The TRPV3 channel of the bovine rumen: localization and functional characterization of a protein relevant for ruminal ammonia transport
The TRPV3 channel of the bovine rumen: localization and functional characterization of a protein relevant for ruminal ammonia transport
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牛瘤胃的 TRPV3 通道：与瘤胃氨转运相关的蛋白质的定位和功能表征
DOI：
10.1007/s00424-020-02393-2
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Stumpff
中科院分区：
文献类型：
--
作者：
Kaessmeyer;Sponder;Stumpff
大量的氨（NH3或NH 4+）从肠道吸收，与肝病中的脑炎，牲畜中蛋白质效率低下以及含氮气候气体的排放有关。确定运输机制似乎迫在眉睫。最近的功能和mRNA数据表明，从牛的前胃吸收氨可能涉及TRPV 3通道。本研究的目的是对牛TRPV 3同源蛋白（bTRPV 3）进行序列测定，并对该蛋白在瘤胃组织中的定位以及NH 4+的转运进行验证。测序后，bTRPV 3在HEK-293细胞和异种卵母细胞中过表达。通过表位筛选选择抗体，并用于检测过表达细胞和牛瘤胃的免疫印迹中的蛋白质，显示预测的~ 90 kDa的信号。仅在瘤胃中，出现了额外的~ 60 kDa条带，其可能代表先前描述的等长度的bTRPV 3剪接变体。免疫组化显示染色从ruminalstratum basaletostratum granulosum。用pH敏感的微电极测量表明，NH 4+酸化异种卵母细胞，bTRPV 3的过表达增强了对NH 4+的渗透性。单通道测量显示，除了在bTRPV 3表达后观察到的四倍大的通道外，异种卵母细胞内源性表达小阳离子通道。内源性和bTRPV 3通道都传导NH 4+、Na+和K+。我们的结论是，bTRPV 3表达的瘤胃上皮细胞的蛋白水平。结合以前的研究数据，Na+，Ca 2+和NH 4+的运输中的作用出现。钙稳态，瘤胃pH值和氮效率在牛的后果进行了讨论。
Large quantities of ammonia (NH3or NH4+) are absorbed from the gut, associated with encephalitis in hepatic disease, poor protein efficiency in livestock, and emissions of nitrogenous climate gasses. Identifying the transport mechanisms appears urgent. Recent functional and mRNA data suggest that absorption of ammonia from the forestomach of cattle may involve TRPV3 channels. The purpose of the present study was to sequence the bovine homologue ofTRPV3(bTRPV3), localize the protein in ruminal tissue, and confirm transport of NH4+. After sequencing, bTRPV3 was overexpressed in HEK-293 cells andXenopusoocytes. An antibody was selected via epitope screening and used to detect the protein in immunoblots of overexpressing cells and bovine rumen, revealing a signal of the predicted ~ 90 kDa. In rumen only, an additional ~ 60 kDa band appeared, which may represent a previously described bTRPV3 splice variant of equal length. Immunohistochemistry revealed staining from the ruminalstratum basaletostratum granulosum. Measurements with pH-sensitive microelectrodes showed that NH4+acidifiesXenopusoocytes, with overexpression of bTRPV3 enhancing permeability to NH4+. Single-channel measurements revealed thatXenopusoocytes endogenously expressed small cation channels in addition to fourfold-larger channels only observed after expression of bTRPV3. Both endogenous and bTRPV3 channels conducted NH4+, Na+, and K+. We conclude that bTRPV3 is expressed by the ruminal epithelium on the protein level. In conjunction with data from previous studies, a role in the transport of Na+, Ca2+, and NH4+emerges. Consequences for calcium homeostasis, ruminal pH, and nitrogen efficiency in cattle are discussed.
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DOI：
10.1152/ajpgi.00418.2004
发表时间：
2005-05-01
影响因子：
4.5
作者：
Handlogten, ME;Hong, SP;Weiner, ID
通讯作者：
Weiner, ID
影响因子：
15.9
作者：
DENNING, GM;OSTEDGAARD, LS;WELSH, MJ
通讯作者：
WELSH, MJ
基底外侧 Mg2/Na 交换通过胞质 Mg2 调节绵羊瘤胃上皮的顶端非选择性阳离子通道。
DOI：
10.1152/ajpgi.00275.2004
发表时间：
2005
期刊：
American journal of physiology. Gastrointestinal and liver physiology
影响因子：
--
作者：
S. Leonhard;F. Stumpff;I. Brinkmann;G. Breves;H. Martens
通讯作者：
H. Martens
禁食和饲喂淡水虹鳟鱼肠道氨转运的体外分析：NKCC、K 通道和 Na、K ATP 酶的作用
DOI：
10.1007/s00360-019-01231-x
发表时间：
2019
期刊：
Journal of Comparative Physiology B
影响因子：
--
作者：
J. Rubino;Jonathan M. Wilson;C. Wood
通讯作者：
C. Wood
印度牛与金牛杂交瘤胃上皮氨转运的比较研究
DOI：
10.1111/asj.13107
发表时间：
2018
期刊：
Animal science journal = Nihon chikusan Gakkaiho
影响因子：
--
作者：
Rabbani;Akhtar;Rosendahl;Grunau;Tietjen;Masood;Kaessmeyer;Gunzel;Rehman;Stumpff
通讯作者：
Stumpff