Abstract
The numerical density of neurons in the CA1 region of the rat dorsal hippocampus has been estimated by a stereological method, the disector, using pairs of video images of toluidine blue-stained, plastic-embedded, 0.5-μm-thick sections, 3 μm distant from each other. The chemical properties of those disector-counted cells were further analyzed by postembedding immunocytochemical methods on adjacent, semithin sections using antibodies against gamma-aminobutyric acid (GABA) and a specific calcium-binding protein, parvalbumin (PV). The density of neurons in the CA1 region was 35.2 × 103/mm3; numerical densities in the stratum oriens (SO), stratum pyramidale (SP), and strata radiatum-lacunosum-moleculare (SRLM) were 11.3 × 103/mm3, 272.4 × 103/mm3, and 1.9 × 103/mm3, respectively. The numerical densities of GABA-like immunoreactive (GABA-LIR) and PV-immunoreactive (PV-IR) neurons were 2.1 × 103/mm3 and 1.1 × 103/mm3, respectively, which were 5.8% and 3.2% of all neurons, respectively. In the CA1 region only about 60% of PV-positive neurons were GABA-LIR. However, taking the previous observation into consideration that almost all hippocampal PV-positive neurons were immunoreactive for the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD), neurons that were immunoreactive to either GABA or PV or both (GABA+ and/or PV + neurons) were regarded as a better representative of GABAergic neurons in this region; thus, the numerical density of these GABA + and/or PV + neurons was 2.5 × 103/mm3 and they were 7.0% of all neurons in the CA1 region. Lamellar analysis showed that the numerical densities of GABA+ and/or PV+, GABA-LIR, and PV-IR neurons were highest in the SP, where they were 8.2 × 103/mm3, 6.2 × 103/mm3, and 5.4 × 103/mm3, respectively. The results of the present study indicate that the proportions of GABAergic neurons and a subpopulation of them, PV-containing GABAergic neurons, to other presumable non-GABAergic neurons are far smaller in the CA1 region of the hippocampus than in several neocortical regions previously reported.
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References
Amaral DG, Ishizuka N, Claiborne B (1990) Neurons, numbers and the hippocampal network. Prog Brain Res 83:1–11
Bendayan M, Zollinger M (1983) Ultrastructural localization of antigenic sites on osmium-fixed tissues applying the protein A-gold technique. J Histochem Cytochem 31:101–109
Bjugn R, Gundersen HJG (1993) Estimate of the total number of neurons and glial and endothelial cells in the rat spinal cord by means of the optical disector. J Comp Neurol 328:406–414
Boss BD, Turlejski K, Stanfield BB, Cowan WM (1987) On the numbers of neurons in fields CA1 and CA3 of the hippocampus of Sprague-Dawley and Wistar rats. Brain Res 406:280–287
Brændgaard H, Gundersen HJG (1986) The impact of recent stereological advances on quantitative studies of the nervous system. J Neurosci Methods 18:39–78
Brændgaard H, Evans SM, Howard CV, Gundersen HJG (1990) The total number of neurons in the human neocortex unbiasedly estimated using optical disectors. J Microsc 157:285–304
Celio MR (1986) Parvalbumin in most γ-aminobutyric acid-containing neurons of the rat cerebral cortex. Science 231:995–997
Demeulemeester H, Arckens L, Vandesande F, Orban GA, Heizmann CW, Pochet R (1991) Calcium binding proteins and neuropeptides as molecular markers of GABAergic interneurons in the cat visual cortex. Exp Brain Res 84:538–544
Gulyás AI, Tóth K, Dános P, Freund TF (1991) Subpopulations of GABAergic neurons containing parvalbumin, calbindin D28k, and cholecystokinin in the rat hippocampus. J Comp Neurol 312:371–378
Gundersen HJG (1977) Notes on the estimation of the numerical density of arbitrary profiles: the edge effect. J Microsc 111:219–223
Gundersen HJG (1986) Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones. In memory of William R. Thompson. J Microsc 143:3–45
Jones EG, Hendry SHC (1986) Co-localization of GABA and neuropeptides in neocortical neurons. Trends Neurosci 9:71–76
Kägi U, Berchtold MW, Heizmann CW (1987) Ca2+ -binding parvalbumin in rat testis: characterization, localization, and expression during development. J Biol Chem 262:7314–7320
Katsumaru H, Kosaka T, Heizmann CW, Hama K (1988a) Immunocytochemical study of GABAergic neurons containing the calcium-binding protein parvalbumin in the rat hippocampus. Exp Brain Res 72:347–362
Katsumaru H, Kosaka T, Heizmann CW, Hama K (1988b) Gap junctions on GABAergic neurons containing the calcium-binding protein parvalbumin in the rat hippocampus (CA1 region). Exp Brain Res 72:363–370
Kawaguchi Y, Katsumaru H, Kosaka T, Heizmann CW, Hama K (1987) Fast spiking cells in rat hippocampus (CA1 region) contain the calcium-binding protein parvalbumin. Brain Res 416:369–374
Kosaka T, Kosaka K, Tateishi K, Hamaoka Y, Yanaihara N, Wu J-Y, Hama K (1985) GABAergic neurons containing CCK-8-like and/or VIP-like immunoreactivities in the rat hippocampus and dentate gyrus. J Comp Neurol 239:420–430
Kosaka T, Heizmann CW, Tateishi K, Hamaoka Y, Hama K (1987a) An aspect of the organizational principle of the γ-aminobutyric acidergic system in the cerebral cortex. Brain Res 409:403–408
Kosaka T, Katsumaru H, Hama K, Wu J-Y, Heizmann CW (1987b) GABAergic neurons containing the Ca2+-binding protein parvalbumin in the rat hippocampus and dentate gyrus. Brain Res 419:119–130
Kosaka T, Wu J-Y, Benoit R (1988) GABAergic neurons containing somatostatin-like immunoreactivity in the rat hippocampus and dentate gyrus. Exp Brain Res 71:388–398
Kosaka T, Isogai K, Barnstable CJ, Heizmann CW (1990) Monoclonal antibody HNK-1 selectively stains a subpopulation of GABAergic neurons containing the calcium-binding protein parvalbumin in the rat cerebral cortex. Exp Brain Res 82:566–574
Matute C, Streit P (1986) Monoclonal antibodies demonstrating GABA-like immunoreactivity. Histochemistry 86:147–157
Ottersen OP, Storm-Mathiesen J (1984) Neurons containing or accumulating transmitter amino acids. In: Björklund A, Hökfelt T, Kuhar MJ (eds) Classical transmitters and transmitter receptors in the CNS, part II. (Handbook of chemical neuroanatomy, vol 3) Elsevier/North-Holland, Amsterdam, pp 141–246
Ren JQ, Aika Y, Heizmann CW, Kosaka T (1992) Quantitative analysis of neurons and glial cells in the rat somatosensory cortex, with special reference to GABAergic neurons and parvalbumin-containing neurons. Exp Brain Res 92:1–14
Somogyi P, Hodgson AJ, Smith AD, Nunzi MG, Gorio A, Wu J-Y (1984) Different populations of GABAergic neurons in the visual cortex and hippocampus of cat contain somatostatin- or cholecystokinin-immunoreactive material. J Neurosci 4:2590–2603
Sterio DC (1984) The unbiased estimation of number and sizes of arbitrary particles using the disector. J Microsc 134:127–136
Tandrup T (1993) A method for unbiased and efficient estimation of number and mean volume of specified neuron subtypes in rat dorsal root ganglion. J Comp Neurol 329:269–276
West MJ (1990) Stereological studies of the hippocampus: a comparison of the hippocampal subdivisions of diverse species including hedgehogs, laboratory rodents, wild mice and men. Prog Brain Res 83:13–36
West MJ, Gundersen HJG (1990) Unbiased stereological estimation of the number of neurons in the human hippocampus. J Comp Neurol 296:1–22
West MJ, Coleman PD, Flood DG (1988) Estimating the number of granule cells in the dentate gyrus with the disector. Brain Res 448:167–172
West MJ, Slomianka L, Gundersen HJG (1991) Unbiased stereological estimation of the total number of neurons in the subdivisions of the rat hippocampus using the optical fractionator. Anat Rec 231:482–497
Woodson W, Nitecka L, Ben-Ari Y (1989) Organization of the GABAergic system in the rat hippocampal formation: a quantitative immunocytochemical study. J Comp Neurol 280:254–271
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Aika, Y., Ren, J.Q., Kosaka, K. et al. Quantitative analysis of GABA-like-immunoreactive and parvalbumin-containing neurons in the CA1 region of the rat hippocampus using a stereological method, the disector. Exp Brain Res 99, 267–276 (1994). https://doi.org/10.1007/BF00239593
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DOI: https://doi.org/10.1007/BF00239593