Cav3 channels consist of three isoforms, Cav3.1 (α1G), Cav3.2 (α1H), and Cav3.3 (α1I), which produce low-threshold spikes that trigger burst firings in nociceptive neurons of the spinal dorsal horn (SDH) and dorsal root ganglion (DRG). Although Cav3.2 plays a crucial role in pathological pain, its distribution in SDH still remains controversial. One study showed that Cav3.2 is ubiquitously expressed in neurons, but another study implied that Cav3.2 is expressed restricted to astrocytes. To unravel these discrepancies, we used methods of immunohistochemistry either with or without antigen retrieval (AR) pre-treatment to detect Cav3 in SDH and DRG from both rats and mice. Moreover, Cav3.2 mRNA was detected in mice SDH using in situ hybridization. We found that the expression pattern of Cav3.2 but not Cav3.1 and Cav3.3 in SDH were largely different with or without AR pre-treatment, which showed a neuron-like and an astrocyte-like appearance, respectively. Double staining further demonstrated that Cav3.2 was mainly co-stained with the neuronal marker NeuN in the presence of AR but was with glial fibrillary acidic protein (GFAP, marker for astrocytes) in the absence of AR pre-treatment. Importantly, Cav3.2 mRNA was mainly co-localized with Cav3.2 but not GFAP. Together, our findings indicate that AR pre-treatment or not impacts the expression pattern of Cav3.2, which may make a significant contribution to the future study of Cav3.2 in SDH.

中文翻译：

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抗原提取预处理导致大鼠和小鼠脊髓背角中Cav3.2的不同表达模式。

Cav3通道由三种同工型Cav3.1（α1G），Cav3.2（α1H）和Cav3.3（α1I）组成，它们产生低阈值尖峰，从而触发脊髓背角（SDH）伤害感受神经元的爆发性放电。和背根神经节（DRG）。尽管Cav3.2在病理性疼痛中起关键作用，但其在SDH中的分布仍存在争议。一项研究表明，Cav3.2在神经元中普遍存在，但另一项研究表明，Cav3.2仅限于星形胶质细胞表达。为了消除这些差异，我们使用了免疫组织化学方法，通过或不采用抗原修复（AR）预处理，来检测大鼠和小鼠SDH和DRG中的Cav3。此外，使用原位杂交在小鼠SDH中检测到Cav3.2 mRNA。我们发现Cav3.2的表达模式，但不是Cav3.1和Cav3的表达模式。SDH中3个在有或没有进行AR预处理的情况下都存在很大差异，分别显示出神经元样和星形胶质样外观。双重染色进一步证明，在AR存在下，Cav3.2主要与神经元标志物NeuN共染色，而在不进行AR预处理的情况下，其与胶质纤维酸性蛋白（GFAP，星形胶质细胞标志物）共染色。重要的是，Cav3.2 mRNA主要与Cav3.2共定位，但不与GFAP共定位。总之，我们的发现表明AR预处理是否会影响Cav3.2的表达模式，这可能对SDH中Cav3.2的未来研究做出重要贡献。2在存在AR的情况下主要与神经元标记NeuN共同染色，而在不进行AR预处理的情况下与神经胶质纤维酸性蛋白（GFAP，星形胶质细胞的标志物）共同染色。重要的是，Cav3.2 mRNA主要与Cav3.2共定位，但不与GFAP共定位。总之，我们的发现表明AR预处理是否会影响Cav3.2的表达模式，这可能对SDH中Cav3.2的未来研究做出重要贡献。2在存在AR的情况下主要与神经元标记NeuN共同染色，而在不进行AR预处理的情况下与神经胶质纤维酸性蛋白（GFAP，星形胶质细胞的标志物）共同染色。重要的是，Cav3.2 mRNA主要与Cav3.2共定位，但不与GFAP共定位。总之，我们的发现表明AR预处理是否会影响Cav3.2的表达模式，这可能对SDH中Cav3.2的未来研究做出重要贡献。