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At least 1,207 records · Page 67Linked to original sources
PubMed1985

Ultrasonic microdissection of the mouse mandible: exposure of the vasculature of alveolar bone and myelinated axons of the pulp.

The present scanning electron microscopic study describes a new method for the exposure of blood vessels of mouse alveolar bone and myelinated nerves of the dental pulp. This technique differs from others because it does not remove the periodontal ligament, allowing study of the vascular continuity between alveolar bone and periodontal ligament. Fixed and demineralized mandibles are digested with bacterial collagenase (1 mg/ml) at 37 degrees C for 12 hours, exposed to buffered osmium tetroxide for 24 hours, and ultrasonicated at 80 kHz for 5 minutes. The technique demonstrates that the vascular distribution of the interdental and interradicular septa is different. Vessels pass horizontally through the interdental septum and are continuous with vessels of the adjacent periodontal ligament. Vessels of the interradicular septum branch from a central vessel, pass toward the adjacent periodontal ligament, and become continuous with its vessels. Thus, the pattern of vessel distribution of the interdental septum of the mouse has little similarity to that of man or of research animals. The present study provides an improved method for demonstration of bone vasculature and pulpal axons while retaining valuable anatomical landmarks.

Alveolar Process↗
PubMed1987

Localization of aromatase in the brain of the male African catfish, Clarias gariepinus (Burchell), by microdissection and biochemical identification.

Aromatase activity was determined in small discrete areas of the brain of the African catfish, Clarias gariepinus, by a radiometric assay. The fore- and midbrain were divided into eighteen 500-microns transverse sections. From these sections several punches (0.3 mg of tissue) were taken and incubated with [19-3H]-androstenedione. The aromatase activity, calculated from the release of tritium label during aromatization, is expressed in pmol mg-1 tissue hour-1. The highest activity (3.7 pmol) was detected in the preoptic region. The more caudally located area tuberalis, including the nucleus lateralis tuberis and the nucleus recessus lateralis, also showed a relatively high activity (2.5 pmol). A similar activity was found in the most rostral part of the telencephalon and the dorsal parts of the mesencephalon, i.e., the tectum opticum and torus semicircularis (2.3 pmol). A moderate aromatase activity was observed in remaining parts of the brain, except the cerebellum and hindbrain, in which aromatase activity was hardly detectable (0.1-0.3 pmol). It is concluded that a high aromatase activity is present in regions known to be involved in the regulation of reproduction. Since both the torus semicircularis and the tectum opticum display a high aromatase activity, it is suggested that also these structures are involved in reproductive processes.

Animals↗
PubMed1990

Comparison of osmium/sonication and EDTA/sonication microdissection techniques in exposing the adepithelial basal lamina surface of developing rat colon.

We have used two epithelial-stripping techniques in our studies of the basal lamina in the developing rat colon. The first involves prolonged osmication followed by sonication; the second uses chelation of calcium by EDTA followed by sonication. Both techniques remove the epithelium from the basal lamina; however, the EDTA/sonication technique appears to produce a cleaner adepithelial surface of the basal lamina. In addition, the fine structure of the basal lamina appears to be better preserved in specimens prepared by the EDTA/sonication technique. In contrast, the basal lamina of specimens prepared by the osmium/sonication technique had a shattered appearance that we believe is due to an increase in the fragility of the delicate fetal basal lamina.

Animals↗
PubMed1993

Biosynthesis of dopamine and serotonin in the rat brain after repeated cocaine injections: a microdissection mapping study.

The purpose of the present study was to examine the effects of chronic cocaine on dopamine (DA) and serotonin (5-HT) synthesis in several rat brain regions implicated in drug reinforcement. Male rats were treated twice daily with cocaine (15 mg/kg, ip) or saline for 1 week. After 42 hr of abstinence, rats were challenged with either cocaine (15 mg/kg, ip) or saline, followed by the aromatic L-amino acid decarboxylase inhibitor 3-hydroxybenzylhydrazine (NSD-1015; 100 mg/kg, ip). Animals were decapitated 30 min after NSD-1015 and discrete brain regions were microdisected from 300 microns frozen sections. Postmortem tissue levels of 3,4-dihydroxyphenylalanine (DOPA) and 5-hyroxytryptophan (5-HTP) were quantified by HPLC with electrochemical detection and used to estimate biosynthesis of DA and 5-HT, respectively. In chronic saline-treated rats, cocaine dramatically suppressed DA and 5-HT synthesis in all forebrain regions examined, including: medial prefrontal cortex, nucleus accumbens, caudate nucleus, olfactory tubercle, and basolateral amygdala. The degree of inhibition ranged from 35-65% and was more pronounced in 5-HT neurons compared to DA neurons in the same tissue sample. In general, chronic cocaine did not significantly alter basal levels of DOPA or 5-HTP; a notable exception was lateral hypothalamus, where chronic cocaine reduced basal DA synthesis to 75% of control. After repeated cocaine injections, the synthesis-inhibiting effect of a challenge injection of cocaine was attenuated in many brain areas. These data suggest that whereas acute cocaine decreases DA and 5-HT synthesis in forebrain, chronic cocaine is not neurotoxic to DA and 5-HT neurons. In addition, the mechanism(s) mediating cocaine-induced suppression of monoamine synthesis may become desensitized by chronic exposure to the drug.

5-Hydroxytryptophan↗
PubMed1994

Mapping of the beta 2 subunit gene (GABRB2) to microdissected human chromosome 5q34-q35 defines a gene cluster for the most abundant GABAA receptor isoform.

The gamma-aminobutyric acid receptor (GABAAR) is a multisubunit Cl- channel that mediates most fast inhibitory synaptic transmission in the central nervous system. Molecular evolution has given rise to many genetic variants of GABAAR subunits, including alpha 1-6, beta 1-4, gamma 1-4, delta, and rho 1-2, suggesting that an enormous number of combinations of subunits are possible. Here we report that the beta 2 gene is located on chromosome 5q34-q35, defining a cluster comprising alpha 1, beta 2, and gamma 2 genes that together code for the most abundant GABAAR isoform. The fact that intron position is conserved in the beta 1-3 genes, taken together with the observation that chromosomes 4 and 15 also contain distinct alpha-beta-gamma gene clusters, strongly suggests that an ancestral alpha-beta-gamma cluster was duplicated and translocated to at least two different chromosomes. This organization of GABAAR gene clusters may have been preserved as linkage provides a mechanism for facilitating coordinate gene expression.

Amino Acid Sequence↗
PubMed1992

A DNA library from an individual Beta patellaris chromosome conferring nematode resistance obtained by microdissection of meiotic metaphase chromosomes.

We have used a standard protocol established for human chromosomes to create a chromosome-specific plasmid library from a Beta patellaris chromosome conferring nematode resistance. A monosomic addition line was chosen carrying 18 sugar-beet (Beta vulgaris L.) and one wild-beet (B. patellaris) chromosome. The wild-beet chromosome can readily be identified as a univalent during metaphase I of meiosis. Highly synchronized meiotic material was used to excise the univalents from four pollen mother cells. The chromatin was lysed in a 1 nl collection drop, the DNA purified and restricted with Rsa I, ligated into a vector containing universal sequencing primers, and amplified by the polymerase chain reaction. The amplified DNA was inserted into a standard plasmid vector and cloned. Approximately 23,000 recombinant plasmids were obtained of which 15,800 could be utilized. Their insert sizes ranged from 80 to 700 bp with an average of 130 bp. 61 clones were tested in more detail by genomic Southern hybridization with sugar-beet and wild-beet DNA. Of these 32 plasmids (52%) contained single-copy inserts, 11 (18%) were specific for wild-beet DNA indicating that the DNA cloned originates in the univalent chromosome. The application of this technique for establishing high-density RFLP maps for discrete regions of plant genomes is discussed.

Animals↗
PubMed1981

Microdissection and cloning of DNA from a specific region of Drosophila melanogaster polytene chromosomes.

Fragments from section 3 of the salivary gland X chromosome of D. melanogaster were dissected with a micromanipulator. The DNA was extracted, cut and ligated to a lambda vector in a volume of a few nanoliters in an oil chamber monitored through a microscope. From about 10 pg of DNA we obtained 80 recombinant clones, a sample of which were analysed and shown to contain Drosophila DNA which hybridises in situ to the region of section 3 of the X chromosome. With this technique we can isolate clones from any desired region as small as 200 kb from the euchromatic arms of polytene chromosomes.

Bacteriophage lambda↗
PubMed1994

Gene mapping from a bovine 1;29 DNA library prepared with chromosome microdissection.

Bovine gene mapping is progressing rapidly using syntenic group mapping based on somatic cell hybrids and linkage, and to a lesser extent on in situ hybridization. Single chromosome DNA libraries are a logical next step, and this was, therefore, the aim of our laboratory. Since we have access to several cattle with t(1;29) and this chromosome is readily distinguishable, we chose this as our first target--recognizing that we would not produce a "single" chromosome library in the strict sense because two autosomes are represented. We utilized an inverted microscope and a micromanipulator fitted with glass instruments pulled specifically to dissect off approximately 100 t(1;29) chromosomes per microdrop. A glass chamber made to accommodate a hanging drop was used to extract the DNA under a dissecting microscope. The DNA was then cleaved with EcoRI and inserted in lambda gtwes arms. Host cells were then infected with these phage and positive clones obtained. The first clone, isolated from this library by hybridization with a human collagen 6A1 cDNA, was mapped by in situ hybridization to bovine Chromosome some (Chr) 1q12-q14, near the centromere. The second clone, an anonymous DNA fragment (D1S11), was mapped to 1q43-q46, near the terminal end.

Animals↗