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Biomedical subjects

B H Hwang

Publications and source records attributed to B H Hwang.

41 records · Page 3Linked to original sources

A method for studying glyoxylic acid induced fluorescence and ultrastructure of monoamine neurons.

Inasmuch as precise correlations of light- and electronmicroscopy are crucial for understanding biostructure, it seemed necessary to bring together the advantages of the glyoxylic acid (GA) method (for inducing monoamine fluorescence) and electron microscopy. A combined fluorescence and electron microscope method using GA is introduced. The brain is perfused by 2% GA in Krebs-Ringer bicarbonate buffer (pH 7.0) and this solution is followed by 4% paraformaldehyde containing 0.5% glutaraldehyde in Sorensen's phosphate buffer (pH 7.4). Sections are cut by cryostat or by vitratome and incubated in 2% GA in phosphate buffer (pH 7.0). Using fluorescence microscopy, features of interest are sketched and/or photographed. Afterwards, the same or subsequent section is processed for electron microscopy. Since axons of catecholamine-containing neurons (as well as their perikarya and terminals) are visualized by GA, the recommended procedure expands the range of studies concerning monoamine neurons that can now be carried out effectively.

Animals↗

Fine structural changes in the hamster pineal gland after blinding and superior cervical ganglionectomy.

Pineal glands of male hamsters 8 weeks after removal of both eyes or both superior cervical ganglia and those of untreated animals were studied by electron microscopy. In the blinded hamsters the reproductive organs were remarkably involuted, whereas the pinealocytes enlarged and were characterized by a tremendous hypertrophy of the smooth-surfaced endoplasmic reticulum, in the mesh of which some dense cored vesicles were distributed. In constrast the pinealocytes of ganglionectomized hamsters atrophied and were noted by a large number of lysosomes and sparsity of the agranular reticulum, the testes being significantly larger than the controls. The findings were interpreted to be compatible with the view that the pineal capability of producing antigonadal substance is augmented by blinding and is suppressed by cervical ganglionectomy due to the impairment of normal functioning of the pineal by denervation. It is assumed that the degree of development of the smooth endoplasmic reticulum in the pinealocyte is parallel to, and reflects that of the pineal activity. Probably the membranous organelle is involved in the elaboration of the pineal antigonadal substance.

Animals↗

G protein activation by endomorphins in the mouse periaqueductal gray matter.

The midbrain periaqueductal gray matter (PAG) is an important brain region for the coordination of mu-opioid-induced pharmacological actions. The present study was designed to determine whether newly isolated mu-opioid peptide endomorphins can activate G proteins through mu-opioid receptors in the PAG by monitoring the binding to membranes of the non-hydrolyzable analog of GTP, guanosine-5'-O-(3-[(35)S]thio)triphosphate ([(35)S]GTPgammaS). An autoradiographic [(35)S]GTPgammaS binding study showed that both endomorphin-1 and -2 produced similar anatomical distributions of activated G proteins in the mouse midbrain region. In the mouse PAG, endomorphin-1 and -2 at concentrations from 0.001 to 10 microM increased [(35)S]GTPgammaS binding in a concentration-dependent manner and reached a maximal stimulation of 74.6+/-3.8 and 72.3+/-4.0%, respectively, at 10 microM. In contrast, the synthetic selective mu-opioid receptor agonist [D-Ala(2),NHPhe(4), Gly-ol]enkephalin (DAMGO) had a much greater efficacy and produced a 112.6+/-5.1% increase of the maximal stimulation. The receptor specificity of endomorphin-stimulated [(35)S]GTPgammaS binding was verified by coincubating membranes with endomorphins in the presence of specific mu-, delta- or kappa-opioid receptor antagonists. Coincubation with selective mu-opioid receptor antagonists beta-funaltrexamine or D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Phe-Thr-NH(2) (CTOP) blocked both endomorphin-1 and-2-stimulated [(35)S]GTPgammaS binding. In contrast, neither delta- nor kappa-opioid receptor antagonist had any effect on the [(35)S]GTPgammaS binding stimulated by either endomorphin-1 or -2. These findings indicate that both endomorphin-1 and -2 increase [(35)S]GTPgammaS binding by selectively stimulating mu-opioid receptors with intrinsic activity less than that of DAMGO and suggest that these new endogenous ligands might be partial agonists for mu-opioid receptors in the mouse PAG.

Analgesics, Opioid↗

Fluorescence microscopy used in conjunction with horseradish peroxidase localization and electron microscopy for studying sympathetic nuclei of the rat spinal cord.

After application of horseradish peroxidase (HRP) to the transected cervical sympathetic trunk, labeled sympathetic neurons were seen in the intermediolateral nucleus (IML), central autonomic nucleus (CAN) and intercalated nucleus of rat spinal cords. As studied by glyoxylic acid-induced histofluorescence microscopy (FM), catecholaminergic (CA) terminals were most densely packed in the IML. There were 60 +/- 2 CA varicosities per 2,200 mu2 area in 20 mu thick sections through the IML. A combined FM/HRP method confirmed that preganglionic sympathetic neurons are heavily innervated by CA terminals. CA boutons were tagged with 5-hydroxydopamine for EM identification, and in the IML, 56% of CA boutons were seen to make synaptic contacts as compared to 60% in the CAN. It seems likely that virtually all CA boutons may form synapses but serial sections through boutons were not studied. It is inferred that preganglionic sympathetic neurons of both IML and CAN are served by CA inputs, and electron microscopy has revealed that most or all of these CA terminals transmit their signals via synapses; which may permit more precise structural sorting than non-synaptic transmission.

Adrenergic Fibers↗

Determination of pyrolysis products of smoked methamphetamine mixed with tobacco by tandem mass spectrometry.

This study examines the pyrolysis products of smoked methamphetamine mixed with tobacco that was trapped with a C8 adsorbent cartridge and then detected by gas chromatography-tandem mass spectrometry. According to the results, the mainstream smoke contains 2-methylpropyl-benzene, 2-chloropropyl-benzene, 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one, 3-ethyl-phenol, methamphetamine, dimethylamphetamine, hydroquinone, 3-methyl-5-(1-methylethyl)-methylcarbamate phenol, N-methyl-N-(2-phenylethyl)-acetamide, 4-(3-hydroxy-1-butenyl)-3,5,5-trimethyl-2-cyclohexene-1-one, propanoic acid, N-acetylmethamphetamine, phenyl ester, and furfurylmethylamphetamine. In addition, the compounds in sidestream smoke are 2-propenyl benzene, phenylacetone, methamphetamine, dimethylamphetamine, benzyl methyl ketoxime, 3,4-dihydro-2-naphthalenone, N-folmyamphetamine, N-acetylamphetamine, bibenzyl, N-folmylmethamphetamine, N-acetylmethamphetamine, N-propionymethamphetamine, and furfurylmethylamphetamine. Moreover, the presence of methamphetamine promotes the oxidation of the tobacco components.

Drug Interactions↗