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B K Hartman

Publications and source records attributed to B K Hartman.

At least 37 records · Page 2Linked to original sources
PubMed1988

Immunohistochemical localization of neuropeptides in the vocal control regions of two songbird species.

Immunohistochemistry was used to map the distribution of four neuropeptides in song control regions of two songbird species, the European starling (Sturnus vulgaris) and the song sparrow (Melospiza melodia). We searched for positively stained cell bodies or apparent terminals containing vasoactive intestinal peptide (VIP), methionine-enkephalin (MET), cholecystokinin (CCK), and substance P (SUB P). Intraventricular colchicine pretreatment was administered to enhance the visualization of peptide-containing cell bodies. Four areas implicated in the central control of song were examined. Three of these areas are sexually dimorphic telencephalic nuclei characteristic of songbirds: the caudal nucleus of the ventral hyperstriatum (HVc), the robust nucleus of the archistriatum (RA), and the magnocellular nucleus of the anterior neostriatum (MAN). The fourth region is the mesencephalic nucleus intercollicullaris (ICo), common to all birds, which contains the dorsomedial nucleus (DM) that appears to be specifically involved in the motor control of song. The pattern of neuropeptide localization was similar between the two species. However, the neuropeptides were heterogeneously dispersed among the four areas. VIP and MET were the most widely distributed, whereas CCK and SUB P were seen only in DM. MAN and HVc revealed remarkably similar patterns of staining for both MET and VIP. Fine varicosities immunolabeled for both these peptides appear to encircle nonreactive somata. In both these nuclei positively stained somata were observed for MET but not for VIP. In RA there was a dense accumulation of MET-positive multipolar cell bodies. VIP-containing neurons were seen in the surrounding archistriatum and caudal neostriatum but not in RA itself. Cell bodies and fibers for all four peptides were observed in DM; in no case were they limited to this subregion, but rather seemed to encompass the surrounding intercollicular area as well. The widespread distribution of VIP and MET strongly suggests a role for these peptides in the acquisition or production of passerine song.

Afferent Pathways↗
PubMed1987

Immunohistochemical staining and enzyme activity measurements show myo-inositol-1-phosphate synthase to be localized in the vasculature of brain.

Rabbit anti-bovine myo-inositol-1-phosphate synthase was used to examine the distribution of that enzyme in perfused and immersion-fixed bovine brain and testis. In brain, intense and specific staining was found in the walls of all the vascular elements including cerebral capillaries. The remainder of brain parenchyma exhibited only low levels of background staining. In testis, an organ rich in the enzyme, blood vessels showed no specific staining. Instead, the enzyme was found in the seminiferous epithelium of the seminiferous tubules, perhaps localized in spermatozoa. To confirm the brain finding, the activity of myo-inositol-1-phosphate synthase was measured in bovine brain microvessel preparations and brain pial vessels. In these preparations the activity of the enzyme was found on average to be 7 and 22 times enriched over that in whole brain, respectively. The activities of two other enzymes of inositol metabolism, myo-inosose reductase and myo-inositol-1-phosphatase, were also examined for their distribution in brain. Those enzymes were found to be generally distributed. The surprising finding of a vascular localization of myo-inositol-1-phosphate synthase in brain raises new questions about the mechanism by which myo-inositol is concentrated to such high cellular levels in the principal substance of that organ.

Animals↗
PubMed1986

Transient midline raphe glial structure in the developing rat.

A major glial structure is present during development within the midline raphe of the midbrain, hindbrain, and cervical spinal cord of the rat. It is composed of great numbers of glial cell bodies lying immediately ventral to the cerebral ventricular system and the large radial processes extending from these cells toward the ventral surface of the brain roughly within the midsagittal plane. There is also a smaller group of glial cells on the dorsal surface of the aqueduct and the central canal whose processes extend to the dorsal surface of the brain. The entire structure exhibits an intensely positive immunoreactivity with the antibody to the S-100 protein, a nervous-system-specific protein found primarily in the cytoplasm of astrocytes. This immunoreactivity makes possible a clear visualization of the extent, magnitude, and continuity of this structure from at least embryonic day 15, the first age examined, until postnatal days 7-8, when it is no longer visible by this technique. This glial structure has several prominent morphological characteristics. During prenatal and early postnatal development the fibers forming the ventral aspect of the structure in the midbrain and hindbrain are formed into two parallel plates on either side of the midline with S-100-negative tissue between the plates. As development progresses, S-100-positive fibers are continually added so that the plates become thicker at the expense of the nonstaining intervening area. By postnatal day 4 only a single midline plate of fibers is visible, occupying the entire midline raphe. In the region of the pontine flexure the entire structure takes on a distinctly pleated configuration. This fact produces a curious "sine wave" appearance when the plane of section crosses these vertical pleats. At postnatal day 5 the structure begins to disappear, and it is no longer visible by 7-8 days postnatal. This glial structure does not stain with antisera to glial fibrillary acidic protein, a protein associated with fibrous astrocytes, or routine cell stains such as cresyl violet. With these techniques the raphe area appears essentially devoid of identifiable cellular elements.

Animals↗
PubMed1986

N-methylaspartate: an effective tool for lesioning basal forebrain cholinergic neurons of the rat.

The ability of the excitotoxin, N-methyl-D,L-aspartic acid (NMA), to destroy basal forebrain cholinergic (BFC) neurons was evaluated. NMA (100 nmol) was directly injected into the peripallidum, a region containing a proportionately large number of cortically-projecting BFC neurons. Cholineacetyltransferase (ChAT) activity 10 days later was markedly and significantly reduced (up to 62%) in the cortex ipsilateral to the lesion. NMA induced a focal lesion affecting BFC neurons without damaging axons of passage or causing lesions distant from the site of injection. ChAT immunohistochemistry (IHC) was used to directly demonstrate loss of ChAT-positive neurons from the lesion site. This loss persisted at all survival times examined, from 2 days to 7.5 months post-injection.

Animals↗
PubMed1984

Hypertension induced changes in cerebral capillary permeability to water are mediated by afferent neuronal connections from the carotid sinus to the brain.

Mild acute hypertension was induced in rats pharmacologically and non-pharmacologically with metaraminol, angiotensin II or bilateral vagotomy. The extraction fraction of water (Ew) was measured in both hypertensive and normotensive animals. It was demonstrated that cerebral blood flow did not change during hypertensive states. Under constant flow conditions Ew becomes a direct index of the cerebral capillary permeability to water. Ew was significantly decreased (P less than 0.001) in all brain regions examined in the hypertensive animals as compared to normotensive controls. However, the decrease Ew induced by hypertension was not observed if the animal had previously received bilateral carotid sinus denervation. This observation indicated that the capillary permeability response induced by hypertension is mediated by a neurogenic mechanism involving information transfer from the peripheral baroreceptors in the carotid sinus along afferent connections into the central nervous system.

Afferent Pathways↗
PubMed1984

Demonstration of retrogradely transported endogenous nerve growth factor in axons of sympathetic neurons.

The presence of retrogradely transported endogenous nerve growth factor (NGF) in sympathetic nerves of the guinea pig was demonstrated directly by fluorescent and peroxidase immunohistochemistry in the ligated superior postganglionic nerve of the superior cervical ganglion. Fixed, frozen sections of previously ligated nerve were incubated with either rabbit antiserum against guinea pig NGF (gpNGF), rabbit antibodies against gpNGF purified on a mouse NGF (mNGF) affinity column, the portion of rabbit antiserum against gpNGF that did not bind to the mNGF affinity column, or nonimmune rabbit serum. Positive staining on the peripheral side of the ligation was obtained only with unfractionated antiserum against gpNGF and with purified antibodies against gpNGF. The staining properties of the various antiserum preparations correlated with their ability to block gpNGF- and mNGF-induced neurite outgrowth in the embryonic chick dorsal root ganglion bioassay and in the PC12 bioassay. Homogenates of superior postganglionic nerve supported growth of embryonic chick dorsal root ganglia and differentiation of PC12 cells. This support was blocked by the specific antisera against NGF used in the immunohistochemistry experiments. These experiments demonstrate that endogenous NGF, presumably released by peripheral target tissues, is retrogradely transported in vivo.

Animals↗
PubMed1983

An endogenous inhibitory factor for choline acetyltransferase.

Chromatography of partially purified choline acetyltransferase (CAT) over carboxymethyl cellulose may result in the loss of up to 95% of the enzyme activity. This loss of activity can be prevented by running the chromatographs at low protein concentration with a large gradient volume suggesting that interactions between CAT and other endogenous proteins are involved in the mechanism of inactivation. Further experiments showed that CM-cellulose chromatography separates an endogenous inhibitory factor(s) and an endogenous activating factor(s) which protects the enzyme from the action of the former. The inhibitory factor elutes with CAT and produces almost complete inactivation unless the protein concentration is maintained below 0.05 mg/ml. Mixing experiments demonstrated that the activating factor is capable of blocking the effect of the inhibitory factor. The low degree of temperature dependence of the inhibitory factor essentially rules out the possibility that the inhibitor is a proteolytic enzyme. The I50 was estimated to be 10(-7) M or less suggesting a possible physiological role of these factors in the regulation of CAT activity.

Animals↗
PubMed1983

Choline acetyltransferase. Purification procedure and factors affecting chromatographic properties and enzyme stability.

Purified choline acetyltransferase had a specific activity of 142 mumol of acetylcholine produced min-1 mg-1 and consisted of two proteic forms with Mr = 72,000 and 76,000 on sodium dodecyl sulfate gel electrophoresis. The separation of more than one peak of enzyme activity on CM-cellulose chromatography was shown to reflect the interaction of choline acetyltransferase with other proteins rather than the resolution of different isoenzymes. Purified choline acetyltransferase exhibited a high degree of stability. Enzyme stability was greatly dependent upon the procedures used to reach a given degree of purity, rather than the degree of purity per se, suggesting that specific proteins may be involved in the mechanism of enzyme inactivation. Use of affinity chromatography over Sepharose-blue dextran early in the preparation produced enzyme which was unstable both to storage and to concentration. Although the addition of other proteins such as bovine serum albumin had no significant stabilizing effect, the presence of acetyl coenzyme A during concentration prevented inhibition. Finally, it was shown that the purified enzyme is representative of the total enzyme present in brain in terms of both specific activity and immunochemical properties.

Animals↗
PubMed1983

Synthesis of a naphthylvinylpyridine derivative and its use for affinity chromatography of choline acetyltransferase.

N-(10-carboxy)decamethylene-4(1-naphthylvinyl)pyridinium chloride, a derivative of the choline acetyltransferase (CAT) inhibitor naphthylvinylpyridine (NVP) was synthesized and used as a ligand for affinity chromatography of choline acetyltransferase. The preparation of this inhibitor included the quaternization of naphthylvinylpyridine with 11-Br-undecanoic acid methyl ester to obtain N-(10-carbomethoxy)decamethylene-4-(1-naphthylvinyl)pyridinium bromide, followed by hydrolysis to free the carboxylic group. This inhibitor (C11-NVP+) had a potency comparable to that of N-methyl-4(1-naphthylvinyl) pyridinium iodide (C1-NVP+) which is the most potent derivative of NVP but which lacks a functional group for conjugation to Sepharose. The C11-NVP+ was then bound through the carboxylic group to aminoalkyl Sepharose by a carbodiimide promoted condensation reaction. Interaction of CAT with the inhibitor retarded its elution from a column of Sepharose-C11-NVP+ and permitted the purification of the enzyme to electrophoretic homogeneity starting from a preparation in which CAT represented about 20% of the total proteins. Conventional procedures of protein purification had previously been unsuccessful in isolating the enzyme in pure form. Inhibition studies showed that CAT could exhibit either a "high" or a "low" sensitivity to inhibition by naphthylvinylpyridine and its derivatives (I50 with C1-NVP+ = 0.57 microM or 5.2 microM). A direct relationship existed between the sensitivity of CAT to these inhibitors and the retention of the enzyme by the affinity column.

Chemical Phenomena↗
PubMed1982

Antidepressants alter cerebrovascular permeability and metabolic rate in primates.

External detection of the annihilation radiation produced by water labeled with oxygen-15 was used to measure cerebrovascular permeability and cerebral blood flow in six rhesus monkeys. Use of oxygen-15 also permitted assessment of cerebral metabolic rate in two of the monkeys. Amitriptyline produced a dose-dependent, reversible increase in permeability at plasma drug concentrations which are therapeutic for depressed patients. At the same concentrations the drug also produced a 20 to 30 percent reduction in cerebral metabolic rate. At higher doses normal autoregulation of cerebral blood flow was suspended, but responsivity to arterial carbon dioxide was normal.

Amitriptyline↗
PubMed1982

A new double labeling technique using tyrosine hydroxylase and dopamine-beta-hydroxylase immunohistochemistry: evidence for dopaminergic cells lying in the pons of the beef brain.

Despite numerous data on the organization of the central catecholaminergic (CA) system, the nature of many CA terminal fields, fiber pathways and cell bodies, remains unknown. The discrepancies between the results obtained by CA histofluorescence and dopamine-beta-hydroxylase (DBH) immunofluorescence method suggest that certain areas described as noradrenergic (NA) may be in fact dopaminergic (DA). A method to solve this problem is to compare the localization of the two CA synthetizing enzymes: tyrosine hydroxylase (TH) and DBH. The first enzyme is a marker of all CA neurons, whereas the second one is a specific marker of noradrenergic and adrenergic neurons. We decided to develop a technique which allows visualization of both antigens on a same section. We chose to combine immunofluorescence and immunoperoxidase techniques which yield very different staining. The optimal conditions for this double labeling technique were developed in bovine brain, the homologous system for our antibodies. The results obtained by this procedure provide evidence that DA cells together with NA cells extend throughout the rostral part of the pons, and outline a successful technique for a more extensive investigation of the DA system. The same method should be applicable to other investigations involving other antigens.

Animals↗
PubMed1982

An intravenous technique for the measurement of cerebral vascular extraction fraction in the rat.

An intravenous injection method to measure cerebral vascular extraction fractions of highly diffusible substances in the rat is described. The brain extraction fractions of 3H-labeled water (Ew) and ethanol (Ee) were defined as the ratio of either of those tracers to the freely diffusible reference tracer, 14C-butanol, in the brain, divided by the ratio of the tracers available for the extraction during the time between simultaneous intravenous injection of the tracers and decapitation of the rat. Ew and Ee were measured in five regions of brain, including brainstem and cerebellum, under PaCO2 conditions ranging from 15 to 85 mm Hg. The extraction fractions for both test tracers were shown to vary with PaCO2-induced flow changes according to the equation, ln(1 - E) = -PS/F. When PS/F values calculated from regional measurements of Ew and Ee were plotted versus PaCO2, least squares regression equations of the plots could be used to compare permeabilities of both tracers at any given PaCO2 value. Ratios of the permeabilities of water and ethanol varied regionally but were relatively constant in a given region under different flow states. This intravenous injection method allows for accurate measurement of the extraction fractions of even highly diffusible tracers under varied flow conditions in all brain regions regardless of arterial blood supply.

Animals↗
PubMed1982

Development and maturation of central nervous system myelin: comparison of immunohistochemical localization of proteolipid protein and basic protein in myelin and oligodendrocytes.

The immunohistochemical localization of two myelin specific proteins-basic protein (BP) and proteolipid protein (PLP)-was compared during the process of myelination. Although both proteins were present in oligodendrocytes, (i) neither protein was observed in oligodendrocytes not already closely associated with nerve fibers exhibiting a fluorescent coating; (ii) in any discrete anatomical area oligodendrocytes were positive for BP before PLP was visible; and (iii) as myelination progressed, immunoreactivity for BP in oligodendrocytes appeared to decrease and simultaneously PLP immunofluorescence became visible in this cell type. During the period of active myelination, fibers exhibited a distinct varicose appearance. As myelination progressed, the myelin sheath increased in thickness and these varicosities became less prominent, eventually completely disappearing. Therefore, the nature and the appearance of varicosities can be used as an index of the relative stage of maturation of myelin in an individual fiber. In general, PLP appeared in fibers at a later stage of maturation than did BP based on the above criteria. However, in a relatively small number of fine fibers PLP was observed at a very early stage. In fully mature myelin, very large fibers were frequently more intensely fluorescent for BP than PLP, whereas fine myelinated fibers were more intensely stained for PLP. These observations are consistent with the following interpretations. (i) Substantial differentiation of oligodendrocytes occurs prior to appearance of either of these proteins by immunofluorescence. (ii) BP is added to the myelin sheath prior to PLP and there appears to be a shift in priority of synthesis from BP to PLP in individual oligodendrocytes during the process of myelination. (iii) Very small fibers often contain low concentrations of BP relative to PLP, and conversely, very large fibers may contain a high concentration of BP relative to PLP. Thus, the relative concentration of these proteins in myelin appears not to be constant but may vary as a function of the size of the myelinated fiber.

Aging↗
PubMed1982

Role of the central adrenergic system in mediating amitriptyline-induced alteration in the mammalian blood-brain barrier in vivo.

All tricyclic antidepressants increase the degree of equilibration of [3H]water across the cerebral capillary (Ew) as measured by a dual-label radioactive tracer technique. By using amitriptyline (AMI) as a prototype, a series of studies was conducted to determine the mechanism for this drug effect. The AMI-induced increase in Ew was blocked by 6-hydroxydopamine ablation of central aminergic neurons and by pretreatment with phenoxybenzamine, an alpha adrenergic antagonist. Pretreatment with propranolol, a beta adrenergic antagonist, did not block the AMI-induced increase. Central serotonergic ablation by p-chloroamphetamine had no effect on the AMI-induced increase. Treatment with atropine and hydroxyzine separately also did not alter Ew. Based on these results, the AMI-induced increase in Ew appears to be mediated by the effect of the drug on central adrenergic neurons. The serotonergic, anticholinergic and antihistaminergic actions of AMI, by themselves, do not appear to play a role in this phenomenon. The results are compatible with the concept that the central adrenergic system functions, in part, to regulate the cerebromicrocirculation.

Amitriptyline↗
PubMed1981

Immunohistochemical localization of catechol-O-methyltransferase in circumventricular organs of the rat: potential variations in the blood-brain barrier to native catechols.

Catechol-O-methyltransferase (COMT) was localized in the organum vasculosum of the lamina terminals, subfornical organ, subcommissural organ and area postrema of rat brain using an indirect immunofluorescence technique. COMT immunofluorescence was apparent in neuroglia within the organum vasculosum and was most intense in the ependyma between this structure and the optic recess of the third ventricle. In both the subfornical organ and the area postrema, COMT was localized in a neuroglial network, but was noticeably absent in the ependymal layer. COMT immunofluorescence in the ependyma of the subcommissural organ was continuous with the more intense immunofluorescence of the cuboidal ependyma of the third ventricle. Each of the circumventricular organs studied, with the exception of the subcommissural organ, lies outside the blood-brain barrier. However, the unique pattern of COMT immunofluorescence in the area postrema and the subfornical organ suggests that these two structures, of all circumventricular organs, are most likely to permit the entry of peripherally circulating catechols to the cerebrospinal fluid.

Animals↗
PubMed1981

Measurement of cerebral vascular extraction fractions in the rat using intracarotid injection techniques.

A small volume (5 microliters) common carotid arterial injection method is described for the quantitation of cerebral vascular extraction fractions (Et) of diffusion limited tracer molecules in the rat. The method is a modification of a technique introduced by Oldendorf and widely used for the study of blood-brain barrier phenomena. While the Oldendorf technique has proven valuable for estimating the relative permeabilities of substances, it is limited in measuring Et under conditions of physiologically or pharmacologically altered permeability or blood flow. The method described in this paper--using a small volume (5 microliters) common carotid injection, a freely diffusible reference tracer, [14C]butanol, and a 5 sec circulation time--allows for measurements of Et that reflect changes in blood flow and small differences in permeability. The modified method is important for the study of the regulation of cerebral vascular permeability and flow in an inexpensive animal model.

Animals↗
PubMed1981

An immunohistochemical study of the organization of catecholaminergic cells and terminal fields in the paraventricular and supraoptic nuclei of the hypothalamus.

The distribution of catecholaminergic fibers and cell bodies in the paraventricular and supraoptic nuclei of the hypothalamus was investigated with immunohistochemical methods in the adult albino rat. Sections through the nuclei were stained with antisera to the catecholamine synthesizing enzymes tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH), and phenylethanolamine-N-methyltransferase (PNMT). The results suggest that adrenergic (PNMT-stained) fibers innervate the entire parvocellular division of the paraventricular nucleus, although the highest density of fibers was found in the medial part of the division. Only widely scattered adrenergic fibers are found in the magnocellular division of the nucleus and in the supraoptic nucleus. Noradrenergic fibers appear to innervate the periventricular zone of the paraventricular nucleus and those parts of the paraventricular and supraoptic nuclei that contain predominantly vasopressinergic neurons in both the normal and in the homozygous Brattleboro rat. Significant numbers--somewhat more than 500--of dopaminergic (TH-stained) neurons are found in the paraventricular nucleus; the cells are distributed throughout the nucleus but are concentrated in the medial and periventricular parts of the parvocellular division. Double-labeling experiments with the retrogradely transported tracer true blue indicate that between 4% and 8% of the dopaminergic neurons in the paraventricular nucleus project to the region of the dorsal vagal complex and/or thoracic levels of the spinal cord. It is concluded that adrenergic inputs to the paraventricular nucleus may influence cells that project to the median eminence and to preganglionic autonomic cell groups in the medulla and spinal cord. Noradrenergic inputs to the supraoptic and paraventricular nuclei may influence primarily vasopressinergic cells that project to the posterior lobe of the pituitary, as well as cells in the periventricular part of the paraventricular nucleus that project to the median eminence.

Animals↗