Immunohistochemical localization of an inositol 1,4,5-trisphosphate receptor, P400, in neural tissue: studies in developing and adult mouse brain

Abstract

The immunohistochemical localization of P400/inositol 1,4,5-trisphosphate (InsP3) receptor protein was studied in developing and adult mouse brain by using monoclonal antibodies. The developmental expression pattern of P400/InsP3 receptor protein differed among different classes of neurons. It was first detected in the somata of immature Purkinje cells at embryonic day 17, in the ventrolateral region of the posterior vermis in the cerebellum. Axonal immunoreactivity within the cerebellar nuclei was first present at postnatal day 3. Neurons in the retrosplenial cortex, the anterior olfactory nucleus, and the CA1 region of the hippocampus expressed immunoreactivity earlier than other regions of the brain. In the adult brain, not only the Purkinje cell but also many other types of cells in many areas of the brain expressed P400/InsP3 receptor, though to a lesser extent. These included the neurons in the striatum, globus pallidus, nucleus accumbens septi, anterior olfactory nucleus, olfactory tubercle, precommissural hippocampus, hippocampus, substantia nigra, cerebral cortex, pons, and certain hypothalamic nuclei. Forebrain cortical regions that receive afferents from the olfactory bulb, such as the anterior olfactory nucleus, olfactory tubercle, prepiriform cortex, entorhinal cortex, and amygdala, exhibited distinct immunoreactivity, while olfactory bulb was almost devoid of staining. Immunoreactivity in the axonal pathways was also found in the limbic-hypothalamic pathways, strionigral projection, and part of the corpus callosum. Results of Western blot analysis and 3H-InsP3 binding assay were consistent with the qualitative regional differences of immunoreactivity demonstrated by immunohistochemical study. The location of InsP3 receptor in the brain correlates well with the InsP3 binding sites demonstrated by an autoradiographic study.

Presynaptic calcium stores underlie large-amplitude miniature IPSCs and spontaneous calcium transients

“…Another explanation comes from evidence for functional intracellular Ca 2+ stores in presynaptic terminals. First, inositoltrisphosphate (InsP 3 ) receptors are immunolocalized in presynaptic terminals of the deep cerebellar nuclei and retina 7,8 . Second, at the frog neuromuscular junction, agents that affect ryanodine-sensitive Ca 2+ stores also regulate presynaptic intracellular Ca 2+ (Ca 2+ i ) rises and acetylcholine release during high-frequency stimulation 9,10 .…”

Monoclonal antibodies distinctively recognizing the subtypes of inositol 1,4,5‐trisphosphate receptor: Application to the studies on inflammatory cells

“…2). It was reported that IP,R type 1 and IP,R type 3 gave specific bands of about 250 kDa on SDS-PAGE using an anti-mouse IP,R type 1 monoclonal antibody 4Cll [18] and an anti-IP,R type 3 polyclonal antibody [13], respectively. The antibody 4Cll was shown to react with the protein with the same migration in this study (data not shown).…”

“…In practice, using monoclonal antibodies against IP3R type 1, tissue and cellular localization, phosphorylation and tetramer formation of the IP,R subtype have been elucidated [2,3,18,271. It was shown by cDNA cloning, Northern blot and in situ hybridization analysis that there are at least three subtypes in the IP3R family and that these subtypes are differently expressed in tissues [1 l-161 and in many specific hematopoietic cells [17].…”

“…These findings prompted us to investigate the difference in functions, and tissue/cellular and subcellular localization of these IP,R subtypes. Immunohistochemical studies using the monoclonal antibodies raised against IP,R type 1 have shown that whilst the subtype is distributed in many areas of the brain, its expression is especially high in the Purkinje cells [18]. However, to date, such studies for IP3R type 2 and type 3 are rather limited [11,12,14] essentially because of the lack of monoclonal antibodies specific to each of IP,R subtypes, especially those to IP,R type 2 and type 3.…”

Prior activation of inositol 1,4,5-trisphosphate receptors suppresses the subsequent induction of long-term potentiation in hippocampal CA1 neurons

“…The type 1 IP 3 R (IP 3 R1) is the major IP 3 R family member in the central nervous system and is predominantly enriched in cerebellar Purkinje cells and hippocampal neurons (Furuichi et al 1989;Nakanishi et al 1991). Activation of IP 3 Rs during, or after, HFS is involved in the mechanism of LTP induction in hippocampal CA1 neurons (Yoshioka et al 2010).…”