Summary
In the rat olfactory bulb, the majority of interneurons in the glomerular layer (GL) are supposed to be generated during first postnatal week. Low and repeated doses of X-rays (200 rad x 4 and 200 rad x 6) were used during this period to impair the development of interneurons. The resulting effects on olfactory bulb neurons were examined stereologically and immunocytochemically in animals of 4 and 12 weeks of age. Quantitative analysis showed that, 1) the volume of the GL decreased to 55% (1200 rad) – 70% (800 rad) of control, 2) numerical cell densities in GL decreased to 40% (1200 rad) – 60% (800 rad) of control, thus resulting in 3) a decrease of the total cell number in GL to 20% (1200 rad) – 40% (800 rad) of control in irradiated olfactory bulbs of animals 4 weeks old. In comparison, mitral cells, which are generated prenatally, were much less affected (total cell number: 70–80% of control), indicating a selective loss of cells generated during the first postnatal week in GL. Effects on somata and processes immunoreactive for GABA, tyrosine hydroxylase (TH), calbindin D-28K and parvalbumin (PV) were examined in irradiated bulbs of both 4 and 12 week-old rats. All of these immunoreactive elements showed a drastic decrease in all layers. Semiquantitative analysis showed that in the GL, calbindin D-28K immunoreactive (calbindin D-28K(+)) neurons decreased more extensively than TH immunoreactive (TH(+)) and GABA-like immunoreactive (GABA(+)) neurons; that is, TH(+) and GABA(+) neurons decreased to 20% (1200 rad) – 40% (800 rad) of control, whereas calbindin D-28K(+) neurons decreased to 10% (1200 rad) – 30% (800 rad) of control in the GL of irradiated bulbs. These findings indicated that larger proportions of calbindin D-28K(+) neurons might be generated during the first postnatal week than those of GABA(+) and TH(+) neurons. Furthermore, in irradiated bulbs the proportion of GABA(-)TH(+) cells in TH(+) cells increased to about twice of control, and the estimated total numbers of GABA(-)TH(+) cells in irradiated rats were 95% (800 rad) and 40% (1200 rad) of control. These observations suggest that the majority of GABA(-)TH(+) neurons were less affected by X-ray irradiation during the first postnatal week and thus that they might be generated in the prenatal period. Since during the first 2 postnatal weeks, neurons showing GABA(-)TH(+) were not seen in GL (Kosaka et al. 1987a), the majority of GABA(-)TH(+) neurons in adult olfactory bulb were assumed to change their phenotype at some postnatal developmental period.
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References
Altman J, Anderson WJ, Wright KA (1968) Differential radiosensitivity of stationary and migratory primitive cells in the brains of infant rats. Exp Neurol 22:52–74
Baimbridge KG, Miller JJ (1982) Immunohistochemical localization of calcium-binding protein in the cerebellum, hippocampal formation and olfactory bulb of the rat. Brain Res 245:223–229
Bayer SA (1983) 3H-thymidine-radiographic studies of neurogenesis in the rat olfactory bulb. Exp Brain Res 50:329–340
Bayer SA, Altman J (1975a) The effects of X-irradiation on the postnatally-forming granule cell populations in the olfactory bulb, hippocampus, and cerebellum of the rat. Exp Neurol 48:167–174
Bayer SA, Altman J (1975b) Radiation-induced interference with postnatal hippocampal cytogenesis in rats and its long-term effects on the acquisition of neurons and glia. J Comp Neurol 163:1–20
Bogan N, Brecha N, Gall C, Karten HJ (1982) Distribution of enkephalin-like immunoreactivity in the rat main olfactory bulb. Neuroscience 7:895–906
Celio MR (1990) Calbindin D-28k and parvalbumin in the rat nervous system. Neuroscience 35:375–475
Davis BJ, Macrides F (1983) Tyrosine hydroxylase immunoreactive neurons and fibers in the olfactory system of the hamster. J Comp Neurol 214:427–440
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
Gundersen HJG, Jensen EB (1987) The efficiency of systematic sampling in stereology and its prediction. J Microsc 147:229–263
Halász N (1986a) Early X-irradiation of rats. 1. Methodological description and morphological observations on the olfactory bulb. J Neurosci Meth 18:255–268
Halász N (1986b) Early X-irradiation of rats. 2. Effect on granule cells and their dendrodendritic synapses in the olfactory bulb. Neuroscience 20:709–716
Halász N (1987) Early X-irradiation of rats. 3. Survival of dopaminergic periglomerular cells of the olfactory bulb. Z Microsk Anat Forsch 101:1027–1034
Halász N, Hökfelt T, Norman AW, Goldstein M (1985) Tyrosine hydroxylase and 28K-Vitamin D-dependent calcium binding protein are localized in different subpopulations of periglomerular cells of the rat olfactory bulb. Neurosci Lett 61:103–107
Halász N, Johansson O, Hökfelt T, Ljungdahl Å, Goldstein M (1981) Immunohistochemical identification of two types of dopamine neuron in the rat olfactory bulb as seen by serial sectioning. J Neurocytol 10:251–259
Halász N, Ljungdahl Å, Hökfelt T, Johansson O, Goldstein M, Park D, Biberfeld P (1977) Transmitter histochemistry of the rat olfactory bulb. 1. Immunohistochemical localization of monoamine synthesizing enzymes. Support for intrabulbar, periglomerular dopamine neurons. Brain Res 126:455–474
Halász N, Ljungdahl Å, Hökfelt T (1978) Transmitter histochemistry of the rat olfactory bulb. 2. Fluorescence histochemical, autoradiographic and electron microscopic localization of monoamines. Brain Res 154:253–271
Halász N, Shepherd GM (1983) Neurochemistry of the vertebrate olfactory bulb. Neuroscience 10:579–619
Hicks SP (1958) Radiation as an experimental tool in mammalian developmental neurology. Physiol Rev 38:337–356
Hsu S-M, Raine L, Fanger H (1981) Use of avidin-biotinperoxidase complex (ABC) in immunoperoxidase techniques. J Histochem Cytochem 29:577–580
Kägi U, Berchtold MW, Heizmann CW (1987) Ca2+-binding parvalbumin in the rat testis: characterization, localization, and expression during development. J Biol Chem 262:7314–7320
Kosaka T, Hataguchi Y, Kama K, Nagatsu I, Wu J-Y (1985a) Coexistence of immunoreactivities for glutamate decarboxylase and tyrosine hydroxylase in some neurons in the periglomerular region of the rat main olfactory bulb: possible coexistence of gamma-aminobutyric acid (GABA) and dopamine. Brain Res 343:166–171
Kosaka T, Kosaka K, Tateishi K, Hamaoka Y, Yanaihara N, Wu J-Y, Hama K (1985b) 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, Nagatsu I, Wu J-Y, Hama K (1986) Use of high concentrations of glutaraldehyde for immunocytochemistry of transmitter-synthesizing enzymes in the central nervous system. Neuroscience 18:975–990
Kosaka K, Hama K, Nagatsu I, Wu J-Y, Ottersen OP, Storm-Mathisen J, Kosaka T (1987a) Postnatal development of neurons containing both catecholaminergic and GABAergic traits in the rat main olfactory bulb. Brain Res 403:355–360
Kosaka T, Kosaka K, Hataguchi Y, Nagatsu I, Wu J-Y, Ottersen OP, Storm-Mathisen J, Hama K (1987b) Catecholaminergic neurons containing GABA-like and/or glutamic acid decarboxylase-like immunoreactivities in various brain regions of the rat. Exp Brain Res 66:191–210
Kosaka T, Kosaka K, Heizmann CW, Nagatsu I, Heizmann CW, Nagatsu I, Wu J-Y, Yanaihara N, Hama K (1987c) An aspect of the organization of the GABAergic system in the rat main olfactory bulb: laminar distribution of immunohistochemically defined subpopulations of GABAergic neurons. Brain Res 411:373–378
Kosaka K, Hama K, Nagatsu I, Wu J-Y, Kosaka T (1988) Possible coexistence of amino acid (γ-aminobutyric acid), amine (dopamine) and peptide (substance P); neurons containing immunoreactivities for glutamic acid decarboxylase, tyrosine hydroxylase and substance P in the hamster main olfactory bulb. Exp Brain Res 71:633–642
Matute C, Streit P (1986) Monoclonal antibodies demonstrating GABA-like immunoreactivity. Histochemistry 86:147–157
Monti Graziadei GA, Margolis FL, Harding JW, Graziadei PPC (1977) Immunocytochemistry of the olfactory marker protein. J Histochem Cytochem 25:1311–1316
Nagatsu I (1983) Immunohistochemistry of biogenic amines and immunoenzyme-histochemistry of catecholamine-synthesizing enzymes. In: Parvez S, Nagatsu T, Nagatsu I, Parvez H (eds) Methods in biogenic amine research. Elsevier/North-Holland, Amsterdam, pp 873–909
Pinching AJ, Powell TPS (1971a) The neuron type of the glomerular layer of the olfactory bulb. J Cell Sci 9:305–345
Pinching AJ, Powell TPS (1971b) The neuropil of the glomeruli of the olfactory bulb. J Cell Sci 9:347–377
Pinching AJ, Powell TPS (1971c) The neuropil of the periglomerular region of the olfactory bulb. J Cell Sci 9:379–409
Pinol MR, Kägi U, Heizmann CW, Vogel B, Sequier J-M, Haas W, Hunziker W (1990) Poly- and monoclonal antibodies against recombinant rat brain calbindin D-28K were produced to map its selective distribution in the central nervous system. J Neurochem 54:1827–1833
Ribak CE, Vaughn JE, Saito K, Barber R, Roberts E (1977) Glutamate decarboxylase localization in neurons of the olfactory bulb. Brain Res 126:1–18
Sterio DC (1984) The unbiased estimation of number and sizes of arbitrary particles using the disector. J Microsc 134:127–136
Tsuruo Y, Hökfelt T, Visser TJ (1988) Thyrotropin-releasing hormone (TRH)- immunoreactive neuron populations in the rat olfactory bulb. Brain Res 447:183–187
West RW (1972) Superficial warming of epoxy blocks for cutting of 25–150 μm sections to be resectioned in the 40–90 nm range. Stain Technol 47:201–204
Yanai J, Rosselli-Austin L (1978) Normal homing behavior in infant rats despite extensive olfactory bulb granule cell losses. Behav Biol 24:539–544
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Kosaka, K., Taomoto, K., Nagatsu, I. et al. Postnatal X-ray irradiation effects on glomerular layer of rat olfactory bulb: quantitative and immunocytochemical analysis. Exp Brain Res 90, 103–115 (1992). https://doi.org/10.1007/BF00229261
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DOI: https://doi.org/10.1007/BF00229261