Nucleotide sequence of 5S rDNA and localization of the ribosomal RNA genes to metaphase chromosomes of the Tilapiine cichlid fish, Oreochromis niloticus

In this study, we report the cloning and nucleotide sequence of PCR-generated 5S rDNA from the Tilapiine cichlid fish, Oreochromis niloticus. Two types of 5S rDNA were detected that differed by insertions and/or deletions and base substitutions within the non-transcribed spacer (NTS). Two 5S rDNA loci were observed by fluorescent in situ hybridization (FISH) in metaphase spreads of tilapia chromosomes. FISH using an 18S rDNA probe and silver nitrate sequential staining of 5S-FISH slides showed three 18S rDNA loci that are not syntenic to the 5S rDNA loci.

Localization of peroxidase activity in blood mononuclear phagocytes in pacu fish (Piaractus mesopotamicus)

The localization of peroxidase activity in different cell regions is used as a criterion for classifying the stage of maturity of mammalian mononuclear phagocytes, with a positive peroxidase reaction indicating the presence of monoblasts, promonocytes, monocytes, and macrophages. Peroxidase activity was observed ultrastructurally in the circulating blood of pacu fish (Piaractus mesopotamicus), identifying monoblasts, promonocytes, monocytes, and macrophages. These observations suggest that differentiation of mononuclear phagocytes occurs in the blood circulation of fish, whereas in mammals, monoblasts and promonocytes are detected in bone marrow, with only monocytes detected in circulating blood and differentiation into macrophages occurring in other body compartments.

Short interspersed repetitive elements (SINEs) from the cichlid fish, Oreochromis niloticus, and their chromosomal localization by fluorescent in situ hybridization

In the present study, we describe the cloning and characterization of a new SINE-like element from O. niloticus (ROn-2) and show the distribution of this SINE and a previously isolated SINE, ROn-1, in the chromosomes of O. niloticus. The ROn-2 element is 359 base pairs (bp) in length, contains short direct terminal repeats, a tRNA-related region similar to tRNA Val and tRNA Arg, a tRNA-unrelated region, and a poly-A tail. Analysis of the chromosomal distribution of ROn-1 and ROn-2 by fluorescent in situ hybridization showed that both SINE sequences are present in all chromosomes of tilapia, and organized in small clusters. The only exception was a large cluster of ROn-1 repeats found in the middle of the long arm of chromosome 1. In view of our data we discuss the hypothesis that the absence of large clusters of SINE sequences and the structural composition of these sequences may explain the absence of base-specific fluorochrome bands in the chromosomes of tilapia.

Molecular characterization and chromosomal localization of two families of satellite DNA in Prochilodus lineatus (Pisces, Prochilodontidae), a species with B chromosomes

Prochilodus lineatus, an abundant species in the Mogi-Guaçu river basin, represents a large part of the region's fishing potential. Karyotypic analyses based on classic cytogenetic techniques have revealed the presence of 54 metasubmetacentric type chromosomes, together with the occurrence of small supernumerary chromosomes with intra and interindividual variations. This paper describes the genomic organization of two families of satellite DNA in the P. lineatus genome. The chromosomal localization these two repetitive DNA families through fluorescence in situ hybridization (FISH) demonstrated that the SATH1 satellite DNA family, composed of approximately 900 bp, was located in the pericentromeric region of a group of chromosomes of the standard complement, as well as on all the B chromosomes. The SATH2 satellite family has a monomeric unit of 441 bp and was located in the pericentromeric regions of some chromosomes of the standard complement, but was absent in the B chromosomes. Double FISH analyses showed that these two families participate jointly in the pericentromeric organization of several chromosomes of this species. The data obtained in this study support the hypothesis that the B chromosomes derive from chromosomes of the standard complement, which are carriers of the SATH1 satellite DNA.

Chromosome localization of the ribosomal genes 18S and 5S in four stocks of rainbow trout (Oncorhynchus mykiss) from cultivated and naturalized stocks in Brazil

In the present study, fluorescence in situ hybridization (FISH) was employed to determine the chromosomal location of genes 18S rDNA and 5S rDNA in four rainbow trout stocks. In specimens from the stocks of Núcleo Experimental de Salmonicultura de Campos do Jordão and Gavião river, 18S genes were located at a subterminal position in the long arms of two submetacentric chromosomes, whereas in specimens from stocks of Mount Shasta and Teresópolis they were found in the short arms. In all analyzed stocks, 5S genes were located in two chromosome pairs. In a subtelocentric pair, 5S genes were present in the short arms and, in the other submetacentric pair, 5S genes were at an interstitial position. In the latter, 18S and 5S genes were contiguous. Taking into account that both 18S and 5S rDNA genes have been localized in the short arm of a submetacentric chromosome in almost all rainbow trout samples so far studied, the presence of such genes in the long arm, as seen in the samples from Núcleo Experimental de Salmonicultura de Campos do Jordão and Gavião river, supports the hypothesis of a pericentric inversion involving this chromosome segment in the ancestor line of these stocks. The observed polymorphism allowed the identification of a very useful genomic marker, and may therefore constitute an important tool in the genetic management of rainbow trout stocks.

Localization of rDNA sites in holocentric chromosomes of three species of triatomines (Heteroptera, Triatominae).

Chromatin organization in the holocentric chromosomes of three triatomines species was cytologically studied by fluorescent in situ hybridization with a 45S rDNA probe of Drosophila melanogaster to localize ribosomal genes. In Triatoma tibiamaculata, metaphases I showed telomeric highlights in a single, larger bivalent. In T. protacta, hybridization was detected in one of the telomeres of an autosomal chromosome. In T. platensis, there were highlights in a single, smaller chromosome (X chromosome). The results obtained did not agree with the expected localization of rDNA genes in the sex chromosomes of triatomines, as demonstrated by silver impregnation, and suggest that the chromosome reorganization that occurred in this group during evolution may be a more important mechanism involved in rDNA distribution.

Rhipicephalus (Boophilus) microplus (Canestrini, 1887) (Acari: Ixodidae): Acid phosphatase and ATPase activities localization in salivary glands of females during the feeding period

This study investigates the presence and the localization of acid phosphatase and ATPase in the salivary glands of Rhipicephalus (Boophilus) microplus female ticks during feeding. Semi-engorged females showed a larger amount of acid phosphatase compared to those at beginning of feeding, localized mainly in the apical portion of the secretory cells, and in the basal labyrinth of the interstitial cells. Ultrastructural observations also demonstrated its presence in secretion granules and inside some nuclei of secretory cells at beginning of feeding. Acid phosphatase in a free form probably has a hemolymph and/or ribosomal origin and participates in salivary gland secretion control. ATPase was detected in basal membrane of all types of acini and/or in the cytoplasm of the secretory cells at both feeding stages. The enzyme activities found strongly suggests that cell death by apoptosis occurs during the degenerative process. © 2006 Elsevier Inc. All rights reserved.

Localization of total proteins and lactate dehydrogenase in hamster epididymis

The concentration of total protein measured by photocolorimetric methodology and reported as units per 100 mg of tissue decreased from the initial segment to the cauda epididymidis of the Golden hamster, being significant the numeric difference observed between these two regions. This observation was related with an increased synthesis and secretion of proteins to the lumen in proximal segments of the epididymidis duct, mainly in initial segment, as proposed for other rodents. LDH activity was higher in initial segment and distal cauda than in the caput and corpus epididymidis, although no significant differences in mean values had been observed. The high LDH activity observed in initial segment and cauda epididymidis of hamster had been related to an expressive epithelium metabolic activity presented in these regions. This metabolic activity help to guarantee the survival of spermatozoa stored in cauda epididymidis. Furthermore, lower LDH activity noted in the caput and corpus epididymidis might be related with a progressive reduction of glycolysis in initial maturation step of spermatozoa mainly verified in corpus epididymidis. © 2007 Sociedad Chilena de Anatomía.

Evaluation of tropospheric and ionospheric effects on the geographic localization of data collection platforms

The Brazilian National Institute for Space Research (INPE) is operating the Brazilian Environmental Data Collection System that currently amounts to a user community of around 100 organizations and more than 700 data collection platforms installed in Brazil. This system uses the SCD-1, SCD-2, and CBERS-2 low Earth orbit satellites to accomplish the data collection services. The main system applications are hydrology, meteorology, oceanography, water quality, and others. One of the functionalities offered by this system is the geographic localization of the data collection platforms by using Doppler shifts and a batch estimator based on least-squares technique. There is a growing demand to improve the quality of the geographical location of data collection platforms for animal tracking. This work presents an evaluation of the ionospheric and tropospheric effects on the Brazilian Environmental Data Collection System transmitter geographic location. Some models of the ionosphere and troposphere are presented to simulate their impacts and to evaluate performance of the platform location algorithm. The results of the Doppler shift measurements, using the SCD-2 satellite and the data collection platform (DCP) located in Cuiabá town, are presented and discussed.

Alkaline phosphatase localization in golden hamster epididymidis: A biochemical and histochemical investigation

The aim of this study was to determine alkaline phosphatase (ALP; E.C. 3.1.3.1) activity and major expression in homogenates obtained from different regions of Golden hamster epididymis, comprising the initial segment, head, body and tail, with concomitant research of this enzyme localization and activity in samples of tissues. These were collected from the same regions and investigated by histochemical conventional study performed on frozen histological sections. No significant differences in mean ALP activity, reported as U.100 mg-1 of tissue, were observed among the biological specimens collected from the epididymidis initial segment (0.92 ± 0.28 U.100 mg-1 tissue), head (1.07 ± 0.67 U.100 mg-1 tissue) and body (0.77 ± 0.23 U.100 mg-1 tissue). However, mean ALP activity was significantly higher in the epididymal tail (8.94 ± 0.40 U.100 mg-1 tissue) compared with the precedent segments. The findings suggested that ALP plays a significant role in the tail of the Golden hamster epididymidis, mediating androgenic segregation necessary to maintain the epithelial integrity. Furthermore, ALP acts on active transport of substances between the luminal fluid and spermatozoon membrane, and contrariwise. Thus, the high concentration of ALP in the epididymal tail helps to indicate the importance of this enzyme in the metabolism and maintenance of spermatozoa maturation and storage into the epididymidis luminal compartment, perhaps directly influencing the normal reproductive morphophysiology.

Localization of myosin-Va in subpopulations of cells in rat endocrine organs

Myosin-Va is a Ca 2+/calmodulin-regulated unconventional myosin involved in the transport of vesicles, membranous organelles, and macromolecular complexes composed of proteins and mRNA. The cellular localization of myosin-Va has been described in great detail in several vertebrate cell types, including neurons, melanocytes, lymphocytes, auditory tissues, and a number of cultured cells. Here, we provide an immunohistochemical view of the tissue distribution of myosin-Va in the major endocrine organs. Myosin-Va is highly expressed in the pineal and pituitary glands and in specific cell populations of other endocrine glands, especially the parafollicular cells of the thyroid, the principal cells of the parathyroid, the islets of Langerhans of the pancreas, the chromaffin cells of the adrenal medulla, and a subpopulation of interstitial testicular cells. Weak to moderate staining has been detected in steroidogenic cells of the adrenal cortex, ovary, and Leydig cells. Myosin-Va has also been localized to non-endocrine cells, such as the germ cells of the seminiferous epithelium and maturing oocytes and in the intercalated ducts of the exocrine pancreas. These data provide the first systematic description of myosin-Va localization in the major endocrine organs of rat. © 2008 Springer-Verlag.

Localization of a Bose-Einstein condensate in a bichromatic optical lattice

By direct numerical simulation of the time-dependent Gross-Pitaevskii equation, we study different aspects of the localization of a noninteracting ideal Bose-Einstein condensate (BEC) in a one-dimensional bichromatic quasiperiodic optical-lattice potential. Such a quasiperiodic potential, used in a recent experiment on the localization of a BEC, can be formed by the superposition of two standing-wave polarized laser beams with different wavelengths. We investigate the effect of the variation of optical amplitudes and wavelengths on the localization of a noninteracting BEC. We also simulate the nonlinear dynamics when a harmonically trapped BEC is suddenly released into a quasiperiodic potential, as done experimentally in a laser speckle potential. We finally study the destruction of the localization in an interacting BEC due to the repulsion generated by a positive scattering length between the bosonic atoms. © 2009 The American Physical Society.

Temporal localization of osteocalcin protein during healing of tooth extraction sockets in rats.

The aim of this study was to evaluate the expression of osteocalcin protein during the alveolar bone healing process in rats. Twenty four rats were used in this study and, after anesthetic induction, they had their right upper incisors extracted. At 7, 14, 21 and 28 days after the tooth extraction, the animals were injected 4% formaldehyde. The histological tissue pieces were colored in hematoxilin and eosin and the immunohistochemistry reaction for osteocalcin was performed. At seven days lesser neoformed trabeculae bone and a small quantity of osteocalcin labeling were observed. At 14 and 21 days a larger quantity of neoformed trabeculae bone and higher osteocalcin values were detected. At 28 days the largest quantity of neoformed trabeculae bone and a decrease on the amount of osteocalcin immunolabelling were noticed. According to our results and considering the limits of the present study it is possible to conclude that a greater osteocalcin expression is observed at 14 and 21 days postoperatively, characterizing the periods when intense mineralization of the bone tissue occurs during the alveolar bone healing process.

Cellular localization of androgen synthesis in equine granulosa-theca cell tumors: Immunohistochemical expression of 17α-hydroxylase/17,20-lyase cytochrome P450

Elevated blood testosterone concentrations, often accompanied by male-typical behaviors, is a common signalment of mares with granulosa-theca cell tumors (GCTCs), but no definitive information exists regarding the cellular differentiation of tumors associated with androgen secretion. This study was conducted to localize and thereby define the cellular expression of 17α-hydroxylase/17,20-lyase cytochrome P450 (P450c17), the enzyme most directly responsible for androgen synthesis, in 30 GTCTs and control tissues (gonads and adrenal glands) using immuno-histochemistry (IHC). Immuno-reactivity for P450c17 was evident in approximately half of 30 specimens examined, was most consistent in the interstitial cells surrounding existing or developing cysts, and was less intense in cells within cysts in the smaller proportion of specimens where this was observed. In control tissues, the expression of P450c17 was localized primarily in theca interna of normal ovarian follicles, in theca-lutein cells of some corpora lutea, but not in granulosa-lutein cells. Testicular interstitial cells and islands of adreno-cortical cells located in the adrenal medulla of the adrenal cortex further established the specificity of the antisera used. These data provided the first substantive evidence that polyhedral cells identified previously in GTCTs by histopathology have the potential to synthesize and secrete androgens, similar to theca interna and theca lutein cells in normal equine ovaries. © 2010 Elsevier Inc.