Semantic Error Chapter 47

Semantic Error Chapter 47 – Despite decades of discussion in the neuroanatomical literature, the role of synaptic “spines” in synaptic development and function remains unclear. Canonically, spinules are finger-like projections arising from presynaptic spines and covered by presynaptic spines. When the presynaptic bouton covers the spine in this way, the membrane spacing between the spine and the surrounding bouton may be much larger than the membrane interface of the synaptic active site. Therefore, spines may represent mechanisms of extrasynaptic communication between neurons and/or serve as structural “anchors” that increase the stability of cortical synapses. Despite their potential to influence synaptic function, we have little information on the percentage of developing and adult cortical bouton populations with spines or the percentage of these cortical spine boutons (CSPs) with spines of distinct neuronal/glial origin. whether activity onset or cortical plasticity is associated with increased cortical SBB prevalence. Here, we used 2D and 3D electron microscopy to characterize the distribution of spines in excitatory presynaptic boutons at key developmental sites in the primary visual cortex (V1) of female and male ferrets. Because the prevalence of SBB in V1 increases during postnatal development, ∼25% of boutons leading to V1 excitability in late adolescence are spiny. Furthermore, the majority of SBB spines during late development originate from postsynaptic spines and adjacent boutons/axons, suggesting that synaptic spines enhance synaptic stability and axo-axonal connections in the mature sensory cortex.

Synaptic spinules are finger-like projections of neurons that are fully embedded in the presynaptic bouton, increasing synaptic connectivity and stability. Although their existence has been debated for decades, their relationship to spinal distribution, origin of projections, and neocortical sensory function is unclear. In this study, we use 2D and 3D electron microscopy techniques to investigate the formation of spiny cortical boutons (SCBs) and their relationship to sensory function and plasticity. Our results show that SBB distribution before neocortical synapses does not coincide with the onset of sensory activity or increased cortical plasticity, suggesting that a quarter of presynaptic boutons are spiking during adolescence. Thus, as these synapses mature, synaptic spines become progressively more important in cortical function.

Semantic Error Chapter 47

Semantic Error Chapter 47

Synapses underlie the sensory, motor, and cognitive functions of the nervous system, and changes in synaptic connectivity and stability directly affect animal behavior and memory (Sherrington, 1906; Liewald et al., 2008; Liu et al., 2012). Furthermore, changes in synaptic morphology can accurately predict changes in synaptic strength and stability (Murthy et al., 2001; Ostroff et al., 2002; Branko et al., 2010; Holderit et al., 2012; Araya et al., 2014). ). ; Meyer et al., 2014; Quinn et al., 2019). However, synaptic spines, a ubiquitous and important synaptic structure, are enigmatic and understudied.

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Spinules are a conserved component of synapses throughout the animal kingdom (Case et al., 1972; Bailey et al., 1979), but their function is unclear. Canonically, spines are thin, finger-like bundles of postsynaptic dendrites that may be covered by other neurites ( Pappas and Purpura, 1961 ; Westrum and Blackstad, 1962 ; Tarrant and Routtenberg, 1977 ; Spack and Harris, 204 ). When neuronal structures such as presynaptic boutons cover spines, the membrane location between the spike discharge structures and the capping bouton increases the synaptic “active zone” in some cases up to 26-fold (Rodriguez-Moreno et al., 2018). Therefore, spines may represent an unknown mechanism of neuronal communication and/or they may serve as structural “anchors” that increase synapse strength and stability. If true, increased dorsal synapses in functionally defined microcircuits would lead to major changes in brain function, such as enhanced memory or improved recovery after injury.

To date, the best anatomically and physiologically characterized types of spines are those of the mammalian hippocampus, particularly the postsynaptic spines of stratum radiata CA1 in adult rats and the dendrites of dentate pyramidal cells (Tarrant and Routtenberg et al., 1977; , 1994; Toney et al., 1994). ., 1999; Spacek and Harris, 2004). Although spines typically arise from a spine and are capped by presynaptic boutons, the term “spine” has been expanded to describe any neurite or glial projection capped by another neuron or glial process (Petralia et al., 2015). In CA1 stratum radiatum, ∼32% of spines project to spines, and most of these spines are covered by presynaptic boutons (∼90%; Spacek and Harris, 2004 ). Interestingly, following increases in neuronal activity during electrical and chemical long-term potentiation (LTP; Geinisman et al.

(Tao-Cheng et al., 2009) and activation of estradiol receptors (Murphy and Andrews, 2000). These data have led to the idea that spines may act as a signaling element for changes in neuronal circuits due to brain activity (Spacek and Harris, 2004) or as a mechanism for restoring the presynaptic membrane during increased activity (Tao-Chen et al.). ). , 2009).

However, generalization of these findings in the hippocampus to other brain regions (eg, neocortex) remains unclear, and whether higher levels of developmental activation/plasticity correlate with higher levels of spinal cord proliferation in vivo is unclear. In addition, several studies have examined the percentage of presynaptic boutons with spines, a key component in understanding the potential of spine synaptic connections and/or stability. For example, a 2D ultrastructural study of thalamocortical (TC) boutons revealed that 28% of TC boutons in layer 4 (L4) of the primary visual cortex (V1) had a 2D spiniform (ie, putative spiniform) profile (Erisir and Dreusicke, 2005). ) and a 3D study of TC boutons in the L4 region of the barrel cortex reported that 13% of regenerated postsynaptic dendritic spines innervate TC boutons ( Rodriguez-Moreno et al., 2018 ). Thus, spines may be a key feature of neocortical synapses.

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Here, we used the broad definition of spines to describe the invaginating projections of neurites or glia covered by excitatory cortical presynaptic boutons. We sought to determine the proportion of boutons with synaptic excitatory spines (SBBs) within the excitatory bouton population in V1 throughout postnatal development and the origin of spines projecting to these SBBs. Furthermore, we investigated whether the onset of perceptual activity or increased levels of cortical plasticity leads to a proportional increase in the proportion of SBB in V1. Using 2D and 3D electron microscopy analysis, we found that (1) the proportion of SBBs increases with strengthening and developmental plasticity at L4 excitatory synapses in V1 and (2) SBBs from postsynaptic spines and adjacent boutons. we found that it covers the spine. /mature axons and (3) ∼25% of excitatory synaptic boutons in late juvenile ferrets are spiny.

= 3). These age periods were chosen because they are key developmental points: before eye opening (ie, before ∼p32), the peak of the critical period of eye dominance and morphological plasticity of TS axons (∼p46), and the end of this critical period in V1 (∼p60). ), age approaching puberty in ferrets (>p90; Isa et al., 1999). For 2D analysis, measurements from animals at similar developmental stages were grouped [ie, p21–p28 (

= 3)], after finding that within-group (i.e., p21 and p28) synaptic length and bouton area measurements were not statistically significant (one-way ANOVA and

Semantic Error Chapter 47

> 0.1; data not shown). All animal procedures and protocols followed NIH guidelines for the humane treatment of animals and were approved by the University of Virginia Animal Care and Use Committee.

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Animals were given an overdose of Nembutal (>50 mg/kg) and transcardially perfused with 4% paraformaldehyde and 0.5% glutaraldehyde in 0.1 m phosphate buffer (PB; pH 7.4). To avoid increased spinal cord diffusion due to long perfusion times, only animals with a time from thoracic incision to fixation <100 s were included in this study ( Tao-Cheng et al., 2009 ). After perfusion, brains were removed and stored in 4% paraformaldehyde at 4 °C overnight. The next day, each brain was dissected, the occipital lobe blocks were placed in vibrato, and coronal sections of V1 were cut at 60 μm. Free-floating coronal sections were immediately treated with 1% sodium borohydride, rinsed 5–6 times in PBS, and free-floating sections were stored at 4°C in PBS containing 0.05% sodium azide.

Sections prepared for TEM analysis were rinsed in 0.1 m PB and then immersed in 1% osmium tetroxide (0.1 m PB) for 1 h. Sections were then rinsed in 0.1 m PB, dehydrated in an ethanol dilution series, and then incubated in 4% uranyl acetate (70% ethanol) overnight at 4 °C. Sections were then dehydrated in acetone and incubated in three mixtures of progressively concentrated acetone-Epon 812 resin (EMS, catalog #RT14120) overnight at room temperature for 4 hours. The parts were then glued flat between two plastic films (EMS, catalog no. 50425-10) and placed overnight in an oven heated to 60 °C. Sections from the fourth layer of endoscopic region V1 of implanted coronal sections were labeled under light microscopy using white matter markers ( Law et al., 1988 ), cut from planar mounts, and mounted.

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