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At least 1,225 records · Page 68Linked to original sources
PubMed1994

Electrophysiological investigation of microdissected gastric glands of bullfrog. I. Basolateral membrane properties in the resting state.

In the present experiments we have made a new attempt to characterize the ion transport properties of H(+)-secreting cells of the gastric mucosa using electrophysiological techniques. Individual gastric glands of bullfrog fundus mucosa were manually dissected, mounted in holding pipettes and superfused with various test solutions while individual cells were punctured with conventional or H(+)-sensitive double-barrelled microelectrodes. All measurements were performed in the resting state (0.1 mmol/l cimetidine). In HCO3(-)-containing control Ringer solution the cell membrane potential (Vb) averaged -45.6 +/- 0.9 mV (+/- SEM, n = 54). From the fast initial Vb responses to changing bath K+, Na+, Cl- or HCO3- concentrations we deduced that the basolateral cell membrane contains conductances for K+, Na+, and Cl- but not for HCO3-, and that a Na(+)-HCO3- cotransporter is not present. The K+ conductance was inhibited by Ba2+ (3 mmol/l), but the Cl- conductance was not inhibited by 4,4' diisothiocyanato-stilbene-2,2' disulphonic acid (DIDS, 0.3 mmol/l), nor selectively inhibited by 5-nitro-2-(3)- phenylpropyl-aminobenzoate (NPPB, 10 mumol/l). In a great number of cells the Vb response to Cl- substitution revealed two components: an initial spiking depolarization which reflected conductive Cl- efflux and a secondary slow hyperpolarization, the origin of which was not immediately evident. Since the latter response could be mimicked by CO2-free perfusion, strongly depressed by Ba2+ and eliminated by DIDS, we conclude that it reflects HCO3- uptake into the cells via a DIDS sensitive Cl-/HCO3- exchanger which alkalinizes the cells and stimulates the basolateral K+ conductance. Our results confirm, revise and extend the results of previous, less direct, investigations of gastric cell ion transport.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1979

The measurement of coenzyme A and a coenzyme A-dependent enzyme. In microdissected epidermal material using coupled enzyme and bioluminescent reactions.

A sensitive micromethod for the determination of Coenzyme A and its esters down to about 0.2 pmol in a volume of 10 microliters and of the activity of citrate synthase is outlined. Epidermal material from healthy and psoriatic skin was utilized in microgram quantity as tissue source. The assay utilizes the ketoglutarate dehydrogenase reaction to yield NADH on addition of free Coenzyme A and the subsequent measurement of NADH by a bioluminescent reaction with Acromobacter fischerii. The total Coenzyme A content in six healthy subjects measured in stratum Malpighii was 1.58 +/- 0.19 mmol per kg dry weight. In six psoriatic patients non-involved and involved epidermis contained 1.51 +/- 0.27 and 1.50 +/- 0.25 mmol/kg, respectively. Long-chain acyl-Coenzyme A comprised about 20% in lesion-free skin and 60% of total content in the involved psoriatic epidermis. The activity of citrate synthase in basal layers of healthy epidermis was 0.30 +/- 0.04 mkat/kg dry weight.

Animals↗
PubMed1989

Cytosolic free calcium in single microdissected rat cortical collecting tubules.

Cytosolic free Ca2+ ([Ca2+]i) was measured in single fragments of rat cortical collecting tubule (CCT) by using fura-2 and a tubule superfusion device. Under basal conditions, i.e. with 1 mM of external Ca2+ ([Ca2+]o), the average steady state [Ca2+]i was 179 +/- 16 nM (n = 44 tubules). Random alterations of [Ca2+]o between 0 mM and 4 mM led to corresponding variations in steady state [Ca2+]i levels, which were linearly correlated with [Ca2+]o (average slope 93 +/- 34 nM [Ca2+]i per 1 mM [Ca2+]o for six tubules). In contrast, [Ca2+]i was little affected by decreasing external Na+ concentration. Cell membrane depolarization with 100 mM of external K+ induced a sustained drop in [Ca2+]i (21% as an average). The data suggest that steady state [Ca2+]i in CCT cells resulted from a non-saturable passive entry of calcium ions across cell membranes balanced with an active extrusion by calcium ATPase (pump and leak mechanism). The passive component cannot be accounted for either by Na+/Ca2+ exchangers nor by voltage-dependent calcium channels; it is best explained by the presence of voltage-independent calcium channels in cell membranes.

Animals↗
PubMed1985

Stimulation of phospholipid turnover by angiotensin II and phenylephrine in proximal convoluted tubules microdissected from mouse nephron.

Angiotensin II and phenylephrine significantly increased phospholipid turnover in cortical tubule suspensions. To further localize this effect we studied the 32P incorporation into phosphatidylcholine (PC) and phosphatidylinositol (PI) in single dissected proximal convoluted tubules of mouse nephron. Both hormones significantly stimulated 32P labeling of PC and PI indicating that the proximal tubule is a target site of angiotensin II and phenylephrine action.

Angiotensin II↗
PubMed1993

The trochlear nerve: anatomy by microdissection.

This work is based on the microscopic study of 30 trochlear nerve trunks (15 heads). In 17 cases, the trunk arose from two nerve bundles, in 8 cases from one bundle, and for the other 5 nerves, three or four bundles. The mean total length of the trochlear nerve was 86 mm. The nerve may be separated into the 3 following parts: infratentorial, intracavernous, intraorbital. In all 30 cases studied, the first part of the nerve was infratentorial, thus leading us to suggest the term "infratentorial part" for this segment of the nerve. In 27 cases, contact was found with the superior cerebellar artery, in the infratentorial part. In the intracavernous part of ten nerves we found two rami tentorii and in eight cases fibers were exchanged with the ophthalmic nerve. In the orbit, 18 trochlear nerves crossed the posterior ethmoidal artery. 23 trochlear nerves ended on the medial face of the superior oblique muscle. The remaining 7 ended at the superior border of the muscle.

Adult↗
PubMed1987

Distribution of 3,4-dihydroxyphenylacetic acid (DOPAC) and 3,4-dihydroxyphenylglycol (DOPEG) in microdissected brain structures and the pituitary gland: metabolite changes in the median eminence in response to hyperprolactinemia and suckling.

Dopamine (DA), norepinephrine (NE), epinephrine (E), 3,4-dihydroxyphenylglycol (DOPEG) and dihydroxyphenylacetic acid (DOPAC) were determined simultaneously by a radioenzymatic, thin-layer chromatographic assay able to detect 1-10 pg of the parent compounds and 80-120 pg of their metabolites. A localization study of these compounds in 20 micro-dissected hypothalamic and limbic structures and the anterior and posterior pituitary glands of male rats was completed. DOPAC was detectable in 14 of 22 structures with the lowest DOPAC/DA ratio being found in the caudate nucleus (7.1%) and the highest in the medial aspect of the ventromedial nucleus of the hypothalamus (422.0%). There was a higher DOPAC/DA ratio in the lateral (21.5%) than in the medial (11.3%) portion of the median eminence suggesting that a greater portion of released DA in the medial median eminence enters the portal circulation. DOPEG was detectable in 6 of 22 structures with DOPEG/NE ratios ranging from 8% (interstitial nucleus of the stria terminalis, ventral aspect) to 32% (medial median eminence). A poor correlation exists between DOPAC and DA concentrations in the various brain regions while there was a stronger relationship between DOPEG and NE concentrations. Male rats were rendered hyper-prolactinemic for 48 hours with injections of ovine prolactin (oPRL) every 8 hours (4 mg/kg body weight sc). In such rats there was a suppression of endogenous rat PRL (rPRL) secretion, the DOPAC/DA ratio increased 2.2-fold in the medial (MEm) and 1.9-fold in the lateral median eminence (MEl), and the DA concentration in the anterior pituitary also increased 2.6-fold. In 10 day postpartum lactating rats, suckling produced marked increases in serum rPRL but no change in DOPAC/DA ratios in the ME or in the DA concentration in the anterior pituitary. The data reveal a wide range of DOPAC/DA ratios (7-422%) in brain regions containing cell bodies, axons and terminals of the different dopaminergic neuronal tracts in brain and pituitary. Considering the DOPAC/DA ratios in the MEm and MEl, it is suggested that a large perturbation of dopaminergic transmission produces a significant ratio change while a smaller perturbation is not detected by this index of neuronal metabolism.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1996

Tetrasomy 18p de novo: identification by FISH with conventional and microdissection probes and analysis of parental origin and formation by short sequence repeat typing.

We report a de novo supernumerary isochromosome 18p in a child with tetrasomy 18p, analyzed by a straightforward combination of cytogenetic and molecular cytogenetic methods. The diagnostic procedure consisted of standard banding techniques and fluorescence in situ hybridization (FISH) with centromere and library DNA probes for chromosome 18, and 18p-specific FISH probes prepared by chromosome dissesction and in vitro amplification. The maternal origin as well as the most probable cell stages of formation of the supernumerary isochromosome were determined by typing of short sequence repeats (SSRs). The pattern of allelic distribution suggests a nondisjunction during meiosis followed by a centromeric misdivision in an early postzygotic mitosis as the most probable mode of isochromosome 18p formation. The combination of the applied methods represents a powerful tool to investigate the nature and the origin of de novo marker chromosomes.

Alleles↗
PubMed1976

Anatomical and functional heterogeneity of nephrons in the rabbit: microdissection studies and SNGFR measurements.

The single nephron glomerular filtration rate (SNGFR) was determined in superficial (S) and juxtamedullary (JM) nephrons of 10 anesthetized rabbits by the 14C ferrocyanide infusion technique. The length of the proximal tubules and the volume of the glomeruli were also determined for the same nephrons. SNGFR was higher in JM than in S: 28.6 +/- 3.4 versus 22.6 +/- 3.0 nl/min, P less than 0.001. In JM nephrons, glomeruli were larger than in S: 1.3 +/- 0.1 versus 0.9 +/- 0.1 nl, P less than 0.001, whereas there was no difference between proximal tubule length in either category (s, 8.7 +/- 0.3 and JM, 8.9 +/- 0.5 mm). In 6 out of 8 animals, SNGFR was significantly correlated to glomerular volume. Lack of correlation was observed between SNGFR and length of proximal tubule in all animals but one. These results show that the rabbit, as well as small rodents and the dog, has a higher SNGFR in juxtamedullary than superficial glomeruli. Although this functional difference is not related to the length of the proximal tubule in each individual animal, the ratio between the mean SNGFR value and the mean length of the proximal tubule in superficial rabbit nephrons is similar to the ratio found in other species.

Animals↗
PubMed2000

Laser capture microdissection: beyond functional genomics to proteomics.

Proteomics will drive biology and medicine beyond genomics, and can have a profound impact on molecular diagnostics. The posttranslational modifications of cellular proteins that govern physiology and become deranged in disease cannot be accurately portrayed by gene expression alone. Consequently, new technology is being developed to discover, and quantitatively monitor, proteomic changes that are associated with disease etiology and progression. In the past, proteomic technologies were restricted to tumor cell lines or homogenized bulk tissue specimens. This source material may not accurately reflect molecular events taking place in the specific cells of the tissue itself. This article describes a completely new class of proteomic-based approaches aimed at the identification and investigation of protein markers in the actual histologically defined cell populations that are immersed in heterogeneous diseased tissue. It is envisioned that these investigations will eventually lead to novel diagnostic, prognostic, or therapeutic markers that can be applied to monitor therapeutic toxicity or efficacy.

DNA, Complementary↗