| Evidence Sentence: |
Compared to sham-MA control without manual stimulation, we find that MA at P6 markedly increases c-Fos positive neurons in a number of the forebrain regions (n=5 in each group). |
| Evidence Sentence: |
In the present study, we find that in the Fos-CreER; Ai9 mice model, MA at P6 markedly increases c-Fos positive neurons in a number of forebrain regions, including somatosensory cortex (S), ectorhinal cortex (Ect), and dorsolateral entorhinal cortex (DLEnt), claustrum (CI), accumbens nucleus (Acb), caudate putamen (CPu), basolateral (BL) and central (Ce) divisions of amygdaloid nucleus, bed nucleus of the stria terminalis (STLD), ventral posterior division of the thalamic nucleus (VP), paraventricular hypothalamic nucleus (PVN), and arcuate hypothalamic nucleus (Arc). |
| Evidence Sentence: |
A microscopic image shows an example of c-Fos expression (indicated by an arrow) in the Arc of a representative mouse following P6 acupuncture treatment. |
| Evidence Sentence: |
The c-Fos expression in the neuron, appearing as red dots, is found bilaterally in many regions across the forebrain in the mice subjected to sham-MA (n=5, Figure 2) or MA (n=5, Figure 3). |
| Evidence Sentence: |
In those areas, no or scarce c-Fos positive neurons are found similarly in both sham-MA and MA group. |
| Evidence Sentence: |
We systematically examine c-Fos expression at the multiple rostro-caudal levels noted above, according to the mouse brain atlas. |
| Evidence Sentence: |
We identify a notable significant increase in c-Fos positive neurons in some specific nuclei of the forebrain in mice treated with MA, described in detail below, compared to sham-MA (Figures 2-4 and Table 1). |
| Evidence Sentence: |
In the cortex, significant increases in c-Fos expression (p<0.05) are observed in the mice treated with MA in the somatosensory cortices (S), including primary (S1) and secondary (S2) somatosensory cortex, ectorhinal cortex (Ect), and dorsolateral entorhinal cortex (DLEnt), compared to control mice (Table 1 and Figure 4). |
| Evidence Sentence: |
In the MA-treated mice, c-Fos expression density in the S tends to be much higher at the rostral levels (Bregma 0.86 through -0.82 mm), followed by a decrease with more caudal levels (Figure 4). |
| Evidence Sentence: |
In the Ect, more c-Fos expression is observed rostrally but Ect Fos expression decreases when approaching the midbrain (Figure 4, Bregma -1.34 through -4.96 mm). |
| Evidence Sentence: |
We find c-Fos positive neurons in the basal nuclei in both groups. |
| Evidence Sentence: |
However, the number of cells expressing c-Fos (about 18 cells per section) in the claustrum (CI) of MA-treated mice is three times more than that in the mice subjected to sham-MA, in which few cells (approximate 6 cells per section) expressing c-Fos are found (Figure 4, Bregma 1.54 through -0.82 mm). |
| Evidence Sentence: |
Most c-Fos expression is observed in the shell of the Acb with scattered expression within its core. |
| Evidence Sentence: |
Despite the presence of c-Fos expression throughout the caudate putamen (CPu) of all mice in two groups, there are more (slightly over double) neurons expressing c-Fos in MA-treated mice than those in mice that underwent sham-MA (Figure 4, Bregma 1.54 through -1.34 mm). |
| Evidence Sentence: |
C-Fos expression is found more frequently in this nucleus, particularly in the Ce of MA-treated mice, compared to mice that underwent sham-MA (Figures 2 and 3). |
| Evidence Sentence: |
There is 24 times more c-Fos expression identified in the Ce of mice with MA treatment than that in sham-MA controls (p<0.05; Table 1, Figure 4, Bregma -0.58 through -2.06 mm). |
| Evidence Sentence: |
In the BL, c-Fos expression is not increased as dramatically as that in the Ce. |
| Evidence Sentence: |
In the bed nucleus of the stria terminalis (BNST), the most expression of c-Fos is identified in the lateral division of BNST (STLD), particularly in the dorsal part of STLD in MA-treated mice. |
| Evidence Sentence: |
In contrast, very few neurons expressing c-Fos are found in the mice that underwent sham-MA (Figures 2 and 3, and Figure 4, Bregma 0.14 mm). |
| Evidence Sentence: |
Such prominent expression of c-Fos is not noted in the other parts of the BNST in mice treated with MA or sham-MA. |
| Evidence Sentence: |
We observe, in the thalamus, a significant increase in c-Fos expression primarily in its ventral posterior division (VP) of the mice subjected to MA treatment, compared to sham-MA controls (p<0.05; Table 1 and Figure 4, Bregma -1.34 through -2.06 mm). |
| Evidence Sentence: |
The expression of c-Fos is identified in the lateral and medial portions of the VP (Figure 4). |
| Evidence Sentence: |
Less c-Fos positive cells are noted in the lateral parts of the VP (VPL) than those in the medial part of the VP (VPM) in both MA and sham-MA groups. |
| Evidence Sentence: |
However, importantly, the c-Fos positive cells are significantly increased in VPL but not in VPM in MA-treated mice, in comparison with controls. |
| Evidence Sentence: |
In the hypothalamus, a marked increase in c-Fos expression is located in the paraventricular nucleus (PVN) and arcuate nucleus (Arc) in the mice treated with MA, compared to sham-MA controls (p<0.05; Table 1, Figures 2-4). |
| Evidence Sentence: |
In the Arc nucleus, while there are some Fos-positive cells noted in the mice treated with sham-MA, obviously more c-Fos expression is observed in MA-treated mice, especially approaching the level at Bregma -2.06 mm as shown in Figures 2-4. |
| Evidence Sentence: |
We note that in the areas of the midbrain, excluding the aforementioned cortex regions, there is no significant difference in c-Fos expression between MA-treated group and control group (Table 1). |
| Evidence Sentence: |
Notably, in the periaqueductal gray (PAG), we observe c-Fos expression primarily located in its lateral portions, with no or little presence in its dorsal parts, approaching the level at Bregma -4.24 mm in both groups. |
| Evidence Sentence: |
Among all regions in which more c-Fos expression is found in MA-treated mice than in controls, there are significant correlations of the numbers of Fos-positive neurons between some nuclei following MA treatment, including PVN and STLD, S and CI, CI and Arc, as well as Ect and STLD (Table 2 and Figure 5). |
| Evidence Sentence: |
Fos-CreER Based Genetic Mapping of Forebrain Regions Activated by Acupuncture |
| Evidence Sentence: |
Thus, we mapped P6 specific neural activation by MA in the forebrain using the Fos-CreER; Ai9 mice model, which allows for enhanced sensitivity and efficiency compared to conventional immunohistochemical staining. |
| Evidence Sentence: |
In the present study, we find that in the Fos-CreER; Ai9 mice model, MA at P6 markedly increases c-Fos positive neurons in a number of forebrain regions, including somatosensory cortex (S), ectorhinal cortex (Ect), and dorsolateral entorhinal cortex (DLEnt), claustrum (CI), accumbens nucleus (Acb), caudate putamen (CPu), basolateral (BL) and central (Ce) divisions of amygdaloid nucleus, bed nucleus of the stria terminalis (STLD), ventral posterior division of the thalamic nucleus (VP), paraventricular hypothalamic nucleus (PVN), and arcuate hypothalamic nucleus (Arc). |
| Evidence Sentence: |
These selected sections are closely matched to corresponding levels of Bregma of the forebrain, which are demonstrated in the Franklin & Paxinos mouse brain atlas as well as in Figure 4 which schematically depicts Fos-positive cells to brain regions. |
| Evidence Sentence: |
We also find that there is no significant difference in the number of Fos-positive cells between the two groups in the majority of forebrain regions (Table 1). |
| Evidence Sentence: |
In the Ect, more c-Fos expression is observed rostrally but Ect Fos expression decreases when approaching the midbrain (Figure 4, Bregma -1.34 through -4.96 mm). |
| Evidence Sentence: |
Unlike the expression observed in the Ect, the majority of Fos expression in the DLEnt, positioned just below the Ect, are found in sections closer to the midbrain (Figure 4, Bregma -2.06 through -2.80 mm). |
| Evidence Sentence: |
Although Fos-positive neurons are identified in the majority of cortex regions, there is no significant difference in the number of these neurons between sham-MA and MA groups. |
| Evidence Sentence: |
Fos-positive neurons are increased significantly in the accumbens nucleus (Acb) of MA-treated mice in comparison with sham-MA mice (p<0.05; Table 1). |
| Evidence Sentence: |
The Fos-positive cells are located mainly in the medial and central parts of the Ce (Figure 4). |
| Evidence Sentence: |
There is a significant difference in the number of Fos-positive cells between these two groups (p<0.05; Table 1). |
| Evidence Sentence: |
Besides, there is no significant difference between the two groups (Table 1) in the number of Fos-positive neurons in the ventrolateral, ventromedial, mediodorsal, central medial, and submedius divisions of the thalamic nucleus, and neither in the paratenial thalamic nucleus and habenular nucleus. |
| Evidence Sentence: |
Notably, we only find very few Fos-positive neurons (about one cell per section) in the PVN of mice underwent sham-MA. |
| Evidence Sentence: |
In contrast, about 13 Fos-positive neurons per section are identified in MA-treated mice (Figures 2-4, Bregma -0.82 mm). |
| Evidence Sentence: |
In the Arc nucleus, while there are some Fos-positive cells noted in the mice treated with sham-MA, obviously more c-Fos expression is observed in MA-treated mice, especially approaching the level at Bregma -2.06 mm as shown in Figures 2-4. |
| Evidence Sentence: |
Among all regions in which more c-Fos expression is found in MA-treated mice than in controls, there are significant correlations of the numbers of Fos-positive neurons between some nuclei following MA treatment, including PVN and STLD, S and CI, CI and Arc, as well as Ect and STLD (Table 2 and Figure 5). |
