Molecular cytogenetics and taxonomy of insects,with particular reference to the coleoptera

We assess and review the impact of the new cytogenetic techniques in insects for their roles in taxonomy and for a better knowledge of their chromosomal structure. Particular emphasis is given to molecular cytogenetics by fluorescent in situ hybridization, localization of AT- or GC-rich regions with fluorochromes, and restriction endonuclease banding in beetle chromosomes. The main features of the cytogenetics of Coleoptera are treated in detail, taking into account the range of variation in chromosome number and genome size, and our in-depth findings on the constitutive heterochromatin and satellite DNAs of tenebrionid beetles. Some other topics of interest for insect cytogenetics, such as the meiotic association of sex chromosomes, the nucleolar organizing regions (NORs), and the molecular constitution of telomeres, are also discussed from the taxonomic and structural viewpoints.     

A light and electron microscope study of the gills of larval lampreys (Geotria australis) with particular reference to the water-blood pathway.

The gills of ammocoetes of the Southern Hemisphere lamprey Geotria australis have been studied using light and electron microscopy. Emphasis has been placed on describing the structures and vessels involved in gaseous exchange, and on providing quantitative data for the water-blood barrier, including diffusion distance, diffusing capacity and the relative volumes of the component tissues. Although lamprey gills lie inside rather than outside the branchial skeleton as in gnathostomatous fishes. the morphology and ultrastructure of the gill filaments and secondary lamellae of G. australis larvae are very similar to those of teleost fishes. The extensive blood spaces within the secondary lamellae are enclosed by pillar cell bodies and pillar cell flanges which support two layers of epithelial cells. The outer surfaces of the epithelial cells are ridged and covered in a flocculent material which probably represents mucus. Differences were observed in the components of the water-blood barrier at the distal edges and at the surface of the secondary lamellae. At the distal edge, the lining of the marginal channel consisted of an endothelial cell rather than the pillar cell flanges which line the blood spaces of other regions. Based on light micrograph measurements, these differences result in a reduction in the arithmetic mean thickness of the water-blood barrier from 3.62 μm over the pillar cells to 2.22 μm over the marginal channel. Using values for the water-blood barrier obtained from light micrographs, the arithmetic and harmonic mean diffusing capacities were calculated as 1.1046 and 1.7589 ml O₂min/mm Hg/Kg.     

The vasculature of the rat embryo: an electron microscope study

The ultrastructure of portions of the arterial and venous systems of the 11.5 day old Wistar rat embryos has been studied by scanning and transmission electron microscopy. The vessels at this stage of development are in the form of capillaries, and the arterial and venous types can be distinguished by the morphology of the endothelial cells by SEM. The endothelial cells of the arterial vessels gave prominent nuclear bulges and numerous microvilli apart from their spindle shape, whilst those of the veins appear flattened, are polygonal in shape, and have few microvilli. Transmission electron microscopy shows that the endothelial cells of the arteries and veins are identical in structure. The ultrastructure of these cells resembles that of endothelial cells at later stages of development including the adult type in that mature forms of cytoplasmic organelles are obtained. In studies on the intercellular junctions and fenestrations with lanthanum nitrate, the impression is formed that the vessels at this stage are impermeable to small molecular size particles, compared with adult capillaries. This suggests that cytoplasmic vesicles must play a major role in the transport of macromolecules in the 11.5 day embryonic vessels.     

The distribution of bacteria and fungal spores in Blelham Tarn with particular reference to an experimental overturn

Summary During an experimental overturn at Blelham Tarn, surface water bacteria and fungal spore contents (determined by plating) ranged from 600–6,000 and from 0.3–30 per ml respectively. Following heavy rainfall however, bacterial numbers rose to over 10,000 and fungal spore numbers to over 100 per ml. These high concentrations normally extended down to the thermocline but not below it. The evidence was that particulate matter washed in by rain, and presumably carrying an attached bacterial and fungal flora, was largely lost to the lake through the outflow stream and did not sediment through to the hypolimnion.     

The nucleus of the basal optic root in the pigeon: an electron microscope study

The ultrastructure of the nucleus of the basal optic root in an avian species (Columba livia) was investigated. The ectomamillary nucleus (EMN) in which terminates the basal optic tract reveals three types of neurons: 1) small round neurons bearing a scanty cytoplasm in organelles, 2) medium-sized neurons, spindle-shaped with a dense population of organelles and 3) large multipolar neurons with well developed perikaryal elements. Some of these neurons have their inner plasma-membrane which fuse to make junctional zones alternating between attachment plates and gap junctions. The analysis of the neuropil displays four types of vesicle-containing profiles (VCP), Type I VCP, identified as optic terminals, are numerous (49%), contain round vesicles (500-550 A) and establish Gray type I contacts principally with dendrites. They also participate in serial and triadic arrangements. Type II VCP have lighter hyaloplasm and are less numerous (6,7%). Rounded vesicles (450-500 A) with a clear content synapse also with Gray type I active zones on dendrites. Some of these profiles have the peculiarity of both a chemical and electrical transmission known as mixed synapses. Type III VCP are larger and contain a mixed population of rounded and flattened vesicles which synapse according to Gray type II. Type IV VCP are characterized by a light hyaloplasm where the microtubules are a predominant organelle. Their active zones are also of Gray type II.     

The occurrence of glycogen-membrane complexes in fungi an electron microscope study

Summary Membrane-particle complexes were studied in the vegetative hyphae ofSclerotinia fructigena. The established techniques of differential staining for electron microscopy and pretreatment with ribonuclease and-amylase indicated that the particles were glycogen.The physiological significance of such structures in fungi is unknown at present, but their resemblance to similar glycogen-membrane complexes existing in animal cells is discussed.     

Sampling strategy related to fish distribution, with particular reference to the Norfolk Broads

Large-scale movements of fish within a complex freshwater system such as the Norfolk Broads can give rise to erroneous sampling data if the extent of these movements is not fully appreciated. The Norfolk Broads habitat is briefly described, and data are presented showing that mean estimates of standing crop in the broads vary from <1gm^-2 in the winter to 9.4 gm^-2 in the summer. The widespread nature of the phenomenon is demonstrated by winter and summer data from five broads, and a more detailed picture of the seasonal changes is shown by an intensive 2-year study on one of these broads. Winter aggregations of fish have been found in sites adjacent to rivers connected with the broads, with densities of up to 36.7 fish m^-2 and a biomass of 1787 gm^-2. Accurate overall estimates of fish standing crop can only be obtained when fish are maximally dispersed during the summer months.     

Studies on the Golgi apparatus by electron microscopy with particular reference to Aoyama's technique

1. Aoyama's silver impregnation method for the Golgi apparatus has been used on exocrine cells of the pancreas of the mouse and studied by electron microscopy in order to determine as precisely as possible where the silver is deposited. Similar cells have also been fixed in buffered osmium tetroxide solution and compared with cells treated by the silver technique. 2. Examination of the Aoyama preparations usually revealed a light deposition of silver in the cytoplasm (hyaloplasm or matrix) and a heavy deposition of silver around a series of closely apposed vacuoles. The heavy deposition of silver was regarded as revealing the chromophilic region of the Golgi apparatus while the vacuoles were identified as the chromophobic component. 3. Comparison of the silver preparations with those fixed in buffered osmium tetroxide solution showed that the silver was primarily deposited in the region of the Golgi membranes.     

Hymenolepis diminuta: an electron microscope study of ion absorption

Ten-day-old Hymenolepis diminuta were incubated in media containing either the divalent cations barium or lead, or the divalent anions sulfate or ferrocyanide. Mitochondria of the tegument, but of no other tissue with the time scale used, accumulated all these ions in conspicuous amounts.By visual inspection mitochondria appeared to accumulate ions in the following order of preference, lead > sulfate > barium > ferrocyanide. These results have been interpreted with reference to Lehninger's theory of ion accumulation in vertebrate mitochondria, in that it is suggested the results depend on the presence or absence of permeant cations. From the viewpoint of ion accumulation, the mitochondria of Hymenolepis diminuta appear to be similar to vertebrate mitochondria.Ion deposits also occurred attached to apical and basal plasma membranes of the tegument and to the membranes of the multitubular complex. Reactions at these sites appear to be due to the properties of their associated glycocalyx. The presence of deposits in and around multitubular complexes also supports the previous suggestion that this structure may play a role in ion or water transport in this tapeworm. The deposits which occurred in the interstitial material appear to be the result of a reaction between acidic groups present in the matrix material of this connective tissue and cations in the incubating and fixing solutions.     

The distribution of capillaries in the somatic musculature of two vertebrate types with particular reference to teleost fish

The distribution of capillaries in teleost and rat striated muscles was investigated using a number of different methods. A new method for directly viewing capillaries was developed. Teleost white muscle has a capillary: fibre (C:F) ratio of between 0.2 and 0.3; and 0.6 to 1.0 peripheral capillaries per muscle fibre. 26-49% of fibres had no peripheral capillaries. Values for the rat gastrocnemius were 1.2, 2.6 and 4.8% respectively which compares well with literature values. Flathead red muscle had a C:F ratio of between 1.9 and 2.5; and between 5.3 and 6.6 peripheral capillaries per muscle fibre depending on the method used. Values for rat soleus were 1.8 and 4.1 respectively. Teleost pink fibres had an intermediate number of capillaries. Rat striated muscle, particularly the gastrocnemius, was found to be heterogeneous with respect to the distribution of capillaries. Flathead red muscle was homogeneous whilst teleost white muscle was only slightly variable. Flathead red muscle fibres are well suppled with subsarcolemmal mitochondria. These show a clumped distribution corresponding to the position of capillaries. In contrast teleost white fibres are almost totally devoid of these and all other mitochondria. No differences were observed in the vascularisation of either muscle type along the length of the fish. The results are discussed in relation to the division of labour between fibre types during swimming.     

Ontogeny of the pig hypothalamic neurosecretory system with particular reference to the distribution of neurophysin

The immunoperoxidase cytochemical reaction was applied to the localization of neurophysin-containing elements in the fetal and adult pig hypothalamus. In the 60 day fetal pig, cells of the supraoptic nucleus (SON) were the only structues in the hypothalamus in which neurophysin was detected. However, by 87 days the cell bodies in both the SON and paraventricular nucleus (PVN) contained neurophysin-like material. The distribution of immunoreactive material in the 111 day fetal animal was similar to that found in the adult pig. In transverse section of the mature pig the SON exists in two discrete components; an antero-lateral group of cells connected by scattered cells to a smaller postero-medial group. Anteriorly, the PVN appears as a line of cells bordering the third ventricle but as we proceed posteriorly the dorsal aspect expands laterally to give a wedge-shaped group of cells. In mid-sagittal sections, the cells of the PVN are distributed over a wide area of the anterior hypothalamus in a triangular profile. The borders between the SON and PVN became more difficult to define in medial sections than in lateral sections. Continuous gradient polyacrylamide gel electrophoresis was carried out on the neural lobe extracts from fetal, newborn and adult pigs. Proteins with an electrophoretic mobility similar to that of porcine neurophysins-I, -II and -III were present in the newborn and 98 day fetal pig. It is concluded that material immunoreactive with anti-neurophysin serum is present in the hypothalamus of the 60 day fetal pig. Furthermore, at late fetal development and during the postnatal period it is tentatively suggested that the neurophysin present in the pituitaries of these animals is chemically identical with that of adult neurophysin.