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Updated: Jul 14, 2026

Methods to Study Epithelial Transport Protein Function and Expression in Native Intestine and Caco-2 Cells Grown in 3D
Published on: March 16, 2017
A functional comparison of recombinant and native somatostatin sst2 receptor variants in epithelia
N D Holliday1, I R Tough, H M Cox
1Wolfson Centre for Age-Related Diseases, King's College London, Hodgkin Building, Guy's Campus, London, UK.
Background And Purpose:
Somatostatin (SRIF-14) exerts broad spectrum antisecretory effects by activating the somatostatin 2 (sst(2)) receptor. The rat (r) sst(2) receptor exists in 'long' (sst(2a)) and 'short' (sst(2b)) forms that differ in their C termini, while a single human (h) sst(2a) exists. This study compares the characteristics of recombinant rsst(2a), rsst(2b) and hsst(2a) activation in human epithelia, and with native sst(2) responses in rat colon.
Experimental Approach:
Epithelial layers of each clone or rat colon were placed in Ussing chambers and short-circuit current (I (SC)) measured in response to SRIF-14 and chosen analogues. The relative potencies and ability to cause desensitization to SRIF-14 were assessed, and the affinities of the sst(2) antagonist, D-Tyr(8) CYN154806 for hsst(2a), rsst(2a) and native rat colon sst(2) receptors were established.
Key Results:
Basolateral SRIF-14 responses were transient in hsst(2a) and rsst(2a) epithelia, but prolonged in rsst(2b)-expressing cells. Activation of rsst(2a) resulted in significant desensitization to SRIF-14 and receptor phosphorylation, whereas the rsst(2b) receptor did neither. Sst(2)-preferred agonists (BIM23190C and BIM23027) reduced I (sc) with similar potency and both caused complete desensitization to SRIF-14. CYN154806 antagonized hsst(2a) and rsst(2a) receptors with pK (B) values of 7.9 and 7.8, respectively. In rat colon mucosa, CYN154806 blocked SRIF-14 responses with a pA (2) value of 8.2, and BIM23190C responses with a pK (B) of 8.4.
Conclusions And Implications:
SRIF-14 caused rapid rsst(2a) receptor phosphorylation and desensitization of epithelial antisecretory responses, neither of which occurred with the rsst(2b) receptor. These mechanisms are most likely to be a prerequisite for sensitivity to sst(2)-analogues with radiotherapeutic potential.
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