Patterns of connexin36 and eGFP reporter expression among motoneurons in spinal sexually dimorphic motor nuclei in mouse.

Prabhisha Silwal, Pratyaksh Singhal, Joanne Mm Senecal, Julie Em Senecal, Bruce D Lynn, James I Nagy
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Abstract

Background: Sexually dimorphic spinal motoneurons (MNs) in the dorsomedial nucleus (DMN) and dorsolateral nucleus (DLN) as well as those in the cremaster nucleus are involved in reproductive behaviours, and the cremaster nucleus additionally contributes to testicular thermoregulation. It has been reported that MNs in DMN and DLN are extensively linked by gap junctions forming electrical synapses composed of connexin36 (Cx36) and there is evidence that subpopulation of MNs in the cremaster nucleus are also electrically coupled by these synapses.

Methodology: We used immunofluorescence methods to detect enhanced green fluorescent protein (eGFP) reporter for Cx36 expression in these motor nuclei.

Results: We document in male mice that about half the MNs in each of DMN and DLN express eGFP, while the remaining half do not. Further, we found that the eGFP+ vs. eGFP- subsets of MNs in each of these motor nuclei innervate different target muscles; eGFP+ MNs in DMN and DLN project to sexually dimorphic bulbocavernosus and ischiocavernosus muscles, while the eGFP- subsets project to sexually non-dimorphic anal and external urethral sphincter muscles. Similarly, eGFP+ vs. eGFP- cremaster MNs were found to project to anatomically distinct portions of the cremaster muscle. By immunofluorescence, nearly all motoneurons in both DMN and DLN displayed punctate labelling for Cx36, including at eGFP+/eGFP+, eGFP+/eGFP- and eGFP-/eGFP- cell appositions.

Conclusions: Most if not all motoneurons in DMN and DLN are electrically coupled, including sexually dimorphic and non-dimorphic motoneurons with each other, despite absence of eGFP reporter in the non-dimorphic populations in these nuclei that have selective projections to sexually non-dimorphic target muscles.

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小鼠脊髓性双态运动核中运动神经元的连接蛋白36和eGFP报告表达模式
背景:背内侧核(DMN)和背外侧核(DLN)以及襞核中的性双态脊髓运动神经元(MNs)参与生殖行为,襞核还对睾丸的体温调节做出了贡献。有报道称,DMN和DLN中的MN通过间隙连接广泛相连,形成由Connexin36(Cx36)组成的电突触,有证据表明,嵴核中的亚群MN也通过这些突触进行电耦合:我们使用免疫荧光方法检测这些运动核中 Cx36 表达的增强型绿色荧光蛋白(eGFP)报告物:结果:我们在雄性小鼠体内发现,DMN和DLN中约有一半的运动核表达eGFP,而其余一半则不表达。此外,我们还发现,这些运动核中eGFP+与eGFP-亚群分别支配不同的靶肌肉;DMN和DLN中eGFP+的MN投射到性双态的球海绵体肌和异海绵体肌,而eGFP-亚群则投射到性非双态的肛门肌和外尿道括约肌。同样,发现 eGFP+ 与 eGFP- 的嵴状肌 MN 投射到解剖学上不同的嵴状肌部分。通过免疫荧光,DMN和DLN中几乎所有的运动神经元都显示出Cx36的点状标记,包括在eGFP+/eGFP+、eGFP+/eGFP-和eGFP-/eGFP-细胞连接处:结论:DMN和DLN中的大多数(如果不是全部的话)运动神经元是电耦合的,包括性双态和非双态运动神经元之间的电耦合。
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