Adult development of the hippocampal-serotonin system of C57BL/6N mice; analysis of high-affinity uptake of 3H-5HT in slices and synaptosomes

Efrain Azmitia, Michael J. Brennan, David Quartermain

Research output: Contribution to journalArticle

Abstract

The saturable and specific high-affinity uptake of [3H]serotonin ([3H]5HT) (5 × 10-8 M) was studied in slices from the hippocampus, parietal cortex, septum-preoptic area, and hypothalamus of male 2, 6, 12 and 24-32 month old C57BL/6N mice. Hippocampal [3H]5-HT uptake showed a significant biphasic relationship to age, with lower values in the 2 and 24-32 month old mice compared to 6 month old mice. No significant age effects were seen in the other regions, or in [3H]norepinephrine high-affinity uptake in the hippocampus. Studies of the high-affinity uptake mechanism in synaptosomal preparations were made in a subgroup of 12 and 24 month old mice. A micro-assay using a tissue-harvester measured high-affinity uptake on 8-30 μl of the P2 suspension (crude-synaptosomal preparation). The high-affinity uptake was linear for 4 min at 37°C and inhibited in both the adult and aged tissue by 10-5 M cold 5-HT (83 and 78% respectively), 10-5 M fluoxetine (85 and 82% respectively) and 10-3 M NaCN (57 and 57% respectively). Kinetic analysis of the [3H]5HT high-affinity uptake in the hippocampus (3 min, 37°C) revealed the same apparent Km for serotonin at both ages (6.7 x 10-8 M), but a 44% decrease in Vmax in the aged hippocampal synaptosomal high-affinity uptake compared to adults (120 vs 215 pmol of 5-HT/g-tissue/3 min). These results are discussed in relationship to the reported age effects on the intrinsic neurons of the hippocampus.

Original languageEnglish (US)
Pages (from-to)39-44
Number of pages6
JournalNeurochemistry International
Volume5
Issue number1
DOIs
StatePublished - 1983

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Synaptosomes
Inbred C57BL Mouse
Serotonin
Hippocampus
Parietal Lobe
Preoptic Area
Fluoxetine
Hypothalamus
Suspensions
Norepinephrine
Neurons

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Cellular and Molecular Neuroscience

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Adult development of the hippocampal-serotonin system of C57BL/6N mice; analysis of high-affinity uptake of 3H-5HT in slices and synaptosomes. / Azmitia, Efrain; Brennan, Michael J.; Quartermain, David.

In: Neurochemistry International, Vol. 5, No. 1, 1983, p. 39-44.

Research output: Contribution to journalArticle

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abstract = "The saturable and specific high-affinity uptake of [3H]serotonin ([3H]5HT) (5 × 10-8 M) was studied in slices from the hippocampus, parietal cortex, septum-preoptic area, and hypothalamus of male 2, 6, 12 and 24-32 month old C57BL/6N mice. Hippocampal [3H]5-HT uptake showed a significant biphasic relationship to age, with lower values in the 2 and 24-32 month old mice compared to 6 month old mice. No significant age effects were seen in the other regions, or in [3H]norepinephrine high-affinity uptake in the hippocampus. Studies of the high-affinity uptake mechanism in synaptosomal preparations were made in a subgroup of 12 and 24 month old mice. A micro-assay using a tissue-harvester measured high-affinity uptake on 8-30 μl of the P2 suspension (crude-synaptosomal preparation). The high-affinity uptake was linear for 4 min at 37°C and inhibited in both the adult and aged tissue by 10-5 M cold 5-HT (83 and 78{\%} respectively), 10-5 M fluoxetine (85 and 82{\%} respectively) and 10-3 M NaCN (57 and 57{\%} respectively). Kinetic analysis of the [3H]5HT high-affinity uptake in the hippocampus (3 min, 37°C) revealed the same apparent Km for serotonin at both ages (6.7 x 10-8 M), but a 44{\%} decrease in Vmax in the aged hippocampal synaptosomal high-affinity uptake compared to adults (120 vs 215 pmol of 5-HT/g-tissue/3 min). These results are discussed in relationship to the reported age effects on the intrinsic neurons of the hippocampus.",
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