Epithelial localization of green fluorescent protein-positive cells in epididymis of the GAD67-GFP knock-in mouse
Epithelial localization of green fluorescent protein-positive cells in epididymis of the GAD67-GFP knock-in mouse
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DOI：
10.2164/jandrol.04157
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发表时间：
2005-09-01
影响因子：
--
通讯作者：
Watanabe, M
中科院分区：
文献类型：
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作者：
Abe, H;Yanagawa, Y;Watanabe, M
γ-氨基丁酸 (GABA) 是大脑中主要的抑制性神经递质，也存在于许多外周非神经元组织中，包括男性生殖器官。然而，GABA能细胞在各个器官中的分布尚不清楚。 GAD67-GFP 敲入小鼠是研究大脑中 GABA 能神经元分布和形态的有用模型。我们通过荧光显微镜检查了GAD67-GFP敲入小鼠的雄性生殖器官，发现具有强绿色荧光蛋白（GFP）信号的细胞仅存在于附睾初始段和近端头的上皮中。特征性的细胞形态表明这些是狭窄的细胞。这些 GFP 阳性窄细胞还表达 GAD67 和 GABA。逆转录聚合酶链反应 (RT-PCR) 分析表明，附睾中表达的主要谷氨酸脱羧酶 (GAD) 异构体是 GAD67。 RT-PCR 分析还表明，编码功能性受体组装所需的 GABA(A) 和 GABA(B) 受体亚基的 mRNA 在附睾中表达。近端附睾中检测到的GABA(A)受体亚基mRNA包括α2、β1、γ1和γ3，并且检测到GABA受体的R1和R2亚基mRNA。 GABA(A)受体亚基蛋白的免疫组织化学分析显示，a2、01和gamma亚基在精子中表达，而我们在上皮细胞中未检测到这些GABA(A)受体亚基。 GABA(B) 受体由 GAD67-GFP 敲入小鼠和野生型 Jcl:ICR 小鼠的窄细胞和精子产生。我们的数据表明，GABA 系统可能在狭窄细胞和管腔中的精子中具有重要的功能作用。
gamma-Aminobutyric acid (GABA), which is a major inhibitory neurotransmitter in the brain, is also found in many peripheral nonneuronal tissues, including male reproductive organs. However, the distribution of GABAergic cells in various organs is not known. The GAD67-GFP knock-in mouse is a useful model for studying the distribution and morphology of GABAergic neurons in the brain. We examined the male reproductive organs of GAD67-GFP knock-in mice by fluorescence microscopy and found cells with strong green fluorescent protein (GFP) signal exclusively in the epithelium of the initial segment and proximal caput of the epididymis. The characteristic cell morphology suggested that these were narrow cells. These GFP-positive narrow cells also expressed GAD67 and GABA. Reverse transcription polymerase chain reaction (RT-PCR) analysis showed that the predominant glutamic acid decarboxylase (GAD) isoform expressed in the epididymis is GAD67. RT-PCR analysis also revealed that mRNAs encoding the GABA(A) and GABA(B) receptor subunits necessary for the assembly of functional receptors are expressed in the epididymis. GABA(A) receptor subunit mRNAs detected in the proximal epididymis included alpha 2, beta 1, gamma 1, and gamma 3, and both the R1 and R2 subunit mRNAs of GABA,, receptors were detected. Immunohistochemical analysis of GABA(A) receptor subunit proteins revealed that a2, 01, and gamma subunits expressed in spermatozoa, whereas we did not detect these GABA(A) receptor subunits in epithelial cells. GABA(B) receptors were produced by narrow cells and spermatozoa of GAD67-GFP knock-in and wild-type Jcl:ICR mice. Our data suggest that the GABA system might have important functional roles in narrow cells and on spermatozoa in the lumen.