دورية أكاديمية
أعرض في EDS

Precise CRISPR-Cas9-mediated mutation of a membrane trafficking domain in the Drosophila vesicular monoamine transporter gene.

التفاصيل البيبلوغرافية
العنوان: Precise CRISPR-Cas9-mediated mutation of a membrane trafficking domain in the Drosophila vesicular monoamine transporter gene.
المؤلفون: Asuncion JD; Medical Scientist Training Program, David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA.; Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA.; UCLA Neuroscience Interdepartmental Program, University of California, Los Angeles, CA, 90095, USA., Eamani A; Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA., Rohrbach EW; Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA.; UCLA Neuroscience Interdepartmental Program, University of California, Los Angeles, CA, 90095, USA., Knapp EM; Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA., Deshpande SA; Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA., Bonanno SL; Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA., Murphy JE; Department of Biological Sciences, Delaware State University, Dover, DE, USA, 19901, USA., Lawal HO; Department of Biological Sciences, Delaware State University, Dover, DE, USA, 19901, USA., Krantz DE; Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA.
المصدر: Current research in physiology [Curr Res Physiol] 2023 Jun 20; Vol. 6, pp. 100101. Date of Electronic Publication: 2023 Jun 20 (Print Publication: 2023).
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Elsevier B.V Country of Publication: Netherlands NLM ID: 101769251 Publication Model: eCollection Cited Medium: Internet ISSN: 2665-9441 (Electronic) Linking ISSN: 26659441 NLM ISO Abbreviation: Curr Res Physiol Subsets: PubMed not MEDLINE
أسماء مطبوعة: Original Publication: [Amsterdam] : Elsevier B.V., [2019]-
مستخلص: Monoamine neurotransmitters such as noradrenalin are released from both synaptic vesicles (SVs) and large dense-core vesicles (LDCVs), the latter mediating extrasynaptic signaling. The contribution of synaptic versus extrasynaptic signaling to circuit function and behavior remains poorly understood. To address this question, we have previously used transgenes encoding a mutation in the Drosophila Vesicular Monoamine Transporter ( dVMAT ) that shifts amine release from SVs to LDCVs. To circumvent the use of transgenes with non-endogenous patterns of expression, we have now used CRISPR-Cas9 to generate a trafficking mutant in the endogenous dVMAT gene. To minimize disruption of the dVMAT coding sequence and a nearby RNA splice site, we precisely introduced a point mutation using single-stranded oligonucleotide repair. A predicted decrease in fertility was used as a phenotypic screen to identify founders in lieu of a visible marker. Phenotypic analysis revealed a defect in the ovulation of mature follicles and egg retention in the ovaries. We did not detect defects in the contraction of lateral oviducts following optogenetic stimulation of octopaminergic neurons. Our findings suggest that release of mature eggs from the ovary is disrupted by changing the balance of VMAT trafficking between SVs and LDCVs. Further experiments using this model will help determine the mechanisms that sensitize specific circuits to changes in synaptic versus extrasynaptic signaling.
Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(© 2023 The Authors.)
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تواريخ الأحداث: Date Created: 20230706 Latest Revision: 20230718
رمز التحديث: 20240628
مُعرف محوري في PubMed: PMC10318446
DOI: 10.1016/j.crphys.2023.100101
PMID: 37409154
قاعدة البيانات: MEDLINE
الوصف
تدمد:2665-9441
DOI:10.1016/j.crphys.2023.100101