CHARACTERIZATION OF DAR & NON DAR CELLS IN THE MOSQUITO RECTUM

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 蚊子是许多病原体的宿主，包括寄生虫，细菌，病毒和真菌，并且在17世纪到20世纪造成的人类疾病和死亡比所有其他原因加起来还要多。 今天，仅疟疾就感染了全世界5亿人，每年至少有200万人死亡。 为了开发有效和具体的控制方法，对蚊子生存至关重要的生物过程有详细的了解至关重要。 在按蚊中，一个长期被忽视的过程是离子调节，其中一个关键器官是直肠。 我试图用自参比离子选择性电极技术表征新鉴定的按蚊直肠幼虫细胞类型（DAR和非DAR细胞）的功能。 生物电流研究中心是这个项目的宝贵资源，为我的研究提供了必要的设备和专业知识。 为了更好地了解按蚊直肠的DAR和非DAR细胞的功能，我测量了在2%（新鲜）或50%人工海水中饲养的白曼按蚊蚊幼虫直肠细胞的基膜处的K+和H+离子通量。 K+离子通量非常低，我无法得出明确的结论，因为我无法区分实际通量和背景。 然而，我发现一个一致的H+离子通量模式，似乎是独立的饲养盐度。 我在DAR细胞中发现了强烈的H+离子流出，而在非DAR细胞中通量明显下降。 此外，乙酰唑胺（一种碳酸酐酶抑制剂）抑制DAR细胞通量，但不抑制非DAR细胞通量。 这支持了先前的数据，即碳酸酐酶差异定位于按蚊直肠的DAR细胞。 来自其他药理学阻断剂的数据暗示离子外排中的V-ATP酶，并且排除K+/H+转运蛋白。 结合这些生理数据和分子免疫学技术，我们可以更清楚地了解按蚊幼虫的离子调节。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Mosquitoes are hosts to a number of pathogens including parasites, bacteria, viruses, and fungi, and were responsible for more human disease and death in the 17th through 20th centuries than all other causes combined. Today malaria alone infects 500 million people worldwide, and kills at least two million people each year. It is critical to have a detailed understanding of the biological processes crucial for mosquito survival in order to develop effective and specific control methods. One process which has long been overlooked in anopheline mosquitoes is ion regulation, a key organ of which is the rectum. I sought to characterize the functions of the newly identified cell types (DAR and non-DAR cells) of the larval anopheline rectum using the self-referencing ion-selective electrode technique. The Biocurrents Research Center was an invaluable resource for this project providing both the equipment and the expertise necessary for my research. To obtain a better understanding of the functions of the DAR and non-DAR cells of the anopheline rectum, I measured K+ and H+ ion flux at the basal membrane of the rectal cells of Anopheles albimanus mosquito larvae reared in either 2% (fresh) or 50% artificial sea water. K+ ion flux was very low and I was unable to draw definitive conclusions because of my inability to distinguish actual flux from background. However, I found a consistent H+ ion flux pattern that seemed to be independent of rearing salinity. I found a strong H+ ion efflux at the DAR cells and a clear drop in flux at the non-DAR cells. Additionally, acetazolamide (a carbonic anhydrase inhibitor) inhibited DAR cell flux, but not non-DAR cell flux. This supports previous data that carbonic anhydrase differentially localizes to the DAR cells of the anopheline rectum. Data from other pharmacological blockers implicates V-ATPase in the ion efflux and excludes K+/H+ transporters. Combining this physiological data with molecular and immunological techniques can afford us a clearer picture of ion regulation in anopheline larvae.