Early development in the annual fish Cynolebias viarius

The detailed characteristics and timing of early development in Cynolebias viarius , an annual fish endemic to Uruguay, were obtained from in vivo observations of embryos cultured under standardized laboratory conditions. The chorion ultrastructure is reported for this species for the first time. Comparison with previous data from other annual fishes showed that C. viarius exhibited the longest early development including cleavage, blastula formation and epiboly.     

An ultrastructural analysis of the dispersed cell phase during development of the annual fish,Cynolebias

Cell ultrastructure was investigated during the dispersion phase of development in the annual fish Cynolebias. Three cellular populations encompass the yolk mass during dispersion, namely, 1) the yolk syncytial layer (YSL) or periblast, which lies directly over the surface of the yolk; 2) the deep blastomeres of the blastoderm, which engage in morphogenetic movements on the surface of the YSL and beneath the enveloping layer prior to forming the future embryo; and 3) the enveloping layer (EVL) of the blastoderm, which is a cohesive epithelium that forms the outermost cell layer of the blastoderm. Deep blastomeres contain numerous mitochondria and scattered glycogen rosettes that appear to function in the utilization of energy reserves. These cells also possess surface extensions such as filopodia and ruffles. Numerous microfilaments running parallel to the plasma membrane occur in cell extensions and in the cortical cytoplasm of neighboring blastomeres. In bleb-like extensions such as ruffles, microfilamentous stress fibers run parallel to the plane of the plasma membrane and prevent cellular organelles from entering the hyaline cap of the ruffle. Deep blastomeres also have basal projections that contain glycogen as well as pits in the basal membrane. Blastomeres move about using the YSL as a substrate. The YSL possesses specializations for nutrient uptake, storage, and transport such as numerous multivesicular bodies and large amounts of glycogen. Glycogen, in the rosette form, occurs in extraordinary amounts, virtually occluding the cytoplasm. Glycogen reserves are postulated to serve as an energy source during diapause. Glycogen is sometimes contained within villous projections that extend from the apical surface of the YSL. This configuration suggests the possibility of glycogen transport to the overlying deep blastomeres. Specializations of the EVL include apical tight junctions and basal lateral zonulae adherentes that interdigitate with those of adjacent EVL cells. The EVL serves as an impermeable membrane that protects the developing egg from the vicissitudes of its environment.     

The relation between pituitary gland and thyroid growth during the lifespan of the annual fish Cynolebias whitei and Nothobranchius korthausae: gonadotropic and thyrotropic cells

Summary In the annual cyprinodont Cynolebias whitei the cell types responsible for the increase of pituitary growth at the onset of maturation and for pituitary hyperplasia in old specimens were identified as gonadotropic cells and thyrotropic cells, respectively. The gonadotropic cells showed a high affinity to anti-carp -GTH serum, both at light- and electron-microscopical levels. The allometric relation of total gonadotropic cell volume to body length, determined for fish from six weeks up to six months of age, showed no inflections. Therefore pituitary growth in maturing fish may be partly a result of proliferation of gonadotropes, although gonadotropic cells do not contribute to pituitary hyperplasia in old fish. Thyrotropic cells showed a weak affinity to anti-carp -GTH serum at light-microscopical level. Under the electron microscope thyrotropic cells displayed signs of activation in maturing fish and signs of proliferation in old fish. The allometric relation of thyroid gland volume to body length paralleled that of pituitary volume to body length. Histologically the thyroid gland showed signs of inactivity in adult fish and of hyperplasia in old fish. The possibility, that gonadal maturation, pituitary thyrotropic activity, and growth of the thyroid in maturing fish are related through the inhibitory action of gonadal steroids on thyroid hormone release, is discussed. Pituitary hyperplasia in old fish is the result of proliferation of thyrotropic cells. Similar hyperplasia of pituiary and thyroid glands was observed in old Nothobranchius korthausae.     

Naturally occurring flavans unsubstituted in the heterocyclic ring

The naturally occurring free and glycosylated flavans are reviewed. The distribution, methods of isolation and structure elucidation of flavans, by comprehensive spectral analyses, are presented. Biological properties of flavans are appraised.     

Environmental influences on prolactin cell development in the cyprinodont fish,Cynolebias whitei

Summary A combined immunocytochemical and morphometric study on the development of the prolactin (PRL) cells of the annual cyprinodont Cynolebias whitei, transferred as newly hatched larvae to water with different salinities and/or Ca²⁺-concentrations, was carried out. The percentage of the pituitary volume occupied by PRL cells and the affinity of PRL cells for immunocytochemical staining were used as criteria for their activity. Exposure of the larvae for one day to salt water (260 mOsm/kg) led to a significant reduction in the pituitary volume occupied by PRL cells, indicating an osmoregulatory function of PRL shortly after hatching. In fish reared in diluted artificial seawater (70 and 260 mOsm/kg) or Na⁺-enriched fresh water the development of PRL cells was significantly retarded, but such an effect was not observed in fish placed in Ca²⁺-enriched fresh water. These experiments show that in C. whitei the development and activity of PRL cells are influenced by changes in environmental osmolarity and not by changes in ambient Ca²⁺-concentration.     

Naturally occurring variation in Arabidopsis: an underexploited resource for plant genetics

The definition of gene functions requires the phenotypic characterization of genetic variants. Currently, such functional analysis of Arabidopsis genes is based largely on laboratory-induced mutants that are selected in forward and reverse genetic studies. An alternative complementary source of genetic variation is available: the naturally occurring variation among accessions. The multigenic nature of most of this variation has limited its application until now. However, the use of genetic methods developed to map quantitative trait loci, in combination with the characteristics and resources available for molecular biology in Arabidopsis, allow this variation to be exploited up to the molecular level. Here, we describe the current tools available for the forward genetic analysis of this variation, and review the recent progress in the detection and mapping of loci and the cloning of large-effect genes.     

Naturally occurring mitochondrial DNA heteroplasmy in the MRL mouse

The MRL/MpJ mouse is an inbred laboratory strain of Mus musculus, known to exhibit enhanced autoimmunity, increased wound healing, and increased regeneration properties. We report the full-length mitochondrial DNA (mtDNA) sequence of the MRL mouse (Accession # EU450583), and characterize the discovery of two naturally occurring heteroplasmic sites. The first is a T3900C substitution in the TPsiC loop of the tRNA methionine gene (tRNA-Met; mt-Tm). The second is a heteroplasmic insertion of 1-6 adenine nucleotides in the A-tract of the tRNA arginine gene (tRNA-Arg; mt-Tr) at positions 9821-9826. The level of heteroplasmy varied independently at these two sites in MRL individuals. The length of the tRNA-Arg A-tract increased with age, but heteroplasmy at the tRNA-Met site did not change with age. The finding of naturally occurring mtDNA heteroplasmy in an inbred strain of mouse makes the MRL mouse a powerful new experimental model for studies designed to explore therapeutic measures to alter the cellular burden of heteroplasmy.     

Mitochondrial genes: signals and noise in the phylogenetic reconstruction of the annual killifish genus Cynolebias (Cyprinodontiformes, Rivulidae)

The genus Cynolebias (Cyprinodontiformes: Rivulidae) is a locally endemic and speciose group of Neotropical fishes with an annual life cycle. Members of the genus vary greatly in morphology and behaviour, and extensive interspecific karyotypic divergence has been documented among species from Uruguay, Argentina and Rio Grande do Sul, Brazil. We present a molecular systematic hypothesis of the relationships between these Cynolebias species based on phylogenetic analysis of a combined dataset containing 1825 base pairs of DNA sequence, representing three mitochondrial genes. The protein-coding cytochrome -b gene, the 12S and 16S rRNA mitochondrial genes, alone and in combination, yield robust support for monophyly within Cynolebias . Furthermore, our analyses identify two major Cynolebias clades, one of which contains at least four monophyletic groups. Corrected mtDNA genetic distances range from 5.2 to 17.5% between Cynolebias species, and application of a molecular clock suggests the occurrence of two pulses of cladogenesis, one in the late Miocene and another in the Pliocene–Pleistocene. © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 76 , 49–59.     

The ultrastructure of prolactin cells in the annual cyprinodont Cynolebias whitei during its life cycle

Summary Prolactin (PRL) cells were studied electron-microscopically and morphometrically in the annual cyprinodont fish, Cynolebias whitei during its life cycle. In prehatching larvae, PRL cells possessed small secretory granules, giant mitochondria and a well-developed Golgi apparatus. During hatching, no changes were observed in the volume density of the secretory granules, indicating that no increased release of PRL occurs at hatching. A significant change in the composition of PRL cells, i.e., the volume densities per cytoplasm volume of the different organelles, occurred between one day and one week of age. Thereafter, only minor differences were observed between age groups, indicating that no major changes occur in PRL cell activity during the lifespan of C. whitei. However, the volume density per cell volume of the nucleus decreased steadily with age during the lifespan. A comparison of the PRL cells in young and adult fish reared in fresh water (FW) with siblings reared from hatching in diluted sea water (1/3 SW) did not reveal any differences with respect to the volume densities of the organelles, including the secretory granules. However, significant differences were observed with respect to the diameter, electron-dense content and affinity to anti- PRL serum of the secretory granules. These differences indicate that, despite the similar volumetric composition of the PRL cells, their secretory granules contain a substantially higher concentration of PRL in FW-reared fish than in 1/3 SW-reared fish.     

Naturally occurring tubulin-containing paracrystals in Allogromia: immunocytochemical identification and functional significance

Bundles of microtubules (MTs) are readily visualized in vivo by videomicroscopy in highly flattened reticulopodia of the foraminiferan protozoan Allogromia sp. strain NF. In this report we use videomicroscopy, immunocytochemistry, and high-voltage electron microscopy to characterize the dynamic changes that occur in this extensive MT cytoskeleton, and in the associated cytoplasmic transport, during induced withdrawal and subsequent reextension of reticulopodia. Within seconds after application of the withdrawal stimulus (seawater substitute made hypertonic with MgCl2) intracellular bidirectional transport along linear MT-containing fibrils ceases and is replaced by an inward, constant-velocity flow of cytoplasm along the fibrils. As withdrawal continues, most fibrils become wavy and coalesce to form phase-dense pools. These wavy fibrils and phase-dense pools contain a paracrystalline material and few if any MTs. Same-section correlative immunofluorescence and high-voltage electron microscopy reveal that the paracrystalline material contains tubulin. During recovery linear fibrils (MTs) rapidly extend from the phase-dense pools (paracrystals), which concurrently shrink in size, thus reestablishing normal network morphology and motility. We conclude that the MT cytoskeleton in Allogromia reticulopodia is transformed during withdrawal into a tubulin-containing paracrystal, which serves as a temporary reservoir of MT protein and an initiation site for MT regrowth.     

Naturally occurring hypertrophic pyloric stenosis in the domestic rabbit.

Hypertrophic pyloric stenosis was observed to occur naturally in laboratory rabbits. Three cases in juvenile and adult rabbits and their antemortem histories were examined. The gross and microscopic pathology were studied. It was shown that not only the pylorus, but the stomach as a whole was increased in size both relatively and absolutely. The thickness of the pylorus was doubled, but its circumference was not increased, in hypertrophic stenosis. The chief histologic finding was hypertrophy of circular and longitudinal smooth muscles and their enveloping fascia. The myenteric plexi were essentially normal in appearance. The literature on a similar condition in man, dog, and cat was reviewed.     

Naturally occurring wounds and wound healing in chick embryo wings

Summary Spontaneous cutaneous wounds occur in avian embryos (chick, duck, quail) in various prominent parts of the body, notably the elbow, the knee and the outer face of feather buds. The frequency and size and the light and electron microscopic morphology of elbow wounds in the chick embryo are described. The cutaneous lesion appears in over 80% of the embryos at around 7 days of incubation, persists through 14 days, and finally heals completely at around 16 days of incubation. No trace of the wound is visible after that age. Wound healing of these spontaneous lesions was analysed with light microscopy (using indirect immunofluorescence for the localization of type I collagen, fibronectin and laminin) and electron microscopy. The main feature of the very slow healing process, as compared with the rapid cicatrization of experimental excision wounds, appears to be a continuous damage of the healing epidermis, until, finally, definitive wound closure occurs between 14 and 16 days of incubation. In the damaged region, where the epidermis is absent, the dermis exhibits an increased density of type I collagen fibres and of fibronectin. The upper face of the bare dermis is deprived of laminin. Spontaneous lesions do not occur in isolated wings explanted on the chick chorioallantoic membrane, where the wings do not become mobile and are not in contact with the amnion. The observations and explantation experiments suggest that the skin damage is caused by friction and abrasion of the bending elbow against the amnion or the amniotic fluid.     

Naturally occurring Tyzzer's disease and intestinal spirochetosis in guinea pigs

Bacillus piliformis infection (Tyzzer's disease) occurred in two young guinea pigs, causing unthriftiness and diarrhea which resulted in death. There was necrosis and inflammation of the ileum, cecum, and colon. Intestinal epithelial cells contained organisms resembling Bacillus piliformis. Spirochetes were found in the cecum and colon, mainly in crypts. Acute diarrhea occurred in another guinea pig which became cachetic and was killed. Histologically, large numbers of spirochetes were present in the wall of both the cecum and colon, and they were associated with severe necrosis and inflammation. Bacillus pilformis was not found in this animal.