were seen only in postnatal tissues. The finding that, in the median eminence, the development of granular vesicles precedes that of agranular vesicles is discussed with reference to the times at which neurosecretory materials and monoamines become detectable in the region.

Notes: Summary The intrinsic innervation of the anterior byssal retractor muscle in Mytilus edulis L. has been demonstrated using Maillet's osmium-zinc iodide technique (ZIO). Grouped and isolated osmiophilic cells, thought to be neurons, and nerve fibers forming neuromuscular contacts and nerve endings, have been observed. When the period of fixation is not overly prolonged, glio-interstitial cells may also be distinguished.

Notes: Summary A study was made of the ultrastructure of the paracervical (Frankenhäuser) ganglion of the newborn rat, using immersion fixation by glutaraldehyde (2.5%) followed by OsO4 (1%), or KMnO4 (3%) fixation. The cells containing dense—core vesicles were divided into three groups: (1) primitive sympathetic cells, (2) cells containing some dense-core vesicles 700–1100 Å in size and structurally resembling sympathetic neurons, called principal neurons, and (3) cells containing many dense-core vesicles with a larger, darker dense core, 800–2000 Å in diameter, called granule-containing cells. Using glutaraldehyde-osmium fixation, the principal neurons were further divided into dark and light cells on the basis of electron opacity of the cytoplasmic matrix. The granule-containing cells were believed to correspond to the small, intensely fluorescent cells (SIF-cells) previously described using the formaldehyde-induced fluorescence technique. On the basis of the amount of granules, the granulecontaining cells were classified as mature or maturing SIF-cells and as more primitive SIF-cells, and developing sympathicoblasts. The development of synapses in autonomic ganglia was discussed.

Notes: Summary A study of the ultrastructure of the developing thymus of the leopard frog (Rana pipiens) revealed that the thymus had undergone all of the major changes which would persist through larval life and metamorphosis by the time that the animals had reached larval stage IV of Taylor and Kollros (1946). These changes included development of an outer, lymphoid cortical region and an inner, essentially nonlymphoid medulla; mitotic activity among lymphoid cell precursors and the formation of the first small lymphocytes; development of complex cysts containing PAS-positive material and the appearance of other signs of secretory activity among epithelial cells of the medulla; and differentiation of large myoid cells containing bundles of striated muscle fibrils. The changes are particularly noteworthy because they first appear during a period in which the animals are known to be developing the capacity to respond immunologically to allografts.

Notes: Summary Glio-vascular relationships were studied in the developing rat cervical spinal cord with electron microscopy. Capillaries were first evident on the eleventh prenatal day and were surrounded by undifferentiated cells and neuroblasts separated from the capillary wall by a perivascular space. This space persisted until day thirteen, when basement membranes began to appear. Small cellular processes containing glycogen were visible adjacent to the capillary basement membrane beginning at day fifteen and became more prominent as development proceeded. The origin of these cellular processes was not apparent, but they were most likely collaterals of epithelial elements. In prenatal animals neuroblasts or neurons were in direct contact with the capillary basement membrane. Cells resembling adult astrocytes were first observed during the nineteenth day. Astrocytic processes completely invested the capillaries, separating them from neurons in the spinal cord of all postnatal animals.

Notes: Summary The ultrastructural alterations in the anterior midgut epithelium of the fly Calliphora were investigated by electron microscopy. Nine developmental stages, from late larval to late adult, were studied. During metamorphosis, a tube is formed by proliferation of undifferentiated cells residing in cell nests in the basal part of the larval epithelium. The process of microvillus formation is described and compared with that in the embryonic stage. Differences between larval and imaginal cells were found in the rate of microvillus formation and in the occurrence of dense vesicles. The formation of a basal labyrinth coincides roughly with microvillus formation. Glycogen deposits occur mainly in the pupal stage and are almost always absent after emergence. Epithelial crypts are formed in late pupae by modification of the existing layer of cells together with a localized development of muscle fibres. In the adult stage changes may take place in the form of the cell apex, which generally assumes a more pointed shape, and in the amount of osmiophilic material, aggregating almost exclusively in the supranuclear part of the cell.

Notes: Summary The ultrastructure of the pars intermedia (PI) of the normal VII +/+ and hereditary nephrogenic diabetes inspidus DI Os/+ mice has been studied with particular reference to the morphology of the glandular cells and their innervation. Four types of cells were observed in both the genotypes of mice, 1) the light glandular cell, 2) the dark cell, 3) a type of cell similar to ependymal cells and 4) a small percentage of typical ACTH cells, observed mostly on the PI border of the cleft and rarely in the centre of PI. The predominant light glandular cells contain mainly two types of membrane bound granules: 1) electron dense core granules, which measure 1500–2500 Å and 2) electron lucent vesicles, which measure 3000–4000 Å in diameter. Granules of intermediate size with various density are also present in both types of mice. The electron dense core granules are predominant in DI Os/+ mice, whereas, electron lucent vesicles are predominant in the normal VII +/+ mice. Similar uniform size membrane bound electron dense granules have been observed in ACTH cells of PI and pars distalis. From earlier experimental evidences and the present observations, it is concluded that the dense core granules in PI may be synthesizing ACTH or ACTH-like substance. It is also discussed that these dense core granules may further mature and give rise to MSH in the form of electron lucent vesicles. If it is so, PI light glandular cells may have dual functions, of producing MSH and ACTH. One of the functions of ependymal-like cells, may be the transport of PI secretion. Three types of nerve endings are observed throughout the PI, making synaptic contact with the predominant cell type. The innervation is more in DI Os/+ mice than in normal mice. The classification of these nerves is according to Bargmann and co-workers 1) peptidergic neurosecretory fibers, contain mainly membrane bound dense core granules, measuring 1200 to 1800 Å, and are the classic neurosecretory granules; 2) adrenergic fibers, measuring 700–900 Å; 3) cholinergic fibers, measuring 300–400 Å. Adrenergic and cholinergic fibers are more towards the hypophysial cleft. The increased innervation, the synaptic contact, the extremely hypertrophied PI and the greater activity of its light glandular cells in the DI Os/+ mice show the PI is under the influence of the nervous system.

Notes: Summary The innervation of the salt gland of the goose, the duck and the swan was investigated by means of electron microscopy. Axonal swellings were observed in relationship to secretory cells as well as to central duct cells. The terminals contain synaptic and densecored vesicles. There are no specialized pre- and postsynaptic membranes.

Notes: Summary Morphological evidence is presented supporting the possibility of basal secretion into hypendymal capillaries of the adult rabbit subcommissural organ (SCO). The synthetic apparatus of the SCO cell is described as well as the heterogeneous granules and vesicles which are concentrated in the basal processes bordering a widened perivascular space. The origin of the electron dense granules, of which two fairly distinct subgroups are found, is discussed. A binding of secretory sacs to the lateral plasma membrane is seen. The possibility of a lateral secretion is supported by the presence of a system of extracellular channels between SCO cells which are filled with a flocculent material resembling that of the secretory sacs. Nerve perikarya which are separated from the SCO by only a few glial fibers are demonstrated. Synapses are described in nerve fascicles bordering on the hypendymal capillaries. The possibility of an innervation of the hypendymal region is discussed as well as possible nervous connections with the pineal gland.

Notes: Summary During the embryonic and larval developmental stages of the frog, Rana temporaria L, anti-β 1–24, α 17–39 corticotropine, α and β MSH antibodies were used to define, with immunofluorescence technique, the appearence of corticotropic and melanotropic cells. A very small number of fluorescent corticotropic cells appears for the first time during the embryonic stage (10 mm), just before the differentiation of the pars intermedia. The cells are small, their large nucleus is surrounded by a fine rim of fluorescent cytoplasm. During premetamorphic stage, the anti-ACTH antibodies (anti-β 1–24 and anti-α 17–39 corticotropine) reveal more fluorescent cells in the whole pars distalis. The pars intermedia cells can also be visualized by both antisera. At the end of prometamorphosis and during climax the corticotropic cells show a more precise localization. As in adult frog pars distalis, they are concentrated in the rostral half of the lobe. With the same technique anti-α and β MSH antibodies reveal only the cells of the pars intermedia. No other cell type of the pars distalis reacts with these antibodies. This technique has the advantage to show that the ACTH and the MSH cells appear very early during the embryonic life.

Notes: Summary Air-dried stretch preparations were used to study adrenergic nerve fibres and catecholamine-containing cells with the formaldehyde-induced fluorescence method in the abdominal tissue block containing the para-aortic paraganglia, in the neurovascular structures of the cervical region including the carotid body, in the bladder, in the ileum, in the mesentery, in the vagus nerve and in the sympathetic ganglia of 5- or 15-day-old rats. The adrenergic nerves and the catecholamine-containing cells were well preserved and showed little or no diffusion of amines. While most intensely fluorescent cells of the main para-aortic body disappeared during the first two postnatal weeks, some such cells survived and they showed long, slender fluorescent processes. Administration of 20 mg/kg of hydrocortisone acetate daily for 5 days after birth caused a striking increase in the number and size of the clusters of the intensely fluorescent cells in the organ of Zuckerkandl, in the sympathetic ganglia and in the bladder, as well as an increase in the fluorescence intensity of the carotid body. In rats treated with hydrocortisone for 5 days and left to recover for 10 days an increased fluorescence was still observed. However, in the organ of Zuckerkandl the intensely fluorescent cells of hydrocortisone-treated 15-day-old rats showed less processes than those of the control rats of the same age.

Notes: Summary The salivary glands of the moth, Manduca sexta (Insecta: Sphingidae), are unlike most other salivary glands in that they are innervated from one source only. Vital staining of nerves with methylene-blue reveals numerous fine nerves extending to the glands from the oesophageal nerve, a part of the stomatogastric or visceral nervous system. Light and electron microscopy confirm that only the fluid-secreting cells, confined to a discrete region in these glands, are innervated. Axons with or without glial wrappings are found in intercellular spaces between fluid-secreting cells. Axons lacking a glial sheath contain, after glutaraldehyde-osmium tetroxide fixation, large granular and small agranular vesicles. In nerve endings in glands fixed with permanganate these smaller vesicles are granular, having the electron-dense cores characteristic of monoamine-containing neurons. These nerve endings with “synaptoid areas” are in close (“direct”) contact with the fluid-secreting cells.

Notes: Summary Electron microscope studies of axons, distributing singly or in small bundles in the human ventricular and atrial myocardium, indicate a few per-cent of the axon profiles to be significantly large in diameter (1.5–3.0 μ). They are characteristically packed with a profuse number of mitochondria along with large granular vesicles, glycogen rosettes, lysosomic bodies; and some of them terminate on a “specific terminal cell” (Knoche and Schmitt). These mitochondria-rich, large axons are assumed to be terminal portions of the cardiac afferents. About half of the axons encountered in the ventricle and 2/3 in the atrium are non-vesiculated, usually less than 0.5 μ. in diameter. The varicosities containing numerous vesicles are mostly 0.5–1.5 μ in diameter and are assumed to be terminal portions of the cardiac efferents. The ratio between the number of axon profiles containing small granular vesicles and that of axon profiles containing agranular vesicles without small granular vesicles is 2∶1 in the ventricular myocardium and 1∶1.7 in the atrial myocardium.

Notes: Summary The hilus cells in the ovary of the pig have been investigated with the electron microscope. These elements are identical with Leydig cells. The hilus cells contain an abundant agranular endoplasmic reticulum, which is either organized as a tubular network or as an onion like system of closely packed flattened cisternae sometimes filling up half of a cell and often being concentrated around lipid droplets and pigment granules. The mitochondria have cristae, tubuli and sacculi and a matrix of variable electron density. The golgi cisternae are poorly developed. The cells have centrioles, and there are hints of an amitotic cell division. The cytoplasm of the hilus cells contains lipid droplets, which have a diameter up to 5 μm. The pigment granules are extremely polymorphic, nearly always surrounded by a single membrane. They possibly may be derived from lipid droplets and mitochondria. Single axons of the hilus nerve occasionally penetrate the basement membrane and come into close contact with the hilus cells. Typical synapses were not observed. The results are discussed on the background of light microscopical, biochemical and pathological findings and compared with those obtained on Leydig cells of different species. Various cell images are interpreted to represent progressive stages of maturation.

Notes: Summary The fine structure of the secretory units of the mouse submandibular gland was studied according to the developmental sequence. The embryonic submandibular gland consists of terminal tubules and ducts. Myoepithelium is associated only with the terminal tubules, and the cells of the primary intercalated ducts show characteristics of the young striated duct cells. The major changes shortly after birth consist of: 1) opening of the secretory lumina, 2) increasing rough ER and its altered configuration, 3) dilatation of Golgi cisternae and 4) changes in the granular structure. These findings suggest that the salivary secretion first occurs after birth, and acinar differentiation or transformation of the secretory cells of the terminal tubules is induced and profoundly affected by the commencement of the secretory activity. In the intercalated ducts this process is somehow inhibited, and the granular cells found in the adult can be considered as the remnants of the secretory cells of the terminal tubules.

Notes: Summary 1. Structure and development of the integument and the larval oenocytes during the pupal and pharate imaginai stage of Culex pipiens L. (Dipt.) are described. 2. Formation and digestion of the pupal cuticle and the formation of the imaginai cuticle take place within temporarily fixed limits. 3. While the pupal endocuticle is digested the lamina of the at times innermost lamella appears as a “ecdysial membrane”. 4. The oenocytes are characterized by their agranular, tubular endoplasmic reticulum (ATER) and its changes. The structure of the oenocytes is comparable to that of vertebrate cells engaged in steroid hormone synthesis. 5. The morphology of the oenocytes shortly after pupal ecdysis changes mainly by the continual raising of so called lipid vesicles. These vesicles arise from dilatations of tubular elements of the agranular endoplasmic reticulum. 6. Studied under the light microscope these changes appear as channel structures (“secretory phase”) shortly after pupal ecdysis; later on the cytoplasma becomes vacuolar in structure (“degeneration phase”). 7. By staining with Sudan black B and Sudan III/IV lipids can be demonstrated in dilatations of the tubules of the agranular endoplasmic reticulum. However they do not stain after fixation with osmium tetroxide. 8. In the oenocytes massive autophagy takes place just before imaginai ecdysis. It is followed by breakdown of the ATER and other parts of the cytoplasma. 9. There is no evidence that the oenocytes are directly responsible for the formation of epicuticular lipids.

Notes: Summary The rabbit SA-node was outlined electrophysiologically and its adrenergic and cholinergic innervation patterns were studied with the electron microscope. Differentiation between adrenergic and cholinergic terminals was achieved by fixation of the specimens in KMnO4 which produces dense-cored synaptic vesicles in adrenergic terminals, whereas synaptic vesicles in cholinergic terminals are empty. It was found that adrenergic and cholinergic nerve terminals often come in close apposition to each other, the distance between adjoining membranes being in the order of 250 Å. At times, faint membrane thickenings could be seen in these places. The available pharmacological, physiological and morphological evidence leaves little room for doubt that cholinergic terminal fibers can influence the adrenergic ones. From mainly morphological evidence it is also postulated that adrenergic terminals influence cholinergic terminals.

Notes: Summary The median eminence and the pars nervosa of Rana esculenta have a different structure. The median eminence has 2 different zones: the outer zone situated near the pars distalis and the inner zone under the ependyme. In the outer zone there are, according to the size and the shape of the granules, 5 types of nerve terminals. 1. Endings containing spherical fine dense granules of 800 to 1000 Å in diameter; 2. Endings with spherical granules from 1000 to 1200 Å in diameter; 3. Endings with granules of irregular shape which are bigger than the former (1200 to 1600 Å); 4. Endings with spherical dense granules of about 1200 to 1800 Å in diameter; 5. A few endings containing only clear vesicles. Type 3 and type 4 endings are probably neurosecretory. The inner zone contains numerous neurosecretory fibres. They are of two types: one with big granules (1600–2400 Å), the second with smaller granules (1300–2000 Å). Non-neurosecretory fibres have also been observed. The pars nervosa contains two principal types of neurosecretory fibres: one with dense granules of 1600 to 2400 Å in diameter, the other with lighter granules of about 1300 to 2000 Å. In the external zone lining the pars intermedia, aminergic fibres with fine granules have been observed.

Notes: Summary The innervation of the portal vein of the white rat was examined with light-, fluorescence-, and electronmicroscopic techniques. The results are as follows: 1. Paraldehyde treated vein preparations (Falck's fluorescence method) demonstrate a predominantly longitudinally orientated external nerve plexus, being situated on the outermost muscle cells. Near the liver the nerve net is characterized by broad meshes, near the intestinal tract by narrow ones. The circular subendothelial inner plexus originates in the outer plexus of the intestinal vascular bed. 2. Nerve bundles in the fibrous adventitia were demonstrated with Gomori's Acethylcholinesterase method. In other respects, the enzyme activity was only observed in the intercellular spaces of the muscle layer. The electronmicroscopic demonstration of Acetylcholinesterase (Karnovsky's method) further showed that the enzyme activity is restricted to the muscle cell membrane. 3. The electronmicroscopic examination verified the results obtained with fluorescence microscopic techniques. a) In the proximity of the liver, only isolated nerve bundles occur on the outer muscle layer. The individual nerves are entirely surrounded by Schwann cells. Only a few of the axons in the vicinity to the muscle cell have agranular vesicles. Evaginations of the vesicular axons occur infrequently. Their distance from the muscle cell amounts to 1000–2000 Å. b) In more than one thousand thin sections, no axons were found inside the thick muscular layer. c) Subendothelial axons (inner plexus) are either partially or totally evaginated from the Schwann cells. They are densely filled with empty vesicles (350–650 Å) and contain a few dense core vesicles of 800–1600 Å in diameter. Synaptic endings were not observed. d) A dense collection of vesicle-containing axons, that were partially in their entirety and partially only from the muscle cell proximal side evaginated from the Schwann cells, were observed in the single muscle layer at the junction of the superior mesenteric and the portal vein. From these bundles, smaller bundles and individual axons pass between the muscle cells and reach the endothelium. Typical synapses were not observed. No vesiclecontaining axon was nearer than 1000 Å to the muscle cell. 4. Those axons possessing vesicles and being evaginated are considered to be vegetative conducting pathways. The excitation of the effector structures by transmitter substances is discussed in connection with the post mortem autonomic vascular contractility.

Notes: Summary 1. There is a gradual proximo-distal increase in the thickness of the muscle coat of the human ductuli efferentes, duetus epididymidis and ductus deferens. Circularly arranged smooth muscle bundles predominate in the ductuli efferentes and ductus epididymidis of the caput section. Scanty strands of longitudinally and obliquely oriented smooth muscle bundles form an additional, incomplete outer muscle layer around the ductus epididymidis of the corpus. Small smooth muscle-like cells constitute the muscle elements of the upper sections of the excretory ducts (from the ductuli efferentes to the midcauda). At the transition of the corpus and cauda epididymidis ordinary large smooth muscle cells join the small contractile cells to form—in more distal sections of the cauda—a composed, thick subepithelial muscle coat. In most distal portions of the cauda, the two-layered muscle coat of the ductus epididymidis is transformed into a three-layered coat, a pattern of construction which is retained in the vas deferens. 2. Electron microscopically, three types of contractile cells are distinguished in the human ductuli efferentes and ductus epididymidis: a) contractile cells of medium transparency containing exclusively thin myofilaments (60 Å in diameter), b) dark contractile cells containing bundles of thin myofilaments (60 Å in diameter) and single coarse filaments (140 Å in diameter), c) light contractile cells with loosely dispersed, interweaving thin and thick myofilaments. Commutual diameter changes at regular intervals are seen in individual myofilaments, giving the impression of structural periodicity not unlike that of filaments of striated muscle. Ordinary smooth muscle cells of the cauda epididymidis and vas deferens are characterized by uniformly sized, closely packed but evenly distributed thin myofilaments with numerous dense patches. 3. Fluorescence microscopy performed on formaldehyde treated freeze dried tissues reveals that the contractile cells of the ductuli efferentes in man and monkey receive a low number of single adrenergic terminal fibres penetrating the depth of the muscle coat. The adrenergic innervation of the ductus epididymidis is restricted to small peritubular nerve fascicles running contiguous to the most superficially located bundles of smooth muscle-like cells. The adrenergic ground plexus is rather wide-meshed in the proximal cauda, becomes increasingly dense in more distal cauda sections and in initial, funicular portions of the vas deferens, and reaches maximum density in abdominal parts of the ductus. Perivascular and adventitial adrenergic plexuses are well developed at arteries of the caput and corpus epididymidis in man, monkey, rabbit, guinea-pig and rat. 4. Electron microscopically, noradrenergic nerves have been identified by the presence of small granular vesicles in preterminal varicose axon dilatations. Nerve fibre swellings filled with small empty spherical vesicles have been considered to belong to “cholinergic” neurons whereas occasional varicosities equipped with some large membrane bound granules and abundant mitochondria may represent local expansions of sensory axons. 5. Neuromuscular relationships in the upper sections of excretory ducts comprise adrenergic synapses by distance (more than 1000 Å), and a few intimate, ensheated close contacts, whereas the main type of contact of nerves to ordinary smooth muscle cells in the lower duct section is by means of close but not intimate approach (500–2000 Å). 6. Adrenergic synapses in the ductus epididymidis and ductus deferens of the monkey resemble—what concerns their morphology, relationship to effectors and distribution pattern—those of man. 7. In accordance with the total number of vascular and non-vascular adrenergic nerves, visualized by fluorescence microscopy, the amount of noradrenaline varied considerably in different sections of the human male internal genital organs: The lowest amounts were estimated in the testis (0.12±0.03 μg/g). Medium to high concentrations were detected in various sections of the caput and corpus epididymidis (ductuli efferentes 0.60±0.09 μg/g; ductuli efferentes and caput 0.72±0.13 μg/g; corpus epididymidis 1.04±0.25 μg/g; proximal cauda 0.95±0.17 μg/g; distal cauda 0.97±0.19 μg/g). The highest noradrenaline content was found in the human vas deferens (prox. vas deferens 1.11±0.21 μg/g; interm. vas deferens 1.20±0.42 μg/g; distal portion 1.43±0.39 μg/g). 8. For comparison, the noradrenaline content of the testis and epididymis of the rhesus monkey, the epididymis of the rabbit and the vas deferens of the rabbit, mouse, guinea-pig and rat has been determined. 9. Adrenaline of exogenous origin was detected in the vas deferens, cauda epididymidis and plexus pampiniformis of two cases who received this catecholamine as part of the local anaesthetic drug mixture. Due to methodological reasons, the presence of small amounts of adrenaline of endogenous source in adrenergic nerves of the human and monkey internal male genital organs cannot be excluded. 10. The differences in motility behaviour of the ductus epididymidis (spontaneous, rhythmic contractions) and ductus deferens (absence of any spontaneous movements under conditions at rest) in vivo and in vitro have been correlated with the occurrence of specialized contractile cells in the upper segment (ductuli efferentes, ductus epididymidis of the caput, corpus and initial cauda) and ordinary large smooth muscle cells in the lower segment (ductus epididymidis of the distal cauda and the vas deferens) and furthermore correlated with differences in the pattern of the adrenergic innervation; the concept is advanced that progressive cytological differentiation of smooth muscle cells and the development of a dense direct adrenergic innervation suppresses autocontractility and, that the reverse condition may favour spontaneous motility of smooth muscle elements.

Notes: Summary Routine electron microscopy and a zinc iodide-osmium tetroxide technique (ZIO), recently found to be specific for synaptic vesicles, were used to study the origin of synaptic vesicles during postnatal development in the lumbosacral enlargement of the albino rat. In immature nervous tissue, a large number of vesicles, indistinguishable from synaptic vesicles (S vesicles), were found in the Golgi apparatus and in different portions of the axon where they were often intermingled with elements of the smooth endoplasmic reticulum (SER). Ten to twenty percent of these S vesicles within the Golgi apparatus as well as the majority of these vesicles in all parts of the axon were positive to ZIO. Much of the SER in axons was also positive. The number of vesicles and elements of the SER showed some decrease in the non-terminal portion of axons on day 21 and even more of a decrease in adult neurons. These data suggest that synaptic vesicles are produced in the Golgi apparatus and SER in immature neurons. The decrease in S vesicles and SER in adult neurons suggests a drop in synaptic vesicle production after synaptogenesis has ended. In addition, the material that has been studied shows that ZIO staining is not limited to synaptic vesicles during development since oligodendroglia and endothelial cells are also stained during this period.

Notes: Summary The glandula infraorbitalis buccalis in rabbits represents the type of a tubuloacinar mucous secreting gland. The secretory product of gland cells is electron lucent. Extrusion generally is characterized by the formation of an opening at the cell surface and the discharge of the secretory material. Intercellular canaliculi are absent. Mitochondria and arrays of granular endoplasmic reticulum are more numerous than in other mucous glands of rabbits. Intercalated ducts are short and connect the tubules with intralobular ducts. Striated ducts are absent in the infraorbital gland. Nerve terminals occur in some instances between gland cells and between glandular and myoepithelial cells.

Notes: Summary 1. Three species of the genusGnathonemus (Gn. petersii, moorii andstanleyanus) as all species of the family Mormyridae possess two pairs of electric organs situated symmetrically on each side of the vertebral column between the caudal, dorsal and anal fins. Each organ is composed of a series of 70–170 electroplaques. 2. The stalks of the electroplaques contain no nerves. Unbranched motor nerve fibres innervate the end sections of the stalks by synaptic knobs inserting into cavities of the electroplasma. 3. The electroplasma membrane surrounds the stalk (also in the region where the stalk penetrates through holes in the body of the electroplaque) and the electroplaque. The membrane is deeply indented on the cranial side as well as on the caudal side of the electroplaque. Through this the surface is greatly enlarged. In the inside of the electroplaque are striated muscles. 4. The motor and sensory nerve fibres are clearly different in diameter. 5. The number of the nerve cells in the spinal ganglions is, in the electric organ, about a multiple larger than in the segments of the body muscular system situated in front of the electric organ. 6. The reconstruction of the peripheral sensory nerves of one segment (in view of electron microscopical analyses), reveals that these, with the exception of two free nerve endings in the integument and the free nerve endings in the dorsal myoseptum, mainly innervate four large tendons on the side of the electric organs. 7. The innervation of the electric organs leads to the hypothesis that the tendons, in connection with the function of the electric organs, possess sensory functions.

Notes: Summary A developmental study at the electron microscopic level was conducted of the fat body cells of Hyalophora cecropia (L.). During the last larval instar the fat body increases in volume and the cells exhibit a well developed rough endoplasmic reticulum and protein bodies of diverse sizes. In the pupal fat body, the protein bodies appear to be enclosed by a double membrane and contain glycogen granules, ribosomes and mitochondrion-like structures. In addition, there are large lipid globules, cytolysomes and rough endoplasmic reticulum. The ultrastructure of the protein bodies suggests the development of large bodies by fusion of smaller protein bodies. Changes in fat body cell ultrastructure were followed during adult development and cytological evidence was obtained for the depletion of protein, glycogen and lipid in the female during this period. The female adult fat body cell contains free ribosomes, protein bodies, many mitochondria, a few lipid globules and glycogen granules. The male moth fat body cells have many mitochondria, a few glycogen granules, essentially no protein bodies, but an abundance of large lipid globules. Studies on the influence of egg maturation on the morphology of the fat body of Hyalophora gloveri (L.) revealed that ovariectomy of pupae yielded adults having more fat body than normal females, and that the fat body cells of the ovariectomized animals contained more glycogen, lipid and protein. Male pupae receiving ovarian implants developed into adults containing eggs and possessed more fat body than normal females but less than normal males. Very few glycogen granules were found in the fat body cells of normal males or males with implanted ovaries.

Notes: Summary 1. 1. The cuticular apparatus of the basiconic sensilla in Necrophorus is built up by four cells: the enveloping cell No. 1 secretes the dendritic sheath, the trichogen cell produces the part of the hair with pores, the tormogen cell the hairbase without pores, and the enveloping cell No. 4 produces the base and the ringwall of the sensillum. 2. 2. The development of the hair starts with the growth of the sheathed dendrite. This growth is homologue to the increase in length of the dendrite during moulting in hemimetabole insects. 3. 3. The trichogen cell surrounds the dendrite only up to 2 μ distal of the epithelial surface and then grows beside the dendrite to the full length of the future hair. After having secreted the cuticulin layer the free ends of sheath and dendrite degenerate. 4. 4. The development of the pore tubules system begins with the secretion of the cuticulin layer. When the hairwall has reached 30% of its thickness the pore tubules are fully developed. The outer liquor space is formed by withdrawal of the trichogen cell from the newly built hair. The dendrite grows into the hair inside the retreating trichogen cell. Few hours after emerging from the pupa the dendrite begins to split into about 50 branches: between the microtubules of the dendrite many vesicles arise which then fuse into continuous interspaces.

Notes: Summary The fine structure of the pancreatic nerves of the domestic fowl has been studied. Naked axon beadings were found in membranous contact with endocrine as well as exocrine cells. From an anatomical point of view it seems reasonable to suggest that the endocrine glands might be subjected to some influence of the autonomic nervous system.

Notes: Summary A fine structural study of sternothyroid muscle spindles of young adult rat demonstrates the rare occurrence of unique nerve endings. These endings are situated in the juxta-equatorial region of nuclear-bag fibers, adjacent to the annulo-spiral sensory endings. They consist of a bundle of terminal axons less than 0.3 μ in diameter and appear to be disposed nearly longitudinally to the axis of the intrafusal fibers. Whereas, the annulo-spiral endings consist of a single axon, coiling around the intrafusal fibers. Transverse sections of these muscle spindles reveal these unique endings scattered around the muscle fibers, fitting into depressions on their surface. The innermost axons directly face the muscle surface and are separated by a narrow gap less than 200 Å in width. No Schwann cell process appears to be associated with these endings. From their unique multi-axonal composition, these endings are termed “bundled endings”. Investigations of developing muscle spindles show the occasional presence of a similar multi-axonal composition of sensory endings in perinatal rats. It is suggested that “bundled endings” are sensory in nature, carrying an immature feature over to adult life.

Notes: Summary An electron-microscopic study has been made of the glial cells in the developing lateral funiculus of the cervical spinal cord in fetal rhesus monkeys. The various macroglial cell types, their precursor cells, and microglia are discussed in detail. An astrocytic lineage is proposed in which glioblasts present in the lateral funiculus give rise to astroblasts that then develop into mature astrocytes. Oligoblasts apparently migrate into the lateral funiculus as such and develop into active oligocytes. The active oligocytes become most predominant during the initial stages of myelinogenesis and are in direct continuity with developing myelin. The active oligocytes develop into mature oligocytes after myelination is completed. Microglia cells are present throughout development as three forms; resting microglia, globose microglia, and active microglia. The globose and active microglia predominates at specific times early in development when degeneration of apparent neuronal processes is taking place. The microglia cells are characterized by dense nuclear chromatin clumps, lipid inclusion bodies, dense vesicles, and, often, intracellular debris.

Notes: Summary The formation and development of synaptic contacts between dissociated chick spinal cord neurons has been investigated. By the 6th day in vitro “immature” profiles with few vesicles were observed. By 14–18 days “mature” types with numerous vesicles were found, indistinguishable from those of newly hatched chick spinal cord. After this period degeneration occurred, and was especially marked in the post-synaptic element. Such degeneration could be postponed by the addition of small numbers of somatic muscle cells. The Kanaseki and Kadota (1969) technique was applied to the study of coated vesicles at various stages of synaptic development.

Notes: Summary Colchicine (0.1 M) or vinblastine (0.01 M) was locally applied on the sciatic nerves of newborn rats. Both colchicine and vinblastine caused reversible disappearance of axonal neurotubules and appearance of increased amounts of neurofilaments at the site of application. Subsequent morphogenesis of myoneural junctions in the tibialis anterior muscle was studied after histochemical demonstration of acetylcholinesterase (AChE; E.C. 3.1.1.7) and non-specific cholinesterase (Ns. ChE; E.C. 3.1.1.8) activity in the myoneural area. Development of the postsynaptic muscle plasma membrane of the myoneural junction was arrested in the ipsilateral, but not in the contralateral control side, for a period of about three weeks following treatment with the test substances. After this delay the myoneural morphogenesis continued normally and neurotubules were seen in the axoplasm. Since disruption of neurotubules is likely to cause blockage of the intratubular axoplasmic transport system, it seems possible that the neurotrophic influence responsible for the development of the postsynaptic muscle membrane is mediated through a secretory product transported along axons intratubularly to the nerve endings.

Notes: Summary The development of the pars tuberalis was studied in the rat fetus from 13 days of gestation to 6 weeks after birth. After the closure of Rathke's pouch, the pars tuberalis anlage is clearly distinguishable from the anlagen of the partes intermedia and distalis. It comprises the entire basal portion of the adenohypophysial anlage; the limit between the anlagen of the pars tuberalis and the pars distalis is defined by Atwell's recess, i.e. the pathway taken by the hypophysial vessels coming from the vascular plexus of the median eminence. At 14 days the pars tuberalis cells are characterized by the presence of glycogen which persists in the adult. Their secretory differentiation (elaboration of granules with a diameter of 100–120 nm) is obvious at 15 days of gestation. It therefore, clearly precedes that of the other hypophysial cell types. Its functional differentiation takes place well before its adhesion to the primary vascular plexus of the portal system. Cystic formations appear just before birth in the pars tuberalis, much later than those of the pars distalis. These observations on the development of the pars tuberalis, together with previous observations on the adult PT in various species, showing that the specific glandular cells of the pars tuberalis are cytologically different from all known adenohypophysial cell types, seem to indicate a specific endocrine function of this lobe.

Notes: Summary The anatomical development of the tympanal organ, the tracheal system and the tympana of Teleogryllus commodus is described. The tympanal organ is undifferentiated until instar 3 when the first scolopales appear. The organ develops in a proximo-distal direction but the more distal groups begin to form before the proximal groups are complete. All groups of scolopidia are represented in instar 7 but numbers of scolopidia continue to increase until the adult stage. Scolopales and scolopale cells are first formed at less than adult size and increase in size during subsequent instars. The subgenual organ is present in instar 1 and is complete in instar 4. The posterior and anterior tympanal trachease develop respectively from the tracheal trunk and tracheal branch, which are present from instar 2. These remain as simple, unconnected tubes until instar 7 and begin to show the adult form during instars 8 and 9, finally enlarging in conjunction with the tympana at the last two moults. The tympana first appear as hairless areas of cuticle in the larval stages: the posterior tympanum in instar 8 and anterior tympanum in instar 10, but the translucent appearance of the adult tympana is not present before the final moult.

Notes: Summary The innervation of the Purkinje fibres in the atrium of the heart of the adult fowl was investigated by light microscopy, using the Champy-Maillet OsO4-ZnI2 technique and the cholinesterase reaction and by electron microscopy. After impregnation of the tissue with OsO4-ZnI2, the dark-stained nerve fibres were clearly visible on the unstained Purkinje fibres. In the upper part of the posterior wall of the right atrium, the diffuse portion of the conducting system is especially richly innervated by varicose and smooth nerve fibres. Some of these fibres are cholinesterase-positive. The terminal axons run in the space between the Purkinje fibres and the fibrocytic envelope. They are either naked or accompanied by Schwann cell processes. In addition to varicosities containing granular vesicles, there are varicosities containing agranular vesicles with oval profiles. In the “en passant” synapses, the width of the synaptic cleft between a varicosity and the Purkinje cell is about 600 Å. The innervation of the Purkinje fibres appears more like the innervation of smooth muscle than that of striated skeletal muscle. The possible role of Purkinje fibres as mechanical receptors is discussed.

Notes: Summary The pathways of axonal transport of secretions from neurosecretory cells (NSC) in the medial group (viz. A-, A1, B-, and C-type NSC) and the lateral group (L-type NSC) are described. Individual axons can be recognized in the electron microscope by the kind of neurosecretory particles they contain. In general, the secretions from the medial NSC are carried to the contralateral nervi corporis cardiaci (NCC), those from the lateral NSC to the ipsilateral NCC. Some axons from the A-type NSC, in addition, may run to ipsilateral NCC. All A-type axons have collaterals which run to the ipsilateral NCC. The medial and lateral bundles of “mixed” axons run through one paired NCC but remain separated spatially. Release of secretion from the C-type NSC can take place before the corpus cardiacum is reached. A- and A1-type NSC have additional collaterals that branch from the proximal part of the axons and penetrate deeply into the neuropile of the protocerebral lobes. Local swellings appear to be closely associated with fibers from non-neurosecretory neurons. The sites of contact are characterized by the accumulation of microvesicles (400 Å) near an electron-dense cleft of 150–200 Å width, and resemble regular chemical synapses. The microvesicles prove to be present within the neurosecretory fiber in most of the specimens studied, and within the non-neurosecretory fiber in only a few cases. It seems most likely that the collaterals in some phases convey afferent signals to the NSC which inhibit the release of an efferent neurochemical “messenger” of unknown nature into the neuropile.

Notes: Summary Vegetative growth of Nosema sp. occurs within the gut submucosal cells of Callinectes sapidus. Vegetative cell morphology is dominated by profiles of endoplasmic reticulum, numerous free ribosomes and aggregates of vesicles enclosed by a membranous sac. The dikaryotic vegetative cell is the earliest stage found in the target area for sporogenesis, the sarcoplasm of the striated muscle cell. The next obvious stage is the sporoblast mother cell; it undergoes karyokinesis without breakdown of the nuclear envelope. Intranuclear mitotic microtubules extend from the chromosomes to the intact nuclear envelope. After repeated nuclear divisions, the sporoblast mother cell undergoes delayed cytokinesis and a series of sporoblast progeny develops. The polar filament is the first visually apparent system to develop during sporogenesis. It appears to be of dual origin: (1) the central core component is condensed in Golgi-like saccules, and (2) the envelopes around the core originate from the endoplasmic reticulum. The polaroplast, which forms after early polar filament development, appears to originate as an elaboration of the endoplasmic reticulum.

Notes: Summary Study of the pineal complex of several Columbiformes demonstrates: a) the presence, in embryos, of numerous evaginations of the diencephalic roof; b) the presence, in adults, of a parenchymal accessory structure, probably a remnant of one or more secondary evaginations; c) the variability, among specimens, of the diencephalic course of the pineal tract fibers; d) the predominant asymmetry of the basal attachment of the pineal stalk to the left of the mid-line.

Notes: Summary The time and place of occurrence of the neurosecretory substance in the hypothalamo-neurohypophysial system of the guinea pig during embryogenesis have been investigated. Use is made of the luminiscence of neurosecretion stained with paraldehydefuchsin when observed in a dark field. It is established that the neurosecretory material occurs first in some cells of the supraoptic nucleus about the 39th–40th day of intrauterine development. In the paraventricular nucleus it is observed about the 44th–45th day. At that time it is seen also in Eminentia mediana and in the neurohypophysis. In the latter, however, it is in a smaller amount than in the areas situated above it. These results are discussed in connection with the transport theory of Bargmann and Scharrer.

Notes: Summary The development of intercellular junctions in the neural retina of the chick embryo between the seventh and nineteenth day of incubation has been studied. The main findings are: 1. The zonulae adhaerentes, which make up the outer limiting membrane of the adult retina, are present throughout the period of development covered by this study. 2. Small intercellular junctions of the macula adhaerens diminuta type appear in large numbers in the plexiform layers of the retina of 10 days incubation and are retained throughout development. 3. Synapse-like structures appear in the inner plexiform layer of the retina after 14 days of incubation. The possible relevance of these intercellular junctions to retinal morphogenesis is discussed.

Notes: Summary All cells in the optic vesicle of Xenopus embryos from stages 27 to 31 have the same ultrastructure. They are elongated and appear to extend from the internal to the external surfaces of the optic vesicle. They are bound together by terminal bars at the internal (lumen) margin, have microvilli and a cilium on the internal margin, and are covered with a basement membrane on the external margin. Their cytoplasm contains abundant free ribosomes, polysomes, mitochondria, yolk and lipid inclusions, and sparse endoplasmic reticulum. Although other studies have shown that retinal ganglion cells originate at stages 29–30 and have their central connections determined before stage 31, these events could not be correlated with any ultrastructural changes. The first sign of differentiation in retinal cells was an increase in endoplasmic reticulum and Golgi apparatus at stage 32. Microtubules and microfilaments appeared at stage 33 in association with the first axonal outgrowth from retinal ganglion cells. Cytodifferentiation proceeded gradually until large areas of Nissl substance had developed by stage 35. At larval stage 48 the ganglion cells resembled those in the adult.

Notes: Summary The nerve supply to the iridic melanophores of the rat was studied with the electron microscope. The adrenergic and cholinergic terminals were identified with the aid of 5-hydroxydopamine, which produces dense-cored 400–800 Å synaptic vesicles in adrenergic axon varicosities, whereas the synaptic vesicles of cholinergic axons remain empty. It was found that both adrenergic and cholinergic terminal axons come in close apposition (200–250 Å) with the melanophores. The appositions have the same appearance as synapses in peripheral tissues. It seems likely that the murine iridic melanophores have a double innervation, although its functional significance is obscure.