Butyrate Permeation across the Isolated Ovine Reticulum Epithelium

Abstract

Simple SummaryShort-chain fatty acids are the main source of energy for ruminants. The effective uptake of these substrates from the forestomach is a prerequisite for the health and performance of these animals. Thus far, the mechanisms of uptake have been investigated almost exclusively in the epithelium of the largest forestomach section, the rumen. Previous research suggests that the reticulum is also involved in the uptake of short-chain fatty acids, but the mechanisms involved have not been studied and may differ from those known from the rumen epithelium due to the different milieu in this compartment. To investigate this, ovine reticulum epithelium was mounted in Ussing chambers, and the transport of radiolabeled butyrate (as a representative of short-chain fatty acids) across the tissue was measured with and without the addition of inhibitors of particular transport proteins. Our results show that butyrate can be taken up effectively across the reticulum epithelium via pathways that are energized by the Na+/K+-ATPase and may involve monocarboxylate transporters, sodium-proton exchangers, and anion channels. However, our results are not completely congruent to those obtained in the rumen epithelium. These modifications could assure the effective uptake of short-chain fatty acids from the reticulum lumen under the particular conditions (p. e. high pH) of this forestomach compartment.We hypothesized that, due to the high pH of this compartment, the reticulum epithelium displays particular features in the transport of short-chain fatty acids (SCFA). Ovine reticulum epithelium was incubated in Ussing chambers using a bicarbonate-free buffer solution containing butyrate (20 mmol L−1). p-hydroxymercuribenzoic acid (pHMB), 5-(N-Ethyl-N-isopropyl)amiloride (EIPA), or ouabain were added to the buffer solution as inhibitors of monocarboxylate transporters, sodium-proton-exchangers, or the Na+/K+-ATPase, respectively. The short-circuit current (Isc) and transepithelial conductance (Gt) were monitored continuously while the flux rates of 14C-labelled butyrate were measured in the mucosal-to-serosal (Jmsbut) or serosal-to-mucosal direction (Jsmbut). Under control conditions, the mean values of Isc and Gt amounted to 2.54 ± 0.46 µEq cm−2 h−1 and 6.02 ± 3.3 mS cm−2, respectively. Jmsbut was 2.1 ± 1.01 µmol cm−2 h−1 on average and about twice as high as Jsmbut. Incubation with ouabain reduced Jmsbut, while Jsmbut was not affected. The serosal addition of EIPA did not affect Jmsbut but reduced Jsmbut by about 10%. The addition of pHMB to the mucosal or serosal solution reduced Jmsbut but had no effect on Jsmbut. Mucosally applied pHMB provoked a transient increase in the Isc. The serosal pHMB sharply reduced Isc. Our results demonstrate that butyrate can be effectively transported across the reticulum epithelium. The mechanisms involved in this absorption differ from those known from the rumen epithelium.