Biomedical subjects

E L Gustafson

Publications and source records attributed to E L Gustafson.

31 records · Page 2Linked to original sources

PubMed1992

Developmental regulation of phosphoprotein gene expression in the caudate-putamen of rat: an in situ hybridization study.

The regional and cellular ontogeny of the mRNA encoding the dopamine- and cAMP-regulated phosphoprotein, DARPP-32, has been studied in rat striatum by quantitative in situ hybridization histochemistry. The mRNA for DARPP-32 exhibited a characteristic developmental profile. The hybridization signal was first visible on the day of birth, at which time DARPP-32 mRNA was concentrated in patches in the caudate-putamen. By the end of the first postnatal week, the majority of neurons in the caudate-putamen expressed the DARPP-32 message. Levels of mRNA per cell increased markedly during the second postnatal week, and peaked around the beginning of the third week. The adult level of DARPP-32 mRNA was lower than that observed at the apex of mRNA expression, on a per cell basis, while the proportion of neurons expressing detectable levels of message remained relatively constant. In the nucleus accumbens and olfactory tubercle, DARPP-32 mRNA development lagged somewhat behind that observed in the caudate-putamen, but was similar in other respects. A non-quantitative study employing an oligonucleotide probe complementary to the mRNA encoding another cAMP-regulated phosphoprotein, ARPP-21, revealed a similar developmental sequence to DARPP-32. The present results suggest that for DARPP-32 mRNA, genetic and, possibly, environmental factors play a role in determining the developmental patterns observed.

Aging↗

PubMed1991

Immunocytochemical localization of phosphatase inhibitor-1 in rat brain.

The localization of phosphatase inhibitor-1 was investigated in rat brain by use of immunocytochemistry. Studies were performed with an affinity purified IgG raised against purified rabbit skeletal muscle inhibitor-1. In rat brain tissue homogenates, this antibody reacted only with a 29 kDa protein corresponding to inhibitor-1. Immunocytochemical studies with this antibody revealed numerous immunoreactive cell bodies and fibers. The highest concentration of immunoreactive perikarya was observed in the caudate-putamen and nucleus accumbens, and these appeared to be exclusively medium-sized neurons. Other areas containing substantial populations of immunoreactive neurons included the suprachiasmatic nucleus of the hypothalamus, lateral hypothalamus, horizontal limb of the diagonal band of Broca, dentate gyrus of the hippocampal formation, habenula, superior colliculus, claustrum, endopiriform nuclei, and neocortex. The distribution of terminals containing inhibitor-1 coincided with the distribution of terminal fields known to originate from the above regions. Thus, plexuses of immunoreactive axons were seen in the globus pallidus, substantia nigra pars reticulata, paraventricular hypothalamus, dorsal thalamus, CA3 region of the hippocampus, and interpeduncular nucleus. These results demonstrate that phosphatase inhibitor-1, a cyclic AMP-regulated inhibitor of phosphatase-1, is differentially distributed in the rat CNS. Given the widespread role of protein phosphorylation and dephosphorylation in intracellular signal transduction, these results suggest that neurons containing high levels of inhibitor-1 may share common, hitherto unrecognized, properties in terms of neurotransmitter regulation and/or responsiveness.

Animals↗

PubMed1990

Localization of DARPP-32 immunoreactive neurons in the bed nucleus of the stria terminalis and central nucleus of the amygdala: co-distribution with axons containing tyrosine hydroxylase, vasoactive intestinal polypeptide, and calcitonin gene-related peptide.

The distribution and morphology of neurons containing the dopamine- and cyclic AMP-regulated phosphoprotein, DARPP-32, were investigated in the bed nucleus of the stria terminalis (BST) and the central nucleus of the amygdala (CeA). DARPP-32 immunoreactive neurons are numerous in both regions, but are restricted to the lateral dorsal and the lateral juxtacapsular subdivisions of the BST, and the central lateral and lateral capsular subdivisions of the CeA. Immunoreactive neurons in the lateral dorsal BST, and the central lateral and lateral capsular CeA are similar morphologically, while those in the juxtacapsular BST appear to be a subpopulation of striatal medium-sized spiny neurons. The distribution of DARPP-32 immunoreactive neurons in the BST and CeA overlaps considerably with axonal plexuses containing tyrosine hydroxylase (TH), vasoactive intestinal polypeptide (VIP), and calcitonin gene-related peptide (CGRP). These studies provide further evidence of the close relationship between the CeA and BST, and also provide anatomical evidence for possible interactions between neurotransmitters, neuropeptides, and phosphoproteins.

Amygdala↗

PubMed1990

Characterization in mammalian brain of a DARPP-32 serine kinase identical to casein kinase II.

DARPP-32, a dopamine- and cyclic AMP-regulated phosphoprotein of Mr 32,000, is phosphorylated in vitro by casein kinase II at a site which is also phosphorylated in intact cells. In the present study, we show that a protein kinase activity, present in caudate-putamen cytosol, phosphorylates DARPP-32 on a seryl residue located on the same thermolytic peptide that is phosphorylated by purified casein kinase II. This DARPP-32 serine kinase was indistinguishable from casein kinase II on the basis of a number of biochemical criteria. Excitotoxic lesions of the caudate-putamen and immunocytochemistry revealed the presence of casein kinase II in the medium-sized striatonigral neurons which are known to contain DARPP-32. Casein kinase II activity was high in all rat brain regions studied, and casein kinase II-like immunoreactivity was detected in most brain neurons, although some neuronal populations (e.g., cortical pyramidal cells and large striatal neurons) were stained more intensely than others. In rat caudate-putamen, 45% of the total casein kinase II activity was in the cytosol and 20% in the synaptosomal fraction. In mouse cerebral cortex and caudate-putamen, casein kinase II activity was high at embryonic day 16, and remained elevated during development. In addition to DARPP-32, several major substrates for casein kinase II were observed specifically in brain, but not in liver extracts. The high activity of casein kinase II in brain from the embryonic period to adult age and the existence of a number of specific substrates suggest that this enzyme may play an important role in both developing and mature brain, possibly in modulating the responsiveness of target proteins to various extracellular signals.

Animals↗

PubMed1990

Immunocytochemical localization of the neuron-specific form of the c-src gene product, pp60c-src(+), in rat brain.

Neurons express high levels of a variant form of the c-src gene product, denoted pp60c-src(+), which contains a 6 amino acid insert in the amino-terminal half of the c-src protein. We have determined the localization of pp60c-src(+) in neurons using an affinity-purified anti-peptide antibody, referred to as affi-SB12, that exclusively recognizes this neuron-specific form of the c-src gene product. Using affi-SB12, we examined the distribution of pp60c-src(+) by immunoperoxidase staining of sections through adult rat brains, pp60c-src(+) was widely distributed in rat brain and appeared to be differentially expressed in subpopulations of neurons. The majority of immunoreactive neurons was found in the mesencephalon, cerebellum, pons, and medulla. Telencephalic structures that contained substantial populations of pp60c-src(+)-immunoreactive neurons included layer V of the cerebral cortex and the ventral pallidum. Within individual neurons, pp60c-src(+) immunoreactivity was localized to the cell soma and dendritic processes, while labeling of axons and nerve terminals (puncta) was not as readily detected. Dense accumulations of immunoreactive axons were rare, being most prominent in portions of the inferior and superior olive, and in the spinal trigeminal nucleus. While the regional distribution of pp60c-src(+) immunoreactivity does not correlate with any specific neuronal cell type or first messenger system, this unique pattern of expression of pp60c-src(+) suggests the existence of a previously uncharacterized functional organization within the brain. Furthermore, the localization of this neuron-specific tyrosine kinase in functionally important areas of the nerve cell, namely, dendritic processes, axons, and nerve terminals, suggests that pp60c-src(+) may regulate pleiotropic functions in specific classes of neurons in the adult central nervous system.

Animals↗

PubMed1990

Differential expression of ARPP-16 and ARPP-19, two highly related cAMP-regulated phosphoproteins, one of which is specifically associated with dopamine-innervated brain regions.

ARPP-16 and ARPP-19 are 2 cAMP-regulated phosphoproteins of Mr = 16,000 and 19,000, respectively, which are identical except for the presence of 15 additional amino acids on the NH2-terminus of ARPP-19. The phosphorylation of these 2 proteins is regulated by cAMP and vasoactive intestinal peptide in reaggregate striatal cultures (Girault et al., 1988). Using immunoblots and immunocytochemistry, we have compared the regional, subcellular, phylogenetic, and ontogenetic distributions of these 2 proteins. ARPP-19 was found in all vertebrate species studied and, at various levels, in all tissues of adult rat. ARPP-19 was also present at high levels in malignant cell lines. During development ARPP-19 concentrations were highest in the embryo and decreased during the pre- and postnatal periods. In contrast, ARPP-16 was detected only in some specific neurons of the dopamine-innervated regions of the basal ganglia and the cerebral cortex, which are known to possess D1 dopamine receptors, in particular the striatonigral neurons. ARPP-16 is phylogenetically recent, being found only in birds and mammals, and appears late in ontogenesis, increasing during the postnatal period. These 2 proteins provide a unique model for studying the specificity of signal transduction and gene expression in dopaminoceptive neurons.

Animals↗

PubMed1989

Spatial relationship of the striatonigral and mesostriatal pathways: double-label immunocytochemistry for DARPP-32 and tyrosine hydroxylase.

Antibodies to tyrosine hydroxylase and DARPP-32 were used to examine the spatial arrangement between mesostriatal dopamine projections and the reciprocal pathway from DARPP-32-containing neurons in the basal forebrain. Use of a double-labeling immunocytochemical procedure demonstrated that the mesostriatal and striatonigral pathways run in close proximity throughout the rostral mesencephalon and basal forebrain. The majority of descending axons immunoreactive for DARPP-32 appear to originate in the striatum, including the nucleus accumbens, and run through the internal capsule to innervate the globus pallidus, entopeduncular nucleus, and all subdivisions of the substantia nigra. The ventral tegmental area is sparsely invested with DARPP-32-immunoreactive axons. At all levels, there are also fascicles of DARPP-32-containing fibers which run ventromedial to the internal capsule in the medial forebrain bundle, and which are coextensive with ascending axons of the mesencephalic dopamine the internal capsule in the medial forebrain bundle, and which are coextensive with ascending axons of the mesencephalic dopamine cell groups. Tyrosine hydroxylase-immunoreactive axons are coextensive with DARPP-32-immunoreactive axons in the internal capsule entopeduncular nucleus, and globus pallidus, as well as much of the remainder of the basal forebrain. Although the main source of descending DARPP-32 immunoreactive axons would appear to be the striatum, other possible sources are also discussed.

Animals↗

PubMed1987

Noradrenaline and neuropeptide Y innervation of the rat hypothalamus are differentially affected by 6-hydroxydopamine.

The neuropeptide Y (NPY) innervation of the hypothalamus is thought to arise largely from noradrenaline (NA) neurons of the medullary tegmentum in the rat. This view was tested in this study by analyzing the effects of intraventricular injections of 6-hydroxydopamine (6-OHDA) on hypothalamic NA and NPY innervation. 6-OHDA markedly depletes or eliminates the NA innervation of the hypothalamus, as demonstrated by dopamine-beta-hydroxylase immunohistochemistry, but does not affect the NPY innervation of the hypothalamus. These results indicate that the hypothalamic NPY innervation arises in large part from intrinsic NPY-producing neurons rather than from medullary neurons in which NA and NPY coexist.

Animals↗

PubMed1987

Noradrenaline neuron plasticity in developing rat brain: effects of neonatal 6-hydroxydopamine demonstrated by dopamine-beta-hydroxylase immunocytochemistry.

The present study was conducted to assess the morphological changes produced by neonatal administration of 6-hydroxydopamine (6-OHDA) in the noradrenergic innervation of the developing and adult rat brain. As demonstrated by dopamine-beta-hydroxylase (DBH) immunohistochemistry, the major alterations are the following. First, neocortical and hippocampal noradrenergic innervation is permanently eliminated by the treatment, with lesser effects on other telencephalic structures. These changes appear by postnatal day 5 and are permanent in nature. In adult treated animals, most thalamic nuclei are hyperinnervated by DBH-immunoreactive axons as are the cerebellum and a number of brainstem nuclei. The hyperinnervation of these structures occurs after postnatal day 20, and is extremely specific, with the pattern of organization and distribution of noradrenergic axons in treated animals identical to that of controls. In contrast, the noradrenergic innervation of the hypothalamus is relatively unaffected by 6-OHDA treatment. The principal exception is the development of an anomalous plexus of DBH immunoreactive axons in the lateral hypothalamus. The timing and organization of the changes produced by neonatal 6-OHDA administration are consistent with the hypothesis that noradrenergic neurons, and particularly those of the locus coeruleus, are programmed to produce a defined amount of axon and terminal field, with any developmental loss resulting in a 'pruning effect' such that the total terminal field appears conserved. Given the specificity of the hyperinnervation, inductive influences from the target nuclei probably play a major role in determining the pattern of the noradrenergic innervation.

Animals↗

PubMed1986

Neuropeptide Y localization in the rat amygdaloid complex.

Neuropeptide Y (NPY)-, avian pancreatic polypeptide (APP)-, and molluscan cardioexcitatory peptide (FMRF)-like immunoreactivity in the amygdaloid complex of the rat was investigated immunohistochemically. The distribution of each of these peptides within the amygdala is identical and cross-blocking studies indicate that all three antisera recognize the NPY antigen. Morphologically distinct populations of NPY immunoreactive neurons are differentially distributed in the medial amygdaloid nucleus and at the base of the stria terminalis. Dense plexuses of immunoreactive axons are present in the medial third of the central nucleus and in the dorsal half of the medial nucleus, with light to moderate fiber plexuses present in the lateral and basolateral nuclei and scattered axons present throughout the remainder of the amygdala. The distribution and appearance of NPY immunoreactive plexuses in the amygdala is similar to that described previously for noradrenergic axons arising from brainstem cell groups (Fallon, Koziell, and Moore: J. Comp. Neurol. 180:509-532, '78). However, injections of the noradrenergic neurotoxin 6-hydroxydopamine into the amygdala result in a complete loss of dopamine-beta-hydroxylase (DBH) immunoreactivity in the amygdala and surrounding cortex but leave much of the NPY immunoreactive plexus intact. Similarly, lesions of the locus coeruleus deplete DBH immunoreactivity, leaving NPY-like immunoreactivity in the amygdala unaffected. These results indicate that much of the NPY immunoreactive plexus observed in the amygdala does not arise from brainstem sources in which NPY and noradrenaline are colocalized. Lesions of the stria terminalis or medial nucleus have no observable effect on the density or distribution of NPY immunoreactive terminal fields in the basal forebrain and hypothalamus, suggesting that immunoreactive neurons in the amygdaloid complex do not contribute significantly to this innervation.

Amygdala↗

PubMed1984

Identical immunoreactivity of afferents to the rat suprachiasmatic nucleus with antisera against avian pancreatic polypeptide, molluscan cardioexcitatory peptide and neuropeptide Y.

Avian pancreatic polypeptide (APP)-like, molluscan cardioexcitatory peptide (FMRF)-like and neuropeptide Y (NPY)-like immunoreactivities were studied in a secondary visual pathway in rat brain. The cell bodies of this pathway are located in the lateral geniculate nucleus and its terminal plexus is found in the suprachiasmatic hypothalamic nucleus (SCN). The neurons and terminal plexus demonstrated by antiserum to each peptide are identical, and immunoreactivity is blocked by preabsorption of each antiserum with a low concentration of the antigen against which it was raised. Immunoreactivity is also blocked by preabsorption of each antiserum with either NPY or APP. In contrast, APP- and NPY-like immunoreactivities are blocked only partially when these antisera are preabsorbed with concentrations of FMRF as high as 100 microM. Since NPY is the only one of these peptides that has been isolated from mammalian brain, we conclude that NPY is the endogenous CNS peptide produced by neurons of the lateral geniculate-SCN projection.

Afferent Pathways↗

PubMedDate not supplied

Localization of 5-HT1B, 5-HT1D alpha, 5-HT1E and 5-HT1F receptor messenger RNA in rodent and primate brain.

In situ hybridization histochemistry (ISHH) was used to study the distribution of various 5-HT1 receptor messenger RNAs (mRNA) in the mammalian nervous system. Since the cDNAs encoding the different 5-HT1 receptors, have not been cloned in one single species, brains of the species appropriate for the 5-HT1 receptor messenger RNA (mRNA) have been used. Thus, 5-HT1B and 5-HT1D alpha mRNA were determined in rat and mouse brain, while 5-HT1E and 5-HT1F mRNA were studied in human (and monkey) and guinea-pig brain, respectively. 5-HT1B and 5-HT1D alpha hybridization signals were predominantly present in caudate-putamen and cortical areas; in addition, 5-HT1B mRNA was also detected in hippocampus, cerebellum and cerebral arteries. In general, the distribution of 5-HT1B mRNA was characterized by high densities, whereas 5-HT1D alpha mRNA was expressed at very low levels. Comparison of the localization of the mRNAs to the regional distributions of the 5-HT1B and 5-HT1D binding sites in rat brain (described in a previous study), revealed that both receptor subtypes could be putative presynaptic heteroreceptors, modulating the release of various neurotransmitters in the central nervous system. The mRNA encoding the recently cloned 5-HT1E receptor, which has low affinity for the 5-HT1 receptor ligand 5-carboxamidotryptamine (5-CT), was localized in human brain. It was found to be present in cortical areas, caudate, putamen and amygdala, areas known to contain 5-CT insensitive 5-HT1 binding sites. The regional distribution of the 5-HT1F mRNA was determined in guinea-pig brain: high densities were observed in various cortical areas, the hippocampal formation and claustrum, which are regions known to contain 5-CT insensitive 5-HT1 or non 5-HT1A/1B/IC/ID [3H]5-HT binding sites. Altogether, this ISHH study describes the distribution of mRNAs of recently cloned 5-HT1 receptors in rodent and primate brain and compares these results to the distribution of the heterogeneous population of 5-HT1 binding sites.

Animals↗

PubMedDate not supplied

The distribution of dopamine-beta-hydroxylase, neuropeptide Y and galanin in locus coeruleus neurons.

The locus coeruleus (LC) is composed of noradrenaline-producing neurons that project widely throughout the neuraxis. Subpopulations of LC neuron perikarya have been shown to contain neuropeptide Y (NPY) and galanin (GAL). In the major terminal fields of LC projections, the cerebral cortex, dorsal thalamus and cerebellar cortex, there are differing plexuses of dopamine-beta-hydroxylase (DBH), NPY and GAL immunoreactive axons. DBH immunoreactive plexuses are found in all areas which conform in appearance to previous demonstrations of noradrenaline localization by fluorescence histochemistry. In contrast, there are few NPY immunoreactive axons in thalamus and cerebellum, and the cortical plexus, while similar to the DBH immunoreactive plexus, is not affected by 6-hydroxydopamine treatment. Similarly, there are few GAL immunoreactive axons in either cerebral cortex, dorsal thalamus or cerebellar cortex. Transection of ascending LC axons results in accumulation of DBH but not NPY or GAL immunoreactivity proximal to the lesion. These observations indicate that NPY and GAL are distributed differently in LC neurons from noradrenaline and DBH.

Animals↗