Insect mucosubstances. 3. Some mucosubstances of the nervous systems of the wax-moth (Galleria mellonella) and the stick insect (Carausius morosus)

Synopsis A mass of connective tissue, continuous with the neural lamella, develops on the dorsal side of the abdominal region of the nerve cord of Lepidoptera during the pupal stage. The mucosubstances of this tissue in the wax-moth,Galleria mellonella, have been characterized histochemically using various techniques involving Alcian Blue binding, periodic acid-Schiff and high iron diamine reactions, and enzyme digestions. The results indicate that this fibrous tissue contains chondroitin and dermatan sulphates and neutral glycoproteins.Thoracic ganglia of adult stick insects,Carausius morosus, were subjected to the same histochemical tests. The neural lamella possesses chondroitin, dermatan and keratan sulphates, while the glial lacunar system contains only hyaluronic acid.     

The glycosaminoglycans of the thoracic ganglia of the nymphal stages of the cockroach, Periplaneta americana, and the locust, Locusta migratoria: A histochemical study

The glycosaminoglycans of the connective tissue matrices of the developing meso- and metathoracic ganglia of locusts and cockroach nymphs have been characterized. The neural lamella contains only chondroitin sulphate in the early nymphs, but gradually keratan sulphate accumulates in the later nymphs. The glial lacunar system cannot be detected histochemically in first instar locust nymphs, but it can be seen in the youngest cockroach nymphs; it is clearly visible in the older nymphs of both species. It contains only hyaluronate.A stereological analysis of the developing meso- and meta-thoracic ganglia of the cockroach shows that the relative volumes occupied by the neurones, neuropile, glial cells and glial lacunar system change during post-embryonic development.The physiological functions of the glycosaminoglycans in the neural lamella and glial system are discussed.     

Enzyme effects on the connective tissues of an insect central nervous system

The effect of various enzymes on the two connective tissue matrices of the cockroach central nervous system were investigated. Removal of the neural lamella, using collagenase, allows some of the cells which form the perineurium to pull out of this cell layer but the perineurial bracelet cells maintain an intact blood-brain barrier. Incubation of the nerve cord with hyaluronidase has little or no effect on the neural lamella and allows the selective removal of the matrix from the glial lacunar system. Partial removal of this matrix appears to have little effect on the ability of the axons to conduct action potentials at high frequencies. In addition to this difference in susceptibility of the neural lamella and lacunar matrices to different enzymes, there appears to be a difference between the lacunar matrix of the connectives and of the ganglia, the latter being more resistant to enzyme attack. There is no such difference in the neural lamella covering the ganglia and connectives.     

The organization of the central nervous connectives in Anodonta cygnea (Linnaeus) (Mollusca : Eulamellibranchia)

The cerebro-visceral connective of Anodonta cygnea comprises a non-cellular neural lamella, a few randomly distributed glial elements, and axons, together with an extensive extracellular system. The axons are relatively small, the majority being less than 2.0 micro in diameter. The neural lamella is underlaid by a thin layer of peripheral glial processes. The membranes of the proximate glial processes do not reveal any junctional complexes at this level. In addition to glycogen and fat deposits, the glial cytoplasm contains abundant 'glial granules' which in histochemical tests react like mucoproteins. The extracellular space appears to be freely accessible, for apart from the neural lamella no other visible structures are interposed between it and the fluid surrounding the connective. These structural findings are discussed in relation to the available evidence concerning the ability of axons to function in preparations bathed in low-sodium media.     

Role of the perineurium and glial lacunar system in nutrition and storage of nutrients in the central nervous system of Spodoptera litura Fabr. (Lepidoptera: Noctulidae) at the time of reorganisation of the neural lamella during metamorphosis

Brains and nerve cords of Spodoptera litura (Fabr.) of various stages during metamorphosis were tested for acid mycopolisaccharides (AMPS), PAS-positive substances, glycogen, proteins and lipids. During reorganisation of the neural lamella in pupal period glycogen and PAS-positive substances are stored in perineurium and AMPS in the glial lacunar system (GIS) and below the perineurium as a thin layer. Lipids and AMPS diffuse in the GIS. The perineurium and GIS serve for the passage as well as storage of nutrients. In neuropile, only proteins are present. Significances of these findings is discussed.     

The ultrastructure of the non-neurosecretory components in the brain of the midge,Chironomus riparius Mg. (Diptera: Nematocera)

The ultrastruct of the neural sheath, glial cells and neurons in the brain of the neoimaginal male Chironomus riparius is described. The neural sheath comprises a neural lamella and underlying perineurium. The neural lamella consists of an amorphous matrix in which fine fibrils occur. The perineurium is composed of two cell types forming a continuous layer around the brain. The subjacent cortical layer, composed of the cell bodies of neurons and glial cells, varies considerably in thickness and surrounds the centrally located neuropiles. Three types of glial cells are distinguished on the basis of their positions and appearances. Five types of neurons are described which differ in size and relative frequency of organelles. Four types of axons, including those of neurosecretory cells, are distinguished by their size and content.     

玉米螟幼虫侧单眼的神经鞘和胶质细胞

用透射电镜观察了欧洲玉米螟Ostrinia nubilalis(Hbner)5龄幼虫侧单眼神经的神经围膜、周神经细胞和其他神经胶质.神经围膜与若干周神经细胞包围若42根轴突.周神经细胞的原生质膜在它们的侧面和内面高度卷曲,并与相邻细胞交错对插,这是细胞与细胞间的特殊连接方式;它们的外面以桥粒和半桥粒固定在神经围膜内面.周神经细胞由神经胶质细胞演化而来,所形成的膜称神经束膜,它与神经围膜组成围在侧单眼神经外面的神经鞘.侧单眼神经内的神经胶质细胞大而平整,具有许多突起物(相当于脊椎动物的少突神经胶质细胞),每一个突起物包被一个感光轴突.神经胶质细胞包被轴突的形式有三种不同的类型:一种是相邻轴突间插入15层神经胶质细胞突起物所形成的普通轴系膜形式,另两种是神经胶质细胞突起物在一个轴突的周围,由一些褶所形成的不同形式.最后,对这些神经胶质细胞以不同形式包被轴突的功能意义进行了讨论.     

Fine structure of the neural sheath,glia and neurons in the brain of the housefly,Musca domestica

Summary The fine structure of the neural sheath, glial cells and nerve cells in the brain of adult male houseflies is described. The neural sheath is composed of neural lamella and perineurium. The neural lamella consists of an external lamina and collagen-like fibrils which are embedded in an amorphous matrix. The perineurial cells form a continuous layer around the brain. On their inner surface, perineurial cells form junctional complexes with glial cell processes. A cortical cellular layer composed of neurons and glial cells surrounds the centrally located neuropil. Three types of glial cells are identified. Glial cells differ in size and in relative development and distribution of organelles. Thin processes of glioplasm completely surround the cell bodies of the neurons. Five types of neurons are described. Most of the neurons are monopolar, a few are bipolar.Supported by a grant from the National Science Foundation     

The envelopes of the nervous system of pseudoscorpions and scorpions

The present paper demonstrates that in pseudoscorpions and scorpions a compact collagen sheath consisting of fibrils with a 720 A period embedded in a neutral mucopolysaccharide surrounds the nervous system. The same material makes up the trabeculae running into the nervous elements of the ganglia and nerves. Both structures are separated from the neurons and their fibres by entire glial cells or by their processes. The neural lamella is surrounded by a thin and continuous wall of cells forming a storage-epithelium. Amoebocytes are able to circulate between the collagen fibrils of the neural lamella.     

Uptake of peroxidose by the cockroach central nervous system

The central nervous system of the cockroach has been incubated with solutions of an exogenous tracer substance, horseradish peroxidase, and the sites of its penetration and uptake have been studied by electron microscopy. When the nervous tissue is intact, or intact but stretched, the peroxidase is taken up throughout the neural lamella and also penetrates short distances into the extracellular space between adjacent perineurial cells. When the ganglia have been desheathed, reaction product for peroxidase is found in the neural lamella, perineurial cells, and within the cytoplasmic substance of the glial cells adjacent to the desheathed area. This uptake of peroxidase by the injured glial cells in desheathed preparations may reflect an alteration in the normal diffusion pathway from the external medium to the axonal surfaces.     

The subgenus Persicargas (Ixodoidea: Argasidae: Argas): A. (P.) arboreus central nervous system anatomy and histology

The anatomy and histology of the adult Argas (Persicargas) arboreus central nervous system are described and compared with these properties in other ticks. The single, integrated, central nerve mass (CNM) is formed by a fused supra-esophageal part (protocerebrum, cheliceral ganglia, palpal ganglia, and stomodeal pons) and a subesophageal part (4 pairs of pedal ganglia and the complex opisthosomatic ganglion). Single peripheral nerves (pharyngeal and recurrent) and paired peripheral nerves (compound protocerebral, cheliceral, palpal, pedal and opisthosomatic) extend from the CNM to body organs and appendages. Optic nerves, described in other Argas species, are not found in A. (P.) arboreus. Histologically, the CNM is enclosed by a thin-walled periganglionic blood sinus and invested by a collagenous neural lamella followed by a perineurial layer composed of glial cells and containing fine reticular spaces, a cortical layer of association, motor and neurosecretory cell bodies and glial cells, and inner neuropile regions of fiber tracts forming 5 horizontal levels of connectives and commissures.     

Blood cells contribute to glial repair in an insect

Using antibodies specific for haemocytes, we have shown that these blood cells penetrate the abdominal nervous connectives of the cockroach following selective disruption of the glia using the DNA-intercalating drug, ethidium bromide, as a glial toxin. Within 4 days post-lesion, the labelled cells formed a mosaic beneath the neural lamella and penetrated deeply among the disrupted subperineurial glia. These observations confirm that exogenous cells are involved in glial repair and support a previous hypothesis that they play critical roles in both structural repair and the recruitment of endogenous reactive cells.     

The ultrastructural organization of peripheral nerves in two insect species (Periplaneta americana and Schistocerca gregaria)

The ultrastructural organization of various peripheral nerves, including the crural nerve, has been investigated in the locust and cockroach. In some cases the larger nerves are ensheathed by a fat body layer which is not always complete. However, like many nervous connectives, they do possess a continuous acellular neural lamella and a perineurial cell layer which surround the glial-axonal mass. Adjacent perineurial cells are associated with one another by septate desmosomes, gap junctions and tight junctions. These last may represent the morphological basis of the ‘blood-brain barrier’ observed electrophysiologically in these peripheral nerves in another report. Very small nerves of the cockroach, however, although lying embedded in a neural lamella, do not possess a specialized perineurial layer displaying junctional complexes, unless they contain one or more large axons. If they have only one or more small axons, these small nerves may either appear naked, or display a single glial cell process loosely enveloping them; in either case there is no structural basis for a ‘barrier’ system. Various comparisons have been made between locust crural nerve and the cockroach central nervous connectives in an attempt to correlate some aspects of their ultrastructural organization with relevant electrophysiological information.     

Ruthenium red staining of the neural lamella of the brain of Galleria mellonella

Summary The outer surface of the neural lamella, the connective tissue ensheathing the brain, shows the ability to bind ruthenium red in the wax moth larva. Ruthenium red-positive material is sensitive to neuraminidase, hyaluronidase and to some extent to phospholipase C, what suggests that the negative charge on the external surface of the neural lamella depends on the presence of the anionic groups of sialic and hyaluronic acids and phospholipids.     

Galleria mellonella nerve cords in vitro: stage-specific survival and differential responsiveness to beta-ecdysone

The survival of ventral nerve cord segments of Galleria mellonella in tissue culture medium, ascertained by histological and biochemical criteria, ability to shorten when transplanted, and responsiveness to β-ecdysone, is correlated with the stage of development of the donor, and with the presence of a connective tissue sheath, the neural lamella, about the segment. After only 18 hr in vitro connectives which have lost the neural lamella during metamorphosis no longer have the capacity to shorten when transplanted or when exposed to β-ecdysone in the culture medium. By contrast, after 7 days in vitro connectives with a neural lamella shorten appreciably when β-ecdysone is added, or when they are exposed to the humoral milieu of a host undergoing metamorphosis. β-Ecdysone stimulates the incorporation of uridine-5-³H in segments both with and without the neural lamella, but only in segments which have previously begun their metamorphosis. Since shortening in response to β-ecdysone occurs only in connectives which have already begun to shorten, β-ecdysone appears to accelerate physiological processes underway before it is added to the medium rather than initiate metamorphosis in cultured nerve cords.     

The effects of an anti-mitotic drug,bleomycin, on glial repair in an insect central nervous system

Summary The DNA-binding drug, bleomycin, has a profound effect on neural repair following selective glial disruption by ethidium bromide. The contribution of the granule-containing cells (which normally appear in the early stages of repair) is greatly reduced, the restoration of the blood-brain barrier is delayed and the ultrastructural organization of the reorganising perineurium is dramatically changed. The aberrant perineurial structure and function observed in the presence of bleomycin are postulated to result from the effects of the drug on haemocytes which, together with endogenous reactive cells, contribute to the normal process of glial repair.     

Intralamellar neurohemal complexes in the cerebral commissure of the leech Macrobdella decora (Say, 1824): An electron microscope study

Electron microscopy of the cerebral ganglionic commissure of the leech Macrobdella decora (Say, 1824) revealed numerous neurosecretory axons terminating in the neural lamella of both the inner and outer capsules, and in the neural lamella deep within the neuropile. The proximal protions of the terminals, with an investment of glial tissue, contain either numerous large homogeneously electron dense granules, or numerous large granules of varying electron density. The distal portions, often devoid of glia, display numerous infoldings, omega profiles, and electron dense focal sites, and contain numerous neurosecretory granules, small lucent vesicles, and, occasionally, acanthosomes. Statistical analysis of the size distribution and morphology of the neurosecretory granules showed that in many individual terminals the granules are not significantly different from those seen within four groups of neurosecretory cells found in the cerebral ganglion. These terminals, because of their diffuse nature, probably represent a neurohemal complex of a primitive nature. The term “intralamellar complexes” is proposed to describe the form and location of these neurosecretory terminals.     

Glial cells in peripheral nerves of the cockroach, Periplaneta americana

The ultrastructural organization and the junctional complexes of peripheral nerves have been investigated in the cockroach Periplaneta americana. Nerve 5 is surrounded by a layer of connective tissue, the neural lamella, beneath which is a layer of perineurial glial cells wrapping the axons. Adjacent perineurial cells are joined to one another by septate, gap and tight junctions. Septate and gap junctions were observed in freeze-fracture replicas of main trunk nerve 5. Septate junctions were found as rows of PF particles mainly in perineurial cell membranes. Gap junctions exhibited EF macular aggregates in perineurial and subperineurial glial cells. During incubations in vivo with extracellularly applied ionic lanthanum, the lanthanum did not penetrate beyond the perineurium. Where nerve 5 branches and contacts the muscle, lanthanum penetrated freely between the muscle fibres and the nerve branches. In small peripheral branches where the axons are surrounded by single a glial layer, lanthanum is unable to penetrate to the axolemma.     

Fine structure of a lepidopteran nervous system and its accessibility to peroxidase and lanthanum

Summary The avascular ventral nerve cord of the moth, Manduca sexta, possesses an extensive dorsal mass of connective tissue in which lie fibroblasts that produce a collagen-like protein. The lateral and ventral surfaces of the nerve cord are ensheathed by an acellular neural lamella. Beneath this lies a layer of microtubule-laden perineurial cells which are separated from one another at their peripheral borders by lacunae containing electron-opaque material to which the cells are attached by hemi-desmosomes. Beyond these spaces, narrow intercellular clefts occur between the interdigitating perineurial plasma membranes; these are then connected by both gap and tight junctions. The axons beneath are surrounded by glia which also contain many microtubules and which are linked to one another by desmosomes and tight junctions.When intact nerve cords are incubated in horseradish peroxidase, reaction product is subsequently found within the neural lamella as well as in the lacunae and clefts between perineurial cells, but not beyond this level. Desheathed preparations, however, contain peroxidase within the cytoplasm of the exposed glial cells. Lanthanum penetrates the neural lamella and the lacunae, clefts and gap junctions between adjacent perineurial cells, but no further. It therefore appears that the tight junctions in the perineurium may be the site of restriction to the entry of ions and molecules, the existence of which has been suggested previously by electrophysiological investigations.I am grateful to Miss Yvonne R. Carter for her invaluable technical assistance and to Dr. J.E. Treherne and Dr. D.B. Sattelle for helpful discussions.