Tick Cell Culture Analysis of Growth Dynamics and Cellular Tropism of Rickettsia buchneri, an Endosymbiont of the Blacklegged Tick, Ixodes scapularis

The blacklegged tick, Ixodes scapularis, a species of significant importance to human and animal health, harbors an endosymbiont Rickettsia buchneri sensu stricto. The symbiont is largely restricted to the ovaries, but all life stages can harbor various quantiti...

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Autores principales:	Cody J. Thorpe, Xin-Ru Wang, Ulrike G. Munderloh, Timothy J. Kurtti
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	tick cell lines <i>Ixodes ricinus</i> cell line IRE11 blacklegged tick <i>Rickettsia buchneri</i> <i>Ixodes scapularis</i> endosymbiont RNA FISH Science Q
Acceso en línea:	https://doaj.org/article/1b6e948ff7224dc6813f788846a563ae
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spelling	oai:doaj.org-article:1b6e948ff7224dc6813f788846a563ae2021-11-25T17:59:09ZTick Cell Culture Analysis of Growth Dynamics and Cellular Tropism of <i>Rickettsia buchneri</i>, an Endosymbiont of the Blacklegged Tick, <i>Ixodes scapularis</i>10.3390/insects121109682075-4450https://doaj.org/article/1b6e948ff7224dc6813f788846a563ae2021-10-01T00:00:00Zhttps://www.mdpi.com/2075-4450/12/11/968https://doaj.org/toc/2075-4450The blacklegged tick, <i>Ixodes scapularis</i>, a species of significant importance to human and animal health, harbors an endosymbiont <i>Rickettsia buchneri</i> sensu stricto. The symbiont is largely restricted to the ovaries, but all life stages can harbor various quantities or lack <i>R. buchneri</i> entirely. The endosymbiont is cultivable in cell lines isolated from embryos of <i>Ixodes</i> ticks. <i>Rickettsia buchneri</i> most readily grows and is maintained in the cell line IRE11 from the European tick, <i>Ixodes ricinus</i>. The line was characterized by light and electron microscopy and used to analyze the growth dynamics of wildtype and GFPuv-expressing <i>R. buchneri</i>. qPCR indicated that the genome copy doubling time in IRE11 was >7 days. Measurements of fluorescence using a plate reader indicated that the amount of green fluorescent protein doubled every 11 days. Two 23S rRNA probes were tested via RNA FISH on rickettsiae grown in vitro and adapted to evaluate the tissue tropism of <i>R. buchneri</i> in field-collected female <i>I. scapularis</i>. We observed strong positive signals of <i>R. buchneri</i> in the ovaries and surrounding the nucleus of the developing oocytes. Tissue tropism in <i>I. scapularis</i> and in vitro growth dynamics strengthen the contemporary understanding of <i>R. buchneri</i> as a transovarially transmitted, non-pathogenic endosymbiont.Cody J. ThorpeXin-Ru WangUlrike G. MunderlohTimothy J. KurttiMDPI AGarticletick cell lines<i>Ixodes ricinus</i> cell line IRE11blacklegged tick<i>Rickettsia buchneri</i><i>Ixodes scapularis</i> endosymbiontRNA FISHScienceQENInsects, Vol 12, Iss 968, p 968 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	tick cell lines <i>Ixodes ricinus</i> cell line IRE11 blacklegged tick <i>Rickettsia buchneri</i> <i>Ixodes scapularis</i> endosymbiont RNA FISH Science Q
spellingShingle	tick cell lines <i>Ixodes ricinus</i> cell line IRE11 blacklegged tick <i>Rickettsia buchneri</i> <i>Ixodes scapularis</i> endosymbiont RNA FISH Science Q Cody J. Thorpe Xin-Ru Wang Ulrike G. Munderloh Timothy J. Kurtti Tick Cell Culture Analysis of Growth Dynamics and Cellular Tropism of <i>Rickettsia buchneri</i>, an Endosymbiont of the Blacklegged Tick, <i>Ixodes scapularis</i>
description	The blacklegged tick, <i>Ixodes scapularis</i>, a species of significant importance to human and animal health, harbors an endosymbiont <i>Rickettsia buchneri</i> sensu stricto. The symbiont is largely restricted to the ovaries, but all life stages can harbor various quantities or lack <i>R. buchneri</i> entirely. The endosymbiont is cultivable in cell lines isolated from embryos of <i>Ixodes</i> ticks. <i>Rickettsia buchneri</i> most readily grows and is maintained in the cell line IRE11 from the European tick, <i>Ixodes ricinus</i>. The line was characterized by light and electron microscopy and used to analyze the growth dynamics of wildtype and GFPuv-expressing <i>R. buchneri</i>. qPCR indicated that the genome copy doubling time in IRE11 was >7 days. Measurements of fluorescence using a plate reader indicated that the amount of green fluorescent protein doubled every 11 days. Two 23S rRNA probes were tested via RNA FISH on rickettsiae grown in vitro and adapted to evaluate the tissue tropism of <i>R. buchneri</i> in field-collected female <i>I. scapularis</i>. We observed strong positive signals of <i>R. buchneri</i> in the ovaries and surrounding the nucleus of the developing oocytes. Tissue tropism in <i>I. scapularis</i> and in vitro growth dynamics strengthen the contemporary understanding of <i>R. buchneri</i> as a transovarially transmitted, non-pathogenic endosymbiont.
format	article
author	Cody J. Thorpe Xin-Ru Wang Ulrike G. Munderloh Timothy J. Kurtti
author_facet	Cody J. Thorpe Xin-Ru Wang Ulrike G. Munderloh Timothy J. Kurtti
author_sort	Cody J. Thorpe
title	Tick Cell Culture Analysis of Growth Dynamics and Cellular Tropism of <i>Rickettsia buchneri</i>, an Endosymbiont of the Blacklegged Tick, <i>Ixodes scapularis</i>
title_short	Tick Cell Culture Analysis of Growth Dynamics and Cellular Tropism of <i>Rickettsia buchneri</i>, an Endosymbiont of the Blacklegged Tick, <i>Ixodes scapularis</i>
title_full	Tick Cell Culture Analysis of Growth Dynamics and Cellular Tropism of <i>Rickettsia buchneri</i>, an Endosymbiont of the Blacklegged Tick, <i>Ixodes scapularis</i>
title_fullStr	Tick Cell Culture Analysis of Growth Dynamics and Cellular Tropism of <i>Rickettsia buchneri</i>, an Endosymbiont of the Blacklegged Tick, <i>Ixodes scapularis</i>
title_full_unstemmed	Tick Cell Culture Analysis of Growth Dynamics and Cellular Tropism of <i>Rickettsia buchneri</i>, an Endosymbiont of the Blacklegged Tick, <i>Ixodes scapularis</i>
title_sort	tick cell culture analysis of growth dynamics and cellular tropism of <i>rickettsia buchneri</i>, an endosymbiont of the blacklegged tick, <i>ixodes scapularis</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/1b6e948ff7224dc6813f788846a563ae
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_version_	1718411726868709376

Tick Cell Culture Analysis of Growth Dynamics and Cellular Tropism of <i>Rickettsia buchneri</i>, an Endosymbiont of the Blacklegged Tick, <i>Ixodes scapularis</i>

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