Siegler Preston Nicole, Shaughnessy Emma K, Horman Brian, Vierling Tia T, King Darron H, Patisaul Heather B, Huhman Kim L, Alexander Georgia M, Dudek Serena M
Neurobiology Laboratory, National Institute of Environmental Health Sciences, NIH, 111 T.W. Alexander Drive, Research Triangle Park, NC 27709 USA.
Neuroscience Center, University of North Carolina, Chapel Hill, NC.
bioRxiv. 2024 Feb 14:2024.02.12.579957. doi: 10.1101/2024.02.12.579957.
Prairie voles () and Syrian, or golden, hamsters () are closely related to mice () and rats (, for example) and are commonly used in studies of social behavior including social interaction, social memory, and aggression. The CA2 region of the hippocampus is known to play a key role in social memory and aggression in mice and responds to social stimuli in rats, likely owing to its high expression of oxytocin and vasopressin 1b receptors. However, CA2 has yet to be identified and characterized in hamsters or voles. In this study, we sought to determine whether CA2 could be identified molecularly in vole and hamster. To do this, we used immunofluorescence with primary antibodies raised against known molecular markers of CA2 in mice and rats to stain hippocampal sections from voles and hamsters in parallel with those from mice. Here, we report that, like in mouse and rat, staining for many CA2 proteins in vole and hamster hippocampus reveals a population of neurons that express regulator of G protein signaling 14 (RGS14), Purkinje cell protein 4 (PCP4) and striatal-enriched protein tyrosine phosphatase (STEP), which together delineate the borders with CA3 and CA1. These cells were located at the distal end of the mossy fiber projections, marked by the presence of Zinc Transporter 3 (ZnT-3) and calbindin in all three species. In addition to staining the mossy fibers, calbindin also labeled a layer of CA1 pyramidal cells in mouse and hamster but not in vole. However, Wolframin ER transmembrane glycoprotein (WFS1) immunofluorescence, which marks all CA1 neurons, was present in all three species and abutted the distal end of CA2, marked by RGS14 immunofluorescence. Staining for two stress hormone receptors-the glucocorticoid (GR) and mineralocorticoid (MR) receptors-was also similar in all three species, with GR staining found primarily in CA1 and MR staining enriched in CA2. Interestingly, although perineuronal nets (PNNs) are known to surround CA2 cells in mouse and rat, we found that staining for PNNs differed across species in that both CA2 and CA3 showed staining in voles and primarily CA3 in hamsters with only some neurons in proximal CA2 showing staining. These results demonstrate that, like in mouse, CA2 in voles and hamsters can be molecularly distinguished from neighboring CA1 and CA3 areas, but PNN staining is less useful for identifying CA2 in the latter two species. These findings reveal commonalities across species in molecular profile of CA2, which will facilitate future studies of CA2 in these species. Yet to be determined is how differences in PNNs might relate to differences in social behavior across species.
草原田鼠()和叙利亚仓鼠(又称金黄仓鼠)()与小鼠()和大鼠(例如)亲缘关系密切,常用于社会行为研究,包括社会互动、社会记忆和攻击行为。已知海马体的CA2区域在小鼠的社会记忆和攻击行为中起关键作用,并且在大鼠中对社会刺激有反应,这可能是由于其催产素和血管加压素1b受体的高表达。然而,CA2在仓鼠或田鼠中尚未被鉴定和表征。在本研究中,我们试图确定是否可以在田鼠和仓鼠中通过分子手段鉴定出CA2。为此,我们使用针对小鼠和大鼠中已知的CA2分子标记物产生的一抗进行免疫荧光,对来自田鼠和仓鼠的海马切片以及小鼠的海马切片进行平行染色。在此,我们报告,与小鼠和大鼠一样,田鼠和仓鼠海马体中许多CA2蛋白的染色揭示了一群表达G蛋白信号调节因子14(RGS14)、浦肯野细胞蛋白4(PCP4)和纹状体富集蛋白酪氨酸磷酸酶(STEP)的神经元,这些蛋白共同勾勒出与CA3和CA1的边界。这些细胞位于苔藓纤维投射的远端,在所有三个物种中都以锌转运体3(ZnT - 3)和钙结合蛋白的存在为标志。除了对苔藓纤维进行染色外,钙结合蛋白还标记了小鼠和仓鼠中一层CA1锥体细胞,但在田鼠中没有。然而,标记所有CA1神经元的沃尔弗勒姆内质网跨膜糖蛋白(WFS1)免疫荧光在所有三个物种中都存在,并与以RGS14免疫荧光为标志的CA2远端相邻。两种应激激素受体——糖皮质激素(GR)和盐皮质激素(MR)受体——的染色在所有三个物种中也相似,GR染色主要在CA1中发现,而MR染色在CA2中富集。有趣的是,尽管已知在小鼠和大鼠中神经元周围网络(PNN)围绕着CA2细胞,但我们发现PNN的染色在不同物种中有所不同,因为在田鼠中CA2和CA3都显示出染色,在仓鼠中主要是CA3显示染色,只有近端CA2中的一些神经元显示染色。这些结果表明,与小鼠一样,田鼠和仓鼠中的CA2可以通过分子手段与相邻的CA1和CA3区域区分开来,但PNN染色对于在后两个物种中鉴定CA2不太有用。这些发现揭示了不同物种间CA2分子特征的共性,这将有助于未来对这些物种中CA2的研究。尚待确定的是PNN的差异可能如何与不同物种间社会行为的差异相关联。