Zebrafish Tric-b is required for skeletal development and bone cells differentiation

TY - JOUR

T1 - Zebrafish Tric-b is required for skeletal development and bone cells differentiation

AU - Tonelli, Francesca

AU - Leoni, Laura

AU - Daponte, Valentina

AU - Gioia, Roberta

AU - Cotti, Silvia

AU - Fiedler, Imke A K

AU - Larianova, Daria

AU - Willaert, Andy

AU - Coucke, Paul J

AU - Villani, Simona

AU - Busse, Björn

AU - Besio, Roberta

AU - Rossi, Antonio

AU - Witten, P Eckhard

AU - Forlino, Antonella

N1 - Copyright © 2023 Tonelli, Leoni, Daponte, Gioia, Cotti, Fiedler, Larianova, Willaert, Coucke, Villani, Busse, Besio, Rossi, Witten and Forlino.

PY - 2023

Y1 - 2023

N2 - INTRODUCTION: Trimeric intracellular potassium channels TRIC-A and -B are endoplasmic reticulum (ER) integral membrane proteins, involved in the regulation of calcium release mediated by ryanodine (RyRs) and inositol 1,4,5-trisphosphate (IP3Rs) receptors, respectively. While TRIC-A is mainly expressed in excitable cells, TRIC-B is ubiquitously distributed at moderate level. TRIC-B deficiency causes a dysregulation of calcium flux from the ER, which impacts on multiple collagen specific chaperones and modifying enzymatic activity, leading to a rare form of osteogenesis imperfecta (OI Type XIV). The relevance of TRIC-B on cell homeostasis and the molecular mechanism behind the disease are still unknown.RESULTS: In this study, we exploited zebrafish to elucidate the role of TRIC-B in skeletal tissue. We demonstrated, for the first time, that tmem38a and tmem38b genes encoding Tric-a and -b, respectively are expressed at early developmental stages in zebrafish, but only the latter has a maternal expression. Two zebrafish mutants for tmem38b were generated by CRISPR/Cas9, one carrying an out of frame mutation introducing a premature stop codon (tmem38b-/- ) and one with an in frame deletion that removes the highly conserved KEV domain (tmem38bΔ120-7/Δ120-7 ). In both models collagen type I is under-modified and partially intracellularly retained in the endoplasmic reticulum, as described in individuals affected by OI type XIV. Tmem38b-/- showed a mild skeletal phenotype at the late larval and juvenile stages of development whereas tmem38bΔ120-7/Δ120-7 bone outcome was limited to a reduced vertebral length at 21 dpf. A caudal fin regeneration study pointed towards impaired activity of osteoblasts and osteoclasts associated with mineralization impairment.DISCUSSION: Our data support the requirement of Tric-b during early development and for bone cell differentiation.

AB - INTRODUCTION: Trimeric intracellular potassium channels TRIC-A and -B are endoplasmic reticulum (ER) integral membrane proteins, involved in the regulation of calcium release mediated by ryanodine (RyRs) and inositol 1,4,5-trisphosphate (IP3Rs) receptors, respectively. While TRIC-A is mainly expressed in excitable cells, TRIC-B is ubiquitously distributed at moderate level. TRIC-B deficiency causes a dysregulation of calcium flux from the ER, which impacts on multiple collagen specific chaperones and modifying enzymatic activity, leading to a rare form of osteogenesis imperfecta (OI Type XIV). The relevance of TRIC-B on cell homeostasis and the molecular mechanism behind the disease are still unknown.RESULTS: In this study, we exploited zebrafish to elucidate the role of TRIC-B in skeletal tissue. We demonstrated, for the first time, that tmem38a and tmem38b genes encoding Tric-a and -b, respectively are expressed at early developmental stages in zebrafish, but only the latter has a maternal expression. Two zebrafish mutants for tmem38b were generated by CRISPR/Cas9, one carrying an out of frame mutation introducing a premature stop codon (tmem38b-/- ) and one with an in frame deletion that removes the highly conserved KEV domain (tmem38bΔ120-7/Δ120-7 ). In both models collagen type I is under-modified and partially intracellularly retained in the endoplasmic reticulum, as described in individuals affected by OI type XIV. Tmem38b-/- showed a mild skeletal phenotype at the late larval and juvenile stages of development whereas tmem38bΔ120-7/Δ120-7 bone outcome was limited to a reduced vertebral length at 21 dpf. A caudal fin regeneration study pointed towards impaired activity of osteoblasts and osteoclasts associated with mineralization impairment.DISCUSSION: Our data support the requirement of Tric-b during early development and for bone cell differentiation.

KW - Animals

KW - Zebrafish/metabolism

KW - Ion Channels/genetics

KW - Calcium/metabolism

KW - Bone and Bones/metabolism

KW - Osteogenesis Imperfecta/genetics

KW - Cell Differentiation/genetics

U2 - 10.3389/fendo.2023.1002914

DO - 10.3389/fendo.2023.1002914

M3 - SCORING: Journal article

C2 - 36755921

VL - 14

JO - FRONT ENDOCRINOL

JF - FRONT ENDOCRINOL

SN - 1664-2392

M1 - 1002914

ER -