Autoradiographic quantification of muscarinic cholinergic synaptic markers in bat, shrew, and rat brain

Abstract

We employed radioligand binding autoradiography to determine the distributions of pre-and postsynaptic cholinergic radioligand binding sites in the brains of two species of bat, one species of shrew, and the rat. High affinity choline uptake sites were measured with [3H]hemicholinium, and presynaptic cholinergic vesicles were identified with [3H]vesamicol. Muscarinic cholinergic receptors were determined with [3H]scopolamine. The distribution patterns of the three cholinergic markers were simitar in all species examined, and identified known major cholinergic pathways on the basis of enrichments in both pre-and postsynaptic markers. In addition, there was excellent agreement, both within and across species, in the regional distributions of the two presynaptic cholinergic markers. Our results indicate that pharmacological identifiers of cholinergic pathways and synapses, including the cholinergic vesicle transport site, and the organizations of central nervous system cholinergic pathways are phylogenetically conserved among eutherian mammals.

Regional vesicular acetylcholine transporter distribution in human brain: A [18F]fluoroethoxybenzovesamicol positron emission tomography study

“…Binding was highest in the striatal complex, followed by portions of the thalamus, amygdala and hippocampal formation, some neocortical regions, rostral brainstem, and some regions of the cerebellar cortex. The overall distribution and regional density of [ 18 F]FEOBV binding is largely consistent with the distribution of cholinergic terminals described in prior human post-mortem and non-human mammal studies (Albin, Howland, Higgins, & Frey, 1994; Barmack, Baughman, & Eckenstein, 1992; Barmack, Baughman, Eckenstein, & Shojaku, 1992; De Lacalle, Hersh, & Saper, 1993; De Lacalle, Lim, Sobreviela, Mufson, Hersh, & Saper, 1994; Emre, Heckers, Mash, Geula, & Mesulam, 1993; Fitzpatrick, Diamond, & Rackowski, 1989; Fukushima, Kitahara, Takeda, Saika, & Kubo, 2001; Gilmor, et al, 1999; Gilmor, et al, 1996; Heckers, Gelua, & Mesulam, 1992; Ichikawa, Ajiki, Matsuura, & Misawa, 1997; Jaarsma, etal., 1997; Levey, Hallanger, & Wainer, 1987; Mahady, Periz, Emerich, Wahlberg, & Mufson, 2016; Manaye, Zweig, Wu, Hersh, De Lacalle, Saper, German, 1999; Mesulam, 2004; Mesulam & Geula, 1988; Mesulam, Geula, Bothwell, & Hesh, 1989; Mesulam, Hersh, Mash, & Geula, 1992; Mesulam, Mash, Hersh, Bothwell, & Geula, 1992; Roghani, Shirzadi, Butcher, & Edwards, 1998; Schafer, Weihe, Erickson, & Eiden, 1996; Schafer, Eiden, & Weihe, 1998; Woolf, 1991; Woolf, Eckenstein, & Butcher, 1984; Woolf & Butcher, 1985, 1986, 1989; Zhang, Zhou, & Yuan, 2016). Our results are consistent as well with prior VAChT ligand human SPECT imaging studies (Lamare et al 2013; Kuhl et al, 1994; Kuhl et al, 1996).…”

“…Prior investigations in other species, including non-human primates, and work with post- mortem human tissues provided considerable information about the organization of human brain cholinergic pathways (Albin, Howland, Higgins, & Frey, 1994; Barmack, Baughman, & Eckenstein, 1992; Barmack, Baughman, Eckenstein, & Shojaku, 1992; De Lacalle, Hersh, & Saper, 1993; De Lacalle, Lim, Sobreviela, Mufson, Hersh, & Saper, 1994Emre, Heckers, Mash, Geula, & Mesulam, 1993; Fitzpatrick, Diamond, & Rackowski, 1989; Fukushima, Kitahara, Takeda, Saika, & Kubo, 2001; Gilmor, et al, 1999; Gilmor, et al, 1996; Heckers, Gelua, & Mesulam, 1992; Ichikawa, Ajiki, Matsuura, & Misawa, 1997; Jaarsma, etal., 1997; Kus, etal., 2003; Levey, Hallanger, & Wainer, 1987; Mahady, Periz, Emerich, Wahlberg, & Mufson, 2016; Manaye, Zweig, Wu, Hersh, De Lacalle, Saper, German, 1999; Mesulam, 2004; Mesulam & Geula, 1988; Mesulam, Geula, Bothwell, & Hesh, 1989; Mesulam, Hersh, Mash, & Geula, 1992; Mesulam, Mash, Hersh, Bothwell, & Geula, 1992; Roghani, Shirzadi, Butcher, & Edwards, 1998; Schafer, Weihe, Erickson, & Eiden, 1996; Schafer, Eiden, & Weihe, 1998; Woolf, 1991; Woolf, Eckenstein, & Butcher, 1984; Woolf & Butcher, 1985, 1986, 1989; Zhang, Zhou, & Yuan, 2016). Working with post-mortem tissues has intrinsic limitations and inferences from studies of other species may be confounded by inter-species differences.…”

Effect of a diet-induced n-3 PUFA depletion on cholinergic parameters in the rat hippocampus

“…Animals were killed by decapitation, and their brains were rapidly removed and frozen in isopentane at Ϫ 35 Њ C. Coronal cryosections (20 m) were cut, thaw-mounted on gelatin-coated microscope slides, and stored at Ϫ 80 Њ C. The [ 3 H]scopolamine binding assay was performed according to Albin et al (27). Sections were incubated for 30 min at room temperature with buffer (pH 7.4) containing 137 mM NaCl, 3 mM KCl, 1 mM EDTA, 8 mM NaHPO 4 , 1.5 mM KH 2 PO 4 , and 5 nM [ 3 H]scopolamine (70 Ci/mmol, Amersham, France) with (nonspecific binding) or The diet provided about 16.5 MJ/kg diet.…”

Near‐complete adaptation of the PRiMA knockout to the lack of central acetylcholinesterase

“…according to the references: 1)(Wolff et al 2008); 2)(Spurden et al 1997); 3)(Perry and Kellar 1995); 4)(Fernagut et al 2003); 5)(Martres et al 1985); 6)(Ayata et al 1997); 7)(Dean et al 1999); 8)(Albin et al 1994); 9)(Araki et al 2000).…”