<svg width="800" height="600" xmlns="http://www.w3.org/2000/svg"> <!-- 标题 --> <text x="400" y="30" font-family="Arial, sans-serif" font-size="20" font-weight="bold" text-anchor="middle">RA-RARα-5HT1A通路在PFC中的细胞特异性作用机制研究技术路线图</text> <!-- 第一部分：探索关键作用的神经元类型 --> <rect x="50" y="70" width="700" height="220" rx="5" ry="5" fill="#f0f8ff" stroke="#4682b4" stroke-width="2"/> <text x="75" y="95" font-family="Arial, sans-serif" font-size="16" font-weight="bold" fill="#2e4b7c">第一部分：探索RA-RARα-5HT1A通路发挥关键作用的神经元类型</text> <!-- 步骤1.1 --> <rect x="100" y="120" width="600" height="40" rx="3" ry="3" fill="#e6f3ff" stroke="#87ceeb" stroke-width="1.5"/> <text x="120" y="145" font-family="Arial, sans-serif" font-size="14" fill="#333">（1）构建与验证 RARα cKO (Nestin-Cre) ASD模型</text> <!-- 步骤1.1子项 --> <g> <rect x="130" y="160" width="180" height="30" rx="2" ry="2" fill="#ffffff" stroke="#ccc" stroke-width="1"/> <text x="140" y="180" font-family="Arial, sans-serif" font-size="12" fill="#555">• 行为学评估 (三箱社交等)</text> </g> <g> <rect x="320" y="160" width="180" height="30" rx="2" ry="2" fill="#ffffff" stroke="#ccc" stroke-width="1"/> <text x="330" y="180" font-family="Arial, sans-serif" font-size="12" fill="#555">• 突触形态学 (高尔基染色)</text> </g> <g> <rect x="510" y="160" width="180" height="30" rx="2" ry="2" fill="#ffffff" stroke="#ccc" stroke-width="1"/> <text x="520" y="180" font-family="Arial, sans-serif" font-size="12" fill="#555">• 电生理记录 (膜片钳)</text> </g> <!-- 步骤1.2 --> <rect x="100" y="200" width="600" height="40" rx="3" ry="3" fill="#e6f3ff" stroke="#87ceeb" stroke-width="1.5"/> <text x="120" y="225" font-family="Arial, sans-serif" font-size="14" fill="#333">（2）单细胞核RNA测序 (snRNA-seq) 锁定关键神经元亚群</text> <!-- 步骤1.2子项 --> <g> <rect x="130" y="240" width="180" height="30" rx="2" ry="2" fill="#ffffff" stroke="#ccc" stroke-width="1"/> <text x="140" y="260" font-family="Arial, sans-serif" font-size="12" fill="#555">• 细胞聚类与图谱构建</text> </g> <g> <rect x="320" y="240" width="180" height="30" rx="2" ry="2" fill="#ffffff" stroke="#ccc" stroke-width="1"/> <text x="330" y="260" font-family="Arial, sans-serif" font-size="12" fill="#555">• 差异表达与通路分析</text> </g> <g> <rect x="510" y="240" width="180" height="30" rx="2" ry="2" fill="#ffffff" stroke="#ccc" stroke-width="1"/> <text x="520" y="260" font-family="Arial, sans-serif" font-size="12" fill="#555">• 鉴定RARα/5HT1A共下调亚群</text> </g> <!-- 连接箭头与输出 --> <path d="M 400 290 L 400 330" stroke="#4682b4" stroke-width="2" fill="none" marker-end="url(#arrowhead)"/> <rect x="350" y="330" width="100" height="40" rx="3" ry="3" fill="#ffebcc" stroke="#ff9900" stroke-width="1.5"/> <text x="400" y="355" font-family="Arial, sans-serif" font-size="14" font-weight="bold" text-anchor="middle" fill="#b35900">输出：关键神经元亚群 “X”</text> <!-- 第二部分：明确作用机制 --> <rect x="50" y="390" width="700" height="180" rx="5" ry="5" fill="#f0fff0" stroke="#32cd32" stroke-width="2"/> <text x="75" y="415" font-family="Arial, sans-serif" font-size="16" font-weight="bold" fill="#2d5c2d">第二部分：明确RA-RARα-5HT1A通路在关键神经元上的作用机制</text> <!-- 步骤2.1 --> <rect x="100" y="440" width="600" height="40" rx="3" ry="3" fill="#e6ffe6" stroke="#90ee90" stroke-width="1.5"/> <text x="120" y="465" font-family="Arial, sans-serif" font-size="14" fill="#333">（1）构建与验证 “X” 神经元特异性 RARα 敲除小鼠</text> <!-- 步骤2.1子项 --> <g> <rect x="130" y="480" width="290" height="30" rx="2" ry="2" fill="#ffffff" stroke="#ccc" stroke-width="1"/> <text x="140" y="500" font-family="Arial, sans-serif" font-size="12" fill="#555">• 多层次表型验证 (行为、形态、功能)</text> </g> <g> <rect x="430" y="480" width="270" height="30" rx="2" ry="2" fill="#ffffff" stroke="#ccc" stroke-width="1"/> <text x="440" y="500" font-family="Arial, sans-serif" font-size="12" fill="#555">• 确认该亚群在表型中的必要性</text> </g> <!-- 步骤2.2 --> <rect x="100" y="520" width="600" height="40" rx="3" ry="3" fill="#e6ffe6" stroke="#90ee90" stroke-width="1.5"/> <text x="120" y="545" font-family="Arial, sans-serif" font-size="14" fill="#333">（2）深度机制探索：靶细胞转录组测序与下游验证</text> <!-- 步骤2.2子项 --> <g> <rect x="130" y="560" width="290" height="30" rx="2" ry="2" fill="#ffffff" stroke="#ccc" stroke-width="1"/> <text x="140" y="580" font-family="Arial, sans-serif" font-size="12" fill="#555">• 差异基因、通路与调控网络分析</text> </g> <g> <rect x="430" y="560" width="270" height="30" rx="2" ry="2" fill="#ffffff" stroke="#ccc" stroke-width="1"/> <text x="440" y="580" font-family="Arial, sans-serif" font-size="12" fill="#555">• 关键下游分子功能挽救实验</text> </g> <!-- 最终目标 --> <path d="M 400 600 L 400 640" stroke="#32cd32" stroke-width="2" fill="none" marker-end="url(#arrowhead)"/> <rect x="300" y="640" width="200" height="40" rx="3" ry="3" fill="#ffccff" stroke="#cc66cc" stroke-width="1.5"/> <text x="400" y="665" font-family="Arial, sans-serif" font-size="14" font-weight="bold" text-anchor="middle" fill="#660066">阐明细胞特异性作用机制<br>为ASD精准干预提供靶点</text> <!-- 定义箭头标记 --> <defs> <marker id="arrowhead" markerWidth="10" markerHeight="7" refX="9" refY="3.5" 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