Page 427 - IJB-10-4

International Journal of Bioprinting Pregabalin impact on 3D neuronal models

of VMDNs, as evidenced by a reduction in ATP secretion. Another similar gene, Gad67, is the main metabolic
Such findings imply that pregabalin acts upon the electron catalyst involved in GABA synthesis. Using single-
60
transport chain, leading to enhanced production of cell PCR, western blotting, and immunohistochemical
ATP. 19,56 According to our analysis of the effects of the methods, it was previously reported that the expression
drug on cortical neuron formation, the length of the total of the GABAergic neuron protein markers GAD65 and
and dominant neurites, as well as the number of neurites GAD67 is significantly reduced in the central nucleus of the
and branches in Tbr1-positive cortical neurons, are not amygdala in spinal nerve-ligated rats, indicating a decrease
substantially changed following pregabalin treatment. in the GABAergic block under neuropathic conditions.
61
Similarly, it does not significantly modify the number of From cell-fate selection to connection, the LIM
neurites, branch numbers, or the length of neurites in homeodomain family of transcription factors is involved in
Tbr1-negative cortical neuron cultures. a wide range of events in the developing CNS. The medial

The transcription factors Dlx-1, Gsx2, and Emx-2 ganglionic eminence of the ventral telencephalon, which
are essential for forebrain specification. Our findings produces the bulk of cortical interneurons, expresses Lhx6,
suggest that pregabalin use during pregnancy is not safe, a transcription factor belonging to the LIM homeodomain
as it may alter gene expression in the cortical region. The family. As it is expressed in GABA-containing medial
development of telencephalic inter-neurons depends on ganglionic eminence cells that are transferred to the cortex,
the gene expression of transcription factors containing Lhx6 may be involved in the neurochemical uniqueness
homeodomains (Dlx-1 and Dlx-2) in the lateral and and relocation of these neurons, either alone or in
62
medial ganglionic eminences and subpallial embryonic conjunction with other transcription factors. According
structures. The development of telencephalic inter- to our findings, pregabalin-treated cultures displayed
neurons depends on the gene expression of transcription significantly downregulated expressions of Otp, Nhlh2,
factors containing homeodomains (Dlx-1 and Dlx-2) in and Gad67. Initially, it was believed that Nhlh2 was largely
the lateral and medial ganglionic eminences and subpallial responsible for neuronal development and pediatric
63
embryonic structures. Inter-neurons progenitors are neuroblastomas. Olig2 is a basic helix–loop–helix
64
derived from the lateral and medial ganglionic eminences transcription factor highly expressed in the CNS and
and then migrate to different forebrain structures, such predominantly controls neurogenesis as an anti-neurogenic
as the cortex, hippocampus, and olfactory bulb. The factor. However, it may also be necessary for precise
manifestation of the olfactory bulb dopamine phenotype neuronal differentiation at certain developmental stages
was previously associated with the Dlx-1 and Dlx-2 genes, of the CNS. 65,66 While Olig2 is primarily associated with
according to studies on the development of telencephalic oligodendrocyte lineage, emerging research has indicated
57
neurons in Dlx-1- and Dlx-2-deficient mice. In contrast its presence and significance in other neural cell types,
to the control cultures, pregabalin-treated cultures including cortical neurons. Past research has identified
displayed a significant shift (downregulation) in Dlx-2 the expression of Olig2 across multiple neuronal types in
expression (Figure 5E). Similarly, Emx2, predominantly human fetal CNS, including olfactory neurons, postmitotic
expressed in this area from the onset of cortical neuronal interneurons in the spinal cord, and neurons undergoing
development, serves as an effective dorsal indicator for migration in the cortical subventricular zone (SVZ). In
the maturing cerebral cortex. Neuroblasts in the process the forebrain, specifically, Olig2 has been detected in early
of proliferation from the neuroepithelium (ventricular MAP2+ neurons, which are likely in the midst of migration
zone), recognized for their role in controlling neuronal within the SVZ. However, it is significantly lacking in the
radial migration, contribute to this cortical expression more differentiated NeuN+ neurons located in the cortical
67
as well. The phenotype of Emx2-deficient embryos was plate. This study demonstrated that pregabalin exposure
examined, and it was discovered that this transcription causes upregulated expressions of Olig2 and Zic1. The
factor is crucial for neuroblast growth and development. alteration in Zic1 gene expression is of significance due to
58
In the ventral telencephalon, the lateral ganglionic its role as a transcription factor involved in various aspects
eminence and two homeobox genes, Gsx1 and Gsx2 of neural development, including cell fate determination,
68
(previously called Gsh1 and Gsh2, respectively), are differentiation, and morphogenesis. A previous study
among the genes encoding the earliest transcription highlighted robust expressions of Zic1 and Zic3 in
factors produced in neural progenitors. The latter two specific medial areas of the brain, such as the septum,
genes have also been recently found to define various medial cerebral cortex, and choroid plexus, providing
lateral ganglionic eminence neuron subtypes at discrete significant insight into forebrain development and genetic
times and are a primary requirement for lateral ganglionic mutations. A deeper understanding of the molecular
69
eminence progenitor cells. 59 mechanisms underlying pregabalin’s effects on neural

Volume 10 Issue 4 (2024) 419 doi: 10.36922/ijb.3010