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Biomedical subjects

C Prasad

Publications and source records attributed to C Prasad.

At least 19 recordsLinked to original sources
PubMed1992

Decreased striatal D2 dopamine receptors in obese Zucker rats: changes during aging.

The specific binding of [3H]YM-09151-2 was used to investigate the possible differences in age-associated changes in striatal D2 dopamine (DA) receptor properties in genetically obese (fa/fa) Zucker rats and their lean (Fa/?) littermates. The maximal binding sites (Bmax) of D2 DA receptors was found to decline with age in both obese and lean rats; the rate of decline in receptor Bmax was slightly higher in lean than obese rats. However, the Bmax of D2 DA receptor in 6-, 12- and 18-month-old obese rats was significantly lower compared to the age-matched lean rats. These data indicate that obesity decreases the number of striatal D2 DA receptors without affecting the rate at which receptor number decreases with age.

Aging↗
PubMed1992

Production and characterization of a monoclonal antibody to dopamine D2 receptor: comparison with a polyclonal antibody to a different epitope.

A monoclonal antibody (Mab) that recognizes the rat dopamine D2 receptor (DAR) has been generated using DAR specific peptide. The Mab, IgM isotype recognizes five proteins (Mr 220, 145, 95, 66 and 47 kDa) in striatal membrane on Western blot. Preincubation of Mab with free peptide blocked the labeling of all five bands. A polyclonal antibody against peptide from a different region of the DAR, reacted with three out of five proteins (220, 66, and 47 kDa) in these membranes. The DAR antagonist NAPS-biotinyl binds to a 220 kDa protein in striatal membrane on ligand blotts; the labeling can be blocked by the addition of 2 microM sulpride. The 220 kDa Mab reactive protein was less in cerebellum and was absent in the liver. Neither the Mab nor polyclonal antibody inhibited binding of a DAR antagonist, [3H]YM09151-2, to the striatal membranes. These antibodies will enable us to study the structure/function and regulation of the synthesis of DAR protein.

Amino Acid Sequence↗
PubMed1992

Localization of dopamine D2 receptor protein in rat brain using polyclonal antibody.

The precise distribution of the dopamine type D2 receptor has been mapped for the first time in rat brain using an antibody to D2 receptor protein. Polyclonal antisera were collected from rabbits inoculated with an undecapeptide identical to residues 24-34 of the D2 protein sequence. Rat brain slices, 40 microns in thickness, were incubated with either primary antiserum, the antiserum plus free peptide antigen, or pre-immune serum. Antibody binding was visualized by peroxidase-antiperoxidase (PAP) reaction followed by light microscopy. PAP complex bound moderately-to-densely throughout the medial forebrain bundle, and was seen in more discrete regions in the midbrain, consistent with the binding of D2 radioligands. There were some unexpected results, namely in the cerebral cortex and nucleus accumbens, there were unexpectedly steep gradients in binding density, decreasing caudally; no binding was detected in the hippocampus or the substantia nigra pars reticulata. In all positive-staining regions examined, the antibody was highly localized to neuronal cell bodies, except in the frontal cortex where antibody was also evident on basilar dendrites. These data confirm that the polyclonal antibody recognized dopamine D2 receptor protein throughout the rat brain, and suggest that the D2 receptor is distributed more abundantly on somata than on cellular processes.

Animals↗
PubMed1992

Alterations in dendritic spine density in the rat brain associated with protein malnutrition.

Rats that consume a high-protein diet are hyperactive, hyperresponsive to noxious stimuli, and demonstrate elevated basal arousal levels. Although the mechanism involved in dietary protein-induced changes in behavior is unclear, it may involve many changes including abnormal dendrite morphology in the brain. Three groups of rats were pair-fed with isocaloric diets containing 8%, 20% and 50% casein for 4 weeks. Their brains were processed for examination of dendritic spine densities in the frontal, parietal, and entorhinal cortices, the striatum, and the septum. Animals on the 50% casein diet showed increased spine densities in all areas investigated (P less than 0.05), compared to the animals on normal (20%) casein. In contrast, animals maintained on the 8% casein diet showed increased spine densities (P less than 0.05) only in the striatum and entorhinal cortex, and exhibited normal densities in the frontal and parietal cortices, and the medial septum.

Animals↗
PubMed1992

Post-transcriptional regulation of loss of rat striatal D2 dopamine receptor during aging.

The mechanism(s) underlying age-associated diminutions in the rat striatal D2 dopamine receptor (D2-receptor) number was investigated. The levels of D2-receptor mRNA in 4-, 12- and 18-month-old rat striata were found not to change. In contrast, the levels of 110 kDa protein, labeled with a D2-receptor specific antibody, decreased in parallel with [3H]YM-09151-2 binding to striatal membranes. These data suggest a role for post-transcriptional mechanism(s) in age-associated decrease in D2-receptor.

Actins↗
PubMed1992

A low protein-high carbohydrate diet decreases D2 dopamine receptor density in rat brain.

The property of D2 dopamine receptors in the rat brain was evaluated after long-term dietary manipulation. Groups of rats were pair-fed with equicaloric diet containing low protein (8%)-high carbohydrate, high protein (52%)-low carbohydrate and normal protein (20%) for 36 weeks. The low protein-high carbohydrate fed rats exhibited a significant decrease in the density (Bmax) of D2 dopamine receptor in the striatum (28%) and the mesolimbic regions (36%) with no apparent change in the receptor affinity (Kd). These findings suggest that long-term consumption of a low protein-high carbohydrate diet, by decreasing D2 dopamine receptor density, may be an important determinant of central dopaminergic function.

Animals↗
PubMed1992

An antibody to dopamine D2 receptor inhibits dopamine antagonist and agonist binding to dopamine D2 receptor cDNA transfected mouse fibroblast cells.

A polyclonal antibody to dopamine D2 receptor (D2-receptor) has been used to examine the immuno-inhibition in the binding of a D2 antagonist, [3H]YM09151-2 and an agonist, PPHT-fluorescein to dopamine receptor DNA transfected mouse fibroblast cells. The specific activity of the [3H]YM09151-2 binding to transfected (Ltk-RGB) cells is 4-5 fold higher than untransfected (Ltk-) cells. The antibody is able to inhibit the [3H]YM09151-2 binding to the cell membranes from Ltk-RGB cells (Bmax 110.56 +/- 5.26 and 76.20 +/- 5.18 fmoles/mg protein in the presence of preimmune and immune sera, respectively, with no change in the Kd). The flow cytometric analysis of the PPHT-fluorescein labeled Ltk- and Ltk-RGB cells indicated that ligand specific fluorescence is associated only with small Ltk-RGB cells (second peak) and autofluorescence with large cells (first peak). Preincubation of the Ltk-RGB cells with antibody, reduced the fluorescence intensity of the PPHT-fluorescein by 20-25% without changing the auto-fluorescence. These results suggest that peptide antibody recognize D2-receptor in both membranes and in intact cells and interact at or near the ligand binding site of the receptor.

Animals↗
PubMed1992

Resistance to insulin-induced hyperphagia in fat-preferring rats.

Outbred male Sprague-Dawley rats were screened for their macronutrient (fat, carbohydrate and protein) preference profile and divided into two groups, the low- and high fat-preferring groups each deriving 23% and 72% of its total caloric intake from fat respectively. Subcutaneous administration of bovine insulin (6U/kg) resulted in an increase in total caloric intake in the low, but not the high fat-preferring group. Furthermore, the increased caloric intake in the low fat-preferring group was entirely due to increased carbohydrate consumption. These data suggest a defect in the action of insulin in fat-preferring rats.

Animals↗
PubMed1992

Transient decrease in rat striatal D2 dopamine receptor mRNA level after acute haloperidol treatment.

Acute haloperidol administration decreases the number of rat striatal D2 dopamine receptor (D2-receptor). The potential involvement of decreased D2-receptor gene transcription in the above process was examined using Northern blot analysis. There was a dose- and time-dependent transient decrement in the level of striatal D2-receptor mRNA after haloperidol. The decrease in transcription may be mediated via blockade of D2-receptor, since S(-)-sulpiride but not the inactive enantiomer R(+)-sulpiride produced the inhibition.

Animals↗
PubMed1992

Food contains the bioactive peptide, cyclo(His-Pro).

Cyclo(His-Pro) (CHP) is a cyclic dipeptide with numerous biological activities. As small di- and tripeptides may be absorbed intact when ingested orally, we were interested in examining several common foods for the presence of cyclo(His-Pro)-like immunoreactivity (CHP-LI). In all foods tested, CHP-LI was found at levels 5-1500 times those previously found in human plasma. This CHP-LI was identical to authentic CHP by immunoidentity and chromatographic behavior. We conclude that 1) CHP-LI is readily detectable in several common food sources; 2) this CHP-LI is indistinguishable from authentic CHP; and 3) it is likely the CHP-LI in foods is absorbed in quantities sufficient to cause elevations of CHP-LI in plasma to biologically significant levels.

Chromatography↗
PubMed1991

Attenuation of pulsatile changes in the density of striatal [3H]GBR-12935 binding sites during chronic ethanol consumption.

Using [3H]GBR-12935 as a selective ligand, we have examined the properties of rat striatal dopamine transporter complex during chronic ethanol consumption. The results of this study show a significant change in the Bmax (maximal binding capacity), but not Kd (affinity) of [3H]GBR-12935 binding sites. The observed changes were pulsatile and they exhibited an increase (144% of control), decrease (80% of control), tendency to increase (111% of control), increase (141% of control), decrease (84% of control), no change (98% of control), and decrease (80% of control) after 1, 2, 3, 4, 6, 8 and 10 weeks of ethanol consumption, respectively. Since the duration of ethanol exposure appears to be an important variable in regulating the properties of dopamine transporter complex, the behavioral changes observed during chronic ethanol intake may vary with time.

Alcoholism↗
PubMed1991

Motor, but not sensory, cortical potentials are amplified by high-protein diet.

Animals fed a high-protein diet (50% casein) are hyperactive and more responsive to nociceptive stimuli than those fed either a normal- or low-protein diet. The mechanisms mediating dietary protein-induced behavior are unknown and may include both central and peripheral neural effects. Adult, Sprague-Dawley rats were fed 50% casein (treatment group) and 24% casein (control group) ad lib for 36-40 weeks. The animals were anesthetized with alpha-chloralose and urethane (50 mg/kg and 1.5 mg/kg, IP). EEG recordings were averaged while the anesthetized animal was conditioned using an alerting stimulus-imperative stimulus (AS-IS) paradigm. AS consisted of a 1.5 kHz, 90 dB tone cue. This was followed 2 seconds later by IS, an electrical tail stimulation (11 V, 1.4 s duration). Two negative deflections (N1 and N2) were generated by the frontal cortex during the AS-IS interstimulus interval. N1, an alerting response, was not different between the two groups. N2 amplitude and peak latency were significantly increased in the high-protein group (205% and 117% of control, respectively; p less than 0.05). N2 represents the activation of cells in the motor cortex. Brainstem auditory-evoked responses and somatosensory-evoked potentials also were recorded, but no differences were observed between the two diet groups. These data suggest that consumption of a high-protein diet results in an increase in central arousal mechanisms (measured by cortical negativity response), specifically involving increased excitability of the motor cortex, that is not associated with a disorder of information processing in the cerebral cortex (measured by brainstem auditory-evoked responses and somatosensory-evoked potentials).

Animals↗
PubMed1991

Could dietary proteins serve as cyclo(His-Pro) precursors?

Cyclic dipeptides or diketopiperazines are readily generated during in vitro hydrolysis of proteins and polypeptides. This led us to examine whether cyclo(His-Pro) (CHP), a diketopiperazine containing histidine and proline, could be formed in vivo from dietary proteins. The data presented here show that at least in rat, neither urinary nor plasma concentration of CHP is elevated by consumption of a diet rich in proteins. Several dietary supplements derived from casein and/or soy protein hydrolysates, however, contain high levels of CHP-LI. Oral intake of one such supplement led to a sharp increase in the plasma level of CHP-LI.

Animals↗
PubMed1991

Increased cerebrospinal fluid cyclo(His-Pro) content in schizophrenia.

Cyclo(His-Pro) (CHP) is a peptide endogenous to human brain and cerebrospinal fluid (CSF). In animal studies administration of exogenous CHP augments dopaminergic neurotransmission. To explore the role of this peptide in schizophrenia, a disease characterized by a hyperdopaminergic state, we have measured CSF CHP levels in control, never-medicated schizophrenics and medicated schizophrenics. Our data show a 53% increase in CSF levels of CHP in never-medicated schizophrenics (p = 0.015), and a 25% increase in medicated schizophrenics when compared to controls. We speculate that CHP may contribute to the expression of hyperdopaminergic symptoms in schizophrenia.

Adult↗
PubMed1991

Relationship between [3H]mazindol binding to dopamine uptake sites and [3H]dopamine uptake in rat striatum during aging.

Certain drugs exhibit a remarkable correlation between their ability to inhibit synaptosomal uptake of dopamine and the binding of [3H]mazindol to striatal membranes. To investigate the role of mazindol binding sites in the dopamine uptake process and the fate of these sites (labeling dopaminergic neurons) during aging, we have examined the properties of mazindol binding and dopamine uptake in individual young and old rats. There was a 48% decrease (p = 0.0001) in the Bmax of mazindol binding and a 23% decrease (p = 0.0166) in the Vmax of dopamine uptake with no apparent change in their affinities with age. Regression analysis of the relationship between Bmax and Vmax exhibited a significant correlation in old (p = 0.0156) but not young rats (p = 0.1398). These data suggest that the number of mazindol binding sites decreases with age and that the number of sites on the dopamine transporter complex far exceeds the number required to elicit maximal dopamine uptake.

Aging↗
PubMed1991

Antibodies against synthetic peptides predicted from the nucleotide sequence of D2 receptor recognize native dopamine receptor protein in rat striatum.

Two peptides corresponding to amino acid sequences predicted from the nucleotide sequence of the dopamine D2 receptor were synthesized. Peptide I (CGSEG-KADRPHYC) and peptide II (NNTDQNECIIY), corresponding to 24-34 and 176-185 from the NH2 terminus, respectively, were conjugated to keyhold limpet hemocyanin and injected into rabbits. Peptide I showed a greater immunogenic response than did peptide II. Both peptide antibodies exhibited high titer for the homologous antigens, but showed little or no cross-reactivity with heterogeneous peptides. Peptide I antibodies reacted with striatal membrane proteins of apparent molecular masses of 120, 90, 85, and 30 kDa on a western blot. Furthermore, the 90-kDa band was identified as denatured D2 receptor by its high affinity for the D2 selective photoaffinity probe 125I-N'-azidospiperone (125I-NAPS). Photoaffinity labeling of the 90-kDa protein by 125I-NAPS was reduced by 40% in the presence of the peptide I antibody. In addition, evidence is also presented to show the low level of 90-kDa protein in cerebellum which contains little or no D2 ligand binding sites. The antibody to peptide I inhibited the binding of [3H]YM-09151-2, a dopamine D2 receptor selective antagonist, to striatal membranes in a concentration-dependent manner; a 50% inhibition was obtained at a 1:500 dilution of the antisera with 20 pM ligand concentration. The data on the equilibrium inhibition kinetics of [3H]YM-09151-2 binding to striatal membranes were examined in the presence of antibody and showed a 25-30% decrease in Bmax (203.5 +/- 11.0 and 164.6 +/- 3.3 fmol/mg of protein in presence of preimmune and immune sera, respectively) with no change in KD.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗