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Biomedical subjects

M Goldstein

Publications and source records attributed to M Goldstein.

At least 343 records · Page 19Linked to original sources

Studies on dopamine-, tyrosine hydroxylase- and aromatic L-amino acid decarboxylase-containing cells in the rat diencephalon: comparison between formaldehyde-induced histofluorescence and immunofluorescence.

The morphology, number and distribution of catecholaminergic neurons, as visualized either with the aluminum-catalysed formaldehyde method for catecholamines or with the immunohistochemical method for the catecholamine-synthesizing enzymes tyrosine hydroxylase and aromatic L-amino acid decarboxylase, respectively, were analysed within the rat dorsal hypothalamus, ventral thalamus and adjoining regions (A11 and A13 cell groups). Both polyclonal rabbit and monoclonal mouse tyrosine hydroxylase antibodies were used in elution-restaining and double-staining experiments, respectively. Some of the animals also received spinal injections of the fluorescent tracer True Blue in order to retrogradely label cells projecting to the spinal cord. With respect to the number and distribution of catecholaminergic neurons in the A11 and medial A13 cell groups, including the spinal-projecting subpopulation, the results obtained with the two methods were very similar, indicating that within these regions of the CNS the two methods in principle visualize identical cell populations. However, the catecholaminergic cells were distinctly larger and their processes appeared more extensive with the immunohistochemical method. Animals processed for immunohistochemistry exhibited a lower total number of retrogradely labelled cells in the A11 area than those analysed with aldehyde-induced fluorescence despite the fact that both methods revealed similar numbers of retrogradely labelled tyrosine hydroxylase-positive and catecholamine-containing cells, respectively. The reason for these discrepancies, which are probably of methodological nature, are discussed. While this study shows that the results obtained with the two methods within the A11 and medial A13 cell group are very similar and thus strengthens the earlier proposed concept of the organization of the diencephalospinal dopaminergic system, it also documents that in intermingling and nearby CNS regions there are cell bodies which cannot be demonstrated with the aldehyde fluorescence method, but which still contain tyrosine hydroxylase and/or aromatic L-amino acid decarboxylase-like immunoreactivity. One explanation is low levels of enzyme and/or dopamine combined with a comparatively low sensitivity of the histochemical method. Thus, neurons containing both enzymes are probably dopaminergic, even if catecholamine fluorescence cannot be demonstrated. Neurons containing tyrosine hydroxylase, but lacking both aldehyde induced fluorescence and aromatic L-amino acid decarboxylase, may also still be dopaminergic.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Time course of MPTP-induced degeneration of the nigrostriatal dopamine system in C57 BL/6 mice.

MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) is a parkinsonism-inducing dopamine (DA) neurotoxin most effective in primates. MPTP also causes a degeneration of both perikarya and axon terminals of the nigrostriatal DA neurons in C57 BL/6 mice. The time courses of the changes in tyrosine hydroxylase immunoreactive objects, endogenous DA concentrations and specifically bound 3H-mazindol as markers of the integrity of DA neurons were studied in substantia nigra and striatum of adult C57 BL/6 mice, after systemic treatment with MPTP or intranigral injections of the catecholamine neurotoxin 6-hydroxydopamine (6-OHDA). A rapid decrease in the three parameters studied was found in the substantia nigra during the first 2 days after MPTP-treatment while the MPTP-induced effects in the striatum were more protracted and maximal reduction was observed 7 days after MPTP. A basically similar pattern was found when studying the 6-OHDA-induced anterograde degeneration of the nigrostriatal system. These results indicate that in C57 BL/6 mice, MPTP primarily destroys the DAergic perikarya with a subsequent anterograde degeneration of the striatal axon terminals, although a limited rapid destruction of some striatal terminals cannot be excluded.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

Ontogeny of the dopamine and cyclic adenosine-3':5'-monophosphate-regulated phosphoprotein (DARPP-32) in the pre- and postnatal mouse central nervous system.

The ontogeny of a dopamine and cyclic adenosine-3':5'-monophosphate-regulated phosphoprotein with an apparent molecular weight of 32 kilodaltons (DARPP-32) has been studied in the central nervous system of the prenatal, newborn and adult mouse. DARPP-32-immunoreactive somata were first identified at day 12 of gestation, in the primary olfactory cortex and in the ventrolateral medulla oblongata. On day 14 of gestation, neurons containing DARPP-32-like immunoreactivity became apparent in the caudate nucleus, olfactory tubercle, nucleus accumbens, frontoparietal cortex and the ventral medulla oblongata. During the period up to and including birth, the number of cell bodies and fibres in all these areas increased markedly. In addition, DARPP-32-positive neurons became visible in the olfactory nucleus, the arcuate nucleus, and DARPP-32-positive cells appeared in the choroid plexus of the lateral, third and fourth ventricles. DARPP-32-containing fibres could be seen in the median eminence, the ventrolateral thalamus, and in the striatonigral projection, descending in the internal capsule to ramify extensively in the substantia nigra. Only in the cerebellum and suprachiasmatic nucleus did the development of DARPP-32-like immunoreactivity occur postnatally. The development of tyrosine hydroxylase, the rate-limiting enzyme for catecholamine synthesis, was simultaneously examined. The arrival of the tyrosine hydroxylase-containing projection to the caudate nucleus, the olfactory tubercle and the nucleus accumbens apparently occurred 1-2 days after the appearance of DARPP-32-immunoreactive cells within these regions. In the ventral and ventrolateral medulla oblongata, and the primary olfactory cortex, no tyrosine hydroxylase innervation was seen near the DARPP-32-positive neurons at days 12-14. The organization of the DARPP-32-containing somata of the caudate nucleus into aggregates of 5-15 neurons was partly paralleled spatially by an increased density of tyrosine hydroxylase-positive fibres. Many DARPP-32-immunoreactive cells in the immature mouse brain are present by the day of birth, particularly in the areas known to receive a dopaminergic innervation. The development of these presumptive dopaminoceptive DARPP-32-containing neurons does not seem to be dependent on the presence, however, of a dopaminergic input, since in all regions examined DARPP-32-LI preceded the appearance of tyrosine hydroxylase-like immunoreactivity by at least 1-2 days. Indeed, the results suggest that the existence of DARPP-32-like immunoreactivity in cell bodies and dendrites may be a pre-requisite for the formation or subsequent stabilization of dopaminergic synapses.

Aging↗

Men with congenital absence of the vas deferens often have seminal vesicles.

The absence of semen fructose and low ejaculate volume in men with congenital absence of the vas deferens have been attributed to an associated agenesis of the seminal vesicles. Because vasography and retrograde seminal vesiculography cannot be performed in these men, this association has never been confirmed reliably. Therefore, we performed computerized tomographic scans of the seminal vesicles on 26 men with absence of the vas deferens. We found seminal vesicles bilaterally in 12 of these men, unilateral hypoplasia or absence in 8 and bilateral hypoplasia or absence in 6. Four men with normal seminal vesicles and 1 with unilateral hypoplasia underwent exploration in an attempt to find reconstructable vasa. We failed to identify a vas deferens in any of these men. These observations suggest that negative semen fructose and low ejaculate volume in men with absence of the vas deferens may be owing to agenesis or obstruction of the ejaculatory ducts rather than absent seminal vesicles.

Adult↗

Correction of severe penile curves with tunica albuginea autografts.

Nesbit's technique of excising ellipses of the tunica albuginea has been effective in correcting penile curvatures. When this procedure is used for severe angulations, penile shortening can be significant. We report the use of Nesbit ellipses autografted to the contralateral corpus for correction of severe penile curvatures in 3 men. This method resulted in a straight penis with minimal shortening.

Humans↗

Serum ferritin levels in preterm infants after multiple blood transfusions.

We have examined the effect on iron stores of blood transfusions given to premature neonates during hospitalization in the neonatal intensive care unit as reflected by serum ferritin levels measured for 6 months after discharge. Premature infants who were transfused with more than 100 ml packed cells (group D; n = 11) had higher ferritin levels for a longer period than premature infants who were transfused with smaller volumes (group c; n = 9) or premature and mature infants who were not transfused at all (group B; n = 24 and group A; n = 21, respectively). At 4-5 months the serum ferritin levels in group D (489.8 +/- 132.1 micrograms/L; mean +/- SEM) were significantly higher (P less than 0.001) than those of the other groups. The level of group A term infants (77.5 +/- 12.5 micrograms/L) was higher than those of group B premature infants who did not receive a blood transfusion (33.0 +/- 7.1 micrograms/L) or group C who received less than 100 ml (36.5 +/- 8.8 micrograms/L packed red blood cells. However, these differences were not statistically significant. Our data demonstrate that very-low-birthweight infants who receive a large volume of packed cells during hospitalization may accumulate iron stores sufficient for red cell production during the first 6 months of life. Administration of large amounts of supplemental iron, in such cases, may be curtailed.

Anemia, Hypochromic↗

Coexistence and gene expression of phenylethanolamine N-methyltransferase, tyrosine hydroxylase, and neuropeptide tyrosine in the rat and bovine adrenal gland: effects of reserpine.

Expression and regulation of the catecholamine-synthesizing enzymes phenylethanolamine N-methyltransferase (PNMTase; S-adenosyl-L-methionine:phenylethanolamine N-methyltransferase, EC 2.1.1.28) and tyrosine hydroxylase [TyrOHase; tyrosine 3-monooxygenase, L-tyrosine, tetrahydropteridine:oxygen oxidoreductase (3-hydroxylating), EC 1.14.16.2] and the coexisting neuropeptide tyrosine (NPY) were studied in rat and bovine adrenal medulla. By using both immunohistochemistry and in situ hybridization, PNMTase- and NPY-positive cells exhibited a close overlap in bovine medulla and were preferentially localized in the outer two-thirds of the medulla. Although TyrOHase and its mRNA were observed in virtually all medullary gland cells, TyrOHase mRNA levels were much higher in the PNMTase- and NPY-positive cells. After administration of the catecholamine-depleting drug reserpine to rats, a brief increase, followed by a dramatic decrease, in the level of PNMTase mRNA was observed in the adrenal medulla. In contrast, mRNA for both TyrOHase and NPY only exhibited an increase, whereby the TyrOHase mRNA peak preceded that of NPY mRNA. Different regulatory mechanisms may thus operate for these three compounds coexisting in the adrenal medulla.

Adrenal Glands↗

Antiserum raised against residues 159-168 of the guanine nucleotide-binding protein Gi3-alpha reacts with ependymal cells and some neurons in the rat brain containing cholecystokinin- or cholecystokinin- and tyrosine 3-hydroxylase-like immunoreactivities.

Antibodies raised against a synthetic deca-peptide corresponding to a specific sequence of Gi3-alpha protein (an inhibitory guanine nucleotide-binding protein) were used to analyze Gi3-alpha-like immunoreactivity in brain sections from colchicine-treated rats by indirect immunofluorescence histochemistry. Gi3-alpha-peptide-positive cell bodies were found in the ventral tegmental area and substantia nigra, and these cells were also cholecystokinin (CCK)- and tyrosine 3-hydroxylase-positive. Gi3-alpha-peptide staining was observed in perikarya in the hippocampus and in fibers in the nucleus accumbens, tuberculum olfactorium, bed nucleus of stria terminalis, and a spino-thalamic tract, where it coexisted with CCK-like immunoreactivity as well. No coexistence with CCK occurred in Gi3-alpha-peptide-positive ependymal cells outlining the aqueduct and ventricles. Preadsorption of Gi3-alpha antibodies with CCK-8 or CCK-33 did not alter Gi3-alpha-peptide staining. The occurrence of Gi3-alpha-peptide-like immunoreactivity in CCK-containing neurons may indicate the presence of Gi3-alpha protein and in CCK/dopamine neurons may indicate an association of this Gi protein with dopamine autoreceptors.

Animals↗

Telencephalic projections of the A8 dopamine cell group.

The telencephalic projections of the A8 dopamine cell group of the rat were assessed using both anterograde and combined retrograde-immunohistochemical methods. The projections of the A8 neurons onto the forebrain were more extensive than hitherto realized, and encompassed striatal, limbic, and allocortical regions. The A8 neurons were shown to contribute to the dopaminergic innervation of the striatum, nucleus accumbens, olfactory tubercles, amygdala, and bed nucleus of the stria terminalis, and also innervate the pyriform and entorhinal cortices. In addition, projections within the midbrain were observed, and suggested that there may be direct interconnections between the dopaminergic neurons of the A8, A9, and A10 cell groups. These data therefore suggest that the A8 dopamine cell group is uniquely situated to modulate functional activity within both nigrostriatal and mesocorticolimbic regions, and further suggests that heterogeneities of the midbrain dopamine neurons are embedded within a larger homogeneous mesotelencephalic dopamine system.

Animals↗

Proximal tubule Na+-K+-ATPase activity is inhibited during high-salt diet: evidence for DA-mediated effect.

Locally produced dopamine (DA) causes a reversible and dose-dependent inhibition in Na+-K+-ATPase activity in rat proximal tubule (PT) segments [A. Aperia, A. Bertorello, and I. Seri. Am. J. Physiol. 252 (Renal Fluid Electrolyte Physiol. 21): F32-F45, 1987.]. To examine whether this effect might be of physiological importance, rats were given normal-salt (NS) or high-salt (HS) diet for 10 days. HS diet significantly increased Na excretion but did not alter glomerular filtration rate (GFR). Benserazide (Bz), an inhibitor of the enzyme L-aromatic amino acid decarboxylase (AADC) that converts L-dopa to DA, significantly attenuated the natriuresis in HS rats but had no effect on GFR. By use of immunofluorescence (IF) studies AADC was localized to the PT. Specific AADC IF was not observed in the medulla. In AADC-positive PT segments, Na+-K+-ATPase activity was significantly lower in HS rats than in NS rats (P less than 0.001). In AADC-negative medullary thick ascending limb, Na+-K+-ATPase activity was the same in NS and HS rats. If HS rats were given Bz just before study, PT Na+-K+-ATPase activity increased significantly and was not different from Na+-K+-ATPase activity in PT segments from NS rats. Bz had no significant effect on PT Na+-K+-ATPase activity in NS rats. In PT segments from Bz-treated rats, DA inhibited Na+-K+-ATPase activity already at a dose of 10(-8) M, whereas in segments from NS rats, significant inhibition of Na+-K+-ATPase activity was not observed until DA was increased to 10(-7) M.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Cerebral cortical blood flow and oxygen metabolism in normocythemic hyperviscous newborn piglets.

Our study tests the hypothesis that hyperviscosity independent of arterial O2 content reduces cerebral cortical blood flow, O2 delivery, and O2 uptake. After baseline determinations, ten 2- to 4-day-old awake spontaneously breathing piglets were given an intravenous infusion (5 ml.kg-1, body weight) of concentrated cryoprecipitate, whereas eight controls received normal saline. Cerebral cortical blood flow, arterial and superior sagittal sinus O2 content, whole blood viscosity, hematocrit, blood gases, and plasma fibrinogen concentrations were measured at baseline and 3 h after infusion. No significant changes were observed in the control group. Three hours after the infusion of concentrated cryoprecipitate the experimental group showed an increase in whole blood viscosity, whereas hematocrit and arterial O2 content were unchanged. There was a decrease in cerebral cortical blood flow and cerebral cortical O2 delivery, whereas cerebral cortical O2 uptake was unchanged. We conclude that hyperviscosity independent of arterial O2 content reduces cerebral cortical blood flow and that although O2 delivery was reduced in the newborn piglet cerebral cortical O2 uptake was maintained.

Animals↗

Sertoli cell-only syndrome produced by cold testicular ischemia.

This report describes a new method for producing Sertoli cell-only testes in the Lewis rat using 90 min of hypothermic testicular ischemia. The method employs selective occlusion of the testicular blood supply using atraumatic microclips applied with the aid of an operating microscope. The testis is packed in ice-cold saline throughout the ischemic interval, and the deferential artery and vein are ligated. Twelve weeks after the ischemic insult, the testes weigh half that of control testes while there were no differences in prostate or seminal vesicle weights. Microscopic examination of the ischemic damaged testes revealed normal-appearing Leydig and Sertoli cells, but complete absence of germ cells. Assays of testicular enzyme activities indicated that lactic dehydrogenase and sorbitol dehydrogenase were reduced, while alpha-glutamyl transpeptidase activity was normal, consistent with the marked reduction of germ cells. Serum androgen binding protein (rABP) levels were elevated relative to nonischemic controls. By contrast, serum concentrations of testosterone, LH, and FSH were normal. In addition, LHRH elicited identical LH and testosterone responses in control and experimental animals. Testicular blood flow measured with 133Xenon was slightly decreased in Sertoli-cell-only testes. Intratesticular temperatures was normal in all groups. These observations in rats with ischemia-induced Sertoli-cell-only testes are strikingly different from those induced by radiation or genetic defects. Animals with these latter disorders have elevated FSH levels, evidence of altered Leydig cell function as evidenced by elevated LH or abnormal response to LHRH; and normal or low serum rABP levels. We conclude that 1) ischemia produces no abnormalities of the pituitary testicular axis in spite of marked germ cell depletion and 2) Sertoli-cell-only testes of different etiologies can have varied patterns of hormone and rABP secretion.

Androgen-Binding Protein↗