Specific Heterochromatic Banding of Metaphase Chromosomes Using Nuclear Yellow (original) (raw)
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Acta Histochemica, 2005
Numerous non-rigid and bow-shaped cationic fluorochromes exist which bind to chromatin DNA, but only some of them are able to selectively label pericentromeric heterochromatin (C-bands) in metaphase chromosomes. Likewise, some DNA ligands allow the recognition of adenine-thymine sequences establishing hydrogen bonds with acceptor atoms in the minor groove. In the present study, we have employed cationic fluorochromes based on a variety of chemical groups to analyze comparatively their potential to demonstrate C-bands, as well as the relationship between this feature and structural parameters of fluorochromes such as curvature radius and hydrogenbonding ability. Only fluorochromes that bind DNA by hydrogen bonds demonstrated the characteristic C-banding pattern in mouse metaphase chromosomes (together with a weak G/Q-banding), whereas the other (non-hydrogen-bonding) cationic fluorochromes produced uniform emission without any visible banding.
Journal of Neuroscience Methods, 1981
Retrograde transport of HRP and the fluorescent dye DAPI were used to double label spinothalamic tract cells with dual projections to thalamus and cerebellum. Cells containing both retrograde tracers were found in the intermediate gray zone and in the lateral spinal nucleus of the rat lumbosacral spinal cord. Double labeled cells were easily identified by the presence of granular HRP reaction product and the blue fluorescence of DAPI. The use of HRP and DAPI provides an effective double labeling technique for the localization of neurons with nonoverlapping projections in the central and/or peripheral nervous systems.
The crossed mesostriatal projection: neurochemistry and developmental response to lesion
Brain Research, 1983
The projections of substantia nigra and ventral tegmental area (SN-VTA) neurons to either the ipsilateral or contralateral caudateputamen (CP) were studied in intact and 6-hydroxydopamine-treated rats, using both retrograde transport of fluorescent dyes and measurements of dopamine concentration. After unilateral CP injections of nuclear yellow or granular blue in intact 6-or 30-day-old rats, approximately 1% of the retrogradely labeled cells were located in the contralaterai SN-VTA. In 6-or 30-day-old rats which had been injected with 6-hydroxydopamine in the left lateral hypothalamic area (LHA) at 1 day of age, the extent of cell labeling of both the uncrossed projection in the right hemisphere and the crossed projection originating in the left SN-VTA was normal. However, the number of labeled cells of the uncrossed pathway of the left side and the crossed pathway originating in the right SN-VTA were markedly decreased by the lesion, indicating that the crossed nigrostriatal cells decussate before they reach the LHA. The 6-hydroxydopamine-induced decreases in SN-VTA labeling of both the crossed and uncrossed pathway closely matched the degree of dopamine depletion in the lesioned CP at 6 days of age and at 30 days of age. A partial recovery of both measures occurred between days 6 and 30. Thus, retrograde labeling of SN-VTA cells with fluorescent dyes provided a quantitative measure of the dopaminergic innervation of the neostriatum in both the uncrossed and crossed pathways. The crossed mesostriatal cells are probably dopaminergic because: (1) they were as susceptible to the neurotoxic effects of 6-hydroxydopamine as the uncrossed dopamine cells; (2) they were located almost exclusively within the SN, pars compacta and VTA; and (3) their morphology resembled that of ipsilaterally projecting dopamine cells labeled with the same dye. Preliminary histofluorescence results also revealed that the crossed SN-VTA cells contained catecholamine.
Collateral projections of neurons of the rat globus pallidus to the striatum and substantia nigra
Experimental Brain Research, 1984
Double retrograde fluorescent tracing techniques were used to evaluate the possibility that ascending and descending projections from the globus pallidus arise from divergent axon collaterals. Appropriately placed injections of different tracers (True Blue, Nuclear Yellow) into the substantia nigra and the striatum resulted in the double labelling of neurons in the globus pallidus. Conversely, simultaneous injection of two different sites within the striatum did not produce significant double labelling of globus pallidus neurons. These results indicate that at least a portion of the neurons of the globus pallidus project to both the striatum and substantia nigra, and that individual pallidal neurons do not have widespread projections to the striatum.
Monoamine innervation of the forebrain: Collateralization
Brain Research Bulletin, 1982
The forebrain is characterized by a dense, localized dopamine (DA) innervation pattern, a diffuse, widespread norepinephrine (NE) innervation pattern, and a serotonin (5HT) innervation intermediate between the DA and NE patterns. These innervation patterns have implied that basic differences exist in the way DA, NE and 5-HT axons collateralize to different brain structures; that is, DA axons are thought to be poorly collateralized and NE and 5-HT axons are presumed to be more highly collateralized. In the present study, we used injections of retrograde labeling fluorescent dyes into various forebrain regions in order to determine axonal branching patterns from nuclei that contain DA, NE and 5-HT neurons, namely the substantia nigra-ventral tegmental area (SN-VTA), locus coeruleus (LC) and raphe nuclei (DR-MR). The results suggest that at least two subpopulations of neurons can be defined in each monoamine nucleus with respect to the way their axons collateralize. Each area contains a centrally located nuclear area with highly collateralized neurons, and more peripherally situated areas with less highly collateralized neurons. Thus, previous suppositions of the branching of monoamine axons must be revised to account for the existence of cells exhibiting totally different collateralization patterns within each monoamine nucleus. METHOD Experimental Animals and Surgical Procedures Sixty-five female and male albino rats (Simonsen, Gilroy, CA) weighing 180-360 grams, were used in the study. The animals were anesthetized with Nembutal (Abbott), 30 mg/kg IP, or ether (Mallincrodt) and prepared for aseptic surgery. Injections were made in a Kopf Stereotoxic Apparatus.