Metabolic and behavioral alterations associated with viral vector-mediated toxicity in the paraventricular hypothalamic nucleus.
Metabolic and behavioral alterations associated with viral vector-mediated toxicity in the paraventricular hypothalamic nucleus.
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与病毒载体介导的下丘脑室旁核毒性相关的代谢和行为改变。
DOI：
10.1042/bsr20231846
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发表时间：
2024-01-16
影响因子：
4
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目的：结合腺体相关病毒（AAV）介导的CRE重组酶与转基因的Floxed Animals的表达是一种强大的方法，是一种强大的方法，用于分析基因在调节行为和代谢中的功能作用。与使用CRE驱动小鼠线交叉点相比，使用AAV-CRE对不同细胞类型和组织的广泛研究表明，它可以节省时间并避免发育补偿。我们最初试图使用成年动物中颅内AAV-CRE注射来研究肾上腺素释放激素（CRH）在旁牙性下丘脑下核（PVN）中消融的影响。
方法：在本研究中，我们在CRH-loxed和野生型小鼠PVN中立体定位注射AAV8-HSYN-CRE或对照AAV8-HSYN-GFP，以评估行为和代谢影响。然后，我们使用免疫组织化学标志物系统地评估下丘脑肽基神经元和神经胶质细胞的密度。
结果：我们发现，在PVN中，AAV8-HSYN-CRE的一种特定制剂的递送导致肥胖，肉质和焦虑样行为的发展。这种效果与性别和野生型小鼠无关。随后，我们发现AAV8-HSYN-CRE导致病毒载体注射部位导致神经元细胞死亡和神经胶质病。这些行为和代谢缺陷取决于注入PVN。或者采购的AAV-CRE不会再现相同的结果。
结论：我们的发现表明，特定批量的AAV-CRE的递送可能导致PVN中细胞毒性和病变，从而导致强大的代谢和行为影响。这些改变会使CRE介导的基因敲除的解释复杂化，并强调对严格控制的需求。
Objective: Combining adeno-associated virus (AAV)-mediated expression of Cre recombinase with genetically modified floxed animals is a powerful approach for assaying the functional role of genes in regulating behavior and metabolism. Extensive research in diverse cell types and tissues using AAV-Cre has shown it can save time and avoid developmental compensation as compared with using Cre driver mouse line crossings. We initially sought to study the impact of the ablation of corticotropin-releasing hormone (CRH) in the paraventricular hypothalamic nucleus (PVN) using intracranial AAV-Cre injection in adult animals. Methods: In the present study, we stereotactically injected AAV8-hSyn-Cre or a control AAV8-hSyn-GFP in both Crh-floxed and wild-type mouse PVN to assess behavioral and metabolic impacts. We then used immunohistochemical markers to systematically evaluate the density of hypothalamic peptidergic neurons and glial cells. Results: We found that delivery of one specific preparation of AAV8-hSyn-Cre in the PVN led to the development of obesity, hyperphagia, and anxiety-like behaviors. This effect occurred independent of sex and in both floxed and wild-type mice. We subsequently found that AAV8-hSyn-Cre led to neuronal cell death and gliosis at the site of viral vector injections. These behavioral and metabolic deficits were dependent on injection into the PVN. An alternatively sourced AAV-Cre did not reproduce the same results. Conclusions: Our findings reveal that delivery of a specific batch of AAV-Cre could lead to cellular toxicity and lesions in the PVN that cause robust metabolic and behavioral impacts. These alterations can complicate the interpretation of Cre-mediated gene knockout and highlight the need for rigorous controls.