Immunocytochemical localization of somatostatin in the brain of the lizard, Ctenosauria pectinata

The brain of the lizard, Ctenosauria pectinata, was studied light microscopically using an immunocytochemical staining method that is specific for neurohypophysial hormones and somatostatin. It was shown that the telencephalon and particularly the diencephalon contain somatostatin-producing perikarya, while somatostatinergic fibers occur in the entire brain. Similar to the situation in other vertebrates, somatostatin neurons in Ctenosauria pectinata form a population distinct from the neurohypophysial hormone-producing neurons. The small-sized somatostatin neurons were found in the cortex and the diencephalon: (1) ventral from, and partially overlapping with, the classical neurosecretory paraventricular nucleus; and (2) in the region of the infundibular (tuberal) nucleus. Somatostatin fibers were found among the classical neurosecretory fibers of the supraoptico-paraventricular system (tract, median eminence, neural lobe), near to and within the epiphysis, in the septum, in the vicinity of the tectum opticum and the cerebellum, and in the tegmentum.     

Regulation of pathways of glucose metabolism in kidney. The effect of experimental diabetes on the activity of the pentose phosphate pathway and the glucuronate-xylulose pathway.

Measurements have been made of the activities of enzymes of the pentose phosphate pathway, the glucuronate-xylulose pathway, hexokinase and phosphofructokinase in kidney of diabetic and normal rats. The activities of these enzymes keep pace with kidney growth, remaining constant per gram tissue but showing a marked increase on the basis of total activity per 100 g body wt. The formation of ¹⁴CO₂ from [1-¹⁴C]glucose and [6-¹⁴C]glucose by kidney slices from diabetic rats was decreased to approximately half the control value; evidence was obtained for an equivalent dilution of the glucose 6-phosphate pool. Correction of the ¹⁴CO₂ yields for the change in specific activity of glucose 6-phosphate yielded values consistent with the enzyme profile. Calculations from specific yields of ¹⁴CO₂ provided evidence for an increased flux of glucose via the pentose phosphate pathway in the kidney in diabetes. The results are discussed in relation to kidney hypertrophy in diabetes and the requirement for ribose 5-phosphate and NADPH for biosynthetic reactions and in relation to the thickening of the basement membrane in diabetes. These results are in accord with the concept of glucose overutilization by non-insulin-requiring tissues.     

Clustering, haplotype diversity and locations of MIC-3: a unique root-specific defense-related gene family in Upland cotton (Gossypium hirsutum L.)

MIC-3 is a recently identified gene family shown to exhibit increased root-specific expression following nematode infection of cotton plants that are resistant to root-knot nematode. Here, we cloned and sequenced MIC-3 genes from selected diploid and tetraploid cotton species to reveal sequence differences at the molecular level and identify chromosomal locations of MIC-3 genes in Gossypium species. Detailed sequence analysis and phylogenetic clustering of MIC-3 genes indicated the presence of multiple MIC-3 gene members in Gossypium species. Haplotypes of a MIC-3 gene family member were discovered by comparative analysis among consensus sequences across genotypes within an individual clade in the phylogram to overcome the problem of duplicated loci in the tetraploid cotton. Deficiency tests of the SNPs delimited six A_t-genome members of the MIC-3 family clustered to chromosome arm 4sh, and one D_t-genome member to chromosome 19. Clustering was confirmed by long-PCR amplification of the intergenic regions using A_t-genome-specific MIC-3 primer pairs. The clustered distribution may have been favored by selection for responsiveness to evolving disease and/or pest pressures, because large variants of the MIC-3 gene family may have been recovered from small physical areas by recombination. This could give a buffer against selection pressure from a broad range of pest and pathogens in the future. To our knowledge, these are the first results on the evolution of clustering and genome-specific haplotype members of a unique cotton gene family associated with resistant response against a major pathogen.     

Ultrastructure of nervous tissue developing in the anterior chamber of the eye. 1. Perikaryons and dendrites

Embryonic tissues of septum and hippocampus were transplanted into the anterior eye chamber (AEC) of adult rats. The morphology of initial embryonic tissues and of transplants within 3 to 4 months of cultivation in AEC was studied. The transplanted tissue consists of neuroblasts and immature neurones: no synaptic contacts are observed. Within 3 to 4 months, highly differentiated neurones establishing synaptic contacts can be seen in the transplants. At the same time the fine structure of perikaryons and dendrites undergoes some changes: increased vacuolization, transformation of ergastoplasm into lamellar bodies. These can be due to an elevated functional activity of some neurones. Another group of morphological abnormalities (increased number of dendrite processes and microphyllopodia, somatic spines, dendrite cones of growth, tight junctions between perikaryons) suggests incomplete tissue maturation. These might be due to the absence of normal afferent and trophic influences in AEC.     

Mineralization of soil nitrogen in three forest communities from the New England region of New South Wales

Nitrogen (N) mineralization rates and the temperature response patterns of mineral N production in surface (0–7.6 cm) soils were compared in laboratory incubation studies based on disturbed, composite samples. Seasonal variation in the field levels of mineral N, and mineralization potential of intact (7.6 × 5.6 cm diameter) soil cores, were also investigated. Ammonification proceeded rapidly in each soil. Nitrification did not occur in grassy forest (GF) soil but was active in both layered forest (LF) and mossy forest (MF) soils, especially the former. Total mineral N production was greatest in MF and least in LF. Ammonification in disturbed samples was maximal at 50°C in all three soils with a secondary peak at 10°C in LF soil. Nitrification in LF and MF soils was most rapid at 25°C. Several species of ammonifying bacteria with different temperature optima were isolated, indicating that the process of ammonification is a composite of the activities of a variety of decomposer microbes. Mean field levels of mineral N and NH₄–N throughout the year were greatest in MF and least in LF. Seasonal fluctuations in NH₄–N were evident, concentrations being universally low in mid-winter (about 1.5 μgg^-1), increasing to a maximum in late summer (about 5 μg g^-1 in LF: 16–18 μg g^-1 in GF and MF). Field levels of NO₃–N were more constant and never more than 5 μg g^-1 in any community. Both total mineralization and ammonification in intact cores were greatest in MF and least in LF while nitrification was greatest in LF and almost negligible in GF, thus confirming the results obtained with disturbed samples. The potential for mineralization was large in mid-winter when the amount of mineral N was very low, and small in late summer when field levels were higher: this is interpreted as indicating that seasonal climatic factors regulate the availability of substrates for decomposers. Spatial variability in field levels of mineral N and mineral N production in the laboratory was evidenced by significant ‘sampling site’ effects in each community: however, at the sampling intensity used, the presence of bark mounds around Eucalyptus saligna trees could not be shown to affect these attributes. The inability of GF soil to nitrify when incubated in the laboratory could not be ascribed to a high C/N ratio, low pH, lack of substrate ammonium, or a low population of autotrophic nitrifying bacteria. No attempt was made to investigate the presence of allelopathic nitrification inhibitors. No evidence was obtained to support the view that nitrification is atypical of climax communities in situ. The most productive forest (LF) had the greatest capacity to nitrify and the least productive community (GF) the smallest capacity to do so.     

An Immunohistochemical Study of Localization of the Calcium-Binding Protein Recoverin in Retina of the Newt Pleurodeles waltl

The presence and localization of the calcium-binding protein recoverin, initially found in photoreceptors of the bovine eye, were immunochemically studied in retina of the new Pleurodeles waltl. Polyclonal monospecific antibodies against recoverin were raised and the methods of immunoblotting and indirect immunofluorescence were used. A protein with an apparent molecular mass of 26 kDa was found in the retina extract, which was specifically stained by the antibodies against recoverin. Localization of recoverin was studied on the retina sections: an intense reaction was found in the inner segments and a weak reaction was found in the basal part of the outer segments of photoreceptors and in Landolt's clubs of displaced bipolars. The results we obtained suggest for the first time the presence of recoverin in the retina of a representative of the Urodeles and indicate to interspecific conservativeness of this protein and differences of its localization in the retina photoreceptors in different species. The data obtained open a possibility of using recoverin as a marker protein of photoreceptors and displaced bipolars in studies of retina regeneration in newts.     

Cryofractographic study of intercellular junctions in the populations of agar-cultivated Bordetella pertussis

The characteristic feature of replicas obtained from the freeze-fractures of B. pertussis unfixed cultures developing on casein charcoal agar for 1-7 days is the associative growth of highly polymorphic cells, ensured by the ramified system of intercellular connections (IC) formed by the derivatives of the outer layers of the cell wall. This proves that the associative location of bacterial cells, linked by numerous IC, in the preparation is not the artefact appearing in the process of their chemical fixation. In replicas obtained from the freeze-fractures of B. pertussis cultures, previously fixed with glutaraldehyde, osmic acid and uranyl acetate, oval cells with the cytoplasm having a relatively homogeneous structure and with the smoothed-out three-layer cell wall prevail. As a rule, IC are limited to the sites of direct contacts between individual cells.